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The document discusses test plans and available data for the chemical dimethyl disulfide as part of the United States Environmental Protection Agency's High Production Volume Challenge Program.

The document appears to be a test plan for the chemical dimethyl disulfide (DMDS) submitted to the United States Environmental Protection Agency (USEPA) to fulfill their High Production Volume (HPV) Challenge Program commitments.

The purpose of the document is to summarize the available data on dimethyl disulfide and identify any additional testing required to meet the HPV Challenge Program requirements.

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201-16767

Appendix II

Dimethyl Disulfide Test Plan

US HPV CHALLENGE

DIMETHYL 201-16767 DISULFIDE

201-16161A
High Production Volume (HPV) Challenge Program

DIMETHYL DISULFIDE (CAW 624-92-0) Test Plan

Arkema Inc. 2000 Market Street 19103 Philadelphia, PA


December 2005

US HPV CHALLENGE EXECUTIVE SUMMARY

DIMETHYL DISULFIDE

201-16767

Arkema Inc has volunteered to sponsor dimethyl disulfide (DMDS, CAS# 624-92-0) in the USEPA HPV program. The DMDS Test Plan is being submitted to fulfill the United States Environmental Protection Agency (USEPA) High Production Volume (HPV) Challenge Program commitment for DMDS. Data from company proprietary files, peer-reviewed literature, and/or calculated endpoints using widely accepted computer modeling programs have been identified for purposes of this program. Robust summaries of the available data are included in the attached IUCLID. The following table summarizes the available data and proposed testing for DMDS. Table 1: Matrix of Available and Adequate Data on DMDS
Data Available Y/N Y Y Y Y Y Y N Y Y N Y Y Y Y Y Y Y Y Y Testing Planned? Y/N N N N N N N Y N N Y N N N N N N N N N

SIDS ENDPOINT Physical and Chemical Data Melting Point Boiling Point Vapor Pressure Partition Coefficient Water Solubility Environmental Fate Photodegradation Stability in Water (Hydrolysis) Transport/Distribution Biodegradation Ecotoxicity Acute/Prolonged Toxicity to Fish Acute Toxicity to Aquatic Invertebrates (Daphnia) Acute Toxicity to Aquatic Plants (Algae) Toxicity Acute Toxicity (Oral) Acute Toxicity (Dermal) Acute Toxicity (Inhalation) Repeated Dose GeneticToxicity in vitro Gene Mutation Genetic Toxicity in vitro Chromosomal Aberration Reproductive Toxicity Developmental Toxicity

Note: The data used to characterize the OECD SIDS endpoints for substances in this Test Plan were identified either in company proprietary files, peer-reviewed literature, and/or calculated using widely accepted computer modelling programs. All data were evaluated for study reliability in accordance with criteria outlined by the USEPA (1999a). Only studies that met the reliability criteria of 1 (reliable without restrictions) or 2 (reliable with restrictions) were used. Additional data are also included in the IUCLID (International Uniform Chemical Information Dataset) attached in Annex I. A more detailed discussion of the data quality and reliability assessment process used to develop this test plan is provided in Annex II.
2

US HPV CHALLENGE 1.1 Physico-Chemical properties

DIMETHYL DISULFIDE

201-16767

DMDS is a pale yellow liquid with a strong garlic like odor. Experimental data for the physical chemical parameters are available and reported in EPIWIN (USEPA, 2004) and are provided in the following table. Table 2. Parameter
Melting Point Boiling Point Vapor Pressure Kow Partition Coefficient Water Solubility (mg/l)

Physicochemical Data Value


-85C 110C
1 1

29.3 hPa 1.77


1 1

2500

EPIWIN v3.12 Syspro database

Conclusion Adequate data are available for the HPV physical/chemical property endpoints. No additional testing for the HPV program is proposed.

GENERAL INFORMATION ON EXPOSURE 1.2 Production Volumes and Use Pattern

DMDS is on EPAs high production volume list indicating it is manufactured and/or imported at greater than 1 million pounds per year according to the toxic inventory update rule (IUR). 1.2.1 Use Pattern:

DMDS has several industrial uses. It is used in the oil industry as a sulfiding/presulfiding agent to activate catalysts of hydrotreating units, to reduce the number of decoking operations in the petrochemical industry, as a chemical intermediate in the fine chemical industry, and as an anti corrosive in metallurgy. 1.3 1.3.1 Environmental Exposure and Fate Photodegradation

The photodegradation of DMDS was evaluated using EPIWIN 3.12. The half life of DMDS was calculated to be 0.565 hours based on the experimental rate constant of 227 x E-12 cm3/molecule sec. Conclusion

US HPV CHALLENGE

DIMETHYL DISULFIDE

Adequate data are available to assess the photodegradation of DMDS. No additional studies are proposed for the HPV program. 1.3.2 Stability in Water

201-16767

EPIWIN was unable to calculate a hydrolysis rate for DMDS. A hydrolysis study is proposed for DMDS. 1.3.3 Transport between Environmental Compartments

The transport of DMDS between environmental compartments was assessed by fugacity modeling using EPIWIN (v3.12). Results are listed in the table below: Table 3. Fugacity Results for DMDS
Compartment Air Water Soil Sediment Mass amount (%) 1.01 58.1 40.8 0.168 Estimated half life (hr) 1.13 360 360 3.24x e003

1.3.4

B iodegradation

DMDS was not readily biodegradable when evaluated according to OECD 301D. The degradation was less than 10% following 28 days exposure. Conclusion Adequate data are available to assess the biodegradation of DMDS. No additional studies are proposed for the HPV program.

2
2.1.1

HUMAN HEALTH HAZARDS Acute Toxicity

Single exposure (acute) studies indicate DMDS is moderately toxic if swallowed (rat; 290 mg/kg < LD50 < 500 mg/kg), no more then slightly toxic if absorbed through skin (rabbit LD50 >2,000 mg/kg), and slightly toxic if inhaled (rat 4-hr LC50 805 ppm). Conclusion Adequate data are available to assess the acute toxicity of DMDS and no additional studies are proposed. 2.1.2 Repeated Dose Toxicity

DMDS was evaluated in a 90-day repeated dose study on rats according to OECD guidelines. This study featured inhalation dosing, measurement of mortality, body weight changes, food consumption, hematological and blood biochemical examinations, urinalysis, organ weights, histopathology and a functional observational battery. Rats were exposed whole body to 0, 10, 50, 150, and 250 ppm DMDS for 6 hours per day for 90 days. Satellite groups were evaluated
4

US HPV CHALLENGE

DIMETHYL DISULFIDE

following a 2 -week recovery period. Results from this study showed decreased body weights, food consumption, hypoactivity, changes in white blood cell counts, reduced thymus gland weight and increased liver weight. Reversible microscopic changes were noted in the nasal mucosa. Conclusion

201-16767

Adequate data are available to assess the reproductive toxicity of DMDS. No additional testing is proposed for purposes of the HPV program. 2.1.3 Mutagenicity

Several reliable genetic toxicity studies are available for DMDS. Predominantly negative results were obtained with DMDS when tested in vitro (negative bacterial and mammalian mutagenicity assays, negative DNA damage and repair, ambiguous positive in vitro chromosome aberration study using human lymphocytes). Negative results were obtained when DMDS was evaluated in vivo (mouse micronucleus, unscheduled DNA synthesis). Conclusion Adequate data are available to assess the genetic toxicity of DMDS. No additional testing is proposed for purposes of the HPV program. 2.1.4 Toxicity for Reproductive/Developmental Toxicity

Reproductive Toxicity The 90 day repeated dose toxicity study will be used to assess the reproductive toxicity of DMDS. Reproductive organs examined in this study included the epididymus, prostate, and testes in males and ovaries and uterus in females. No lesions were reported. Developmental Toxicity A Developmental Toxicity test was completed for DMDS in Sprague -Dawley rats following OECD Guideline 414 Teratogenicity. DMDS was administered by inhalation to 0, 5, 15, and 50 ppm on gestation days 6 to 15. Maternal toxicity was noted at 15 and 50 ppm. No evidence of developmental toxicity was observed. No additional studies are proposed. Conclusion Adequate data are available to assess the reproductive and developmental toxicity of DMDS. No additional testing is proposed for the HPB program.

HAZARDS TO AQUATIC O RGANISMS

DMDS has been evaluated in an acute daphnia immobilization and algal growth inhibition studies. DMDS is moderately toxic to daphnia with a 48 hour EC50 value of 7 mg/l. DMDS is slightly toxic to Selenastrum capricornutum alga with a 72 hour EC50 of 35 mg/l. No data are available for acute fish and alga. No data are available to assess the acute fish toxicity and an acute fish toxicity (OECD guideline 203) is proposed for DMDS. Conclusion Adequate data are available to assess the aquatic toxicity of DMDS to daphnia and alga but not fish. An acute fish toxicity study is proposed (OECD guideline 203) for DMDS.
5

US HPV CHALLENGE References

DIMETHYL DISULFIDE

201-16767

Atofina, 2005. IUCLID Data Set, CAS No. 624-92-0 dimethyldisulfide. Atofina, Paris, France. Klimisch, H.J., E. Andreae, and U. Tillmann. 1997. A systematic approach for evaluating the quality of experimental and ecotoxicological data. Reg. Tox. and Pharm . 25: 1 -5.
Organisation for Economic Co-operation and Development (OECD) Secretariat. 2002. Manual for
Investigation of HPV Chemicals (November 2002).
U.S. Environmental Protection Agency (USEPA), Office of Pollution Prevention and Toxics. 1998. Guidance for Meeting the SIDS Requirements: Chemical Right-to-Know Initiative. USEPA, Office of Pollution Prevention and Toxics. 1999b. Draft Determining the Adequacy of Existing Data. USEPA, Office of Pollution Pre vention and Toxics and Syracuse Research Corporation. 2004. EPI Suite v 3.12.

US HPV CHALLENGE ANNEX I: DIMETHYL DISULFIDE IUCLID See attached IUCLID documents. ANNEX II: DATA QUALITY ASSESSMENT

DIMETHYL DISULFIDE

201-16767

Available environmental, ecotoxicity, and mammalian toxicity studies were reviewed and assessed for reliability according to standards specified by Klimisch et al., (1997), as recommended by the USEPA (1999a) and the OECD (OECD, 2002). The following reliability classification (Klimisch rating) has been applied to each study assessed:

1 = Reliable without Restriction Includes studies that comply with USEPA- and/or OECDaccepted testing guidelines and were conducted using Good Laboratory Practices (GLPs) and for which test parameters are complete and well documented; 2 = Reliable with Restriction Includes studies that were conducted according to national/international testing guidance and are well documented. May include studies that were conducted prior to establishment of testing standards or GLPs but meet the test parameters and data documentation of subsequent guidance; also includes studies with test parameters that are well documented and scientifically valid but vary slightly from current testing guidance. Also included in this category were physical -chemical property data obtained from reference handbooks, as well as environmental endpoint values obtained from an accepted method of estimation (e.g., USEPAs EPIWIN estimation program); 3 = Not Reliable Includes studies in which there are interferences in either the study design or results that provide scientific uncertainty or in which documentation is insufficient; and, 4 = Not Assignable This designation is used in this dossier for studies that appear scientifically valid but for which insufficient information is available to adequately judge robustness.

Those studies receiving a Klimisch rating of 1 or 2 are considered adequate to support data assessment needs in this dossier. Those key studies selected for inclusion are considered typical of the endpoint responses observed in other studies of a similar nature and design that were identified during our search of the literature.

201-16767A

HPV Challenge Program Submission

Disulfide Oil

Table of Contents

Tables ................................................................................................................................. 3
Figures................................................................................................................................ 3
Appendices......................................................................................................................... 3
Background ....................................................................................................................... 4
Executive Summary .......................................................................................................... 4
1. Introduction ................................................................................................................ 7
2. Physical Chemical Properties.................................................................................. 12
A. B. C. D. E. Melting Point ................................................................................................... 12 Boiling Point .................................................................................................... 13 Vapor Pressure ................................................................................................ 13 Partition Coefficient........................................................................................ 13 Water Solubility .............................................................................................. 14



3. Environmental Fate.................................................................................................. 15
A. B. C. D. Photooxidation................................................................................................. 15 Water Stability ................................................................................................ 15 Biodegradation ................................................................................................ 16 Environmental Distribution ........................................................................... 17


4. Ecotoxicity................................................................................................................. 19
5. Health Effects............................................................................................................ 21
A. B. C. D. Acute Toxicity.................................................................................................. 23 Repeated Dose Toxicity .................................................................................. 24
Mutagenicity.................................................................................................... 26 Reproductive and Developmental Toxicity .................................................. 27

6. Conclusions ............................................................................................................... 28
7. References ................................................................................................................. 34

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List of Tables, Figures, and Appendices


Tables
Table 1. Data Avaialability, Type, and Acceptability for Disulfide Oil ...................... 6
Table 2. Identity and Concentration of the Individual Disulfides in
Disulfidppendeciese Oil ........................................................................................... 10
Table 3. Estimated Physiochemical Constants from EPI Suite ................................. 14
Table 4. Estimated Environmental Fate Parameters from EPI Suite....................... 17
Table 5. Estimated Environmental Distribution from EPI Suite .............................. 18
Table 6. A Comparison of Actual and Estimated Ecotoxicity Values for DMDS.... 20
Table 7. Data Matrix for Disulfide Oil......................................................................... 29

Figures

Figure 1. Flow Diagram for a Merox Unit Producing Disulfide Oil from LPG....... 8
Figure 2. Typical Pathways for the In Vivo Metabolism of Dialkyl Disulfide .......... 21
Figure 3. Mechanism of Redox Cycling and Free Radical Formation from Alkyl
Disulfide Metabolism ..................................................................................... 22

Appendices
Appendix I. Analytical Results: Composition of Disulfide Oil Appendix II. Dimethyl Disulfide Test Plan Appendix III. Dimethyl Disulfide Robust Summaries Appendix IV. IUCLID 5 Report for Disulfide Oil

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Background
An initial data assessment containing reference to disulfide oils, Test Plan for Reclaimed Substances: Streams Containing Naphthenic Acids, Phenolics, Disulfides, Acids or Caustics, was posted to EPAs website on January 20, 2004. This assessment has been revised in response to the EPA and public comment and has been modified so that individual categories or streams of reclaimed substances are addressed separately.

This data review and assessment document examines one member of the originally proposed disulfide category. Originally, it was thought that the disulfide category could be addressed as a technical letter. After more investigation and review of the manufacturing status of the original category members, the Testing Group withdrew sponsorship of three of the substances in the category and determined that a separate assessment on the remaining substance, diethyl and diphenyl disulfide, naptha sweetening (CAS 68955-96-4) was warranted.

Executive Summary
Diethyl and diphenyl disulfide, naphtha sweetening (CAS# 68955-96-4) is primarily composed of low molecular weight dialkyl disulfides that are extracted from C4 to C5 light hydrocarbon streams during the refining of petroleum. The disulfide substance, commonly known as disulfide oil or DSO, can be composed of up to 17 different dialkyl disulfides with alkyl chain lengths no greater than C4. Although the exact composition and concentrations vary depending upon the type of organo-sulfur compounds being extracted, ten disulfides tend to predominate the substance and are representative of the types and amounts of disulfides in DSO.

On the whole, the dialkyl disulfides in DSO constitute a homologous series of chemicals that are perfectly suited for examination using structure-activity analyses (SAR). Although some data are available for DSO, the majority of the testing needs for this substance have been satisfied using SAR and the read across of information available for dimethyl disulfide (DMDS), which is present in DSO in high amounts and is the lowest member of the homologous series. Use of DMDS as a surrogate for DSO in a Page 4

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Disulfide Oil

read across manner is supported by a common mechanism of action that all disulfides exhibit when eliciting harmful systemic effects. This mechanism, which involves the generation of free radical intermediates and the initiation of a redox cycle after an initial disulfide bond cleavage, has been shown to be less active in disulfides that are more highly substituted. Consequently, the toxic potency of dialkyl disulfides decreases as the chain length increases, and the effects observed with DMDS provide a good worst case estimate of the toxicity associated with the remaining members of the series.

In the HPV guidance, the EPA included a provision for the use of SAR to reduce testing needs (USEPA, 1999a). In the guidance, a chemical category is a group of chemicals whose physicochemical and toxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity (USEPA, 1999b). The goal of developing a chemical category is to use interpolation and/or extrapolation to assess chemicals rather than conducting additional testing. It is believed that this analysis provides a good example of how SAR can be effectively used to identify the health hazards associated with structurally similar substances. The advantages afforded by the use of SAR and a read-across extrapolation from DMDS to DSO eliminates the need for redundant testing of a substance that is not released to the environment nor found in the marketplace.

As summarized in Table 1, adequate data are believed to exist for DSO in eighteen of the twenty test categories examined. These testing needs were filled either by actual testing of DSO, by the use of SAR programs and techniques, or by analogy with DMDS, which has previously been reviewed under the HPV Challenge Program. As such, the robust summary for DMDS has been included with this submission. The test areas where DMDS, and by analogy DSO, lack adequate information (water stability and chronic fish toxicity) are scheduled to be filled under voluntary agreement with the HPV sponsor for DMDS. In conclusion, evidence is available showing that the health and environmental hazards associated with DSO has been sufficiently evaluated and no further testing is deemed necessary for this material.

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Table 1. Data Availability, Type, and Acceptability for Disulfide Oil Information Available Results Acceptable Y/N
Y Y Y Y Y Y N Y Y Y N Y Y Y Y Y Y Y Y Y

Estimated Value

DSO Category

Y/N
Physiochemical melting point boiling point vapor pressure partition coefficient water solubility Environmental Fate photooxidation water stability biodegradation distribution Ecotoxicity acute fish chronic fish acute invertebrate acute algae terrestrial Toxicity acute (oral) acute (dermal) acute (inhalation) repeated dose mutagenicity reproductive/developmental Y Y Y Y Y Y Y N Y Y Y Y N Y Y Y Y Y Y Y

Y/N

Y/N

Y/N

Y/N

N N N N N

N N N N N

Y Y Y N Y

N N N Y N

N N N N

N N N N

N N N N

Y N Y Y

N N N N N

N N N N N

N N N N N

Y Y Y Y Y

N N N Y Y Y

Y Y Y N N N

N N N N N N

N N N Y Y Y

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Testing Necessary Y/N


N N N N N N N N N N N N N N N N N N N N

OECD Study

Other Study

GLP Study

HPV Challenge Program Submission

Disulfide Oil

1. Introduction
The High Production Volume Challenge Program has identified diethyl and diphenyl disulfides, naphtha sweetening (CAS# 68955-96-4) as a candidate category based on production volume estimates obtained through the TSCA Inventory Update Rule.

Commonly known as disulfide oil or DSO, this substance is produced by a single company as a byproduct of the petroleum refining process. The substance is not sold commercially nor is it used directly in any downstream products. DSO is a product of mercaptan removal from selected C4 to C5 light hydrocarbon streams by a process known as sweetening, since it removes the sour smelling sulfides from crude petroleum. The mercaptans are extracted from this feedstock in an entirely closed system referred to as a Merox unit, which can be designed to operate with any of a variety of petroleum streams including liquefied petroleum gas (LPG), naphtha, or any other hydrocarbon fraction (see Figure 1).

The Merox unit uses a basic solution of caustic soda as the extracting solvent, which is recycled and reused in a continuous loop following each use. Once removed, the mercaptans are oxidized to disulfides, which are separated from the caustic soda solution. The final disulfide oil is then either disposed of on site or processed as: i) an internal fuel, ii) a feedstock for sulfuric acid production, or iii) an agent for conditioning refinery catalysts.

The initial step in the extraction sequence is depicted by the following reaction equation, with R representing a short chain alkyl group: RSH + NaOH NaSR + H2O The second step is referred to as regeneration and it involves heating and oxidizing the caustic solution leaving the extractor. The oxidation converts the extracted alkyl mercaptans (RSH) to organic disulfides (RSSR), which are less water soluble than

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the initial mono-sulfides thereby facilitating separation and removal from the aqueous caustic solution. The reaction that takes place in the regeneration step is: 4 NaSR + O2 + 2 H2O 2 RSSR + 4 NaOH Figure 1. Flow Diagram for a Merox Unit Producing Disulfide Oil from LPG

The net overall Merox reaction covering the extraction and regeneration steps may be expressed as: 4 RSH + O2 2 RSSR + 2 H2O After decantation of the disulfide oil, the regenerated caustic solution is recirculated back to the top of the extractor in a continuous loop to extract additional mercaptans. Extraction equilibrium is favored by lower molecular weight mercaptans Page 8

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and by lower temperatures. Consequently, the disulfide oil is generally rich in alkyl disulfides with small chain lengths, but the exact chemical composition can vary depending on types of sulfur contaminants in the treated feedstock.

The compositional information in Table 2 was extracted from a recently completed chemical analysis of DSO (Appendix I) and is representative of the types of disulfides found in DSO. The analysis reveals that ten dialkyl disulfides comprise approximately 87% of the total weight in disulfide oil. These disulfides range in molecular weight from 94 to 150 amu and are remarkably similar in chemical structure with each possessing a characteristic disulfide linkage attached to a C1 to C4 alkyl group. Despite the official nomenclature, DSO does not contain appreciable amounts of diphenyl disulfides. The full analytical report presented in Appendix I shows that less than 0.5% of the oil is composed of hydrocarbon solvents and that the balance is composed of low molecular weight mono and trisulfides that generally comprise less than 2% of the total weight percentage. The exception is diisopropyl sulfone, which is present at levels of about 5% by weight. Because sulfones of this type have been shown to lie in the metabolic pathways for dialkyl disulfides (see Health Effects, section 5), its presence in DSO at relatively high amounts does not pose any particular toxicological concern and it can be assumed to act in the same fashion as the disulfide from which it is derived. The benzene levels in DSO have been reduced in recent years and are currently present at concentrations less than 0.1%. Past samples used in the acute toxicity testing performed over fifteen years ago contained benzene levels up to 1.0%.

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Table 2. Identity and Concentration of the Individual Disulfides in Disulfide Oil


Disulfide Constituent Chemical Structure
H3C S S CH3

CAS Number

Chemical Formula

Mol. Wt.

Conc. DSO (% w/w)

dimethyl disulfide methyl ethyl disulfide methyl isopropyl disulfide diethyl disulfide methyl n-propyl disulfide

624-92-0

C2H6S2

94.22

12.0

H3C S
H3C S S CH3 H3C
H3C S S CH3

S CH3

20333-39-5

C3H8S2

108.25

18.2

40136-65-0

C4H10S2

122.28

14.4

110-81-6

C4H10S2

122.28

11.2

H3C S S CH3

2179-60-4

C4H10S2

122.28

7.7

H3C

ethyl isopropyl disulfide ethyl n-propyl disulfide


H3C

S CH3 H3C

53966-36-2

C5H12S2

136.31

11.6

S CH3

30453-31-7

C5H12S2

136.31

7.0

CH3

diisopropyl disulfide

H3C S S CH3 H3C


H3C

4253-89-8

C6H14S2

150.34

2.0

ethyl n-butyl disulfide


H3C

S S CH3

63986-03-8

C6H14S2

150.34

0.5

dipropyl disulfide

S CH3

629-19-6

C6H14S2

150.34

2.5

Total

87.1

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Consistent with published guidelines for identifying and establishing a categorical approach for a chemical mixture, DSO is deemed to meet all of the requirements for consideration as a chemical category. The hallmarks of the DSO series are: i) the regular and predictable fashion in which the alkyl groups affect physical properties and environmental attributes and ii) the pivotal role played by the disulfide bridge in eliciting a toxic response. As such, to the extent possible, information has been assembled for all ten of the disulfide constituents. When information was unavailable, however a surrogate approach has been applied. Since the DSO is essentially a homologous series of chemicals with dimethyl disulfide (DMDS) occupying the lowest position, structureactivity methods provide an acceptable approach for evaluating the properties and fulfilling the testing needs of the entire category. DSO is also well suited for the application of a read across approach for predicting the health and environmental impacts of DSO, where modeling might not meet data needs. To the extent possible, information has been assembled for the primary disulfide constituents of DSO. When, structure-activity data is absent or missing, the effects of DMDS are offered as a reasonable alternative to testing the entire category. This is a well-reasoned decision that was heavily influenced by a common mechanism of action for the environmental and mammalian toxicity of dialkyl disulfides and by systematic knowledge of the impact of carbon chain length on the toxic potency of disulfides. Because DMDS is a well studied chemical that has previously been examined under the HPV Challenge Program, the available test data provide a source of surrogate information for DSO. An examination of the test plan (Appendix II) and robust summary for DMDS (Appendix III) reveal that it has few data deficiencies and those gaps that exist are scheduled to be addressed under a voluntary agreement recently approved and accepted by the US EPA (USEPA, 2008). The data presented in this data assessment for DSO and DMDS were identified either in company proprietary files, peer-reviewed literature, the DMDS IUCLID data set, and/or calculated using accepted computer modeling programs. A robust summary has been prepared in IUCLID 5 format (Appendix IV) that describes the data used in support of this submission on DSO. All data were evaluated for study reliability in accordance with criteria outlined by Klimisch

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et al., (1997) and recognized by the USEPA. Only studies that met the reliability criteria of 1 (reliable without restrictions) or 2 (reliable with restrictions) are presented. In some cases, test data has been extracted from MSDSs because the original reports were either inaccessible or unavailable. These data were judged to be reliable with restriction for the purposes of this data review based on their plausibility and recent release.

2. Physical Chemical Properties


DSO is an extremely flammable substance with a relatively high vapor pressure and low water solubility. At room temperature, the material exists as a yellow liquid with an extremely foul and obnoxious odor. The physical and environmental properties for the individual DSO disulfides have been estimated using EPAs EPI Suite software package and are presented in Table 3 (USEPA, 2007). To assess their validity, the physical property estimates for the individual disulfides were averaged using the additivity rule for ideal mixtures, which provided a rough but effective approximation for comparing the actual measured values for DSO with a calculated estimate. Experimental data was located in the published literature for three of the ten component disulfides in DSO. As shown in Table 1, approximately 64% of DSO is composed of five dialkyl disulfides with an alkyl carbon number of C4 or less. Consequently, the chemical and physical properties associated with these disulfides will exert a disproportionate impact on the properties of the substance.

A. Melting Point
Estimated melting points for the disulfides in DSO were derived using the MPBPWIN (v 1.42) module in the EPI Suite program. Table 3 shows that the estimated values follow a regular progression as a function of carbon number, with the melting points increasing as the carbon content rises. Actual experimental values were located for three chemicals: DMDS, diethyl disulfide (DEDS), and dipropyl disulfide (DPDS). A comparison of the actual measurements against the predicted values for these three chemicals show reasonable agreement with some tendency for the estimation routine to over predict the actual value (predicted values of -69.7, -45.2, and -21.8 C for DMDS, DEDS, and DPDS, respectively). A fractionally-weighted compositional average was

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calculated using the estimated values for all ten disulfides, which were then compared to the actual value for DSO. The fractionally-weighted average of -44.3 C compares well with actual DSO value of -54 C (-65 F) (ST Laboratories, 2008).

B. Boiling Point
Boiling points were estimated using the same software module used to estimate melting points. The predicted values ranged from 100 to 200 C and increased in a direct relationship to molecular weight. Estimated values for DMDS, DEDS, and DPDS show good agreement with the actual measurements (109.8, 154.1, and 193.5 C for DMDS, DEDS, and DPDS, respectively), differing by only a few degrees. The weighted average for the estimated boiling points of all ten disulfides was 131.3 C, which is consistent with the reported boiling point range of 111-174 C for DSO (ST Laboratories, 2008).

C. Vapor Pressure
The ten disulfides in DSO display a considerable range in volatility. Using the MPBPWIN (v 1.42) module, the vapor pressure was estimated to range from 24.5 mmHg for DMDS to 0.50 mmHg for DPDS. These values are in excellent agreement with the actual measured values for these two compounds. The Reid vapor pressure of DSO was determined to be 1.1 psia at 100 F (37.7 C) (ST Laboratories, 2008). This pressure is approximately equal to a true vapor pressure of about 1.1 psi or 57 mmHg at 25 C. By comparison, the fractionally-weighted vapor pressure for the disulfides in DSO was calculated to be 5.87 mmHg at 25 C. The difference between the two values is likely due to the relatively high volatility of the non-disulfide chemicals in DSO and their disproportionate contribution to the overall volatility of the substance.

D. Partition Coefficient
Octanol/water partition coefficients were estimated using the KOWIN (v 1.67) module within EPI Suite. The values in Table 3 are generally similar for all ten disulfides and show no more than a two-fold range in variation from the lowest (DMDS) to highest (DPDS) members of the series. The estimated log Kow value of 1.87 for DMDS agrees well with the actual measured value of 1.77. The fractionally-weighted average value of 2.40 for all ten disulfides was not appreciably different from the value for DMDS, which supports the use of this chemical as a surrogate for the entire blend. Page 13

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E. Water Solubility
The disulfides in DSO show a relatively large range in water solubility. Using the WSKOW (v 1.41) subroutine in EPI Suite, water solubility estimates of 3.74 g/L (DMDS) to 0.04 g/L (DPDS) were calculated. The actual experimental value of 2.5 g/L for DMDS shows good agreement with the estimated value of 2.9 g/L. The fractionallyweighted average of 0.80 g/L for the ten disulfides is also reasonably close to a measured value that was less than 0.01% by weight (<0.1 g/L) for the solubility of DSO in water (ST Laboratories, 2008). Table 3. Estimated Physiochemical Constants from EPI Suite
Melting Point (C) Boiling Point (C) Vapor Octanol/Water Pressure Partition Coeff. (mmHg 25 C) (log Kow) 1.87 (1.77*) 2.36 2.78 2.86 2.86 3.27 3.35 3.76 3.84 3.84 1.77# Water Solubility (g/L) 3.7 (2.5*) 1.06 0.41 0.36 0.36 0.14 0.12 0.05 0.04 0.04 <0.1

Disulfide

dimethyl disulfide methyl ethyl disulfide methyl isopropyl disulfide diethyl disulfide methyl n-propyl disulfide ethyl isopropyl disulfide ethyl n-propyl disulfide diisopropyl disulfide ethyl n-butyl disulfide dipropyl disulfide DSO

-69.7 (-85*) 113.6 (109.8*) 24.5 (21.98*) -57.3 -56.6 136.7 145.6 7.40 4.92 3.31 (4.28) 2.65 1.77 0.96 0.64 0.35 0.50 (0.51) 57

-45.2 (-102) 158.8 (154.1) -45.2 -44.6 -33.4 -32.9 -21.8 158.8 167.4 180.1 188.2 200.4

-21.8 (-86) 200.4 (193.5) -65 111-174

* Actual measured value taken from DMDS test plan (2005). Actual measured value taken from EPIWIN Suite v 3.20 (USEPA 2007).
Actual reported or converted value taken from a certificate of analysis (ST Laboratories, 2008).
# Estimated value.

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3. Environmental Fate
The environmental fate of DSO has not been examined; however, structureactivity information and suggestive anecdotal test data is available for DMDS and the remaining disulfides in the mix. Many of the disulfides in DSO are naturally found in the environment either as ingredients in vegetables, especially garlic and onions, or as products of the microbial oxidation of assimilated mercaptans (TGSC, 2008). Preliminary studies with DMDS and DPDS have shown that these two disulfides are relatively stable in soil and water (Arnault et al., 2004). DMDS, in particular, has been found in many environmental compartments and is considered to have an integral role in the global sulfur cycle (Caron and Kramer, 1994). Natural background concentrations of DMDS have been measured in a wide variety of media including air, surface waters, sediment, wastewater effluent, vegetation, and expired human air (HSDB, 2005). Interestingly, DMDS has been shown to be absorbed from air into moist and dry soils at a rate that was influenced by the presence of soil microbes, which facilitated the uptake into moist soil only (Bremner and Banwart, 1976). This may be an important environmental process for the disulfides in DSO because of their tendency to partition into the soil compartment.

A. Photooxidation
The atmospheric photodegradation of the disulfides in DSO was estimated using the AopWIN (v 1.92) subroutine in the EPI Suite program. As shown in Table 4, the rate of tropospheric photooxidation by reaction with hydroxyl radicals is nearly identical for the ten disulfides in DSO. The atmospheric half-life of each disulfide is approximately 30 min, which meets the definition of a rapidly removed VOC. The estimated rates of DMDS hydroxyl radical reactivity also compared well with the actual value (0.56 versus 1.2 hr).

B. Water Stability
None of the disulfides could be evaluated for aqueous stability because the HYDROWIN algorithm has only been validated for use with a limited number of chemical classes. Available information for DMDS indicates, however, that aqueous hydrolysis at ambient temperature is too slow to be an important environmental fate Page 15

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process when the pH is less than 12 (Bentvelzen et al., 1975). This conclusion is consistent with the relative stability of the disulfide bridge to acid base hydrolysis and reported claims that DMDS slowly hydrolyzes to non-volatile methane sulfinic acid in water at pH 11-12. In addition, voluntary testing of the aqueous stability of DMDS has been agreed to in a previously submitted test plan for this chemical and the information will provide a reasonable surrogate for the water stability of DSO.

C. Biodegradation
The biodegradability of the ten DSO disulfides was examined using the BIOWIN (v 4.00) subroutine in the EPI Suite program. The BIOWIN routine uses eight different methods to evaluate the biological degradation of a target chemical under either aerobic or anaerobic conditions. Although several of the methods suggest that the probability of disulfide biodegradation is relatively high, it is believed that the most reliable information comes from the results for DMDS itself and from those models indicating a lack of ready biodegradability (see Table 4). A closed bottle ready biodegradability test performed with DMDS indicated that less than 10% of the material was degraded over a 28-day period (ELF ATOCHEM, 1995). Ready biodegradability, as defined in accordance with OECD guidelines, only occurs when at least 70% of a chemical is biologically removed from the environment within the 28-day period. Accordingly, DSO is expected to fail the biodegradability test and these conclusions are in agreement with actual test data for DMDS.

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Table 4. Estimated Environmental Fate Parameters from EPI Suite


Photo oxidation (KOH t hrs) 0.56 (1.2*) 0.55 0.53 0.54 0.54 0.52 0.53 0.51 0.53 0.52 1.2# Ready Biodegradation Probability linear non-linear 0.43 0.44 0.30 0.45 0.45 0.31 0.46 0.31 0.46 0.46 0.43# 0.46 0.47 0.26 0.47 0.47 0.26 0.48 0.27 0.49 0.49 0.46#

Disulfide

Water Stability ND ND ND ND ND ND ND ND ND ND ND

Readily Biodegradable no (no) no no no no no no no no no no#

dimethyl disulfide methyl ethyl disulfide methyl isopropyl disulfide diethyl disulfide methyl n-propyl disulfide ethyl isopropyl disulfide ethyl n-propyl disulfide diisopropyl disulfide ethyl n-butyl disulfide dipropyl disulfide DSO

* Actual measured value taken from Finlayson-Pitts and Pitts (2000). Actual measured degradation of 10% over 28-days (DMDS test plan, 2005).
Not determined.
# Estimated value.

D. Environmental Distribution
The environmental distribution of the composite disulfides in DSO is presented in Table 5. The estimated percent distribution in the four environmental media were determined using a Level III multi-media media fugacity model (LEV3EPI) imbedded within the EPI Suite software package and based on the work of Mackay et al. (1996). A level 1 fugacity analysis performed using the EQC (Equilibrium Criterion Model, v2.02) revealed that virtually 100% of each disulfide distributed to the air compartment, which is inconsistent with known partitioning behavior of DMDS in the environment (Farwell et al., 1979; Richards et al., 1991). All ten disulfides show a preference for water or soil with the distribution shifting from water to soil as the dialkyl carbon number increases

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from C2 to C6. The tendency for the disulfides to concentrate in soil warranted an evaluation of terrestrial effects in the following ecotoxicity section of the document.

The estimated half-life for all ten disulfides was identical with values of 1.1 hr, 360 hr, 720 hr, and 135 days for air, water, soil, and sediment, respectively. Except for sediment, which was not identified as a major disulfide reservoir, these half-life estimates do not indicate environmental persistence in any media. The overall persistence in the environment ranged from 119 to 350 hrs and the fractionally-weighted additive contribution for all ten disulfides in DSO was calculated to be 184 days.

Table 5. Estimated Environmental Distribution from EPI Suite Overall Persistence (hrs)
119 160 206 220 221 275 290 338 350 350 119-350#

Disulfide air
dimethyl disulfide methyl ethyl disulfide methyl isopropyl disulfide diethyl disulfide methyl n-propyl disulfide ethyl isopropyl disulfide ethyl n-propyl disulfide diisopropyl disulfide ethyl n-butyl disulfide dipropyl disulfide DSO
#

Environmental Distribution (%) water soil sediment


58.1 41.9 31.9 29.7 29.7 23.3 22.1 18.7 18.1 18.1 18.1-58.1# 40.8 57.2 67.3 69.4 69.5 75.7 76.9 79.6 80.0 80.0 40.8-80.0# 0.2 0.2 0.4 0.4 0.4 0.7 0.7 1.4 1.6 1.6 0.2-1.6#

1.0 0.7 0.5 0.5 0.5 0.3 0.3 0.2 0.2 0.2 0.2-1.0#

Estimated value.

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4. Ecotoxicity
Evidence suggests that the aquatic and terrestrial toxicity of DSO mimics the effects observed with DMDS. Initial modeling of DMDS and the remaining disulfide constituents of DSO using EPAs ECOSAR software package (Meylan and Howard, 1998) reveled that the ecotoxicity of the disulfides increased as a function of alkyl chain length. Although this finding is consistent with the observed increase in octanol/water partition coefficients for these disulfides, the results are inconsistent with available test data and knowledge of disulfide metabolism. The modeling results, therefore, have not been utilized since the assumed mode of action, non-polar narcosis, is most likely incorrect, a condition that often occurs when this endpoint is invoked indiscriminately (de Roode et al., 2006).

The underpinnings for the SAR routines used in the ECOSAR program assume that non-polar narcosis is the operant mode of action for the disulfides; but this class of chemicals is not explicitly represented in the training sets used to develop the mathematical relationships. In fact, disulfides are more likely to operate in terrestrial and aquatic organisms by the same mode of action observed in mammals, which involves disulfide bond cleavage and redox cycling of the free radical intermediates (Mnchberg et al., 2007; Lesser, 2006). The reactive oxygen species produced in this reaction can lead to oxidative stress and protein interactions that are typically more severe and less consistent across species than those elicited by narcotic chemicals (Jager et al., 2007). This lack of applicability is evident when test data for DMDS are compared to the estimates obtained using ECOSAR (see Table 6). The toxicity of DMDS is generally under predicted by a factor 10-200 fold, which signals that a mode of action other than narcosis is in effect.

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Table 6. A Comparison of Actual and Estimated Ecotoxicity Values for DMDS


Estimated Toxicity (mg/L) 92.51 11.67 98.24 60.96 635.4 Actual Toxicity (mg/L) 0.97* - 7 35# 32*

Ecotoxicity Endpoint Acute Fish 96-hr LC50 Chronic Fish 30-day Acute Invertebrate 48-hr EC50 Acute Plant 96-hr EC50 Earthworm 14-day LC50

* Actual measured value taken from Arkema, Inc. (2007). Actual measured value taken from ELF ATOCHEM (1996). # Additional test results for algae (ErC50, EbC50, NOECr, and NOECb) are available (ELF ATOCHEM, 2000).

Additional support for the use of DMDS as a surrogate for the disulfides in DSO comes from available test data for higher homologs in the series. When the acute toxicity of DMDS to fish (0.97 mg/L) is compared to the LC50 results obtained with DEDS (7.43 mg/L), DPDS (2.62 mg/L), and diisopropyl disulfide (8.31 mg/L), there is no apparent increase in toxicity as a function of chain length (NITE, 2003; Chevron Phillips Chemical Company, 2005; Russom et al., 1997). In addition, the 24-hr EC50 value for DEDS (14.5 mg/L) in Daphnia magna is nearly 2-fold greater than the 48-hr value for DMDS (7 mg/L) (Glli et al., 1994). Taken together, these data are consistent with the expected change in potency for the oxidative stress caused by disulfide bond cleavage (see section 5), and confirms that DMDS is the most toxic member of the disulfide series in DSO. Additional testing with DSO is not expected to result in effect concentrations less than those observed DMDS, and therefore no further testing can be justified for the endpoints listed. Despite the lack of DMDS test data for chronic fish toxicity, testing will not be performed within the context of this submission because it will be completed in conjunction with the previously submitted test plan for DMDS. When this voluntary testing is completed, a full complement of ecotoxicity data will be available for DMDS and by analogy DSO.

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5. Health Effects
Sufficient information is available to make reliable and sensible determinations of the health effects of DSO. Whereas some test data is available on the oil itself, the majority of information can be extracted from the robust summary and test plan for DMDS (DMDS Robust Summary, 2005). The rationale and justification for using the health effects data of DMDS as a substitute for the disulfides in DSO are based on sound scientific principles and a plethora of mechanistic information showing that all of the dialkyl disulfides in DSO operate through a common toxic mechanism. This mechanism, which has been well studied and clearly elucidated in the published literature, focuses on the unique characteristics of the disulfide bridge and the ease with which free radical intermediates can be formed once the bridge is cleaved.

Figure 2. Typical Pathways for the In Vivo Metabolism of Dialkyl Disulfides


S S CH3 H3C SH

H3C

H3C

dipropyl disulfid e

propyl mercaptan

R = glutathione

O H3C S S CH3

H3C

CH3 S

propyl methyl sulfide

dipropyl thiosulfinate
CH3 S O O H3C S S O CH3

H3C

propyl methyl sulfoxide

dipropyl sulfone

O H3C S O CH3

propyl methyl sulfone

The metabolism of many, if not all, disulfides is initiated by a thiol-disulfide exchange reaction that substitutes the sulfhydral group of glutathione for a mercaptide

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fragment within the disulfide molecule. This reaction is depicted in Figure 2 for DPDS, whose in vivo metabolism has been examined in the greatest detail of the ten DSO disulfides (Germain et al., 2008; Teyssier and Siess, 2000). Evidence shows that this same initial glutathione exchange reaction also takes place for a host of alkyl, alkenyl, phenyl, and branched chain disulfides (Bach et al., 2008; Munday and Manns, 1994; Munday, 1989; Nishikawa et al., 1987). Using an expert knowledge based system for predicting the metabolic reactions that take place in vivo (Meteor, v 9.0.0), the disulfides in DSO were all predicted to undergo reductive cleavage of the disulfide bond with a high degree of probability (Greene et al., 1999).

The exchange reaction with glutathione is catalyzed by a thioltransferase, also known as glutaredoxin, which is widely distributed in nature and shows high levels of activity in the tissues and organs affected by alkyl disulfide toxicity (Lillig and Holmgren, 2007). This reaction is the key step in the toxic mechanism for dialkyl disulfides. The activation mechanism has been associated with the initiation of a redox cycle that generates excessive quantities of highly reactive free radical intermediates that are capable of interacting with tissue macromolecules at or near the site where they are formed. In some cases this site has been the red blood cell and in other cases the liver depending on the species examined (Munday, 1989). The sequence of reactions in the redox cycling of alkyl disulfide is depicted generically in Figure 3. The first product of the initial thioltansferase exchange reaction is an alkyl mercaptan that undergoes a oneelectron oxidation to a free radical intermediate following ionization. This intermediate is the proximal toxicant, responsible for producing a continuous supply of hydroxyl radicals and other reactive oxygen species that can sustain the redox cycling, cause oxidative stress, and precipitate tissue injury at sites where it is formed.

Importantly, the reactivity of the mercaptans formed in the exchange reaction is directly affected by the length and branching pattern of the attached alkyl substitutents, with longer chain lengths leading to reduced radical stabilization and lower oxidation rates (Munday, 1989). In addition, the reactivity and toxicity of alkyl disulfides has been shown to decrease in the following order n > sec > tert due to the influence of steric

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factors on thioltransferase activity. These data indicate that DMDS will be the most reactive member of the series with the longer chain lengths and higher branching patterns of the remaining homologs ameliorating the toxicity by affecting the rate of formation and ultimate stabilization of the free radical intermediates. This fact provides strong justification for the use of DMDS as a surrogate for the higher chain length disulfides in DSO and validates its use in a read across transfer to other disulfides in the category. The test data for DMDS is therefore offered as a reasonable and mechanistically supportable substitute for DSO, since it represents the most toxic member of the disulfide series. As such, the existing information on DSO and the disulfide category is deemed sufficient, and no further testing is needed nor required to assess the health hazards associated with this category of reclaimed substances.

Figure 3. Mechanism of Redox Cycling and Free Radical Formation from Alkyl Disulfide Metabolism (Munday and Manna, 1994)

2 GSH + RSSR GSSG + 2 RSH RSH RS + H+ (Hb)Fe3O2 + RS + 2H+ (Hb)Fe3 + RS + H2O2


RS + RS (RSSR)
(RSSR) + 02 RSSR + 02
RSH + 02+- H+ RS + H2O2

A. Acute Toxicity
Oral, dermal, and inhalation studies have all been performed with DSO and the results are described in detail in the robust summaries presented in Appendix IV. The oral LD50 value was 1590 mg/kg in female rats and 1700 mg/kg in males (FurediMachacek, 1991a). Gross necropsy on dead and moribund animals revealed intestines filled with red fluid and tan-colored lungs. Darkly colored spleens were noted upon sacrifice of all female rats, with all animals displaying enlarged spleens. In an initial acute oral screening LD50 study on the same material, both female and male rats were

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administered 5000 mg/kg, after which all the animals died (Furedi-Machacek, 1991b). The 4-hr inhalation LC50 value was found to be greater than 4.84 mg/L in male and female rats (Drummond, 1991). The dermal LD50 value was greater than 1800 mg/kg in rabbits (Furedi-Machacek, 1991c). Mild to moderate irritation was observed in a Draize rabbit skin test and the same material was determined to be minimally irritating in rabbit eyes (Furedi-Machacek, 1991d,e). It was negative in a guinea pig sensitization test (Furedi-Machacek, 1991f).

Comparable studies with DMDS revealed an oral LD50 value for rats of 190 mg/kg (Penwalt, 1985a), a dermal LD50 value for rabbits that was greater than 2,000 mg/kg (Penwalt, 1985b), and a 4-hr inhalation LC50 value for rats of 805 ppm (3.10 mg/L) (Tansy et al., 1981). These data suggest that DMDS is more toxic than DSO. By comparison, a single rat oral LD50 value of greater than 2000 mg/kg has been reported for DPDS (Chevron Phillips Chemical Company, 2005). Some disulfides, in particular DMDS and DPDS have been shown to cause mild to severe red blood cell hemolysis in cats, dogs, and a variety of livestock animals following oral ingestion (Gruhzit, 1931; Munday, 1989). Interestingly, vegetables containing relatively high amounts of these and other disulfides have long been associated with hemolytic anemia following accidental or intentional ingestion by dogs and farm animals (Munday and Manns, 1994; Yamato, et al., 2005). Rats, however, are more resistant to dialkyl, but not diaryl, disulfide induced hemolytic damage (Munday and Munday, 2003).

B. Repeated Dose Toxicity


No studies have been reported on the repeat dose effects of disulfide oil, but studies are available for both DMDS and DPDS. DMDS was examined in five separate, well-designed, oral, dermal, or inhalation studies. In the first inhalation study, summarized in the IUCLID dataset for DMDS, male and female rats were exposed to 10, 50, 150, and 250 ppm (0.04, 0.19, 0.58, and 0.96 mg/L) DMDS for 6 hr/day for 90 days (ELF ATOCHEM, 1992). Findings included decreases in body weight and food consumption, reduced thymus gland weights, and increased liver weights. Possible reductions hemoglobin, red blood cell count, and packed cell volume were observed at the highest concentration. Histopathological changes were noted in the nose and spleen. Page 24

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Treatment-related changes in alanine aminotransferase, alkaline phosphatase and total bilirubin indicated some degree of liver involvement. The NOAEL for this study was 10 ppm (0.04 mg/L). In the second inhalation study, rats were exposed for 13 weeks to 5, 25, or 125 ppm (0.02, 0.10, or 0.48 mg/L) DMDS for 6 hr/day (Kim et al., 2006). A treatment-related decrease in body weight gain, food consumption, and thymus weight were observed along with an increase in adrenal gland weight. Histopatholgy did not reveal any increase in the incidence or severity of abnormal tissue alterations relative to controls. Statistically significant decreases were also noted in serum aspartate aminotransferase, blood urea nitrogen, and creatine phosphokinase levels. The NOAEL was 5 ppm (0.02 mg/L) for male rats and 25 ppm (0.10 mg/L) for female rats.

The two dermal studies were performed in male and female New Zealand rabbits treated with DMDS for 6 hr/day by applying the neat material under an occlusive bandage (DMDS Robust Summary, 2005). In the first range-finding study, animals treated with DMDS levels of 0.1, 0.5, or 1 mL/kg/day (106, 505, or 1063 mg/kg/day) for 14 days caused dose-related lethargy or unconsciousness in all treatment groups that dissipated by the end of the day (ELF ATOCHEM, 1989). Severe treatment-related skin lesions were also observed in all three treatment groups. The NOAEL and LOAEL for this study were determined to be less than 106 mg/kg/day and 106 mg/kg/day, respectively. In the second study, the rabbits were treated dermally at levels of 0.01, 0.1, or 1 mL/kg/day (10.6, 106.3, or 1063 mg/kg/day) for 28 days (ATOCHEM, 1989a). Consistent with the range-finding studies, dose-related changes in lethargy and skin irritation were also observed in the more prolonged study. After 13 days, mortality was observed in the rabbits in the high dose group and treatment was terminated in this dose group. The male rabbits in the high dose group also displayed some abnormal changes in hematology and clinical chemistry measurements that were not observed in the female rabbits. Histopathologic examination and organ-weight measurements failed to reveal any treatment-related changes in the adrenals, brain, heart, kidneys, liver, lungs, ovaries, testis, thyroid, or thymus. The NOAEL for systemic effects was 10.6 mg/kg/day and the NOAEL for localized dermal irritation was less than 10.6 mg/kg/day.

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Finally, a 90-day oral feeding study with DPDS failed to show any toxic effects following the dietary administration of 7.3 mg/kg/day or 8.2 mg/kg/day to male or female rats, respectively (Posternak et al., 1969). Food consumption and body weights were recorded weekly and hematological examinations and blood urea nitrogen measurements were performed on half the animals at 7 weeks and on all animals at 13 weeks. A slight non-statistical increase in blood urea nitrogen was observed at end of the study. The organ weight measurements, gross examinations, tissue histopathology performed at necropsy failed to show any treatment-related effects. The dietary NOAELs for DPDS can be roughly equated to inhalation values of 3.1 and 4.6 ppm using standard route-to route extrapolation techniques that assume 100% absorption by both routes (Rennen et al., 2004). Recognizing that only a single dose was administered in the DPDS study, the route conversion still provides a consistency check for the toxic potency of the lowest and highest disulfide members of the DSO category. Since the threshold for DPDS toxicity is essentially equal to, and quite possibly higher than, the threshold value DMDS, the data are consistent with the argument that DMDS is the most toxic member dialkyl disulfide series. In fact, the World Health Organization has determined that the information on DPDS oral toxicity was sufficient for use as a surrogate for evaluating the food safety of a host of related alkyl and allyl disulfides (WHO, 2000).

C. Mutagenicity
Although there are no results available for DSO, DMDS has been examined in a variety of in vivo and in vitro genetic toxicology screening assays (DMDS Robust Summary, 2005). The test results revealed that DMDS was negative in bacterial mutagenicity assays (Penwalt, 1985c), negative in mammalian mutagenicity tests (ELF ATOCHEM, 1990a), negative for DNA damage and repair (ELF ATOCHEM, 1990b), and ambiguously positive in a chromosomal aberration study using human lymphocytes (ELF ATOCHEM, 1990c). Except for the DNA damage and repair assay, these tests were all performed in the presence and absence of metabolic activation. Similarly, negative results were obtained when DMDS was evaluated in vivo in a mouse micronucleus assay at inhalation concentrations of 250 and 500 ppm (ATOCHEM, 1989b), and did not cause unscheduled DNA synthesis in the hepatocytes of rats exposed to 500 ppm (ATOCHEM, 1990). By comparison, DPDS did not cause any reverse Page 26

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mutations in an Ames S. typhimurium assay using strain TA98 (Tsai et al., 1996). None of the disulfides in DSO were judged to be genotoxic by an expert knowledge based system used to predict the health effects of untested chemical substances (Derek, v 9.0.0) (Greene et al., 1999).

D. Reproductive and Developmental Toxicity


Although no studies have been reported on the reproductive or developmental toxicity of disulfide oil, studies performed with DMDS are offered as a reasonable surrogate for the disulfides in DSO. An evaluation of developmental effects was examined in a series of inhalation exposure studies performed in rats with DMDS (DMDS Robust Summary, 2005). In an initial range finding study, pregnant dams were exposed for 6 hr/day on days 6 through 15 of gestation to DMDS concentrations of 10, 50, or 250 ppm (0.04, 0.19, or 0.96 mg/L) (ATOCHEM, 1991a). Treatment-related reductions in body weight gain and food consumption were observed in all treatment groups, but pregnancy incidence, intrauterine death incidence, pre-implantation loss, litter size, sex ratio, and the incidence of malformations were all within the expected range. Mean fetal weights showed an exposure-related reduction in all treatment groups that was considered to be an equivocal finding. The maternal NOAEL was determined to be less than 10 ppm (0.04 mg/L).

In a more detailed study, three groups of 30 mated female rats were exposed to DMDS by whole body exposure at 5, 15 or 50 ppm (0.02, 0.06, or 0.19 mg/L) for 6 hours daily from day 6 to day 15 of gestation (ATOCHEM, 1991b). A similar group of 30 rats, exposed to filtered air only over the same period, served as controls. All animals were maintained until day 20 of gestation, and then sacrificed. No deaths were observed or unusual lesions were observed, but a higher incidence of rough hair coat was seen at 50 ppm (0.19 mg/L). Clinical condition at 5 and 15 ppm (0.02 and 0.06 mg/L) did not differ from controls. Treatment-related reductions in weight gain were observed at 15 and 50 ppm (0.06 and 0.19 mg/L). Food intake was lower than controls at 50 ppm (0.19 mg/L), but comparable at 5 or 15 ppm (0.02 and 0.06 mg/L). There was no effect of treatment on pre or post-implantation loss, litter size or sex ratio. Maternal toxicity was noted at 15 and 50 ppm (0.06 and 0.19 mg/L), but there was no evidence of developmental effects. Page 27

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Litter and fetal weights were reduced at 50 ppm (0.19 mg/L). At 5 and 15 ppm (0.02 and 0.06 mg/L) these parameters were comparable to controls. No malformations were observed in fetuses from the treated groups. A slightly higher incidence of retarded ossification was observed at 50 ppm (0.19 mg/L), which indicated delayed maturation as a result of the lower fetal weight, rather than teratogenicity. The NOAELs for maternal toxicity, teratogenicity, and fetotoxicity were 5, 50, and 15 ppm (0.02, 0.06, and 0.19 mg/L), respectively.

The effects of DMDS on reproductive organs were assessed in male and female rats exposed to 10, 50, 150, or 250 ppm (0.04, 0.19, 0.58, or 0.96 mg/L) DMDS for 6 hr/day for 90 days (ELF ATOCHEM, 1992). Tissue histopathology did not reveal any lesions or damage to the epididymus, prostrate, or testes of the male rats, nor ovaries or uterus of female rats.

6. Conclusions
The preceding examination of the physical properties, health effects, and mode of action of the disulfides in DSO demonstrates that DMDS can be used as reasonable worst case surrogate for this substance. The analysis provides strong and consistent mechanistic evidence that DMDS is the most potent member of the alkyl disulfide series, and that the higher molecular weight members found in DSO do not pose a greater health threat or environmental hazard. Accordingly, the available test data for DMDS, a chemical previously reviewed under the HPV Challenge Program, are offered as a justifiable substitute for DSO. The summary of available findings for DMDS and DSO presented in Table 7 show that all of the testing requirements have met or will be met once the DMDS testing is completed for water stability and chronic fish toxicity. Testing for these endpoints will be completed under a voluntary agreement recently approved and accepted by the US EPA. In conclusion, the data review indicates that DMDS can be used a surrogate for DSO and that all of the necessary testing requirements under the HPV Challenge Program have been satisfied.

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HPV Challenge Program Submission Table 7. Data Matrix for Disulfide Oil
Endpoint Sponsored Substance Supporting Chemical Disulfides, Diethyl and Diphenyl, Dimethyl Disulfide Naphtha Sweetening (624-92-0) (68955-96-4) Summary of Physical and Chemical Properties -54 -102 -21.8 (est) Boiling Point (C) 111 174 109.8 200.4 (est) Vapor Pressure (mmHg at 25C) 57 0.35 24.5 (est) Log Kow 1.77 (Read Across) Water Solubility (mg/L at 25C) < 100 40 3740 (est) 2500 3740 (est) 360 (est) 1.77 1.87 (est) 2.86 (est) 21.98 24.5 (est) 4.28 3.31 (est) 109.8 113.6 (est) 154.1 158.8 (est) -85 -69.7 (est) Supporting Chemical Diethyl Disulfide (110-81-6)

Disulfide Oil

Supporting Chemical Dipropyl Disulfide (629-19-6)

Melting Point (C) -102 -45.2 (est)

-86
-21.8 (est) 193.5 200.4 (est) 0.51 0.50 (est) 3.84 (est) 40 (est)

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HPV Challenge Program Submission Table 7. Data Matrix for Disulfide Oil (contd)
Endpoint Sponsored Substance Supporting Chemical Disulfides, Diethyl and Diphenyl, Dimethyl Disulfide Naphtha Sweetening (624-92-0) (68955-96-4) Summary of Environmental Fate Data Supporting Chemical Diethyl Disulfide (110-81-6)

Disulfide Oil

Supporting Chemical Dipropyl Disulfide (629-19-6)

Indirect (OH-) Photodegradation Half-life (t1/2) 1.2 (Read Across) 1.2 0.56 h (est) 0.54 0.52

Stability in Water (Hydrolysis) Half-life (t1/2) Fugacity (Level III Model) Air (%) Water (%) Soil (%) Sediment (%) Biodegradation at 28 days (%)

Read Across

TBD

0.2 1.0 (est) 18.1 58.1 (est) 40.8 80 (est) 0.2 1.6 (est) < 10 (Read Across)

1.0 (est) 58.1 (est) 40.8 (est) 0.2 (est) < 10

0.5 (est) 29.7 (est) 69.4 (est) 0.4 (est) -

0.2 (est) 18.1 (est) 80.0 (est) 1.6 (est) -

Summary of Environmental Effects Aquatic and Terrestrial Toxicity Data Fish (acute) 96-h LC50 (mg/L) Fish (chronic) ChV 30-day (mg/L) Aquatic Invertebrates 48-h EC50 (mg/L) Aquatic Plants 72-h EC50 (mg/L) (growth) Earthworm 14-day LC50 (mg/kg) 0.97 (Read Across) Read Across 7 (Read Across) 35 (Read Across) 32 (Read Across) 0.97 TBD 7 35 32 7.43 14.5 (24-hr) 2.62 -

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HPV Challenge Program Submission Table 7. Data Matrix for Disulfide Oil (contd)
Endpoint Sponsored Substance Supporting Chemical Disulfides, Diethyl and Diphenyl, Dimethyl Disulfide Naphtha Sweetening (624-92-0) (68955-96-4) Summary of Human Health Data (rat) (rat) 1590 1700 190 (rabbit) (rabbit) > 1800 hdt > 2000 hdt (rat) (rat) > 4.84 hdt 3.1 Supporting Chemical Diethyl Disulfide (110-81-6)

Disulfide Oil

Supporting Chemical Dipropyl Disulfide (629-19-6)

Acute Oral Toxicity LD50 (mg/kg-bw) Acute Dermal Toxicity LD50 (mg/kg-bw) Acute Inhalation Toxicity LC50 (mg/L) Repeated-Dose Toxicity NOAEL/LOAEL Oral (mg/kg-bw/day) Repeated-Dose Toxicity NOAEL/LOAEL Dermal (mg/kg-bw/day) Repeated-Dose Toxicity NOAEC/LOAEC Inhalation (mg/L/day)

(rat) > 2000 hdt (rat) NOAEL = 7.3 - 8.2 (hdt)

(rabbit) NOAEL = 10 (Read Across) LOAEL = 100 (Read Across) (rat) NOAEC = 0.019 - 0.096 (Read Across) LOAEC = 0.096 0.482 (Read Across)

(rabbit) NOAEL = 10 LOAEL = 100) (rat) NOAEC = 0.019 - 0.096 LOAEC = 0.096 0.482

Reproductive Toxicity NOAEC/LOAEC Inhalation (ppm)

No effects were seen following evaluation of reproductive organs in the two 13-week inhalation repeated dose toxicity studies in rats (Read Across)

No effects were seen following evaluation of reproductive organs in the two 13-week inhalation repeated dose toxicity studies in rats

Page 31

HPV Challenge Program Submission Table 7. Data Matrix for Disulfide Oil (contd)
Endpoint Sponsored Substance Supporting Chemical Disulfides, Diethyl and Diphenyl, Dimethyl Disulfide Naphtha Sweetening (624-92-0) (68955-96-4) Summary of Human Health Data Supporting Chemical Diethyl Disulfide (110-81-6)

Disulfide Oil

Supporting Chemical Dipropyl Disulfide (629-19-6)

Developmental Toxicity NOAEC/LOAEC Inhalation (mg/L/day) Maternal Toxicity

(rat) NOAEC = 0.019 (Read Across) LOAEC = 0.058 (Read Across)

(rat) NOAEC = 0.019 LOAEC = 0.058

Developmental Toxicity Genetic Toxicity Gene Mutation In vitro (bacterial) In vitro (mammalian) Genetic Toxicity Chromosomal Aberrations In vitro Genetic Toxicity Chromosomal Aberrations In vivo Genetic Toxicity Other In vitro Unscheduled DNA synthesis Genetic Toxicity Other In vivo Unscheduled DNA synthesis

NOAEC = 0.058 (Read Across) LOAEC = 0.193 (Read Across)

NOAEC = 0.058 LOAEC = 0.193

Negative (Read Across) Negative (Read Across)

Negative Negative

Negative -

Ambiguous (Read Across) (mouse) Negative (Read Across) Ambiguous (mouse) Negative -

Negative (Read Across) (male rat) Negative (Read Across) Negative (male rat) Negative -

Page 32

HPV Challenge Program Submission

Table 7. Data Matrix for Disulfide Oil (contd)

Disulfide Oil

Endpoint

Sponsored Substance Supporting Chemical Disulfides, Diethyl and Diphenyl, Dimethyl Disulfide Naphtha Sweetening (624-92-0) (68955-96-4) Summary of Human Health Data Mildly to moderately irritating Minimally irritating Negative Slightly irritating Slightly irritating Negative

Supporting Chemical Diethyl Disulfide (110-81-6)

Supporting Chemical Dipropyl Disulfide (629-19-6)

Other Information Dermal irritation Other Information Eye irritation Other Information Sensitization
1

This table includes measured and predicted SIDS values for the sponsored substance and three of its components for which measured data were identified and used to meet or support the sponsored substance data requirements. Predicted physical-chemical and environmental fate values for seven other disulfide components also used to support the sponsored substance requirements are presented elsewhere in the document.

(-) Indicates that endpoint was not addressed for this chemical.
(est) indicates estimated.
(TBD) indicates data to be developed.
(hdt) indicates highest dose tested.
Bold values represent measured data.

Page 33

HPV Challenge Program Submission

Disulfide Oil

7. References
Arkema, Inc (2007). Dimethyl disulfide. Material Safety Data Sheet # 000719. Arkema, Inc. Philadelphia, PA. http://www.arkema-inc.com/msds/885.pdf Arnault, I., Mondy, N., Diwo, S., and Auger, J. (2004). Soil behavior of sulfur natural fumigants used as methyl bromide substitutes. Int. J. Environ. Anal. Chem. 84:75-82. ATOCHEM (1989a). Repeated-Dose (28-Day) Dermal Toxicity Study with Dimethyl Disulfide (DMDS) in Rabbits. TNO-CIVO Institutes, Report V 89.371/280554 (as cited in DMDS Robust Summary, 2005). ATOCHEM (1989b). Examination of Dimethyl Disulfide in the Micronucleus Test. TNO-CIVO, Report V 89.366 (as cited in DMDS Robust Summary, 2005). ATOCHEM (1990). An in vivo/in vitro rat hepatocyte DNA-repair assay with dimethyldisulfide (DMDS). TNO-CIVO, Report V 90.082 (as cited in DMDS Robust Summary, 2005). ATOCHEM (1991a). Dimethyl Disulfide (DMDS): Inhalation Range-Finding Study in the Pregnant Rat. Hazleton-UK study no. 6142-514/8 (as cited in DMDS Robust Summary, 2005). ATOCHEM (1991b). Dimethyl Disulfide (DMDS): Inhalation Teratology Study in the Rat, Hazleton UK, Report 6205-514/9 (as cited in DMDS Robust Summary, 2005). Bach, R.D., Dmitrenko, O., and Thorpe, C. (2008). Mechanism of thiolate-disulfide interchange reactions in biochemistry. J. Org. Chem. 73:12-21. Bentvelzen, J.M., McKean, W.T., Gratzl, J.S., Lin, S.Y., and Tucker, W.P. (1975). Kinetics of methyl mercaptan oxidation and dimethyl disulfide hydrolysis in alkaline solutions. Tappi 58:102-105. Bremner, J.M. and Benwart, W.L. (1976). Sorption of sulfur gases by soil. Soil Biol. Biochem. 8:79-83. Caron, F. and Kramer, J.R. (1994). Formation of volatile sulfides in freshwater environments. Sci. Total Environ. 153:177-194. Chevron Phillips Chemical Company (2005). Di-n-propyl disulfide. Material Safety Data Sheet # 74480. Chevron Phillips Chemical Company. The Woodlands, TX. http://www.cpchem.com/enu/msds_unsecured/Import_74480_MSDS_O_ENGLISH_A_ ENGLISH_A_N.pdf

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HPV Challenge Program Submission

Disulfide Oil

De Roode, D., Hoekzema, C., de Vries-Buitenweg, S., van de Waart, B., and van der Hoeven, J. (2006). QSARs in ecotoxicological risk assessment. Reg. Toxicol. Pharmacol. 45:24-35 Drummond, J.G. (1991). Acute Inhalation Toxicity Study of Dialkyl Disulfide Material in Rats. Project No. L08310. IIT Research Institute, Chicago, IL. DMDS Test Plan (2005). Dimethyl Disulfide (CAS# 624-92-0) Test Plan. High Production Volume (HPV) Challenge Program. Arkema Inc. Philadelphia, PA. DMDS Robust Summary (2005). IUCLID Data Set Dimethyl Disulphide CAS No. 624 92-0. Atofina Chemicals Inc. Paris, France. ELF ATOCHEM (1989). Repeated-Dose (14-Day) Dermal Toxicity Range-Finding Study with Dimethyl Disulfide (DMDS) in Rabbits. TNO-CIVO Institutes, Report V 89.058/280553 (as cited in DMDS Robust Summary, 2005). ELF ATOCHEM (1990a). In Vitro Assay for the Induction of Point Mutations in the HGPRT-Locus of Chinese Hamster Ovary Cells by Dimethyldisulfide (DMDS), TNO CIVO Institute, Report V 89.257 (as cited in DMDS Robust Summary, 2005). ELF ATOCHEM (1990b). In Vitro DNA Repair Test on Rat Hepatocytes in Primary Culture. SANOFI, Report RA860891026/PN1 (as cited in DMDS Robust Summary, 2005). ELF ATOCHEM (1990c). Chromosome Analysis of Cultured Human Lymphocytes Following In Vitro Treatment with DMDS. TNO-CIVO Institute, Report V 89.045 (as cited in DMDS Robust Summary, 2005). ELF ATOCHEM (1992). DMDS: 90-Day Inhalation Toxicity Study in the Rat with a 4-Week Recovery Period. Hazelton UK, Report 6491-514/7 (as cited in DMDS Robust Summary, 2005). ELF ATOCHEM (1995). Dimethyl Disulfide. Dtermination de la biodgradabilit facile. Essai en fioles fermes. Ref 95/SAEk/0415/NM (as cited in DMDS Robust Summary, 2005). ELF ATOCHEM (1996). Disulfure De Dimethyle. Toxicit aigu vis--vis des daphnies.Rapport N2606/95/A (as cited in DMDS Robust Summary, 2005). ELF ATOCHEM (2000). Centre d'application de Levallois. Disulfure De Dimethyle. Inhibition de la croissance des algues.Etude N 504/99/A (as cited in DMDS Robust Summary, 2005). Farwell, S.O., Sherrard, A.E., Pack, M.R., and Adams, D.F. (1979). Sulfur compounds volatilized from soils at different moisture contents. Soil Biol. Biochem. 11:411-415.

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HPV Challenge Program Submission

Disulfide Oil

Finlayson-Pitts, B.J. and Pitts, J.N. (2000). Acid deposition. Formation and fates on iorganic acids in the troposphere. In: Chemistry of the Upper and Lower Atmosphere. Theory, Experiments, and Applications. Chpt. 8, p. 328. Academic Press, San Diego, CA. Furedi-Machacek, E.M. (1991a). Acute Oral Toxicity of Dialkyl Disulfides in Rats. Project No. L08309 Study No. 6. IIT Research Institute, Chicago, IL. Furedi-Machacek, E.M. (1991b). Acute Oral Toxicity of Dialkyl Disulfides in Rats (Limit Test). Project No. L08309 Study No. 5. IIT Research Institute, Chicago, IL. Furedi-Machacek, E.M. (1991c). Acute Dermal Toxicity Test of Dialkyl Disulfides in Rabbits. Project No. L08309 Study No. 2. IIT Research Institute, Chicago, IL. Furedi-Machacek, E.M. (1991d). Acute Dermal Irritation/Corrosivity Testing of Dialkyl Disulfides in Rabbits. Project No. L08309 Study No. 3. IIT Research Institute, Chicago, IL. Furedi-Machacek, E.M. (1991e). Primary Eye Irritation Testing of Dialkyl Disulfides in Rabbits. Project No. L08309 Study No. 4. IIT Research Institute, Chicago, IL. Furedi-Machacek, E.M. (1991f). Dermal Sensitization Study of Dialkyl Disulfides in Guinea Pigs using the Modified Buehler Method. Project No. L08309 Study No. 1. IIT Research Institute, Chicago, IL. Glli,R., Rich, H.W., and Scholtz, R. (1994). Toxicity of organophosphate insecticides and their metabolites to the water flea Dapnia magna, the Microtox test and an acetylcholinesterase inhibition test. Aquatic Toxicol. 30:259-269. Germain, E., Semon, E., Siess, M.-H., and Teyssier, C. (2008). Disposition and metabolism of dipropyl disulphide in vivo in rat. Xenobiotica 38:8797. Greene, N., Judson, P.N., Langowski, J.J., and Marchant, C.A. (1999). Knowledge-based expert systems for toxicity and metabolism prediction: DEREK, StAR and METEOR. SAR QSAR Environ. Res. 10:299-314. Gruhzit, O.M. (1931). II. Anemia in dogs produced by feeding disulphide compounds. Am. J. Med. Sci. 181:815-820. HSDB (2005). Methyl Disulfide. Hazardous Substances Data Bank. Toxicology Data Network. National Library of Medicine. Bethesda. MD. http://toxnet.nlm.nih.gov/cgi-bin/sis/search/r?dbs+hsdb:@term+@rn+@rel+624-92-0 Jager, T., Posthuma, L., de Zwart, D., and van de Meent, D. (2007). Novel view on predicting acute toxicity: Decomposing toxicity data in species vulnerability and chemical potency. Ecotoxicol. Environ. Saf. 67:311-322.

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Disulfide Oil

Kim, H.Y., Lee, S.B., Chung, Y.H., Yu, I.J., Park, S.C., Shin, J.Y., Kim, S.H., Shin, D.H., and Kim, J.C. (2006). Evaluation of subchronic inhalation toxicity of dimethyl disulfide in rats. Inhal. Toxicol. 18:395-403. Klimisch, H. J., Andreae, M., and Tillman, U., (1997). A systematic approach for evaluating the quality of experimental toxicological and ecotoxicological data. Reg. Toxicol. Pharmacol 25:1-5. Lesser, M.P. (2006). Oxidative stress in marine environments: Biochemistry and physiological ecology. Ann Rev. Physiol. 68:253-278. Lillig, C.H. and Holmgren, A. (2007). Thioredoxin and related molecules from biology to health and disease. Antioxid. Redox Signal. 9:25-47. Mackay, D., Di Guardo, A., Paterson, S., and Cowan, C.E. (1996). Evaluating the environmental fate of a variety of types of chemicals using the EQC model. Environ. Toxicol. Chem. 15:1627-1637. Meylan, W.M. and Howard, P.H. (1998). Users Guide for the ECOSAR Class Program. U.S. Environmental Protection Agency, Washington, D.C. http://www.epa.gov/oppt/newchems/tools/manual.pdf Mnchberg, U., Anwar, A., Mecklenburg, S., and Jacob, C. (2007). Polysulfides as biologically active ingredients of garlic. Org. Biomol. Chem. 5:1505-1518. Munday, R. (1989) Toxicity of thiols and disulphides: Involvement of free-radical species. Free Radic. Biol. Med. 7:659-673. Munday, R. and Manns, E. (1994). Comparative toxicity of prop(en)yl disulfides derived from Alliaceae: Possible involvement of 1-propenyl disulfides in onion-induced hemolytic anemia. J. Agric. Food Chem. 42:959-962. Munday, R., Munday, J.S., and Munday, C.M. (2003). Comparative effects of mono-, di-, tri-, and tetrasulfides derived from plants of the Allium family: Redox cycling in vitro and hemolytic activity and phase 2 enzyme induction in vivo. Free Radic. Biol. Med. 34:1200-1211. Munday, R. and Munday, J.S. (2003). Comparative haemolytic activity of bis(phenylmethyl) disulphide, bis(phenylethyl) disulphide and bis(phenylpropyl) disulphide in rats. Food Chem. Toxicol. 41:1609-1615. Nishikawa, K., Ziro, S., and Numata, M. (1987). Further studies on the urinary metabolites of thiamine propyl disulfide in rats. J. Pharmacol. Exp. Therap. 157:589-598.

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Disulfide Oil

NITE (2002). Biodegredation and Bioconcentration of existing chemical substances under the chemical substances control law Diethyl disulfide. Chemical Management Center, National Institute of Technology and Evaluation. Tokyo, Japan. http://www.safe.nite.go.jp/english/kizon/KIZON_start_hazkizon.html Pennwalt (1985a). Dimethyl Disulfide. EPA Acute LD50. Products Safety Labs, Report T-5147A (as cited in DMDS Robust Summary, 2005). Pennwalt (1985b). Dimethyl Disulfide. EPA Acute Dermal Toxicity Limit Test. Products Safety Labs, Report T-5150 Pennwalt (1985c). Dimethyl Disulfide, Ames Salmonella/Microsome Plate Test (EPA/OECD). Pharmakon Research International, Inc. Report PH 301-PW-003-85, 31 (as cited in DMDS Robust Summary, 2005). Posternak, J.M., Linder, A., and Vodoz, C.A. (1969). Summaries of toxicological data. Toxicological tests on flavoring matters. Food Cosmet. Toxicol. 7:405-407. Rennen, M.A.J., Bouman, T., Wilschut, A., Bessems, J.G.M., and De Heer, C. (2004). Oral-to-inhaltion route extrapolation in occupational risk assessment: A critical assessment. Reg. Toxicol. Pharmacol. 39:5-11. Richards, S.R., Kelly, C.A., and Rudd, J.W.M. (1991). Organic volatile sulfur in lakes of the Canadian Shield and its loss to the atmosphere. Limnol. Oceanogr. 36:468-482. Russom, C.L., Bradbury, S.P., Broderius, S.J., Hammermeister, D.E., and Drummond, R.A. (1997). Predicitng modes of toxic action from chemical structure: Acute toxicity in the fathead minnow (Pimephales promelas). Environ. Toxicol. Chem. 16:948-967. ST Laboratories (2008). Certificate of analysis: Disulfide Oil. Lab No. C0808-1501, ST Laboratories, Inc. Channelview, TX. Tansy, M.F., Kendall, F.M., Fantasia, J., Landin, W.E., Oberly, R., and Sherman, W. (1981). Acute and subchronic toxicity studies of rats exposed to vapors of methyl mercaptan and other reduced-sulfur compounds. J. Toxicol. Environ. Health 8:71-88. Teyssier, C. and Siess, M.-H. (2000). Metabolism of dipropyl disulfide by rat liver phase I and phase II enzymes and by isolated perfused rat liver. Drug Metab. Dispos. 28:648-654. TGSC (2008). Name information list for chemical substances. The Good Scents Company. http://www.thegoodscentscompany.com. Tsai, S.-J., Jenq, S.N., and Lee, H. (1996). Naturally occurring dialllyl disulfide inhibits the formation of carcinogenic heterocyclic aromatic amines in boiled pork juice. Mutagenesis 11:235-240.

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USEPA (1999a). The Use of Structure-Activity Relationships (SAR) in the High Production Volume Chemicals Challenge Program. US Environmetnal Protection Agency, Office of Pollution Prevention and Toxics. Washington, DC. http://www.epa.gov/hpv/pubs/general/sarfinl1.htm USEPA (1999b). Development of Chemical Categories in the HPV Challenge Program. US Environmetnal Protection Agency, Office of Pollution Prevention and Toxics. Washington, DC. http://www.epa.gov/hpv/pubs/general/categuid.htm USEPA. (2007). Estimation Programs Interface Suite for Microsoft Windows, v 3.20. United States Environmental Protection Agency, Washington, DC, USA. http://www.epa.gov/oppt/exposure/pubs/episuite.htm USEPA (2008). Letter from Mark Townsend, EPA Comments on Chemical RTK HPV Challenge Submission: Dimethyl Disulfide, United States Environmental Protection Agency, Washington, DC, USA. http://www.epa.gov/HPV/pubs/summaries/dimthdsl/c16161ct.pdf WHO (2000). Safety Evaluation of Certain Food Additives and Contaminants. Simple Aliphatic and Aromatic Sulfides and Thiols. WHO Food Additive Series 44. International Program on Chemical Safety. World Health Organization, Geneva, Switzerland. http://www.inchem.org/documents/jecfa/jecmono/v44jec09.htm Yamato, O., Kasai, E., Katsura, T., Takahashi, S., Shiota, T., Tajima, M., Yamasaki, M., and Maede, Y. (2005). Heinz body hemolytic anemia with eccentrocytosis from ingestion of Chinese chive (Allium tuberosum) and garlic (Allium sativum) in dogs. J. Am. Animal Hosp. Assoc. 41:68-73.

Page 39

Appendix I

Analytical Results
Composition of Disulfide Oil

Appendix II

Dimethyl Disulfide Test Plan

US HPV CHALLENGE

DIMETHYL DISULFIDE

201-16161A
High Production Volume (HPV) Challenge Program

DIMETHYL DISULFIDE (CAW 624-92-0) Test Plan

Arkema Inc. 2000 Market Street 19103 Philadelphia, PA


December 2005

US HPV CHALLENGE EXECUTIVE SUMMARY

DIMETHYL DISULFIDE

Arkema Inc has volunteered to sponsor dimethyl disulfide (DMDS, CAS# 624-92-0) in the USEPA HPV program. The DMDS Test Plan is being submitted to fulfill the United States Environmental Protection Agency (USEPA) High Production Volume (HPV) Challenge Program commitment for DMDS. Data from company proprietary files, peer-reviewed literature, and/or calculated endpoints using widely accepted computer modeling programs have been identified for purposes of this program. Robust summaries of the available data are included in the attached IUCLID. The following table summarizes the available data and proposed testing for DMDS. Table 1: Matrix of Available and Adequate Data on DMDS
Data Available Y/N Y Y Y Y Y Y N Y Y N Y Y Y Y Y Y Y Y Y Testing Planned? Y/N N N N N N N Y N N Y N N N N N N N N N

SIDS ENDPOINT Physical and Chemical Data Melting Point Boiling Point Vapor Pressure Partition Coefficient Water Solubility Environmental Fate Photodegradation Stability in Water (Hydrolysis) Transport/Distribution Biodegradation Ecotoxicity Acute/Prolonged Toxicity to Fish Acute Toxicity to Aquatic Invertebrates (Daphnia) Acute Toxicity to Aquatic Plants (Algae) Toxicity Acute Toxicity (Oral) Acute Toxicity (Dermal) Acute Toxicity (Inhalation) Repeated Dose GeneticToxicity in vitro Gene Mutation Genetic Toxicity in vitro Chromosomal Aberration Reproductive Toxicity Developmental Toxicity

Note: The data used to characterize the OECD SIDS endpoints for substances in this Test Plan were identified either in company proprietary files, peer-reviewed literature, and/or calculated using widely accepted computer modelling programs. All data were evaluated for study reliability in accordance with criteria outlined by the USEPA (1999a). Only studies that met the reliability criteria of 1 (reliable without restrictions) or 2 (reliable with restrictions) were used. Additional data are also included in the IUCLID (International Uniform Chemical Information Dataset) attached in Annex I. A more detailed discussion of the data quality and reliability assessment process used to develop this test plan is provided in Annex II.
2

US HPV CHALLENGE 1.1 Physico-Chemical properties

DIMETHYL DISULFIDE

DMDS is a pale yellow liquid with a strong garlic like odor. Experimental data for the physical chemical parameters are available and reported in EPIWIN (USEPA, 2004) and are provided in the following table. Table 2. Parameter
Melting Point Boiling Point Vapor Pressure Kow Partition Coefficient Water Solubility (mg/l)

Physicochemical Data Value


-85C 110C
1 1

29.3 hPa 1.77


1 1

2500

EPIWIN v3.12 Syspro database

Conclusion Adequate data are available for the HPV physical/chemical property endpoints. No additional testing for the HPV program is proposed.

GENERAL INFORMATION ON EXPOSURE 1.2 Production Volumes and Use Pattern

DMDS is on EPAs high production volume list indicating it is manufactured and/or imported at greater than 1 million pounds per year according to the toxic inventory update rule (IUR). 1.2.1 Use Pattern:

DMDS has several industrial uses. It is used in the oil industry as a sulfiding/presulfiding agent to activate catalysts of hydrotreating units, to reduce the number of decoking operations in the petrochemical industry, as a chemical intermediate in the fine chemical industry, and as an anti corrosive in metallurgy. 1.3 1.3.1 Environmental Exposure and Fate Photodegradation

The photodegradation of DMDS was evaluated using EPIWIN 3.12. The half life of DMDS was calculated to be 0.565 hours based on the experimental rate constant of 227 x E-12 cm3/molecule sec. Conclusion

US HPV CHALLENGE

DIMETHYL DISULFIDE

Adequate data are available to assess the photodegradation of DMDS. No additional studies are proposed for the HPV program. 1.3.2 Stability in Water

EPIWIN was unable to calculate a hydrolysis rate for DMDS. A hydrolysis study is proposed for DMDS. 1.3.3 Transport between Environmental Compartments

The transport of DMDS between environmental compartments was assessed by fugacity modeling using EPIWIN (v3.12). Results are listed in the table below: Table 3. Fugacity Results for DMDS
Compartment Air Water Soil Sediment Mass amount (%) 1.01 58.1 40.8 0.168 Estimated half life (hr) 1.13 360 360 3.24x e003

1.3.4

B iodegradation

DMDS was not readily biodegradable when evaluated according to OECD 301D. The degradation was less than 10% following 28 days exposure. Conclusion Adequate data are available to assess the biodegradation of DMDS. No additional studies are proposed for the HPV program.

2
2.1.1

HUMAN HEALTH HAZARDS Acute Toxicity

Single exposure (acute) studies indicate DMDS is moderately toxic if swallowed (rat; 290 mg/kg < LD50 < 500 mg/kg), no more then slightly toxic if absorbed through skin (rabbit LD50 >2,000 mg/kg), and slightly toxic if inhaled (rat 4-hr LC50 805 ppm). Conclusion Adequate data are available to assess the acute toxicity of DMDS and no additional studies are proposed. 2.1.2 Repeated Dose Toxicity

DMDS was evaluated in a 90-day repeated dose study on rats according to OECD guidelines. This study featured inhalation dosing, measurement of mortality, body weight changes, food consumption, hematological and blood biochemical examinations, urinalysis, organ weights, histopathology and a functional observational battery. Rats were exposed whole body to 0, 10, 50, 150, and 250 ppm DMDS for 6 hours per day for 90 days. Satellite groups were evaluated
4

US HPV CHALLENGE

DIMETHYL DISULFIDE

following a 2 -week recovery period. Results from this study showed decreased body weights, food consumption, hypoactivity, changes in white blood cell counts, reduced thymus gland weight and increased liver weight. Reversible microscopic changes were noted in the nasal mucosa. Conclusion Adequate data are available to assess the reproductive toxicity of DMDS. No additional testing is proposed for purposes of the HPV program. 2.1.3 Mutagenicity

Several reliable genetic toxicity studies are available for DMDS. Predominantly negative results were obtained with DMDS when tested in vitro (negative bacterial and mammalian mutagenicity assays, negative DNA damage and repair, ambiguous positive in vitro chromosome aberration study using human lymphocytes). Negative results were obtained when DMDS was evaluated in vivo (mouse micronucleus, unscheduled DNA synthesis). Conclusion Adequate data are available to assess the genetic toxicity of DMDS. No additional testing is proposed for purposes of the HPV program. 2.1.4 Toxicity for Reproductive/Developmental Toxicity

Reproductive Toxicity The 90 day repeated dose toxicity study will be used to assess the reproductive toxicity of DMDS. Reproductive organs examined in this study included the epididymus, prostate, and testes in males and ovaries and uterus in females. No lesions were reported. Developmental Toxicity A Developmental Toxicity test was completed for DMDS in Sprague -Dawley rats following OECD Guideline 414 Teratogenicity. DMDS was administered by inhalation to 0, 5, 15, and 50 ppm on gestation days 6 to 15. Maternal toxicity was noted at 15 and 50 ppm. No evidence of developmental toxicity was observed. No additional studies are proposed. Conclusion Adequate data are available to assess the reproductive and developmental toxicity of DMDS. No additional testing is proposed for the HPB program.

HAZARDS TO AQUATIC O RGANISMS

DMDS has been evaluated in an acute daphnia immobilization and algal growth inhibition studies. DMDS is moderately toxic to daphnia with a 48 hour EC50 value of 7 mg/l. DMDS is slightly toxic to Selenastrum capricornutum alga with a 72 hour EC50 of 35 mg/l. No data are available for acute fish and alga. No data are available to assess the acute fish toxicity and an acute fish toxicity (OECD guideline 203) is proposed for DMDS. Conclusion Adequate data are available to assess the aquatic toxicity of DMDS to daphnia and alga but not fish. An acute fish toxicity study is proposed (OECD guideline 203) for DMDS.
5

US HPV CHALLENGE References

DIMETHYL DISULFIDE

Atofina, 2005. IUCLID Data Set, CAS No. 624-92-0 dimethyldisulfide. Atofina, Paris, France. Klimisch, H.J., E. Andreae, and U. Tillmann. 1997. A systematic approach for evaluating the quality of experimental and ecotoxicological data. Reg. Tox. and Pharm . 25: 1 -5.
Organisation for Economic Co-operation and Development (OECD) Secretariat. 2002. Manual for
Investigation of HPV Chemicals (November 2002).
U.S. Environmental Protection Agency (USEPA), Office of Pollution Prevention and Toxics. 1998. Guidance for Meeting the SIDS Requirements: Chemical Right-to-Know Initiative. USEPA, Office of Pollution Prevention and Toxics. 1999b. Draft Determining the Adequacy of Existing Data. USEPA, Office of Pollution Pre vention and Toxics and Syracuse Research Corporation. 2004. EPI Suite v 3.12.

US HPV CHALLENGE ANNEX I: DIMETHYL DISULFIDE IUCLID See attached IUCLID documents. ANNEX II: DATA QUALITY ASSESSMENT

DIMETHYL DISULFIDE

Available environmental, ecotoxicity, and mammalian toxicity studies were reviewed and assessed for reliability according to standards specified by Klimisch et al., (1997), as recommended by the USEPA (1999a) and the OECD (OECD, 2002). The following reliability classification (Klimisch rating) has been applied to each study assessed:

1 = Reliable without Restriction Includes studies that comply with USEPA- and/or OECDaccepted testing guidelines and were conducted using Good Laboratory Practices (GLPs) and for which test parameters are complete and well documented; 2 = Reliable with Restriction Includes studies that were conducted according to national/international testing guidance and are well documented. May include studies that were conducted prior to establishment of testing standards or GLPs but meet the test parameters and data documentation of subsequent guidance; also includes studies with test parameters that are well documented and scientifically valid but vary slightly from current testing guidance. Also included in this category were physical -chemical property data obtained from reference handbooks, as well as environmental endpoint values obtained from an accepted method of estimation (e.g., USEPAs EPIWIN estimation program); 3 = Not Reliable Includes studies in which there are interferences in either the study design or results that provide scientific uncertainty or in which documentation is insufficient; and, 4 = Not Assignable This designation is used in this dossier for studies that appear scientifically valid but for which insufficient information is available to adequately judge robustness.

Those studies receiving a Klimisch rating of 1 or 2 are considered adequate to support data assessment needs in this dossier. Those key studies selected for inclusion are considered typical of the endpoint responses observed in other studies of a similar nature and design that were identified during our search of the literature.

Appendix III

Dimethyl Disulfide Robust Summaries

201-16161A

IUCLID Data Set


Existing Chemical CAS No. EINECS Name EC No. TSCA Name Molecular Formula Producer related part Company Creation date Substance related part Company Creation date Status Memo Printing date Revision date Date of last update Number of pages Chapter (profile) Reliability(profile) Flags (profile)
: Chapter:1,2,3,4,5,6,7,8,10 : Reliability: without reliability, 1, 2, 3, 4 : Flags: without flag, confidential, non confidential, WGK (DE), TALuft (DE), Material Safety Dataset, Risk Assessment, Directive 67/548/EEC, SlDS

: : : : : :

lD:624-924 624-92-0 dimethyl disulphide 210-871-0 Disulfide, dimethyl C2H6S2

: ATOFINA Chemicals Inc. : 27.12.2005 : ATOFlNA Chemicals Inc. : 27.12.2005

1. General Information

Id 624-9 2-0 Date 31.12.2005

1.0.1

APPLICANT AND COMPANY INFORMATION manufacturer ARKEMA 4-8, cours Michelet La Dfense 10 95091 Paris La Dfense Cedex France +33 1 49 00 80 80

Type Name Contact person Date Street Town Country Phone Telefax Telex Cedex Email Homepage Source 14.12.2005 Type Name Contact person Date Street Town Country Phone Telefax Telex Cedex Email Homepage Remark Source 31.12.2005 1.0.2

: : : : : : : : : : : : : :

Atofina Paris La Dfense Cedex importer of product ARKEMA Chemicals Inc. 2000 Market Street Philadelphia United States

: : : : : : : : : : : : : : :

formerly ATOFINA Inc. Atofina Paris La Dfense Cedex

LOCATION OF PRODUCTION SITE, IMPORTER OR FORMULATOR

1.0.3

IDENTITY OF RECIPIENTS

1.0.4

DETAILS ON CATEGORY/TEMPLATE

1.1.0

SUBSTANCE IDENTIFICATION

IUPAC Name Smiles Code Molecular formula Molecular weight Petrol class Source 23.12.2005

:
:
: C2-H6-S2
: 94.2
:
: Atofina Paris La Dfense Cedex 2 / 51

1. General Information

Id 624-9 2-0 Date 31.12.2005

1.1.1

GENERAL SUBSTANCE INFORMATION typical for marketed substance


organic
liquid
> 99.5 % w/w
Light yellow
Strong garlic odour
ARKEMA, Paris-la-Dfense, France (JFR) Atofina Paris La Dfense Cedex

Purity type Substance type Physical status Purity Colour Odour Source 23.12.2005 1.1.2 SPECTRA

: : : : : : :

1.2

SYNONYMS AND TR ADENAMES

DMDS
2,3-Dithiabutane
Dimethyl disulfide
Dimethyldisulfide
Disulfide, dimethyl
Methyldisulfide
Methyldithiom ethane
Source 27.12.2005
1.3 IMPURITIES : ARKEMA, Paris-la-Dfense, France
Atofina Paris La Dfense Cedex

1.4

ADDITIVES

1.5

TOTAL QUANTITY

1.6.1

LABELLING

1.6.2

CLASSIFICATION

1.6.3

PACKAGING

1.7

USE PATTERN industrial


Chemical industry: used in synthesis
3 / 51

Type of use Category

: :

1. General Information

Id 624-9 2-0 Date 31.12.2005

Source 23.12.2005 Type of use Category Source 23.12.2005 1.7.1

Atofina Paris La Dfense Cedex industrial other: Sulphurization agent (Petrochemical) ARKEMA, Paris-la-Dfense, France (JFR) Atofina Paris La Dfense Cedex

: : :

DETAILED USE PATTERN

1.7.2

METHODS OF MANUFACTURE

1.8

REGULATORY MEASURES

1.8.1

OCCUPATIONAL EXPOSURE LIMIT VALUES

1.8.2

ACCEPTABLE RESIDUES LEVELS

1.8.3

WATER POLLUTION

1.8.4

MAJOR ACCIDENT HAZARDS

1.8.5

AIR POLLUTION

1.8.6

LISTINGS E.G. CHEMICAL INVENTORIES EINECS 210 -871-0 Atofina Paris La Dfense Cedex

Type Additional information Source 23.12.2005 1.9.1

: : :

DEGRADATION/TRANSFORMATION PRODUCTS

1.9.2

COMPONENTS

1.10

SOURCE OF EXPOSURE

4 / 51

1. General Information

Id 624-9 2-0 Date 31.12.2005

1.11

ADDITIONAL REMARKS

1.12

LAST LITERATURE SEARCH Internal and External 3, 4, 5 23.12.2005 ARKEMA, Paris-la-Dfense, France (JFR) Atofina Paris La Dfense Cedex

Type of search Chapters covered Date of search Source 23.12.2005 1.13 REVIEWS

: : : :

5 / 51

2. Physico-Chemical Data

Id 624-9 2-0 Date 31.12.2005

2.1

MELTING POINT -85 C (2) valid with restrictions Critical study for SIDS endpoint (18) : : : : : : : = 84.7 C

Value Reliability
Flag
27.12.2005 Value
Sublimation
Method
Year
GLP
Test substance
Source 15.11.1993 2.2 BOILING POINT

: : :

no data Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (28)

Value
Decomposition
Method
Year
GLP
Test substance
Remark

: : : : : : :

= 109.6 C at 1013 hPa yes no data Start of Decomposition: 390 degree C Decomposition products: Hydrogen sulphide, Dimethyl sulphide and methanethiol Similar result (109.6C) reported in Epiwin 3. 12 syspro experimental database Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (2) valid with restrictions Critical study for SIDS endpoint (28)

Source Reliability
Flag
31.12.2005 2.3 DENSITY

: : :

Type
Value
Method
Year
GLP
Test substance
Source 15.11.1993 2.3.1 GRANULOMETRY

: : : : : : :

density = 1.063 g/cm at 20 C no data Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (28)

6 / 51

2. Physico-Chemical Data

Id 624-9 2-0 Date 31.12.2005

2.4

VAPOUR PRESSURE = 29.3 hPa at 20 C

Value Decomposition Method Year GLP Test substance Source Reliability Flag 27.12.2005 Value Decomposition Method Year GLP Test substance Source 15.11.1993 2.5

: : : : : : : : :

no data Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (2) valid with restrictions Critical study for SIDS endpoint (32) = 38 hPa at 25 C

: : : : : : :

no data Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (28)

PARTITION COEFFICIENT octanol-water = 1.77 at C other (measured) no data Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (2) valid with restrictions Critical study for SIDS endpoint (20) : : : : : : : : octanol-water = 1.87 at C other (calculated)

Partition coefficient Log pow pH value Method Year GLP Test substance Source Reliability Flag 31.12.2005 Partition coefficient Log pow pH value Method Year GLP Test substance Source 04.12.2001

: : : : : : : : : :

Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (31)

2.6.1 SOLUBILITY IN DIFFERENT MEDIA Water 7 / 51

Solubility in

2. Physico-Chemical Data
: = 2500 mg/l at 20 C
:
: at C
:
:
: at 25 C
:
:
:
:
:
: no data
:
: : : :

Id 624-9 2-0 Date 31.12.2005

Value pH value concentration Temperature effects Examine different pol. pKa Description Stable Deg. product Method Year GLP Test substance Remark Source Reliability Flag 31.12.2005 2.6.2 SURFACE TENSION

Unit of water solubility: ppm Similar data (3000 mg/l) reported in EPIWIN v3.12 experimental database Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (2) valid with restrictions Critical study for SIDS endpoint (32)

2.7

FLASH POINT = 16 C
closed cup
other
no data
Method: ASTM D 93 Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (28)

Value Type Method Year GLP Test substance Remark Source 15.11.1993 2.8

: : : :
: :
: :

AUTO FLA MMABILITY

2.9

FLAMMABILITY : flammable
:
:
: no data
:
: Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (28)

Result Method Year GLP Test substance Source 15.11.1993 2.10

EXPLOSIVE PROPERTIES

8 / 51

2. Physico-Chemical Data
: other
:
:
: no data
:
: : Explosive limits of vapours: 1.1 to 16.1 %v/v in air Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex

Id 624-9 2-0 Date 31.12.2005

Result Method Year GLP Test substance Remark Source 15.11.1993 2.11

(28)

OXIDIZING PROPERTIES

2.12

DISSOCIATION CONSTANT

2.13

VISCOSITY

2.14

ADDITIONAL REMARKS

9 / 51

3. Environmental Fate and Pathways

Id 624-9 2-0 Date 31.12.2005

3.1.1

PHOTODEGRADATION air nm based on intensity of sunlight OH = .000000000227 cm/(molecule*sec) = 50 % after .6 hour(s)

Type Light source Light spectrum Relative intensity INDIRECT PHOTOLYSIS Sensitizer Conc. of sensitizer Rate constant Degradation Result

: : : : : : : :

: AOP Program (v1.91) Results: =========================== SMILES : S(SC)C CHEM : Disulfide, dimethyl MOL FOR: C2 H6 S2 MOL WT : 94.19 ------------------- SUMMARY (AOP v1.91): HYDROXYL RADICALS ----------- ------Hydrogen Abstraction = 2.1216 E-12 cm3/molecule-sec Reaction with N, S and -OH = 225.0000 E -12 cm3/molecule-sec Addition to Triple Bonds = 0.0000 E-12 cm3/molecule-sec Addition to Olefinic Bonds = 0.0000 E-12 cm3/molecule-sec Addition to Aromatic Rings = 0.0000 E -12 cm3/molecule-sec Addition to Fused Rings = 0.0000 E-12 cm3/molecule-sec OVERALL OH Rate Constant = 227.1216 E -12 cm3/molecule-sec HALF-LIFE = 0.047 Days (12-hr day; 1.5E6 OH/cm3) HALF-LIFE = 0.565 Hrs ------------------- SUMMARY (AOP v1.91): OZONE REACTION ----------------- ---****** NO OZONE REACTION ESTIMATION ****** (ONLY Olefins and Acetylenes are Estimated) Experimental Database Structure Match: Chem Name : Dimethyl disufide CAS Number: 000624-9 2-0 Exper OH rate constant : 227 E-12 cm3/molecule-sec Exper OH Reference: KWOK,ESC & ATKINSON,R (1994) Exper Ozone rate constant: --- cm3/molecule -sec Exper NO3 rate constant : 7 E-13 cm3/molecule-sec : (2) valid with restrictions Acceptable calculation method based on experimental rate constant. : Critical study for SIDS endpoint

Reliability Flag 31.12.2005 3.1.2

STABILITY IN WATER abiotic at C at C at C

Type t1/2 pH4 t1/2 pH7 t1/2 pH9 Remark

: : : :

: Hydrolysis at ambient temperature and pH<12 is too slow to be an important environmental fate process. 10 / 5 1

3. Environmental Fate and Pathways


Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex (2) valid with restrictions Critical study for SIDS endpoint

Id 624-9 2-0 Date 31.12.2005

Source Reliability Flag 27.12.2005 3.1.3 STABILITY IN SOIL

: : :

(7)

3.2.1

MONITORING DATA

3.2.2

FIELD STUDIES

3.3.1

TRANSPORT BETWEEN ENVIRONMENTAL COMPARTMENTS fugacity model level III


1.01 % (Fugacity Model Level I)
58.1 % (Fugacity Model Level I)
40.8 % (Fugacity Model Level I)
% (Fugacity Model Level II/III)
.165 % (Fugacity Model Level II/III)
other: model
Level III Fugacity Model (Full-Output):
======================================= Chem Name : Disulfide, dimethyl Molecular Wt: 94.19 Henry's LC : 0.00121 atm -m3/mole (Henry database) Vapor Press : 24.5 mm Hg (Mpbpwin program) Log Kow : 1.77 (Kowwin program) Soil Koc : 24.1 (calc by model) Mass Amount Half - Life Emissions (percent) (hr) (kg/hr) Air 1.01 1.13 1000 Water 58.1 360 1000 Soil 40.8 720 1000 Sediment 0.165 3.24e+003 0 Fugacity Reaction Advection Reaction Advection (atm) (kg/hr) (kg/hr) (percent) (percent) Air 9.37e-012 2.21e+003 36.1 73.8 1.2 Water 1.34e-008 400 208 13.3 6.93 Soil 1.17e-007 141 0 4.69 0 Sediment 1.2e-008 0.126 0.0118 0.00421 0.000394 Persistence Time: 119 hr Reaction Time: 130 hr Advection Time: 1.47e+003 hr Percent Reacted: 91.9 Percent Advected: 8.14 Half-Lives (hr), (based upon Biowin (Ultimate) and Aopwin): Air: 1.131 11 / 5 1

Type Media Air Water Soil Biota Soil Method Year Result

: :
: : : : : : :
:

3. Environmental Fate and Pathways


Water: 360 Soil: 720 Sediment: 3240 Biowin estimate: 2.991 (weeks Advection Times (hr): Air: 100 Water: 1000 Sediment: 5e+004 (2) valid with restrictions Critical study for SIDS endpoint

Id 624-9 2-0 Date 31.12.2005

Reliability Flag 31.12.2005 3.3.2 DISTRIBUTION

: :

(19)

3.4

MODE OF DEGRADATION IN ACTUAL USE

3.5

BIODEGRADATION aerobic
< 10 () % after 28 day(s)
other: not readily biodegradable
7 day(s) = .3 %
14 day(s) = 1.1 % 20 day(s) = 1.9 % 28 day(s) < 0 % %
Benzoic acid, sodium salt
14 day(s) = 86.1 %
28 day(s) = 84.5 %
not measured
OECD Guide-line 301 D "Ready Biodegradability: Closed Bottle Test"
1992
no
as prescribed by 1.1 - 1.4
O2 dissolved (mg/l) 0 d 7 d 14 d 20 d 28 d 1- Medium + inoculum mean 8.41 8.26 8.12 7.64 7.32 2- Medium + inoculum + test substance mean 8.42 8.24 8.05 7.51 7.44 3- Medium + inoculum + test substance + reference substance mean 8.37 5.55 5.43 4.79 4.74 4- Medium + inoculum + reference substance mean 8.41 2.61 2.37 2.09 1.68

Type Inoculum Contact time Degradation Result Kinetic of testsubst.

: :
:
: : :

Control substance Kinetic Deg. product Method Year GLP Test substance Result

: : : : : : : :

BOD (O2 mg/mg substance) 12 / 5 1

3. Environmental Fate and Pathways


0d 7d 14 d 20 d 28 d

Id 624-9 2-0 Date 31.12.2005

serie 2 (substance) 0.00 0.01 0.02 0.04 -0.04 serie 3 (inhibition control) 0.00 0.76 0.76 0.80 0.73 serie 4 (reference) 0.00 1.41 1.44 1.39 1.41

BIODEGRADATION (%) 0d 7d 14 d 20 d 28 d

Source Reliability Flag 31.12.2005 3.6

: : :

serie 2 (substance) 0 0.3 1.1 1.9 -1.8


serie 3 (inhibition
control) 0 40.1 39.9 42.2 38.2
serie 4 (reference) 0 84.5 86.1 83.1 84.5
Atofina, Paris-la -Dfense, France.
Atofina Paris La Dfense Cedex
(2) valid with restrictions
Guideline study without detailed documentation. Critical study for SIDS endpoint (8)

BOD5, COD OR BOD5/COD RATIO

3.7

BIOACCUMULA TION

3.8

ADDITIONAL REMARKS

13 / 5 1

4. Ecotoxicity

Id 624-9 2-0 Date 31.12.2005

4.1

ACUTE/PROLONGED TOXICITY TO FISH

4.2

ACUTE TOXICITY TO AQUATIC INVERTEBRATES static


Daphnia magna (Crustacea)
48 hour(s)
mg/l
=7
> 13.4
yes
OECD Guide-line 202
1996
yes
other TS: DMDS, Atofina, 98.93% purity

Type Species Exposure period Unit EC50 EC50, 24 h Analytical monitoring Method Year GLP Test substance Result

: : : : : : : : : : :

: - Biological observations
20 daphnia per concentration
mg/I %Immo nominal 1 13.4 10.6 9.5 7.8 6.3 5.5 4.7 3.8 3.3 85 75 70 60 50 45 20 10 10 10 1 1 2 3 3 3 4 4 5 5 2 1 2 2 2 2 3 4 5 5 3 0 1 1 2 3 3 4 4 4 4 4 1 1 1 1 2 2 4 5 4 5 total 3 5 6 8 10 11 16 18 18 4 18

0 tmoin

Source Test condition

: :

- EC50, 48h : 7 mg/l ; 95% CI : 6.5 - 7.6 mg/l


Atofina, Paris-la -Dfense, France.
Atofina Paris La Dfense Cedex
- Test organisms Daphnia magna Straus Clone A from INERIS, France. Breeding colony realized in the laboratory in an Elendt M7 medium, supplemented with algal based feed. Organisms are selected by sieving. Age at study initiation : < 24h old, laboratory bred - A stock solution is prepared before the beginning of the test, by mixing 100 mg of the substance with 1 liter of dilution water. Test temperature range : 20-21C Exposure vessel type : Closed flasks (120 ml) as test glassware entirely filled with test solutions and stoppered with PTFE bungs and sealed with aluminum caps. -Dilution water is prepared in the laboratory using pure water and salts according to ISO 6341. 25 ml/l of the below solutions , aerated up to oxygen 14 / 5 1

4. Ecotoxicity
saturated

Id 624-9 2-0 Date 31.12.2005

11.76 g CaCl2, 2 H2O /l ultrapure water 4.93 g MgSO4, 7 H2O /l ultrapure water 2.59 g NaHCO3 /l ultrapure water 0.23 g KCl /l ultrapure water - Dilution water chemistry According to ISO 6341
Ca+Mg ions = 2.5 mmol/l.
Ca/Mg = 4
Na/K = 10
pH 7.8 0.2
- incubation of test flasks in darkness. - Water chemistry in test : C nominal (mg/l) 0 3.3 4.0 4.8 5.8 6.9 8.3 10.0 12.0 14.4 02 at 48h (mg/l) 8.3 8.2 8.2 8.3 8.3 8.3 8.3 8.4 8.3 8.3
pH at 48 h
7.89 7.90 7.88 7.88 7.95 7.93 7.96 8.01 8.03 8.00 - Test design Nominal Concentration
Measured
Initial Final Final/Initial
mg/l mg/l %
3.3 3.8 4.7 5.5 6.3 7.8 9.5 10.6 13.4 3.6 4.1 5.2 5.3 6.6 8.2 9.9 11.8 13.7 109.1 107.9 110.6 96.4 104.8 105.1 104.2
111.3
102.2

3.3 4.0 4.8 5.8 6.9 8.3 10 12 14.4

Reliability Flag 27.12.2005 Type Species Exposure period Unit EC50 Analytical monitoring Method Year GLP Test substance Method

- Analytical monitoring Gas chromatography/FID - 5 individuals per replicate : (1) valid without restriction : Critical study for SIDS endpoint (10) : : : : : : : : : : static Daphnia pulex (Crustacea) 4 hour(s) mg/l = 21.4 no other 1963 no no data

: Groups of 3-5 daphnia were dispensed into glass sample vials, each of which containing 5.0 ml of a biological harmless "culture water" at 21C. 15 / 5 1

4. Ecotoxicity

Id 624-9 2-0 Date 31.12.2005 15.0 ml of toxic solution were added.


The vials were transported in the darkness of a covered ,
thermostatically controlled water-bath (21+-0.05C).
The vials were set up in triplicate.
There were 6 concentrations per chemical.
The concentration series was progressively adjusted so as to
approach the 50% mortality.
Controls were included in each experiments to give an
estimate of control-mortality.
Atochem Paris la Defense Atofina Paris La Dfense Cedex (33) : : : : : : : : : : : static
Daphnia pulex (Crustacea)
48 hour(s)
mg/l
=4
= 15
no
other
1970
no
no data

Source 04.12.2001 Type Species Exposure period Unit EC50 EC50, 24h Analytical monitoring Method Year GLP Test substance Remark

Source

Test condition 04.12.2001 4.3

: Method according to: WERNER, A.E.: Sulphur compounds in


kraft pulp mill effluents.Can. Pulp paper Ind., 1963, 16, 3,
35-43.
: Atochem Paris la Defense
EUROPEAN COMMISSION - European Chemicals Bureau Ispra (VA)
Atofina Paris La Dfense Cedex
: The test was made in glass cylinder of 110 ml capacity.The
volume of the test solution was 100 ml.The temperature was
about 20C.
(29)

TOXICITY TO AQUATIC PLANTS E.G. ALGAE Selenastrum capricornutum (Algae)


growth rate
72 hour(s)
mg/l
= 10.43 measured/nominal
= 9.3 measured/nominal
= 35 measured/nominal
yes
OECD Guide-line 201 "Algae, Growth Inhibition Test"
2000
yes
other TS: DMDS, Atofina, 99.65% purity
- Values (mg/l)
ErC50, 72h = 35
ErC10, 72h = 9.3
EbC50, 72h = 11
EbC10, 72h = 10.43
NOECb : 10.43 NOECr : 10.43 16 / 5 1

Species Endpoint Exposure period Unit NOEC EC10 EC50 Limit test Analytical monitoring Method Year GLP Test substance Result

: : : : : : : :
: : : : : :

4. Ecotoxicity

Id 624-9 2-0 Date 31.12.2005

- control response satisfactory : yes - BIOLOGICAL OBSERVATIONS +Cell density at each flask at each measuring point algal conc. (Cell/ml) Sample Replicat N T0 T24h T48h T72h mg/l nom 0 mean 1.00E+04 5.00E+04 2.34E+05 3.28E+06 100 mean 1.00E+04 8.33E+03 2.00E+04 4.23E+04 55.56 mean 1,00E+04 9.00E+03 3.57E+04 2.00E+05 30.86 mean 1.00E+04 1.80E+04 1.08E+05 6.97E+05 17.15 mean 1.00E+04 3.30E+04 2.32E+05 1.68E+06 9.53 mean 1.00E+04 1.60E+04 2.63E+05 1.91E+06 5.29 mean 1.00E+04 4.50E+04 2.87E+07 2.35E+06 +Percent biomass/growth rate inhibition per concentration sample mean Inhibition % integral blomass growth rate

Source Test condition

: :

nominal (mg/l) IAI (%) Ii (%) 0 0.00 0.00 5.29 22.03 5.78 9.53 35.27 9.36 17.15 40.10 11.55 30.86 76.32 26.74 55.56 93.70 48.29 100 98.71 75.09 Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex - Static test Test temperature range : 24 1 C Growth/test medium chemistry Prepared according to 1.6.1.2 of C.3. method (Annex 5 of 92/69/EEC Directive) pH 8 Dilution water source See above Exposure vessel type 120 ml glass bottles completely filled with test solution and stoppered with PTFE bungs and sealed with aluminum caps 17 / 5 1

4. Ecotoxicity

Id 624-9 2-0 Date 31.12.2005 Water chemistry in test (pH and O2 dissolved mg/l)) C% vol T0 0 5.29 9.53 17.15 30.86 55.56 100 7.31 7.03 7.01 7.00 7.00 7.00 7.09 T72h 7.67 7.46 7.46 7.43 7.36 7.27 7.18 T0 7.7 7.4 7.5 7.8 7.5 7.6 8.1 T72h 11.2 10.0 11.1 10.7 9.6 9.4 8.4

Stock solutions preparation Ultrapure water (ultrafiltration, active carbon, ions exchange, 0.22 m filter) Stock solution prepared 1 h before the beginning of the test, by adding 94l of substance in 1 l of dilution water, stirred during 1h. Light levels and quality during exposure Constantly illuminated between 6000 to 10000 lx. - Test design 3 replicates at each test concentration 7 concentrations (nominal) : 0, 5.29, 9.53, 17.15, 30.86, 55.56,100 mg/l (1) valid without restriction Critical study for SIDS endpoint (9)

Reliability Flag 31.12.2005 4.4

: :

TOXICITY TO MICROORGANISMS E.G. BACTERIA

4.5.1

CHRONIC TOXICITY TO FISH

4.5.2

CHRONIC TOXICITY TO AQUATIC INVERTEBRATES

4.6.1

TOXICITY TO SEDIMENT DWELLING ORGANISMS

4.6.2

TOXICITY TO TERRESTRIAL PLANTS

4.6.3

TOXICITY TO SOIL DWELLING ORGANISMS

4.6.4

TOX. TO OTHER NON MAMM. TERR. SPECIES

4.7

BIOLOGICAL EFFECTS MONITORING

18 / 5 1

4. Ecotoxicity

Id 624-9 2-0 Date 31.12.2005

4.8

BIOTRANSFORMATION AND KINETICS

4.9

ADDITIONAL REMARKS

19 / 5 1

5. Toxicity

Id 624-9 2-0 Date 31.12.2005

5.0

TOXICOKINETICS, METABOLISM AND DISTRIBUTION

5.1.1

ACUTE ORAL TOXICITY

Type Value Species Strain Sex Number of animals Vehicle Doses Method Year GLP Test substance Method

: : : : : : : : : : : : :

LD50 290 - 500 mg/kg bw rat Sprague-Dawley male/female 60 other: polyethylene glycol 300 0, 100, 290, 350, 500 and 5300 mg/kg Directive 84/449/EEC, B.1 "Acute toxicity (oral)" 1986 yes other TS DIMETHYL DISULFIDE was administered undiluted at a volume of 5 ml/kg bw, or as a suspension (10 ml/kg) in polyethylene glycol 300 at the dose levels of 100, 170, 290, 350 and 500 mg/kg. Clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test item. All animals were subjected to necropsy. Mortality: - 100 and 170 mg/kg : none - 290 mg/kg : 30 % - 350 mg/kg : none - 500 mg/ kg : 100 % Clinical signs: Sedation, hypotonia, dyspnea, piloerection and coma, appeared just after the administration and disappeared after 24 hours. Body weight: No effect was noted on the body weight gain of the surviving rats. Macroscopic examination: Haemorragic stomachs was observed at the macroscopic examination of the rats dead on the first day (290 and 500 mg/kg). ARKEMA, Paris-la-Dfense, France (JFR). Atofina Paris La Dfense Cedex TEST ORGANISMS: - Adaptation period: 7 days - Number of animals: 5 males + 5 females / dose - Controls: no HOUSING The animals were housed 5 of the same sex per polycarbonate cages ADMINISTRATION: - Exposure route: gavage - Volume administered: see freetext ME - Post dose observation period: 14 days 20 / 5 1

Result

Source Test condition

: :

5. Toxicity

Id 624-9 2-0 Date 31.12.2005

Test substance

Conclusion Reliability Flag 31.12.2005 Type Value Species Strain Sex Number of animals Vehicle Doses Method Year GLP Test substance Method

: : :

EXAMINATIONS: clinical observations, body weight, mortality and necropsy Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Purity: no data The oral LD50 of DIMETHYL DISULFURE in rats is lower than 500 mg/kg but higher than 290 mg/kg. (1) valid without restriction Material Safety Dataset, Directive 67/548/EEC, Critical study for SIDS endpoint (30) LD50 = 190 m g/kg bw rat Wistar male/female 50 CMC 125, 188, 250, 375 and 500 mg/kg other: EPA 40 CFR 163.81-1 yes other TS DIMETHYL DISULFIDE w as administered as a suspension in 3% carboxymethyl cellulose at the dose levels of 125, 188, 250, 375 and 500 mg/kg. Clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test item. All animals were subjected to necropsy. Group Dose Mortality Mortality % g/kg Male Female 1 0.125 0/5 1/5 10 2 0.188 5/5 1/5 60 3 0.250 3/5 4/5 70 4 0.375 5/5 5/5 100 5 0.50 5/5 5/5 100 LD50 = 0.19 (0.15 -0.24) g/kg Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex TEST ORGANISMS: - Adaptation period: 14 days - Number of animals: 5 males + 5 females / dose - Controls: no ADMINISTRATION: - Exposure route: gavage - Volume administered: no data - Post dose observation period: 14 days EXAMINATIONS: clinical observations, body weight, mortality and necropsy

: : : : : : : : : : : : :

Result

Source Test condition

: :

Test substance

STATISTICAL DETERMINATION OF THE LD50: - Litchfield-Wilcoxon method of probit analysis. : Test substance: D imethyl disulfide C AS no.: 624-9 2-0 21 / 5 1

5. Toxicity
Purity: no data Acute Oral Defined LD50: 0.19 g/kg (1) valid without restriction Critical study for SIDS endpoint

Id 624-9 2-0 Date 31.12.2005

Conclusion Reliability Flag 31.12.2005 5.1.2

: : :

(26)

ACUTE INHALATION TOXICITY LC50


= 805 ppm
rat
Sprague-Dawley
male/female
100
0, 500, 700, 775, 800, 840, 875, 950, 1100 and 1581 ppm
4 hour(s)
other: comparable to OECD Guide-line 403
no
other TS

Type Value Species Strain Sex Number of animals Vehicle Doses Exposure time Method Year GLP Test substance Result

: : : : : : :
: : : :
: :

: MORTALITY:
See the attached table
CLINICAL SIGNS: No data MACROSCOPIC OBSERVATION: No data LC50 = 805 (776-835) ppm : Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex : Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Aldrich Batch: no data Purity: no data : TEST ORGANISMS: - Adaptation period: >= 7 days - Number of animals: 5 males + 5 females - Controls: no HOUSING The animals of the same sex were housed 5 per cage ADMINISTRATION: - Exposure : whole-boby inhalation - Analytical control of the concentration: no data EXAMINATIONS: - Clinical observations, mortality and necropsy - Post dose observation period: 14 days STATISTICAL DETERMINATION OF THE LC50: - Litchfield-Wilcoxon method of probit analysis.
Tansy table.bmp
22 / 5 1

Source Test condition

Test substance

Attached document

5. Toxicity

Id 624-9 2-0 Date 31.12.2005

Reliability Flag 31.12.2005 5.1.3

: :

(2) valid with restrictions Critical study for SIDS endpoint (21)

ACUTE DERMAL TOXICITY LD0 >= 2000 mg/kg bw rabbit New Zealand white male/female 10 other: none 2000 mg/kg other: EPA 40 CFR 163.81-2 yes other TS

Type Value Species Strain Sex Number of animals Vehicle Doses Method Year GLP Test substance Method

: : : : : : : : : : : :

Result

: Adaptation period of at least 7 days, five male and five female rabbits. A non-permeable patch containing 2 g/kg body weight of the test material (applied neat) was placed over a 4 -5 cm2 area on each rabbit. After 24 hours exposure to the test material, the patches were removed and the exposed surface was wiped clean of any residual test material using a damp cloth. The rabbits were observed for gross toxicity and mortality at least twice daily for a period of 14 days. Since there were no mortalities, gross necropsies were performed on all survivors at terminal sacrifice. The body weights were recorded on the day of dosing and at 7 and 14 days. : All rabbits appeared active and healthy throughout the test period. There were no overt signs of gross toxicity nor was there any evidence of severe skin lesions. Eight rabbits gained weight over the 14 day observation period and two remained the same. Gross necropsies were unrevealing. All organs and tissues appeared normal. : Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex : TEST ORGANISMS: - Adaptation period: at least 7 days - Number of animals: 5 males + 5 females - Controls: no ADMINISTRATION: - Exposure route: dermal, under a non-permeable patch, over 10% of the body surface - Volume administered: no data 23 / 5 1

Source Test condition

5. Toxicity

Id 624-9 2-0 Date 31.12.2005

Test substance

Conclusion Reliability Flag 31.12.2005 Type Value Species Strain Sex Number of animals Vehicle Doses Method Year GLP Test substance Result

: : :

EXAMINATIONS: - Clinical observations, body weight, mortality and necropsy - Post dose observati on period: 14 days Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Pennwalt Corp. Batch: no data Purity: no data The acute dermal toxicity of Dimethyl Disulfide is > 2.0 g/kg body weight. (1) valid without restriction Material Safety Dataset, Directive 67/548/EEC, Critical study for SIDS endpoint (25) LD0 >= 2000 mg/kg bw rabbit New Zealand white male/female 10 other: none 2000 mg/kg other: Directive 79/831/EEC Annexe V no No mortality was observed. Apathy and prostration were noted in most of the animals between 15 minutes and 3 hours after the application of the product. An increase in the spontaneous activity was noted for some animals the first day of treatment. The behavior of the animals during the remainder of the period of observation was considered normal. No macroscopic lesion was observed. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex TEST ORGANISMS: - Acclimatation period: no data - Number of animals: 5 males + 5 females - Controls: no ADMINISTRATION: - Exposure route: dermal, under a non-permeable patch, over 10% of the body surface - Volume administered: no data EXAMINATIONS: - Clinical observations, body weight, mortality and necropsy - Post dose observation period: 15 days Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: SNEA(P) Batch: A1 Purity: no data (2) valid with restrictions Critical study for SIDS endpoint (12) 24 / 5 1

: : : : : : : : : : : : :

Source Test condition

: :

Test substance

Reliability Flag 31.12.2005

: :

5. Toxicity

Id 624-9 2-0 Date 31.12.2005

5.1.4

ACUTE TOXICITY, OTHER ROUTES

5.2.1

SKIN IRRITATION rabbit undiluted Semiocclusive 4 hour(s) 6 slightly irritating not irritating OECD Guide-line 404 " Acute Dermal Irritation/Corrosion" 1982 no other TS Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex DMDS, purity 98.98%. (2) valid with restrictions Material Safety Dataset, Directive 67/548/EEC (15) : : : : : : : : : : : : : : : rabbit undiluted Occlusive 24 hour(s) 6 1.1 slightly irritating not irritating other: EPA 40 CFR 163.81-5 yes Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex TEST ORGANISMS: - Adaptation period: 8 weeks - Number of animals: 4 m ales + 2 females - Controls: no Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Pennwalt Corp. Batch: no data Purity: no data Based on the average Primary Skin Irritation Score at 48 hours (2.02) and the average score over 14 days (1.10), Dimethyl Disulfide is considered to be a mild primary skin irritant. (1) valid without restriction (23)

Species Concentration Exposure Exposure time Number of animals Vehicle PDII Result Classification Method Year GLP Test substance Source Test substance Reliability Flag 31.12.2005 Species Concentration Exposure Exposure time Number of animals Vehicle PDII Result Classification Method Year GLP Test substance Source Test condition

: : : : : : : : : : : : : : : : :

Test substance

Conclusion

Reliability 31.12.2005

25 / 5 1

5. Toxicity

Id 624-9 2-0 Date 31.12.2005

5.2.2

EYE IRRITATION rabbit undiluted .1 ml 24 hour(s) not rinsed 6 irritating irritating OECD Guide-line 405 "Acute Eye Irritation/Corrosion" 1982 no other TS Mean scores (24+48+72 hours) for the 6 rabbits: - Chemosis: 1.89 - Enanthema: 1.33 - Iris: 1.0. - Cornea: 0.83 Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex DMDS, purity 98.98%. (2) valid with restrictions Material Safety Dataset, Directive 67/548/EEC (15) : : : : : : : : : : : : : : rabbit undiluted .1 ml other: not rinsed for 6 rabbits, rinsed after 20 -30 sec. for 3 rabbits 9 slightly irritating not irritating other: EPA-40 CFR 163-81-4 yes as prescribed by 1.1 - 1.4 The average 24 hour maximum mean total score (MMTS) for the unwashed eyes was 14.8 (minimally irritating.). For the washed eyes the 24 hour MMTS was 6 (minimally irritating). Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex TEST ORGANISMS: - Adaptation period: 7 days - Number of animals: 4 males + 5 females - Controls: no Dimethyl Disulfide is considered to be minimally irritating to both the unwashed and the washed eye. (1) valid without restriction (22)

Species Concentration Dose Exposure time Comment Number of animals Vehicle Result Classification Method Year GLP Test substance Result

: : : : : : : : : : : : : :

Source Test substance


Reliability
Flag
31.12.2005 Species Concentration Dose Exposure time Comment Number of animals Vehicle Result Classification Method Year GLP Test substance Result

: : : :

Source Test condition

: :

Conclusion Reliability 31.12.2005 5.3 SENSITIZATION

: :

26 / 5 1

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 Buehler Test guinea pig st 1 : Induction undiluted occlusive epicutaneous nd 2 : Challenge undiluted occlusive epicutaneous rd 3 : 20 not sensitizing not sensitizing other: EPA-40 CFR 163-81-6 1985 yes

Type Species Concentration

: : :

Number of animals Vehicle Result Classification Method Year GLP Test substance Result

: : : : : : : :

: In the preliminary screen, no erythema was observed at any of the concentrations of test material applied to the skin over a 48 hour period. The test material was therefore tested neat in the full scale sensitization study. After the initial and second challenge applications, the guinea pigs did not exhibit any erythema and were considered non sensitized. Expected responsed were noted in the positive control animals. The data validates the responsiveness of the guinea pigs to DNCB. : Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex : TEST ORGANISMS: - guinea pigs - Weight at study initiation: 256-424 g - Adaptation period: 10 days - Number of animals: 10 males for the test substance 10 males for the positive control (DNCB 0.3%) METHOD - Induction: 10 applications every 2 days (excluding week-end) - duration of the application: 6 hours/day - Challenge test: 10 days after the last induction application - Scoring local reaction: 24 and 48 hours after each induction application and after the challege application Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Pennwalt Corp. Batch: no data Purity: no data Dimethyl Disulfide is a non (contact) sensitizer. (1) valid without restriction Material Safety Dataset, Directive 67/548/EEC (24)

Source Test condition

Test substance

Conclusion Reliability Flag 30.12.2005 5.4

: : :

REPEATED DOSE TOXICITY

Type Species Sex Strain Route of admin.

: : : : :

rat male/female Sprague-Dawley inhalation 27 / 5 1

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 90 days 6 h/day; 5 d/week 4 weeks 10, 50, 150, 250 ppm yes, concurrent vehicle ca. 10 ppm = 50 ppm OECD Guide-line 413 "Subchronic Inhalation Toxicity: 90-day Study" 1981 yes

Exposure period Frequency of treatm. Post exposure period Doses Control group NOAEL LOAEL Method Year GLP Test substance Method

: : : : : : : : : : :

Result

: Four groups of 20 male and 20 female Sprague -Dawley were exposed 6 hours/day, 5 days/week to 0, 10, 50, 150, or 250 ppm DMDS. The exposure of the 150 ppm group was terminated after 6 weeks and its treatment-free subgroup necropsied 2 weeks later. The remaining groups received a 13 week exposure period followed by four weeks for the treatment-free subgroups. : MORTALITY There was no treatment-related mortality. CLINICAL SIGNS
The only clinical signs attributable to treatment were
salivation, lacrimation or reduced activity during exposure
1 and 2 of the 150 and 250 ppm groups and a low incidence of dyspnoea
or wheezing in the early part of the study,
particularly in the 250 ppm animals at week 1.
FOB
Functional observation tests indicated no evidence of neurotoxicity.
BOBY WEIGHT
There was a dosage-related decrease in body weight gain over the
treatment period in treated groups compared with
controls.
FOOD CONSUMPTION
Differences in food consumption paralleled those of body
weight gain and were not statistically significant in the 50 ppm males or the
10 ppm groups.
OPHTHALMOSCOPY
The eyes of the animals were unremarkable.
H AEMATOLOGY
Haematological profiles suggested a possible small reduction in Hb, RBC
and PCV in the 250 ppm female group only.
BOOLD CHEMISTRY
Blood chemistry examinations showed treatment-related
changes in ALT, alkaline phosphatase and bilirubin.
ORGAN WEIGHTS
There were no changes in organ weights that were considered
to be treatment-related.
MACROSCOPIC OBSERVATIONS
There were no treatment-related macroscopic abnormalities at necropsy.
MICROSCOPIC OBSERVATIONS
28 / 5 1

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 In the 10, 50 and 250 ppm animals examined microscopically there was a dose-related effect on nasal mucosa. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex TEST ORGANISMS: - Number of animals: 100 rats : 20 males + 20 females / dose group (4 dose groups + 1 control group) - Aclimatation period: 14 days ADMINISTRATION: - Type of inhalation study: whole body
- Production of test atmospheres:
Five horizontal flow, recirculating exposure chambers were
used.
- Vehicle: filtered air
- Exposure chamber test article concentration
* Measured concentration
Samples for analysis were withdrawn from the exposure
chambers twice hourly.
SATELLITE GROUPS: none RECOVERY GROUPS 10 rats/sex/group were allowed to recover for 4 weeks after termination of the main study animals in groups 1, 2, 3 and 5 and for 2 weeks for group 4 animals. CLINICAL OBSERVATIONS AND FREQUENCY: - Clinical observations
* Morbidity and mortality
* Clinical signs
* Functional observation tests
* Body weight
* Food consumption
* Ophthalmoscopy
- Laboratory investigations
* Haematology:
Haemoglobin, mean cell volume, red blood cell count and
indices: mean cell haemoglobin, mean cell haemoglobin
concentration packed cell volume, total and differential
white blood cell count platelet count.
* Clinical chemistry:
aspartate aminotransferase, alanine aminotransferase,
alkaline phosphatase, sodium, potassium, chloride, calcium
inorganic phosphorus, glucose, urea, total bilirubin,
creatinine, total protein, albumin, albumin/globulin ratio
total cholesterol.
- Pathology
* Necropsy
Full internal and external examination at sacrifice
* Organ weights
* Histology
- Statistical evaluation * ANOVA, T-test Body weight: week 0 * ANOVA, Regression and Dunnett's 29 / 5 1

Source Test condition

: :

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 * ANCOVA, Dunnett's * Kruskal -Wallis, Terpstra-Jonckheere, Wilcoxon Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% Clear treatment-related effects were seen at 50 and 250 ppm and were present to a marginal degree at 10 ppm. It was concluded that the effect level was 50 ppm. The no -effect level was in the region of, but less than, 10 ppm due to the reversible changes in the nasal mucosa (1) valid without restriction Material Safety Dataset, Critical study for SIDS endpoint (11) : : : : : : : : : : : : : : : : : rabbit male/female New Zealand white dermal 28 days 6 h/day no 0.01, 0.1, 1 ml/kg/day (10.63, 106.3 and 1063 mg/kg bw/d) other: sham treated with the occlusive dressing = 10.63 mg/kg bw = 106.3 mg/kg bw OECD Guide-line 410 "Repeated Dose Dermal Toxicity: 21/28-day Study" 1981 yes DMD S was administered daily, by dermal occlusive application (6 hours daily) to four groups of albino rabbits. The dose levels equivalent to 0, 10.63, 106.3, and 1063 mg/kg body weight/day, respectively. The control and 1.0 ml/kg/d group consisting of 10 males and 10 females, and the 0.01 and 0.1 ml/kg/d group consisting of 5 males and 5 females. The animals of the 0.01 and 0.1 ml/kg/d group were treated five days a week during a fourweek period, whereas animals of the 1 ml/kg/d group were treated with DMDS for 2 1/2 weeks (i.e. 13 days of treatment). CLINICAL SIGNS: During daily treatment with DMDS, lethargy was observed in a dose related manner in the mid and high dose group. No treatment-related clinical signs were observed in the animals of the low dose group or in the controls. MORTALITY: During the second and third week of the study treatment-related mortality occurred in males and females of high dose group and treatment was suspended after 13 days of treatment. SKIN REACTIONS: Repeated dermal administration of DMDS caused severe, dose-dependent skin irritation in all dose groups. BLOOD EXAMINATIONS: Haematology and clinical chemistry examinations revealed differences in some blood paremeters and clinical chemistry in the high dose group m ales. No treatment related changes were observed in females. PATHOLOGY: The absolute and relative organ weights measured at autopsy did not show statistically significant differences. Macroscopic examination 30 / 5 1

Test substance

Conclusion

Reliability Flag 31.12.2005 Type Species Sex Strain Route of a dmin. Exposure period Frequency of treatm. Post exposure period Doses Control group NOAEL LOAEL Method Year GLP Test substance Method

: :

Result

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 at autopsy did not reveal any treatment-related changes other than the dermal lesions induced during the treatment with DMDS. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex TEST ORGANISMS: - Number of animals: The control and top-dose group comprised 10 males and 10 females, whereas the low - and mid-dose group comprised 5 males and 5 females. - Aclimatation period: 13 days ADMINISTRATION: - Route: dermal Doses were applied by volume. The respective amounts of the test substance were applied topically to the intact, shaven skin. The test site was covered with porous gauze dressing fixed onto a non-irritating tape. The entire trunk was wrapped to maintain the gauze dressing in position and to retard evaporation of volatile substances. The animals of the con trol group were sham-treated with the patches only. CLINICAL OBSERVATIONS AND FREQUENCY: - Clinical signs: twice a day on exposure days and once a day on non-exposure days. - Mortality: twice a day. - Dermal reactions: At the start of the study and prior to each daily administration. - Body weight: - Food consumption: - Blood examinations: haematology and clinical chemistry determinations were conducted in blood or plasma of the animals * Haematology: Hemoglobin, hematocrit, red blood cell count, white blood cell count, differential leukocyte count, platelet count, mean cell volume, mean cell haemoglobin concentration, mean cell haemoglobin * Biochemistry: . Electrolytes: calcium, chloride, phosphorous, potassium, sodium, . Enzymes: alkaline phosphatase, alanine -aminotransferase, aspartate-aminotransferase, gamma-glutamyl -transferase . Other: albumin, blood creatinine, blood urea nitrogen, albumin/globulin, glucose, total bilirubin, total cholesterol, total serum protein, bile acids ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC): - Weighed organs: adrenals, brain, heart, kidneys, liver, lungs, ovaries, spleen, testes, thyroid and thymus. - Microscopic examinations: Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% The NOAEL of DMDS for systemic toxicity is 10.63 mg/kg bw/d. For local skin effects, the NOAEL is lower than 10.63 mg/kg bw/d. (1) valid without restriction Material Safety Dataset, Critical study for SIDS endpoint (6) 31 / 5 1

Source Test condition

: :

Test substance

Conclusion Reliability Flag 31.12.2005

: : :

5. Toxicity

Id 624-9 2-0 Date 31.12.2005

Type Species Sex Strain Route of admin. Exposure period Frequency of treatm. Post exposure period Doses Control group NOAEL LOAEL Method Year GLP Test substance Method

: : : : : : : : : : : : : : : :

rabbit male/female New Zealand white dermal 14 days 6 h/day no 0.1, 0.5 and 1 ml/kg/day (106, 503 and 1063 mg/kg/day) other: sham treated with the occlusive dressing < .1 mg/kg = .1 mg/kg other: range findi ng study yes other TS

Result

Source Test substance

Reliability 31.12.2005 5.5

: In this range -finding study, DMDS was administered to a restricted number of albino rabbits by dermal occlusive application, daily, during a two-week period. The dose levels applied were 106.3, 531.5, and 1063 mg DMDS/kg body weight/day, repectively, and the daily exposure period was 6 hours. The control group was sham treated with the occlusive dressing only. : During exposure temporary signs slight lethargy in the low-dose group, distinct lethargy in the mid-dose group, and unconscinousness in the high-dose group. At the end of each daily exposure, these effects were no longer observed. During the entire test period of the study, the controls did not show any signs of abnormal beha viour after treatment with the patches only. Repeated dermal administration of DMDS caused severe skin lesions in all three dose groups. : Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex : Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% : (1) valid without restriction
(17)

GENETIC TOXICITY IN VITRO Salmonella typhimurium reverse mutation assay Strains: TA 1535, TA 1537, TA 1538, TA 98, TA 100 0, 5, 50, 150, 500, 1500, and 5000 g/plate >= 5000 g/plate with and without negative OECD Guide-line 471 1983 yes

Type System of testing Test concentration Cycotoxic concentr. Metabolic activation Result Method Year GLP Test substance Method

: : : : : : : : : :

: PRELIMINARY TOXICITY ASSAY The preliminary toxicity assay was used to establish the dose range over which the test article would be assayed. MUTAGENICITY ASSAY 32 / 5 1

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 - Five dose levels of test article along with appropriate vehicle control and positive controls were plated with overnight cultures of TA98, TA100, TA1535, TA1537 and TA1538 on selective agar in the presence and absence of Aroclor induced rat liver S9. All dose levels of test article, vehicle control and positive controls were plated in triplicate. - Second mutation test The procedure was repeated at a later date. EVALUATION OF RESULTS The mean number of revertant colonies for all treatment groups is compared with those obtained for negative and positive control groups. The effect of metabolic activation is assessed by comparing the results obtained both in the presence and absence of the liver microsomal fraction for each treatment group. A compound is deemed to provide evidence of mutagenic potential if (1) a statistically s ignificant dose-related increase in the number of revertant colonies is obtained in two separate experiments, and (2) the increase in the number of revertant colonies is at least twice the concurrent solvent control value. The positive controls responded as expected. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex CONTROL MATERIALS - Negative: culture medium - Solvent: Dimethylsulphoxide - Positive: * With S-9 mix 2-Aminoanthracene at 2 g/plate for strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100. * Without S-9 mix 2-Nitrofluorene at 10 g/plate for strains TA 1538 and TA 98. 9-Aminoacridine at 20 g/plate for strain TA 1537. Sodium azide at 5 g/plate for strains TA 1535 and TA 100. ACTIVATION - S9 derived from Sprague -Dawley rats induced with a single intraperitoneal injection of Aroclor 1254, 500 mg/kg, five days prior to sacrifice. - S9 mix composition: Component S9 Sodium phosphate buffer (pH 7.4) gluc ose 6 -phosphate N ADP KCl MgCl2 Concentration 10% (v/v) 100 mM 5 mM 4 mM 33 mM 8 mM

Remark Source Test condition

: : :

TEST ORGANISMS - Salmonella typhimurium strains: TA98, TA100, TA1535, TA1537 and a 1538 - test organisms were properly maintained and were checked for appropriate genetic markers (rfa mutation, R factor) TEST CONCENTRATIONS (a) Preliminary cytotoxicity assay: Plate incorporation assay: 0, 5, 50, 500 and 5000 g per 33 / 5 1

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 plate were evaluated with and without S9 activation in all strains. A single plate was used, per dose, per condition. (b)Mutation assays: Plate incorporation assay: 50, 150, 500, 1500 and 5000 g per plate were evaluated in triplicate in the presence and absence of S9 activation; all test strains were used. Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Purity 98.98% (1) valid without restriction Material Safety Dataset, Critical study for SIDS endpoint (1) : : : : : : : : : : : Salmonella typhimurium reverse mutation assay Strains: TA 1535, TA 1537, TA 1538, TA 98, TA 100 50, 166, 500, 1666, 5000 g/plate 5000 g/plate with and without negative OECD Guide-line 471 1983 yes PRELIMINARY TOXICITY ASSAY The preliminary toxicity assay was used to establish the dose range over which the test article would be assayed. MUTAGENICITY ASSAY - Five dose levels of test article along with appropriate vehicle control and positive controls were plated with overnight cultures of TA98, TA100, TA1535, TA1537 and TA1538 on selective agar in the presence and absence of Aroclor induced rat liver S9. All dose levels of test article, vehicle control and positive controls were plated in triplicate. - Second mutation test The procedure was repeated at a later date. TEST PROCEDURE - Without metabolic activation 0.1 ml aliquots of bacterial suspension is added to each of one set of sterile tubes. 0.1 ml of the test compound is added to cultures at five concentrations. The negative control is the chosen solvent. The appropriate positive control is also included. - With metabolic activation Methodology is as described above except that 0.5 ml of liver homogenate S-9 mix is added to the tubes in place of sterile buffer. EVALUATION OF RESULTS The mean number of revertant colonies for all treatment groups is compared with those obtained for negative and positive control groups. The effect of metabolic activation is assessed by comparing the results obtained both in the presence and absence of the liver microsomal fraction for each treatment group. A compound is deemed to provide evidence of mutagenic potential if (1) a statistically significant dose-related increase in the number of revertant colonies is obtained in 34 / 5 1

Test substance

Reliability Flag 30.12.2005 Type System of testing Test concentration Cycotoxic concentr. Metabolic activation Result Method Year GLP Test substance Method

: :

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 two separate experiments, and (2) the increase in the number of revertant colonies is at least twice the concurrent solvent control value. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex CONTROL MATERIALS - Negative: culture medium - Solvent: Dimethylsulphoxide - Positive: * With S-9 mix 2-Aminoanthracene at 5 g/plate for strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100. * Without S-9 mix 2-Nitrofluorene at 5 g/plate for strains TA 1538 and Ta98 9-Aminoacridine at 150 g/plate for strain TA 1537. Sodium azide at 10 g/plate for strains TA 1535 and TA 100. ACTIVATION - S9 derived from Sprague -Dawley rats induced with a single intraperitoneal injection of Aroclor 1254, 500 mg/kg, five days prior to sacrifice. - S9 mix composition: Component volume S9 100 l Sodium phosphate buffer 0.2M (pH 7.4) 500 l glucose 6 -phosphate 5 l N ADP 0.1 M 40 l KCl 1.65 M 20 l MgCl2 0.4 20 l TEST ORGANISMS - Salmonella typhimurium strains: TA98, TA100, TA1535, TA1537 and a 1538 - test organisms were properly maintained and were checked for appropriate genetic markers (rfa mutation, R factor) TEST CONCENTRATIONS (a) Preliminary cytotoxicity assay: Plate incorporation assay: 0, 50, 144, 500, 1444 and 5000 g per plate were evaluated without S9 activation with strains TA100 and TA 1538. Two plate was used, per dose, per condition. (b)Mutation assays: Plate incorporation assay: 0, 50, 166, 500, 1666 and 5000 g per plate were evaluated in triplicate in the presence and absence of S9 activation; all test strains were used. Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Purity: no data Dimethyldisulfide was negative in the Ames/Salmonella tester strains TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation preparation over the dose range 50-5000 g/plate. (1) valid without restriction Critical study for SIDS endpoint (27) : : : : Chromosomal aberrati on test Human Lymphocytes 3.7; 11.1; 33.3; 100; 300 g/ml >= 300 g/ml 35 / 5 1

Source Test condition

: :

Test substance

Conclusion

Reliability Flag 31.12.2005 Type System of testing Test concentration Cycotoxic concentr.

: :

5. Toxicity
with and without ambiguous OECD Guide-line 473 1983 yes

Id 624-9 2-0 Date 31.12.2005

Metabolic activation Result Method Year GLP Test substance Method

: : : : : :

: - Preliminary Cytotoxicity Assay: The dose levels used in the chromosome aberration assay were established on the basis of the results of a preliminary toxicity test carried out with 6 concentrations of the test substance (ranging from 0.5 to 1000.0 g/ml), both in the absence and in the presence of the metabolic activation system (S -9 mix). The highest concentration for the toxicity test was determined by the limit of the solubility of the test substance in the tissue culture medium. - Cytogenetic Assay: * Cell Treatment After 48 h of incubation, the cultures were centrifuged. The cell pellets were resuspended in tissue culture medium supplemented with 20 mM HEPES (and 10% S-9 mix, for the test with metabolic activation) and appropriate test solutions. An untreated culture and a culture receiving DMSO served as negative controls. For each concentration of the test substance and for the controls one culture was used. Without S9, the cultures were incubated in closed tubes for another 24 hours including a 2 hour colcemid treatment. With S-9 mix, the exposure of the cells to the test substance was reduced to only 2 hours, because of the toxicity of the S -9 mix for the cells. After the 2 hour incubation period, the cells washed and supplied with freshly prepared culture medium. The cells were incubated for a further 22 hours (including a 2 hour colcimid treatment. * Cell harvesting: Two hours before the end of the total incubation period the cells were arrested in the metaphase stage of the mitosis by the addition of colcemid. The cells were harvested, treated with a hypotonic solution, fixed three hours, and transferred to clean microscope slides. Two slides were prepared from each culture. The slides were stained 1000 stimulated lymphocytes were examined (500 from each slide) to determine the mitotic index (percentage of cells in mitosis). * Metaphase analysis:
From each culture, 100 well-spread metaphases (each
containing 46 chromosomes) were analysed by microscopic
examination for a wide range of structural chromosome
aberrations (gaps, breaks, fragments, dicentrics, exchanges
etc.) and other anomalies (endoreduplication, polyploidy),
according to the criteria recommended by Savage (1975).
- Evaluation criteria:
The major crite rion to designate the results of a chromosome
aberration test as positive is a dose-related, statistically
significant increase in the number of cells with structural
chromosome aberrations. However, a clear dose-response
relationship can be absent because the yield of chromosome
aberrations can vary markedly with post-treatment sampling
time of an asynchronous population and because increasing
doses of clastogens can induce increasing degrees of mitotic
delay. A test substance producing neither a dose-related,
36 / 5 1

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 statistically significant increase in the number of cells
with structural chromosome aberrations, nor a statistically
significant and reproducible positive response at any of the
doses is considered non-clastogenic in this system.
: The test substance did not induce a statistically significant increase in the number of cells with structural chromosome aberrations at non toxic concentrations, both in the absence and in the presence of the S-9 mix. At the very toxic concentration of 300.0 g/ml, both in the absence and in the presence of the S-9 mix, the test substance induced a statistically significant increase in the number of cells with structural chromosome aberrations. The positive control substances, mitomycin C and
cyclophosphamide, induced the expected increase in the
incidence of structural chromosome aberrations.
: Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex : Control Materials: Negative: DMSO Solvent: The test article (dissolved in DMSO) was soluble in culture medium at a maximum concentration of 1 mg/mL
Positive: -S9: mitomycin C (MMC) 0.05 g/mL
+S9: cyclophosphamide (CP) 25 g/mL
Activation:
S9 derived from adult male Wistar rats (Aroclor 1254 induced
rat liver). The composition of the rat liver S9 reaction mix
was: 8 mM magnesium chloride, 33 mM potassium chloride, 5 mM
glucose-6-phosphate, 4 mM nicotinamide adenine dinucleotide
phosphate (NADP), 100 mM sodium phospahte and 40% S9.
Culture Medium:
RPMI 1640 medium supplemented with heat -inactivated foetal
calf serum, 100 units penicillin/mL, 100 g streptomycin/mL,
2 mM L -glutamine and 25 l phytohaemagglutinin/ml
Test compound concentrations used:
Treatment Treatment Recovery Dose levels
condition time time (g/mL) -S9 24hr 24 hr 3.7, 11.1, 33.3 100, 300 +S9 2 hr 24 hr 3.7, 11.1, 33.3 100, 300 : Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.98% : (1) valid without restriction : Material Safety Dataset, Critical study for SIDS endpoint (14) : : : : : : : : : : Mammalian cell gene mutation assay HGPRT assay on CHO cells 0.46; 1.37; 4.12; 12.3; 37.0; 74.0; 111; 333; 667 and 1000 g/ml 74.0-1000 g/ml with and without negative OECD Guide-line 476 1984 yes 37 / 5 1

Result

Source Test condition

Test substance

Reliability Flag 31.12.2005 Type System of testing Test concentration Cycotoxic concentr. Metabolic activation Result Method Year GLP Test substance

5. Toxicity

Id 624-9 2-0 Date 31.12.2005

Method

The dose levels used in the HGPRT assay were established on the basis of the results of a preliminary solubility test. A final concentration of 1,000 g/ml was chosen as highest concentration for the HGPRT assays. The two independent HGPRT-assays were carried out with single cultures for each concentration of the test substance and for the negative and positive controls. In the absence of the S -9 mix, the test substance induced neither a concentration-related increase in the mutant frequency nor a reproducible positive response at one of the test concentrations. In the presence of a metabolic activation system, DMDS induced a slight increase in mutant frequency at several concentrations, in both HGPRT assays. These increases were neither concentration-related nor clearly reproducible. In both HGPRT assays, the test substance appeared to be highly toxic to CHO cells at a concentration range from 74.0-1,000 g/ml. The positive control substances, ethylmethanesulfonate and dimethylnitrosamine, induced the expected increase in the mutant frequency. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex - Control Materials: * Negative: DMSO * Solvent: The test article (dissolved in DMSO) was soluble in culture medium at a maximum concentration of 1 mg/mL * Positive: -S9: Ethylmethanesulfonate 0.2 ml/L +S9: Dimethylnitrosamine 2 or 4 ml/L - Activation: S9 derived from adult male Wistar rats - Culture Medium: Ham's F-12 medium supplemented with 10% heat-inactivated foetal calf serum, 50 g gentamicin/mL and 2 mM L-glutamine. - Evaluation of the results: The following criteria were used to evaluate the data obtained in the HGPRT assay (Li et al. 1987) a) the survival (absolute cloning efficiency) of the negative controls should not be less than 50%, b) the mean mutant frequency of the negative controls should fall within the range of 0-20 6 -TG resistant mutants per 10e6 clonable cells, c) the positive controls must induce a response of a magnitude appropriate for the mutagen under the experimental conditions applied, d) the highest test substance concentration should, if possible, result in a clear cytotoxic response (e.g. 10-30% of the relative initial survival). Any apparent increase in mutant frequency at concentrations of the test substance causing more than 90% toxicity is considered to be an artifact and not indicative of genotoxicity. Genotoxicity of the test substance was evaluated using the following criteria (Li et al. 1987): a) a concentration-related increase in mutant frequency, b) a reproducible positive response for at least one of the test substance concentrations (e.g. the mean mutant 38 / 5 1

Result

Source Test condition

: :

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 frequency should be more than 20 mutants per 10e6 clonable cells). Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% No evidence for a genotoxic effect of DMDS was found in cultured CHO cells, under the conditions used in the HGPRT assay. (1) valid without restriction Material Safety Dataset, Critical study for SIDS endpoint (13) : : : : : : : : : : : DNA damage and repair assay Rat hepatocytes in primary culture 1; 5; 10; 50; 100; 200 and 300 g/ml >= 100 g/ml without negative OECD Guide-line 482 1986 yes other TS - Cytotoxicity evaluation: The test compound cytotoxicity was assessed for both DNA repair studies at the end of the treatment: Each concentration of Dimethyldisulfide was tested in triplicate. - Autoradiography: Autoradiographs were prepared by dipping slides in a photographic emulsion then developed. Slides were stained in hematoxylin -phloxin. - Slide assessment: For each cell, following nuclear grain court, cytoplasmic count was performed on 3 areas of the same size as the nucleus and adjacent to it. - Data interpretation The test compound is considered positive when the mean nuclear grain court is statisticaly greater than that of the control, the mean net nuclear grain court is above 3 grains per nucleus, and the percentage of treated cells in repair is significantly different from that of the controls. In addition, the effect must be shown to be reproducible between experiments. Results - Cytotoxic at 100, 200 and 300 g/ml IC50 evaluated by LDH release: 98 g/ml (2nd study) - not genotoxic at concentrations of 10, 50, 100 and 200 g/ml The positive controls responded as expected. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex - Control Materials: * Negative: pyrene 1 M * Solvent: DMSO The test article was soluble in culture medium at a maximum 39 / 5 1

Test substance

Conclusion

Reliability Flag 31.12.2005 Type System of testing Test concentration Cycotoxic concentr. Metabolic activation Result Method Year GLP Test substance Method

: :

Result

Source Test condition

: :

5. Toxicity
concentration of 100 g/mL * Positive: . 7,12-DMBA (10 M) . 2-aminofluorene (0.1 and 0.5 M)

Id 624-9 2-0 Date 31.12.2005

Test substance

Conclusion Reliability Flag 31.12.2005 5.6

: : :

- Number of cultures/concentration/study: 3 Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% Not genotoxic in vitro in the DNA repair test. (1) valid without restriction Material Safety Dataset, Critical study for SIDS endpoint (16)

GENETIC TOXICITY IN VIVO Micronucleus assay mouse male/female Swiss inhalation 6 h/day for 4 days 0 , 250 and 500 ppm negative OECD Guide-line 474 "Genetic Toxicology: Micronucleus Test" 1983 yes other TS

Type Species Sex Strain Route of admin. Exposure period Doses Result Method Year GLP Test substance Method

: : : : : : : : : : : :

: Three groups of mice were exposed during 6 hours a day for 4 consecutive days (days 0 through 3) to atmospheres containing 0 ppn (5/sex), 250 ppm (5/sex) and 500 ppm DMDS (10/sex). The positive control group (5/sex) was treated once intraperitoneally, 24 hours before sacrifice, with 1.5 mg Mitomycin C per kg body weight. Bone marrow cells were collected from the femur and processed into smears for microscopic examination. One smear from each animal was examined for the presence of micronucleated poly- and normochromatic erythrocytes, (abbreviated MPE and MNE, respectively), and the total numbers of poly- and normochromatic erythrocytes (PE and NE) in a total of at least 2000 erythrocytes (E) in such a way that a minimum of 1000 PE was observed. : Exposure to DMDS resulted in clear signs of intoxication both at the 250 ppm and the 500 p pm level. Mortality was observed in some animals at 500 pmm group. Exposure to 250 ppm and 500 ppm DMDS resulted in body weight loss both in males and females. There were no indications for increases in the incidences of MPE, MNE or ME attributable to treatment with the test material. Mean numbers of PE per 1000 E were slightly lower in mice exposed to 500 ppm DMDS, both in males and females (0.001<P<0.01) pointing to slight cytotoxic effects on bone marrow cells. 40 / 5 1

Result

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 Animals treated with the mutagen Mitomycin C showed an increased incidence of MPE. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex * CONTROL MATERIALS - Positive : Mitomycin C, single ip administration, 1.5 mg/kg Test substance: D imethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% It was concluded that the results of the micronucleus test did not provide any indication of chromosomal damage and/or damage to the mitotic apparatus in bone marrow cells of mice exposed to DMDS. (1) valid without restriction Material Safety Dataset, Critical study for SIDS endpoint (5) Unscheduled DNA synthesis rat male Wistar inhalation 4 hours 0 and 500 ppm negative other: OECD Guide-line 482 1986 yes other TS Dimethyldisulfide (DMDS) was examined for its potential to induce unscheduled DNA synthesis (UDS) in primary rat hepatocytes after short-term exposure of male wistar rats to the test substance by inhalation. For the genotoxicity assay male rats were exposed by inhalation for a period of 4 h to one high concentration of 500 ppm DMDS (m aximally tolerated concentration). Immediately after exposure and after subsequent non-exposure periods of 16 and 24 h, animals were sacrificed for isolation of hepatocytes. The DNA-repair activities were examined by autoradiography in monolayer cultures o f hepatocytes, incubated in the presence of [methyl-3H]thymidine. The hepatocarcinogen 2-acetylaminofluorene (2 AAF), was used as a positive control in the in vivo/in vitro DNA -repair assay and in the in vitro DNA-repair assay (2 AAF). Hepatocytes isolated from animals exposed to air only served as negative controls. DMDS did not induce DNA-repair activities in hepatocytes, either during the 4 h exposure period or during the subsequent 16 h or 24 h after the exposure period. The positive control substance, 2-AAF, induced the expected increase in DNA-repair activities. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex * CONTROL MATERIALS - Positive : . in vivo: 2-AAF, 50 mg/kg single oral administrati o n . in vitro: 2-AAF, 10e -4M 41 / 5 1

Source Test condition

: :

Test substance

Conclusion

Reliability Flag 31.12.2005 Type Species Sex Strain Route of admin. Exposure period Doses Result Method Year GLP Test substance Method

: :

: : : : : : : : : : : : :

Result

Source Test condition

: :

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% It was concluded that DMDS did not induce DNA-repair in rat hepatocytes. (1) valid without restriction Material Safety Dataset, Critical study for SIDS endpoint (2)

Test substance

Conclusion Reliability Flag 31.12.2005 5.7 CARCINOGENICITY

: : :

5.8.1

TOXICITY TO FERTILITY

5.8.2

DEVELOPMENTAL TOXICITY/TERATOGENICITY rat


female
other: Crl: CD(SD)BR
inhalation
day 6 to day 15 of gestation
6 h/day
up to gestation day 20
5; 15; 50 ppm
yes, concurrent no treatment
= 5 ppm
= 50 ppm
= 15 ppm
OECD Guide-line 414 "Teratogenicity"
1981
yes
Three groups of 30 mated female rats were exposed to DMDS by whole body exposure at 5, 15 or 50 ppm for 6 hours daily from day 6 to day 15 of gestation. A similar group of 30 rats, exposed to filtered air only over the same period, served as controls. All animals were maintained until day 20 of gestation, killed and their uterine content assessed. The chamber concentrations of the test article were close to target values throughout the exposure period. There were no deaths. A higher incidence of rough haircoat was observed at 50 ppm. Clinical condition at 5 and 15 ppm did not differ from controls. Dosage-related reductions in weight gain were observed at 15 and 50 ppm. Food intake was lower than controls at 50 ppm but comparable at 5 or 15 ppm. No unusual lesions were observed at necropsy. There was no effect of treatment on pre or post-implantation loss, litter size or sex ratio. Litter and foetal weights were reduced at 50 ppm. At 5 and 15 ppm these parameters were comparable to controls. No malformations were observed in foetuses from the treated groups. A slightly higher incidence of retarded ossification was observed at 50 ppm but was considered to indicate delayed maturation, as a result of the lower foetal weight, rather than a teratogenic 42 / 5 1

Species Sex Strain Rout e of admin. Exposure period Frequency of treatm. Duration of test Doses Control group NOAEL maternal tox. NOAEL teratogen. NOAEL Fetotoxicity Method Year GLP Test substance Method

: : : : : : : : : : : : : : : :
:

Result

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 effect. Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex TEST ORGANISMS: - Number of animals: 100 rats : 25 females / dose group (3 dose groups + 1 control group) - Aclimatation period: no data ADMINISTRATION: - Type of inhalation study: whole body - Vehicle: filtered air - Exposure chamber test article concentration * Measured concentration Samples for analysis were withdrawn from the exposure chambers twice hourly. EXPERIMENTAL OBSERV ATION - Morbidity and mortality All females were examined twice daily to detect any which were dead or moribund. - Clinical observations All females were examined daily from day 3 to day 20 of gestation. Any abnormalities of appearance or behaviour or other signs of reaction to treatment or ill health were recorded. - Body weight The body weight of each female was recorded - Food intake The amount of food consumed by each cage of females was recorded daily from day 3 to day 20 of gestation and reporte d on the body weight intervals. - Terminal studies * Necropsy All females were killed on day 20 of gestation, in random group order and examined macroscopically. * Uterine/implantation data pregnancy status number of corpora lutea number and intrauterine position of implantations subdivided into: live foetuses early intrauterine deaths late intrauterine deaths dead foetuses - Foetal data Foetuses were weighed individually, examined externally and sexed. The viscera of approximately one half of the foetuses in each litter were examined. The skeleton was examined and preserved and stored in absolute glycerol (containing thymol crystals). The remaining foetuses were placed in Bouin's fluid for at least two weeks then transferred to 70% industrial methylated spirit. Foetal abnormalities were recorded as malformations (rare and/or potentially lethal defects) and variations (cormnonly occurring non lethal abnormalities). Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% Exposure to DMDS at 50 ppm elicited maternal toxicity, with 43 / 5 1

Source Test condition

: :

Test substance

Conclusion

5. Toxicity

Id 624-9 2-0 Date 31.12.2005 associated fetal growth retardation (demonstrated by low
weight and retarded ossification). There was no indication
of a teratogenic effect. At 15 ppm, less marked maternal
toxicity was observed and there were no fetal effects.
There was no adverse effect of treatment, maternal or
fetal, at 5 ppm.
(1) valid without restriction Material Safety Dataset, Critical study for SIDS endpoint (3) : : : : : : : : : : : : : : rat female other: Crl: CD(SD)BR inhalation day 6 to day 15 of gestation 6 h/day up to gestation day 20 10, 50 and 250 ppm yes, concurrent no treatment < 10 ppm other: range-finding study yes other TS

Reliability Flag 31.12.2005 Species Sex Strain Route of admin. Exposure period Frequency of treatm. Duration of test Doses Control group NOAEL maternal tox. Method Year GLP Test substance Method

: :

Result

: Three groups of 7 time-mated female rats were exposed by inhalation (whole body) to concentrations of 10, 50 or 250 ppm of DMDS daily from day 6 to day 15 of gestation. A similar group of animals exposed to filtered air by the same route and over the same period acted as controls. All animals were maintained to day 20 of gestation when they were killed and their uterine contents assessed. : All animals survived to day 20 of gestation. Comnon clinical signs were observed at an incidence which increased with dose, in the treated groups only. Dosage -related reductions in body weight gain were apparent in all treated groups over the exposure period. Dosage -related reductions in food intake were apparent in all treated groups over the exposure period. In the intermediate and high dose groups the lower intake persisted until termination. Pregnancy incidence was within the expected range in all groups. Pre-implantation loss was within the expected range in all treated groups. There was no adverse effect of treatment on the incidence of intrauterine deaths. Litter size was within the expected range in all treated groups. Sex ratio was within the expected range in all groups. Mean litter weight was higher than controls in all treated groups. Mean foetal weight showed a dosage -related reduction in the treated groups, but was considered an equivocal result as values for the control and low dose groups exceeded normal limits. No malformations were observed at external examination of foetuses and the incidence of variations did not indicate an adverse effect of treatment. : Atofina, Paris-la -Dfense, France. Atofina Paris La Dfense Cedex : Test substance: Dimethyl disulfide C AS no.: 624-9 2-0 Source: Atochem Purity: 99.88% : (1) valid without restriction 44 / 5 1

Source Test substance

Reliability

5. Toxicity
31.12.2005 5.8.3 TOXICITY TO REPRODUCTION, OTHER STUDIES

Id 624-9 2-0 Date 31.12.2005 (4)

5.9

SPECIFIC INVESTIGATIONS

5.10

EXPOSURE EXPERIENCE

5.11

ADDITIONAL REMARKS

45 / 5 1

6. Analyt. Meth. for Detection and Identification

Id 624-9 2-0 Date 31.12.2005

6.1

ANALYTICAL METHODS

6.2

DETECTION AND IDENTIFICATION

46 / 5 1

7. Eff. Against Target Org. and Intended Uses

Id 624-9 2-0 Date 31.12.2005

7.1

FUNCTION

7.2

EFFECTS ON ORGANISMS TO BE CONTROLLED

7.3

OR GANISMS TO BE PROTECTED

7.4

USER

7.5

RESISTANCE

47 / 5 1

8. Meas. Nec. to Prot. Man, Animals, Environment

Id 624-9 2-0 Date 31.12.2005

8.1

METHODS HANDLING AND STORING

8.2

FIRE GUIDANCE

8.3

EMERGENCY MEASURES

8.4

POSSIB. OF RENDERING SUBST. HARMLESS

8.5

WASTE MANAGEMENT

8.6

SIDE-EFFECTS DETECTION

8.7

SUBSTANCE REGISTERED AS DANGEROUS FOR GROUND WATER

8.8

REACTIVITY TOWARDS CONTAINER MATERIAL

48 / 5 1

9. References

Id 624-9 2-0 Date 31.12.2005

(1)

ATOCHEM. Ames metabolic activation test to assess the potential mutagenic effct of dimethyl disulphide. HRC report no. ATO 10/85427, 15 May 1985. ATOCHEM. An in vivo/in vitro rat hepatocyte DNA-repair assay with dimethyldisulfide (DMDS). TNO-CIVO, Report V 90.082, May 1990. ATOCHEM. Dimethyl Disulfide (DMDS), Inhalation Teratology Study in the Rat, Hazleton UK, Report 6205-514/9, December 1991. ATOCHEM. DIMETH YL DISULFIDE (DMDS): Inhalation renge-finding study in the pregnant rat. Hazleton-UK study no. 6142-514/8, May 1991. ATOCHEM. Examination of dimethyl disulfide in the micronucleus test, TNO-CIVO, Report V 89.366, October 1989. ATOCHEM. Repeated-dose (28 -day) dermal toxicity study with Dimethyl Disulfide (DMDS) in rabbits, TNO-CIVO Institutes, Report V 89.371/280554, February 1989. BENTVELZEN, J.M. et al, 1975.Tappi, 58, 102-5. Kinetics of methyl mercaptan oxidation and dimethyl disulfide hydrolysis in alkaline solutions. ELF ATOCHEM S.A., 1995.DIMETHYL DISULFURE.Dtermination de la biodgradabilit facile.Essai en fioles fermes. Ref 95/SAEk/0415/NM. Elf Atochem S.A., 2000.Centre d'application de Levallois. DISULFURE DE DIMETHYL E.Inhibition de la croissance des algues.Etude N 504/99/A. ELF ATOCHEM SA, 1996.DISULFURE DE DIMETHYLE. Toxicit aigu vis- -vis des daphnies.Rapport N2606/95/A. ELF ATOCHEM, DMDS: 90 day inhalation toxicity study in the rat with a 4 week recovery period, Hazelton UK, Report 6491-514/7, January 1992. ELF ATOCHEM, Dtermination de la toxicit par voie percutane chez le lapin, Hazelton - IFT, Report 505207, 2 May 1985. ELF ATOCHEM, In Vitro assay for the induction of point m utations in the HGPRT -locus of Chinese hamster ovary cells by dimethyldisulfide (DMDS), TNO-CIVO Institute, Report V 89.257, May 1990. ELF ATOCHEM. Chromosome analysis of cultured human lymphocytes following in vitro treatment with DMDS. TNO-CIVO Institute, Report V 89.045, March 1990. ELF ATOCHEM. DISULFURE DE DIMETHYLE. Tests de tolrance locale chez le lapin. Hazleton-IFT, Report 503398, 21 March 1985.

(2)

(3)

(4)

(5) (6)

(7) (8)

(9)

(10) (11)

(12)

(13)

(14)

(15)

49 / 5 1

9. References
(16)

Id 624-9 2-0 Date 31.12.2005

ELF ATOCHEM. In Vitro DNA Repair Test on Rat Hepatocytes in Primary Culture, SANOFI, Report RA860891026/PN1, 22 february 1990. ELF ATOCHEM. Repeated-dose (14 -day) dermal toxicity range-finding study with Dimethyl Disulfide (DMDS) in rabbits. TNO-CIVO Institutes, Report V 89.058/280553, July 1989. Epiwin v 3.12, syspro exp erimental database EPIWIN v3.12 Hansch,C et al. (1995) M.F. Tansy et al., Acute and Subchronic Toxicity studies of Rats Exposed to Vapors of Methyl Mercaptan and Other Reduced-Sulfur Compounds, J. Toxic. Environm. Health 1981, 8, 71-88. Pennwalt Corp. DIMETHYL DISULFIDE, EPA primary eye irritation. Products Safety Labs, Report T-5148, 24 June 1985. Pennwalt Corp. DIMETHYL DISULFIDE, EPA primary skin irritation. Products Safety Labs, Report T-5149, June 24 1985. Pennwalt Corp. DIMETHYL DISULFIDE, Guinea pig sensitization (Buehler). Products Safety Labs, Report T-5151, 30 August 1985. Pennwalt Corp., DIMETHYL DISULFIDE. EPA acute dermal toxicity limit test. Products Safety Labs, Report T-5150, 24 June 1985. Pennwalt Corp., DIMETHYL DISULFIDE. EPA acute LD50. Products Safety Labs, Report T-5147A, 14 August 1985. Pennwalt Corporation. DIMETHYL DISULFIDE, Ames Salmonella/Microsome Plate Test (EPA/OECD). Pharmakon Research International, Inc. Report PH 301-PW-003-85, 31 May 1985. Safety Data Sheet Elf Atochem January 1988 SEPPOVAARA, O. and HYNNINEN, P., 1970.On the toxicity of sulfate -mill condensates.Papper och Tr, 1, 11-23. SNEAP, Dimethyl disulfure (DMDS) - Evaluation de la toxicit aige chez le rat par voie orale, CIT Report 2064 TAR, June 1986. Syracuse Research Corporation (SRC) and EPIWIN v 3.12. Technical Data Sheet A-1130-401 Elf Atochem January 1993 WERNER, A.E.: Sulphur compounds in kraft pulp mill effluents.Can. Pulp paper Ind., 1963, 16, 3, 35-43.

(17)

(18) (19) (20) (21)

(22)

(23)

(24)

(25) (26) (27)

(28) (29) (30)

(31) (32) (33)

50 / 5 1

10. Summary and Evaluation

Id 624-9 2-0 Date 31.12.2005

10.1

END POINT SUMMARY

10.2

HAZARD SUMMARY

10.3

RISK ASSESSMENT

51 / 5 1

Appendix IV

IUCLID 5 Report for Disulfide Oil

Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and di-... Page 1 of 58

Printing Date

2008-11-10 14:42:46 CET

Restriction of specific regulatory purposes

Confidentiality

Name Legal entity owner

Disulfides, dialkyl and di-Ph, naphtha sweetening Lyondell Chemie Nederland B.V. / Rotterdam / Netherlands

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening


UUID Dossier UUID Author Date Remarks

IUC5-30cf773d-ab97-43e9-b5de-19496d0ddae4 0 jrfowles / Lyondell Chemie Nederland B.V. / Rotterdam / Netherlands 2008-11-10 11:55:21 CET

0 Related Information 0.1 Templates 0.2 Categories 0.3 Mixtures 1 General Information 1.1 Identification
Substance identification
Chemical name Legal entity

Disulfides, dialkyl and di-Ph, naphtha sweetening Lyondell Chemie Nederland B.V. / Rotterdam / Netherlands

Reference substance
Reference substance

Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and di-Ph, naphtha sweetening / 68955-96-4
EC number EC name

CAS number

CAS name

68955-96-4 Disulfides, dialkyl and di-Ph, naphtha sweetening


IUPAC name

Type of substance
Composition Origin

UVCB petroleum product

Trade names
Name Name

DSO Disulfide oil

1.2 Composition
Substance composition
Name Brief description

Disulfides, dialkyl and di-Ph, naphtha sweetening Reclaimed substancesextracted from light hydrocarbon streams during petroleum refining

Degree of purity
87.1 % (w/w)

Constituents

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Reference substance

dimethyl disulphide / dimethyl disulfide / 624-92-0


EC number EC name

210-871-0 dimethyl disulphide


CAS number CAS name

624-92-0
IUPAC name

dimethyl disulfide
Typical concentration Reference substance

12 % (w/w) methyl ethyl disulphide / methyldisulfanylethane / 20333-39-5


EC number EC name

CAS number

CAS name

20333-39-5
IUPAC name

methyldisulfanylethane
Typical concentration Reference substance

18.2 % (w/w) isopropyl methyl disulphide / 2 (methyldisulfanyl)propane / 40136-65-0


EC number EC name

254-808-5
CAS number

isopropyl methyl disulphide


CAS name

40136-65-0
IUPAC name

2-(methyldisulfanyl)propane
Typical concentration Reference substance

14.4 % (w/w) diethyl disulphide / 1,1' disulfanediyldiethane / 110-81-6


EC number EC name

203-805-7 diethyl disulphide


CAS number CAS name

110-81-6
IUPAC name

1,1'-disulfanediyldiethane
Typical concentration

11.2

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Reference substance

methyl propyl disulphide / 1-(methyldisulfanyl) propane / 2179-60-4


EC number EC name

218-551-2 methyl propyl disulphide


CAS number CAS name

2179-60-4
IUPAC name

1-(methyldisulfanyl)propane
Typical concentration Reference substance

7.7 % (w/w) ethyl isopropyl disulphide / 2-(ethyldisulfanyl) propane / 53966-36-2


EC number EC name

CAS number

CAS name

53966-36-2
IUPAC name

2-(ethyldisulfanyl)-propane
Typical concentration Reference substance

11.6 % (w/w) ethyl n-propyl disulphide / 1-(ethylsulfanyl) propane / 30453-31-7


EC number EC name

CAS number

CAS name

30453-31-7
IUPAC name

1-(ethylsulfanyl)-propane
Typical concentration Reference substance

7 % (w/w) diisopropyl disulphide / 2-propan-2 yldisulfanylpropane / 4253-89-8


EC number EC name

224-225-0 diisopropyl sulphide


CAS number CAS name

4253-89-8
IUPAC name

2-propan-2-yldisulfanylpropane
Typical concentration

2 % (w/w)

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Reference substance

ethyl n-butyl sulphide / 1-(ethylsulfanyl) butane / 63986-03-8


EC number EC name

CAS number

CAS name

63986-03-8
IUPAC name

1-(ethylsulfanyl)butane
Typical concentration Reference substance

0.5 % (w/w) dipropyl disulphide / 1,1' disulfanediyldipropane / 629-19-6


EC number EC name

211-079-8 dipropyl disulphide


CAS number CAS name

629-19-6
IUPAC name

1,1'-disulfanediyldipropane
Typical concentration

2.5 % (w/w)

Impurities
Reference substance

benzene / benzene / 71-43-2


EC number EC name

200-753-7 benzene
CAS number CAS name

71-43-2
IUPAC name

benzene
Typical concentration Remarks

< 0.1 % (w/w) The benzene levels in DSO have been reduced in recent years and are currently present at concentrations less than 0.1%. Past samples used in the acute toxicity testing performed over fifteen years ago contained benzene levels up to 1.0%.

1.3 Identifiers
1.4 Analytical information
1.5 Joint submission
1.6 Sponsors

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1.7 Suppliers 1.8 Recipients 1.9 Product and process oriented research and development 2 Classification and Labelling 2.1 GHS 2.2 DSD - DPD 3 Manufacture, use and exposure 3.1 Technological process 3.2 Estimated quantities 3.3 Sites 3.4 Form in the supply chain 3.5 Identified uses and exposure scenarios 3.6 Uses advised against 3.7 Waste from production and use 3.8 Exposure estimates 3.9 Biocidal information 3.10 Application for authorisation of uses 4 Physical and chemical properties 4.5 Particle size distribution (Granulometry)
Particle size distribution (Granulometry).001
UUID Dossier UUID Author Date Remarks

IUC5-869e6464-28f9-405a-9661-6c34be94e8e3 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-09-25 16:10:13 CEST

Administrative Data
Purpose flag Data waiving Justification for data waiving

( ) robust study summary ( ) used for classification ( ) used for MSDS study scientifically unjustified Test material is a liquid

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5 Environmental fate and pathways


Environmental fate and pathways
UUID Dossier UUID Author Date Remarks

IUC5-05cdc3e8-d73e-4892-a303-d8fe2b5199c9 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-10-01 14:02:31 CEST

Administrative Data Discussion


The environmental fate of DSO has not been examined; however, structure- activity information and suggestive anecdotal test data is available for DMDS and the remaining disulfides in the mix. Many of the disulfides in DSO are naturally found in the environment either as ingredients in vegetables, especially garlic and onions, or as products of the microbial oxidation of assimilated mercaptans (TGSC, 2008). Preliminary studies with DMDS and DPDS have shown that these two disulfides are relatively stable in soil and water (Arnault et al., 2004). DMDS, in particular, has been found in many environmental compartments and is considered to have an integral role in the global sulfur cycle (Caron and Kramer, 1994). Natural background concentrations of DMDS have been measured in a wide variety of media including air, surface waters, sediment, wastewater effluent, vegetation, and expired human air (HSDB, 2005). Interestingly, DMDS has been shown to be absorbed from air into moist and dry soils at a rate that was influenced by the presence of soil microbes, which facilitated the uptake into moist soil only (Bremner and Banwart, 1976). This may be an important environmental process for the disulfides in DSO because of their tendency to partition into the soil compartment.

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5.2 Biodegradation 5.2.1 Biodegradation in water: screening tests


Biodegradation in water: screening tests.001
UUID Dossier UUID Author Date Remarks

IUC5-a1ec8565-ab04-451c-8f39-7883ead6ddee 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-10-01 13:59:44 CEST

Administrative Data
Purpose flag Study result type Reliablility Rationale for reliability

key study (X) robust study summary ( ) used for classification ( ) used for MSDS
read-across from supporting substance (structural analogue or surrogate)
2 (reliable with restrictions)
study conducted to OECD methods but with no GLP

Data source
Reference
Reference type Author Title

other: HPV submission HPV testing group, API (submitter)


Year

2008

US EPA HPV Challenge Program Data Review and Assessment for Reclaimed Substances: Disulfides, Diethyl and Diphenyl, Naphtha Sweetening (aka Disulfide Oil) CAS # 68955-96-4

Bibliographic
source
Testing laboratory Owner company Company study no.
Reference type
Author Title Bibliographic
source
Testing laboratory Owner
company
Company study no.
Report
date
Report no. Report
no.

American Petroleum Institute


Report date

2008-09-10

study report
ELF ATOCHEM
Year

1995

Dimethyl disulfide. Dtermination de la biodgradabilit facile.Essai en fioles fermes.

Ref 95/SAEk/0415/NM (as cited in DMDS robust summary, 2005)

Data access data submitter is data owner

Materials and methods

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Test type ready biodegradability Test guideline


Qualifier Guideline Deviations

according to OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)

GLP compliance no

Test materials
Test material identity
Identifier

CAS number

Identity 624-92-0 Details on test material

Dimethyl disulfide

Study design

Duration of test (contact time)


28 d Parameter followed for biodegradation estimation O2 consumption Reference substance benzoic acid, sodium salt

Results and discussions


% Degradation of test substance
% Degr.
St.
dev. Parameter Sampling time
Remarks

< 10

O2 consumption 28 d

Applicant's summary and conclusion


Interpretation of results under test conditions no biodegradation observed Conclusions Not readily biodegradable Executive summary

The biodegradability of the ten DSO disulfides was examined using the BIOWIN (v 4.00) subroutine in the EPI Suite program. The BIOWIN routine uses eight different

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methods to evaluate the biological degradation of a target chemical under either aerobic or anaerobic conditions. Although several of the methods suggest that the probability of disulfide biodegradation is relatively high, it is believed that the most reliable information comes from the results for DMDS itself and from those models indicating a lack of ready biodegradability (see table below). A closed bottle ready biodegradability test performed with DMDS indicated that less than 10% of the material was degraded over a 28-day period (ELF ATOCHEM, 1995). Ready biodegradability, as defined in accordance with OECD guidelines, only occurs when at least 70% of a chemical is biologically removed from the environment within the 28-day period. Accordingly, DSO is expected to fail the biodegradability test and these conclusions are in agreement with actual test data for DMDS.
Ready Biodegradation Probability Disulfide dimethyl disulfide methyl ethyl disulfide methyl isopropyl disulfide diethyl disulfide methyl n-propyl disulfide ethyl isopropyl disulfide ethyl n-propyl disulfide diisopropyl disulfide ethyl n-butyl disulfide dipropyl disulfide DSO
Actual #Estimated

Ready Biodegradation Probability non-linear

Readily Biodegradable

linear 0.43 0.44 0.30 0.45 0.45 0.31 0.46 0.31 0.46 0.46 0.43# 0.46 0.47 0.26 0.47 0.47 0.26 0.48 0.27 0.49 0.49 0.46# no (no) no no no no no no no no no no#

measured degradation of 10% over 28-days (DMDS test plan, 2005). value.

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6 Ecotoxicological Information 6.1 Aquatic toxicity


Aquatic toxicity
UUID Dossier UUID Author Date Remarks

IUC5-2a436f97-8317-4414-8f3b-f45a41c1a9b2 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-10-01 11:07:53 CEST

Administrative Data Discussion


Evidence suggests that the aquatic and terrestrial toxicity of DSO mimics the effects observed with DMDS. Initial modeling of DMDS and the remaining disulfide constituents of DSO using EPAs ECOSAR software package (Meylan and Howard, 1998) reveled that the ecotoxicity of the disulfides increased as a function of alkyl chain length. Although this finding is consistent with the observed increase in octanol/water partition coefficients for these disulfides, the results are inconsistent with available test data and knowledge of disulfide metabolism. The modeling results, therefore, have not been utilized since the assumed mode of action, non-polar narcosis, is most likely incorrect, a condition that often occurs when this endpoint is invoked indiscriminately (de Roode et al., 2006). The underpinnings for the SAR routines used in the ECOSAR program assume that non-polar narcosis is the operant mode of action for the disulfides; but this class of chemicals is not explicitly represented in the training sets used to develop the mathematical relationships. In fact, disulfides are more likely to operate in terrestrial and aquatic organisms by the same mode of action observed in mammals, which involves disulfide bond cleavage and redox cycling of the free radical intermediates (Mnchberg et al., 2007; Lesser, 2006). The reactive oxygen species produced in this reaction can lead to oxidative stress and protein interactions that are typically more severe and less consistent across species than those elicited by narcotic chemicals (Jager et al., 2007). This lack of applicability is evident when test data for DMDS are compared to the estimates obtained using ECOSAR (see Table 6). The toxicity of DMDS is generally under predicted by a factor 10-200 fold, which signals that a mode of action other than narcosis is in effect.

Table 6. A Comparison of Actual and Estimated Ecotoxicity Values for DMDS Actual Ecotoxicity Endpoint Acute Fish Estimated Toxicity (mg/L) Toxicity (mg/L)

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96-hr LC50 Chronic Fish 30-day Acute Invertebrate 48-hr EC50 Acute Plant 96-hr EC50 Earthworm 14-day LC50
*
#

92.51

0.97*

11.67

--

98.24

60.96

35#

635.4

32*

Actual measured value taken from Arkema, Inc. (2007). Actual measured value taken from ELF ATOCHEM (1996). Additional test results for algae (ErC50, EbC50, NOECr, and NOECb) are available (ELF ATOCHEM, 2000).

Additional support for the use of DMDS as a surrogate for the disulfides in DSO comes from available test data for higher homologs in the series. When the acute toxicity of DMDS to fish (0.97 mg/L) is compared to the LC50 results obtained with DEDS (7.43 mg/L), DPDS (2.62 mg/L), and diisopropyl disulfide (8.31 mg/L), there is no apparent increase in toxicity as a function of chain length (NITE, 2003; Chevron Phillips Chemical Company, 2005; Russom et al., 1997). In addition, the 24-hr EC50value for DEDS (14.5 mg/L) in Daphnia magna is nearly 2-fold greater than the 48-hr value for DMDS (7 mg/L) (Glli et al., 1994). Taken together, these data are consistent with the expected change in potency for the oxidative stress caused by disulfide bond cleavage (see section 5), and confirms that DMDS is the most toxic member of the disulfide series in DSO. Additional testing with DSO is not expected to result in effect concentrations less than those observed DMDS, and therefore no further testing can be justified for the endpoints listed. Despite the lack of DMDS test data for chronic fish toxicity, testing will not be performed within the context of this submission because it will be completed in conjunction with the previously submitted test plan for DMDS. When this voluntary testing is completed, a full complement of ecotoxicity data will be available for DMDS and by analogy DSO.

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7 Toxicological information 7.1 Toxicokinetics, metabolism and distribution


Toxicokinetics, metabolism and distribution
UUID Dossier UUID Author Date Remarks

IUC5-061810ab-28f3-480c-b273-04a0f2b19afd 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-10-01 17:40:41 CEST

Administrative Data Discussion


Sufficient information is available to make reliable and sensible determinations of the health effects of DSO. Whereas some test data is available on the oil itself, the majority of information can be extracted from the robust summary and test plan for DMDS (DMDS Robust Summary, 2005). The rationale and justification for using the health effects data of DMDS as a substitute for the disulfides in DSO are based on sound scientific principles and a plethora of mechanistic information showing that all of the dialkyl disulfides in DSO operate through a common toxic mechanism. This mechanism, which has been well studied and clearly elucidated in the published literature, focuses on the unique characteristics of the disulfide bridge and the ease with which free radical intermediates can be formed once the bridge is cleaved. The metabolism of many, if not all, disulfides is initiated by a thiol-disulfide exchange reaction that substitutes the sulfhydral group of glutathione for a mercaptide fragment within the disulfide molecule. The reaction for DPDS, whose in vivo metabolism has been examined in the greatest detail of the ten DSO disulfides (Germain et al., 2008; Teyssier and Siess, 2000). Evidence shows that this same initial glutathione exchange reaction also takes place for a host of alkyl, alkenyl, phenyl, and branched chain disulfides (Bach et al., 2008; Munday and Manns, 1994; Munday, 1989; Nishikawa et al., 1987). Using an expert knowledge based system for predicting the metabolic reactions that take place in vivo (Meteor, v 9.0.0), the disulfides in DSO were all predicted to undergo reductive cleavage of the disulfide bond with a high degree of probability (Greene et al., 1999). The exchange reaction with glutathione is catalyzed by a thioltransferase, also known as glutaredoxin, which is widely distributed in nature and shows high levels of activity in the tissues and organs affected by alkyl disulfide toxicity (Lillig and Holmgren, 2007). This reaction is the key step in the toxic mechanism for dialkyl disulfides. The activation mechanism has been associated with the initiation of a redox cycle that

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generates excessive quantities of highly reactive free radical intermediates that are capable of interacting with tissue macromolecules at or near the site where they are formed. In some cases this site has been the red blood cell and in other cases the liver depending on the species examined (Munday, 1989). The sequence of reactions in the redox cycling of alkyl disulfide is depicted generically in Figure 3. The first product of the initial thioltansferase exchange reaction is an alkyl mercaptan that undergoes a one-electron oxidation to a free radical intermediate following ionization. This intermediate is the proximal toxicant, responsible for producing a continuous supply of hydroxyl radicals and other reactive oxygen species that can sustain the redox cycling, cause oxidative stress, and precipitate tissue injury at sites where it is formed. Importantly, the reactivity of the mercaptans formed in the exchange reaction is directly affected by the length and branching pattern of the attached alkyl substitutents, with longer chain lengths leading to reduced radical stabilization and lower oxidation rates (Munday, 1989). In addition, the reactivity and toxicity of alkyl disulfides has been shown to decrease in the following order n > sec > tert due to the influence of steric factors on thioltransferase activity. These data indicate that DMDS will be the most reactive member of the series with the longer chain lengths and higher branching patterns of the remaining homologs ameliorating the toxicity by affecting the rate of formation and ultimate stabilization of the free radical intermediates. This fact provides strong justification for the use of DMDS as a surrogate for the higher chain length disulfides in DSO and validates its use in a read across transfer to other disulfides in the category. The test data for DMDS is therefore offered as a reasonable and mechanistically supportable substitute for DSO, since it represents the most toxic member of the disulfide series. As such, the existing information on DSO and the disulfide category is deemed sufficient, and no further testing is needed nor required to assess the health hazards associated with this category of reclaimed substances. Figure 3. Mechanism of Redox Cycling and Free Radical Formation from Alkyl Disulfide Metabolism (Munday and Manna, 1994)

2 GSH + RSSR GSSG + 2 RSH RSH RS + H+ (Hb)Fe3O2 + RS + 2H+ (Hb)Fe3 + RS + H2O2 RS+ RS (RSSR) (RSSR) + 02 RSSR + 02

RSH + 02+- H+ RS + H2O2

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7.1.1 Basic toxicokinetics


Basic toxicokinetics
UUID Dossier UUID Author Date Remarks

IUC5-0b007d33-7e62-4d14-8deb-058c31d9f6f0 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-10-01 17:35:09 CEST Created

Administrative Data

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7.2 Acute Toxicity 7.2.1 Acute toxicity: oral


Acute toxicity: oral - supporting study on DMDS.003
UUID Dossier UUID Author Date Remarks

IUC5-301e61a2-a179-4390-8bc6-0dde2d34f070 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-07 16:47:11 CET

Administrative Data
Purpose flag

supporting study ( ) robust study summary ( ) used for classification ( ) used for MSDS experimental result 1 (reliable without restriction)
study conducted to EPA method and GLP

Study result type Reliablility Rationale for reliability

Data source
Reference
Reference type
Author Title Bibliographic
source
Testing laboratory Owner company Company study no.

study report
Penwalt Corp.
Year

1985

Dimethyl Disulphide EPA Acute LD50

Products Safety Labs Penwalt Corp.

Report no.

T-5150

Report date

1985-06-24

Data access data submitter is data owner

Materials and methods


Test guideline
Qualifier Guideline

other guideline: EPA 40 CFR 163.81-1

Deviations

GLP compliance yes

Test materials
Test material equivalent to submission substance identity

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yes Test material identity


Identifier Identity

CAS number

624-92-0 Details on test material Dimethyl siulfide

Test animals
Species rat Strain Wistar Sex male/female

Administration / exposure
Route of administration oral: gavage Doses 0, 125, 188, 250, 375 and 500 mg/kg No. of animals per sex per dose 5

Results and discussions


Effect levels
Sex Endpoint Effect level
95% CL
Remarks

male/female LD50 190 mg/kg bw 150 240

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7.5 Repeated dose toxicity 7.5.2 Repeated dose toxicity: dermal


Repeated dose toxicity: dermal - supporting study.002
UUID Dossier UUID Author Date Remarks

IUC5-02163804-7758-4960-b365-31029fa602b8 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-07 17:22:37 CET

Administrative Data
Purpose flag

supporting study ( ) robust study summary ( ) used for classification ( ) used for
MSDS
experimental result 1 (reliable without restriction) range-finding study to full OECD study with GLP

Study result type Reliablility Rationale for reliability

Data source
Reference
Reference type
Author Title

study report
ELF Atochem
Year

1989

Repeated-dose (14-day) dermal toxicity range-finding stuyd with Dimethyl Disulfide (DMDS) in rabbits.

Bibliographic
source
Testing laboratory Owner company Company study no.

TNO-CIVO Institutes ELF Atochem

Report no.

V 89.058/280553

Report date

1989-07-31

Data access data submitter is data owner

Materials and methods


Test guideline
Qualifier Guideline Deviations

other guideline: range-finding study

Test materials
Test material equivalent to submission substance identity yes Test material identity

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Identifier

CAS number

Identity 624-92-0 Details on test material

Dimethyl disulfide

Test animals
Species rabbit Strain New Zealand White Sex male/female

Administration / exposure
Type of coverage occlusive Details on study design 0, 106, 503 and 1063 mL/kg/day

Results and discussions


Effect levels
Endpoint Effect level Sex Basis for effect level / Remarks Endpoint Effect level Sex Basis for effect level / Remarks

NOAEL < 103 mg/kg bw/day (nominal) male/female

LOAEL 103 mg/kg bw/day (nominal)

Observations
Remarks on results including tables and figures

Dose-related lethargy or unconsciousness was observed in all treatment groups that dissipated by the end of the day

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7.6 Genetic toxicity


Genetic toxicity
UUID Dossier UUID Author Date Remarks

IUC5-e5cc156b-75e5-4e83-8056-a891b8376c9c 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-10-01 16:15:21 CEST

Administrative Data Key parameter (optional)


Genetic toxicity negative

Discussion
Although there are no results available for DSO, DMDS has been examined in a variety of in vivo and in vitro genetic toxicology screening assays (DMDS Robust Summary, 2005). The test results revealed that DMDS was negative in bacterial mutagenicity assays (Penwalt, 1985c), negative in mammalian mutagenicity tests (ELF ATOCHEM, 1990a), negative for DNA damage and repair (ELF ATOCHEM, 1990b), and ambiguously positive in a chromosomal aberration study using human lymphocytes (ELF ATOCHEM, 1990c). Except for the DNA damage and repair assay, these tests were all performed in the presence and absence of metabolic activation. Similarly, negative results were obtained when DMDS was evaluated in vivo in a mouse micronucleus assay at inhalation concentrations of 250 and 500 ppm (ATOCHEM, 1989b), and did not cause unscheduled DNA synthesis in the hepatocytes of rats exposed to 500 ppm (ATOCHEM, 1990). By comparison, DPDS did not cause any reverse mutations in an S. typhimurium assay using strain TA98 (Tsai et al., 1996). None of the disulfides in DSO were judged to be genotoxic by an expert knowledge based system used to predict the health effects of untested chemical substances (Derek, v 9.0.0) (Greene et al., 1999).

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7.6.1 Genetic toxicity in vitro


Genetic toxicity in vitro - Chromosome aberration.001
UUID Dossier UUID Author Date Remarks

IUC5-c0639a16-2dd3-4480-8d55-9deb7d6caefe 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-03 14:34:07 CET

Administrative Data
Purpose flag Study result type Reliablility Rationale for reliability

key study (X) robust study summary ( ) used for classification ( ) used for MSDS
read-across from supporting substance (structural analogue or surrogate)
1 (reliable without restriction)
study conducted to OECD guidelines with GLP

Data source
Reference
Reference type
Author Title

study report
ELF ATOCHEM
Year

1990

Chromosome analysis of cultured human lymphocytes following in vitro treatment with DMDS

Bibliographic
source
Testing laboratory Owner
company
Company study no.
Report
date

TNO-CIVO

Report no.

V 89.045

Materials and methods


Type of genotoxicity chromosome aberration Type of study in vitro mammalian chromosome aberration test Test guideline
Qualifier Guideline Deviations

according to OECD Guideline 473 (In vitro Mammalian Chromosome Aberration Test)

GLP compliance yes

Test materials
Test material identity

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Identifier Identity

CAS number 624-92-0

Method
Species/strain
Species/strain Details on mammalian cell lines (if applicable) Additional
strain
characteristics
Metabolic activation Metabolic activation
system

lymphocytes:

with and without Aroclor-induced rat (wistar) liver S9 fraction

Test concentrations 3.7; 11.1; 33.3; 100; 300 g/ml Vehicle dimethyl sulphoxide Controls
Negative
controls
Solvent / vehicle
controls
True
negative
controls
Positive controls

yes

yes

Positive cyclophosphamide control


substance
Remarks Negative
controls
Solvent / vehicle
controls
True
negative
controls
Positive controls

+S9

yes

yes
C

Positive mitomycin control


substance
Remarks

-S9

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Details on test system and conditions - Preliminary Cytotoxicity Assay: The dose levels used in the chromosome aberration assay were established on the basis of the results of a preliminary toxicity test carried out with 6 concentrations of the test substance (ranging from 0.5 to 1000.0 g/ml), both in the absence and in the presence of the metabolic activation system (S -9 mix). The highest concentration for the toxicity test was determined by the limit of the solubility of the test substance in the tissue culture medium. - Cytogenetic Assay: * Cell Treatment After 48 h of incubation, the cultures were centrifuged. The cell pellets were resuspended in tissue culture medium supplemented with 20 mM HEPES (and 10% S-9 mix, for the test with metabolic activation) and appropriate test solutions. An untreated culture and a culture receiving DMSO served as negative controls. For each concentration of the test substance and for the controls one culture was used. Without S9, the cultures were incubated in closed tubes for another 24 hours including a 2 hour colcemid treatment. With S-9 mix, the exposure of the cells to the test substance was reduced to only 2 hours, because of the toxicity of the S -9 mix for the cells. After the 2 hour incubation period, the cells washed and supplied with freshly prepared culture medium. The cells were incubated for a further 22 hours (including a 2 hour colcimid treatment. * Cell harvesting: Two hours before the end of the total incubation period the cells were arrested in the metaphase stage of the mitosis by the addition of colcemid. The cells were harvested, treated with a hypotonic solution, fixed three hours, and transferred to clean microscope slides. Two slides were prepared from each culture. The slides were stained 1000 stimulated lymphocytes were examined (500 from each slide) to determine the mitotic index (percentage of cells in mitosis). * Metaphase analysis: From each culture, 100 well-spread metaphases (each containing 46 chromosomes) were analysed by microscopic examination for a wide range of structural chromosome aberrations (gaps, breaks, fragments, dicentrics, exchanges etc.) and other anomalies (endoreduplication, polyploidy), according to the criteria recommended by Savage (1975). Evaluation criteria - Evaluation criteria: The major crite rion to designate the results of a chromosome aberration test as positive is a dose-related, statistically significant increase in the number of cells with structural chromosome aberrations. However, a clear dose-response relationship can be absent because the yield of chromosome aberrations can vary markedly with post-treatment sampling time of an asynchronous population and because increasing doses of clastogens can induce increasing degrees of mitotic delay. A test substance producing neither a dose-related, statistically significant increase in the number of cells with structural chromosome aberrations, nor a statistically significant and reproducible positive response at any of the doses is considered non clastogenic in this system.

Results and discussions


Test results
Species/strain Metabolic activation Test system Genotoxicity Cytotoxicity

lymphocytes: with and without

no

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Vehicle controls valid Negative controls valid Positive controls valid

Additional information on results cytotoxic concentration 300 g/ml

Overall remarks, attachments


Overall remarks

The test substance did not induce a statistically significant increase in the number of cells with structural chromosome aberrations at non toxic concentrations, both in the absence and in the presence of the S-9 mix. At the very toxic concentration of 300.0 g/ml, both in the absence and in the presence of the S-9 mix, the test substance induced a statistically significant increase in the number of cells with structural chromosome aberrations.

Applicant's summary and conclusion


Interpretation of results negative Conclusions dimethyl disulfide was negative for chromosome aberrations in a cultured human lymphocyte test Executive summary

DSO is prediceted to be similarly negative for chromosome aberrations

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Genetic toxicity in vitro Mammalian cell gene mutation assay.002


UUID Dossier UUID Author Date Remarks

IUC5-096ba069-4515-4d76-bae3-7f7f70ceaf17 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-03 13:53:18 CET

Administrative Data
Purpose flag Study result type Reliablility Rationale for reliability

key study (X) robust study summary ( ) used for classification ( ) used for MSDS
experimental result
1 (reliable without restriction)
study conducted to OECD guidelines with GLP

Data source
Reference
Reference type Author Title

ELF ATOCHEM

Year

1990

In Vitro assay for the induction of point mutations in the HGPRT-locus of Chinese hamster ovary cells by dimethyldisulfide (DMDS)

Bibliographic source Testing laboratory Owner company Company study no. Report date

TNO-CIVO

Report no.

V 89.257

Materials and methods


Type of genotoxicity gene mutation Type of study mammalian cell gene mutation assay Test guideline
Qualifier Guideline Deviations

according to OECD Guideline 476 (In vitro Mammalian Cell Gene Mutation Test)

Principles of method if other than guideline The dose levels used in the HGPRT assay were established on the basis of the results of a preliminary solubility test. A final concentration of 1,000 g/ml was chosen as highest concentration for the HGPRT assays. The two independent HGPRT-assays were carried out with single cultures for each concentration of the test substance and for the

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negative and positive controls. GLP compliance yes

Test materials
Test material equivalent to submission substance identity no Test material identity
Identifier Identity

CAS number

624-92-0 Details on test material Dimethyl disulfide

Method
Species/strain
Species/strain Details on mammalian cell lines (if applicable) Additional
strain
characteristics
Metabolic activation
Metabolic activation
system

Chinese hamster Ovary (CHO)

with and without


S9 derived from adult male Wistar rats

Test concentrations 0.46; 1.37; 4.12; 12.3; 37.0; 74.0; 111; 333; 667 and 1000 g/ml Vehicle Dimethyl sulphoxide Controls
Negative
controls
Solvent / vehicle
controls
True
negative
controls
Positive controls

yes

yes

Positive ethylmethanesulphonate control


substance
Remarks Negative

-S9

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controls Solvent / vehicle


controls
True
negative
controls
Positive controls

yes

yes

Positive N-dimethylnitrosamine control


substance
Remarks +S9 Details on test system and conditions

- Activation:
S9 derived from adult male Wistar rats
- Culture Medium:
Ham's F-12 medium supplemented with 10% heat-inactivated
foetal calf serum, 50 g gentamicin/mL and 2 mM L-glutamine.
Evaluation criteria - Evaluation of the results: The following criteria were used to evaluate the data obtained in the HGPRT assay (Li et al. 1987) a) the survival (absolute cloning efficiency) of the negative controls should not be less than 50%, b) the mean mutant frequency of the negative controls should fall within the range of 0-20 6 -TG resistant mutants per 10e6 clonable cells, c) the positive controls must induce a response of a magnitude appropriate for the mutagen under the experimental conditions applied, d) the highest test substance concentration should, if possible, result in a clear cytotoxic response (e.g. 10-30% of the relative initial survival). Any apparent increase in mutant frequency at concentrations of the test substance causing more than 90% toxicity is considered to be an artifact and not indicative of genotoxicity. Genotoxicity of the test substance was evaluated using the following criteria (Li et al. 1987): a) a concentration-related increase in mutant frequency, b) a reproducible positive response for at least one of the test substance concentrations (e.g. the mean mutant frequency should be more than 20 mutants per 10e6 clonable cells).

Results and discussions


Test results
Species/strain Metabolic activation
Test
system
Genotoxicity Cytotoxicity

Chinese hamster Ovary (CHO) with and without

negative

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Vehicle controls valid Negative controls valid Positive controls valid

yes

yes

Remarks on results including tables and figures

In the absence of the S -9 mix, the test substance induced neither a concentrationrelated increase in the mutant frequency nor a reproducible positive response at one of the test concentrations. In the presence of a metabolic activation system, DMDS induced a slight increase in mutant frequency at several concentrations, in both HGPRT assays. These increases were neither concentration-related nor clearly reproducible. In both HGPRT assays, the test substance appeared to be highly toxic to CHO cells at a concentration range from 74.0-1,000 ug/ml. The positive control substances, ethylmethanesulfonate and dimethylnitrosamine, induced the expected increase in the mutant frequency.

Applicant's summary and conclusion


Interpretation of results negative Conclusions No evidence for a genotoxic effect of DMDS was found in cultured CHO cells, under the conditions used in the HGPRT assay. Executive summary

DSO is expected to be similarly negative in an in vitro mammalian cell gene mutation assay

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Genetic toxicity in vitro - DNA damage and repair.003


UUID Dossier UUID Author Date Remarks

IUC5-e029b412-545c-4e3f-ae2d-910f2073f3b8 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-03 16:29:27 CET

Administrative Data
Purpose flag Study result type Reliablility Rationale for reliability

key study (X) robust study summary ( ) used for classification ( ) used for MSDS experimental result 1 (reliable without restriction) study conducted to OECD guidelines with GLP

Data source
Reference
Reference type Author Title Bibliographic source Testing laboratory Owner company Company study no. Report date

ELF ATOCHEM

Year

1990

In Vitro DNA Repair Test on Rat Hepatocytes in Primary Culture

Sanofi

Report no.

RA860891026/PN1

Materials and methods


Type of genotoxicity DNA damage and/or repair Type of study DNA damage and repair assay, unscheduled DNA synthesis in mammalian cells in vitro Test guideline
Qualifier Guideline

according to OECD Guideline 482 (Genetic Toxicology: DNA Damage and Repair, Unscheduled DNA Synthesis in Mammalian Cells In Vitro)

Deviations

GLP compliance yes

Test materials
Test material equivalent to submission substance identity no

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Test material identity


Identifier Identity

CAS number

624-92-0 Details on test material Dimethyl sulphoxide

Method
Species/strain
Species/strain Details on mammalian cell lines (if applicable) Additional
strain
characteristics
Metabolic activation
Metabolic
activation
system

hepatocytes: rat

without

Test concentrations 1; 5; 10; 50; 100; 200 and 300 g/ml Vehicle dimethyl sulphoxide Controls
Negative controls Solvent / vehicle
controls
True
negative
controls
Positive controls

yes pyrene 1 uM yes

yes

Positive 7,12-dimethylbenzanthracene control


substance
Remarks Negative controls Solvent / vehicle
controls
True
negative
controls
Positive controls

10 uM yes pyrene 1 uM yes

yes

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Positive other: control


substance
Remarks

2-aminofluorene

0.1 and 0.5 uM Details on test system and conditions - Cytotoxicity evaluation: The test compound cytotoxicity was assessed for both DNA repair studies at the
end of the treatment:
Each concentration of Dimethyldisulfide was tested in triplicate.
- Autoradiography: Autoradiographs were prepared by dipping slides in a photographic emulsion then
developed. Slides were stained in hematoxylin -phloxin.
- Slide assessment:
For each cell, following nuclear grain court, cytoplasmic count was performed on 3 areas of the same
size as the nucleus and adjacent to it.
Evaluation criteria - Data interpretation The test compound is considered positive when the mean nuclear grain court is
statisticaly greater than that of the control, the mean net nuclear grain court is above 3 grains per
nucleus, and the percentage of treated cells in repair is significantly different from that of the controls. In
addition, the effect must be shown to be reproducible between experiments.

Results and discussions


Test results
Species/strain Metabolic activation
Test
system
Genotoxicity Cytotoxicity Vehicle controls valid Negative controls valid Positive controls valid

hepatocytes: rat without

negative yes

yes

Remarks on results including tables and figures

Results - Cytotoxic at 100, 200 and 300 ug/ml IC50 evaluated by LDH release: 98 ug/ml (2nd study) - not genotoxic at concentrations of 10, 50, 100 and 200 ug/ml

Applicant's summary and conclusion


Interpretation of results negative without metabolic activation Conclusions DMDS was not genotoxic in vitro in the DNA repair test.

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Executive summary

DSO is expected to have similar behaviour to dimethyl disulfide and be negative in a DNA repair test

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Genetic toxicity in vitro - Ames test supporting study.004


UUID Dossier UUID Author Date Remarks

IUC5-519a6563-4433-4106-ae8c-811430c0931f 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-07 17:09:15 CET

Administrative Data
Purpose flag

supporting study ( ) robust study summary ( ) used for classification ( ) used for MSDS experimental result 1 (reliable without restriction)
Conducted to OECD method and GLP

Study result type Reliablility Rationale for reliability

Data source
Reference
Reference type
Author Title Bibliographic
source
Testing laboratory Owner company Company study no.

study report
Penwalt Corp
Year

1985

Dimethyl disulfide, Ames Salmonella/Microsome Plate Test (EPA/OECD)

Pharmakon Research International Inc. Penwalt Corp.

Report no.

PH 301-P-W-003-85

Report date

1985-05-31

Data access data submitter is data owner

Materials and methods


Type of genotoxicity gene mutation Type of study bacterial reverse mutation assay (e.g. Ames test) Test guideline
Qualifier Guideline Deviations

according to OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance yes

Test materials

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Test material equivalent to submission substance identity yes Test material identity
Identifier Identity

CAS number

624-92-0 Details on test material Dimethyl disulfide

Method
Species/strain
Species/strain Details on mammalian cell lines (if applicable) Additional
strain
characteristics
Metabolic activation
Metabolic activation
system

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

with and without


Aroclor derived rat liver S9

Test concentrations 0, 50, 166, 500, 1666, 5000 ug/plate

Results and discussions


Test results
Species/strain Metabolic activation
Test
system
Genotoxicity Cytotoxicity Vehicle controls valid Negative controls valid Positive controls valid

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 with and without

negative

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7.6.2 Genetic toxicity in vivo


Genetic toxicity in vivo - UDS.001
UUID Dossier UUID Author Date Remarks

IUC5-0614d85d-1a0f-4eb6-93b6-5d8f70ae3e83 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-03 16:31:17 CET

Administrative Data
Purpose flag Study result type Reliablility Rationale for reliability

key study (X) robust study summary ( ) used for classification ( ) used for MSDS
experimental result
1 (reliable without restriction)
conducted to OECD guidelines with GLP

Data source
Reference
Reference type
Author Title Bibliographic source Testing laboratory Owner
company
Company study no.
Report
date

study report
ELF ATOCHEM
Year

1990

An in vivo/in vitro rat hepatocyte DNA-repair assay with dimethyldisulfide (DMDS)

TNO-CIVO

Report no.

V 90.082

Materials and methods


Type of genotoxicity DNA damage and/or repair Type of study unscheduled DNA synthesis Test guideline
Qualifier Guideline Deviations

according to other guideline: OECD 482

GLP compliance yes

Test materials
Test material equivalent to submission substance identity no

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Test material identity


Identifier Identity

CAS number

624-92-0 Details on test material dimethyl disulfide

Test animals
Species rat Strain Wistar Sex male

Administration / exposure
Route of administration inhalation Duration of treatment / exposure 4 hours Doses / concentrations 500 ppm nominal conc. Control animals yes Positive control(s) The hepatocarcinogen 2-acetylaminofluorene (2 AAF), was used as a
positive control in the in vivo/in vitro DNA-repair assay and in the in vitro
DNA-repair assay (2 AAF). Hepatocytes isolated from animals exposed to
air only served as negative controls.

Basis

Examinations
Tissues and cell types examined The DNA-repair activities were examined by autoradiography in monolayer
cultures o f hepatocytes, incubated in the presence of [methyl-3H]thymidine.
Any other information on materials and methods incl. tables

Dimethyldisulfide (DMDS) was examined for its potential to induce unscheduled DNA synthesis (UDS) in primary rat hepatocytes after short-term exposure of male wistar rats to the test substance by inhalation. For the genotoxicity assay male rats were exposed by inhalation for a period of 4 h to one high concentration of 500 ppm DMDS (maximally tolerated concentration). Immediately after exposure and after subsequent non-exposure periods of 16 and 24 h, animals were sacrificed for isolation of hepatocytes.

Results and discussions

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Test results
Sex Genotoxicity Toxicity Vehicle controls valid Negative controls valid Positive controls valid

male negative

Remarks on results including tables and figures

DMDS did not induce DNA-repair activities in hepatocytes, either during the 4 h exposure period or during the subsequent 16 h or 24 h after the exposure period. The positive control substance, 2-AAF, induced the expected increase in DNA-repair activities.

Applicant's summary and conclusion


Interpretation of results negative Conclusions It was concluded that DMDS did not induce DNA-repair in rat
hepatocytes.
Executive summary

DSO is expected to be similarly negative in an in vivo UDS study

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7.8 Toxicity to reproduction


Toxicity to reproduction
UUID Dossier UUID Author Date Remarks

IUC5-608b5c61-d410-4905-b9cb-b4a1e4421f0e 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-10-01 17:29:19 CEST

Administrative Data Effects on fertility


Discussion Although no studies have been reported on the reproductive or developmental toxicity of disulfide oil, studies performed with DMDS are offered as a reasonable surrogate for the disulfides in DSO. An evaluation of developmental effects was examined in a series of inhalation exposure studies performed in rats with DMDS (DMDS Robust Summary, 2005). In an initial range finding study, pregnant dams were exposed for 6 hr/day on days 6 through 15 of gestation to DMDS concentrations of 10, 50, or 250 ppm (0.04, 0.19, or 0.96 mg/L) (ATOCHEM, 1991a). Treatment-related reductions in body weight gain and food consumption were observed in all treatment groups, but pregnancy incidence, intrauterine death incidence, pre-implantation loss, litter size, sex ratio, and the incidence of malformations were all within the expected range. Mean fetal weights showed an exposure-related reduction in all treatment groups that was considered to be an equivocal finding. The maternal NOAEL was determined to be less than 10 ppm (0.04 mg/L). In a more detailed study, three groups of 30 mated female rats were exposed to DMDS by whole body exposure at 5, 15 or 50 ppm (0.02, 0.06, or 0.19 mg/L) for 6 hours daily from day 6 to day 15 of gestation (ATOCHEM, 1991b). A similar group of 30 rats, exposed to filtered air only over the same period, served as controls. All animals were maintained until day 20 of gestation, and then sacrificed. No deaths were observed or unusual lesions were observed, but a higher incidence of rough hair coat was seen at 50 ppm (0.19 mg/L). Clinical condition at 5 and 15 ppm (0.02 and 0.06 mg/L) did not differ from controls. Treatment-related reductions in weight gain were observed at 15 and 50 ppm (0.06 and 0.19 mg/L). Food intake was lower than controls at 50 ppm (0.19 mg/L), but comparable at 5 or 15 ppm (0.02 and 0.06 mg/L). There was no effect of treatment on pre or post-implantation loss, litter size or sex ratio.

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Maternal toxicity was noted at 15 and 50 ppm (0.06 and 0.19 mg/L), but there was no evidence of developmental effects. Litter and fetal weights were reduced at 50 ppm (0.19 mg/L). At 5 and 15 ppm (0.02 and 0.06 mg/L) these parameters were comparable to controls. No malformations were observed in fetuses from the treated groups. A slightly higher incidence of retarded ossification was observed at 50 ppm (0.19 mg/L), which indicated delayed maturation as a result of the lower fetal weight, rather than teratogenicity. The NOAELs for maternal toxicity, teratogenicity, and fetotoxicity were 5, 50, and 15 ppm (0.02, 0.06, and 0.19 mg/L), respectively. The effects of DMDS on reproductive organs were assessed in male and female rats exposed to 10, 50, 150, or 250 ppm (0.04, 0.19, 0.58, or 0.96 mg/L) DMDS for 6 hr/day for 90 days (ELF ATOCHEM, 1992). Tissue histopathology did not reveal any lesions or damage to the epididymus, prostrate, or testes of the male rats, nor ovaries or uterus of female rats.

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7.8.2 Developmental toxicity / teratogenicity


Developmental toxicity / teratogenicity - supporting study.002
UUID Dossier UUID Author Date Remarks

IUC5-01a7a762-2cca-4878-97dd-f854bd73a746 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-07 17:34:27 CET

Administrative Data
Purpose flag

supporting study ( ) robust study summary ( ) used for classification ( ) used for MSDS experimental result 1 (reliable without restriction) range-finding study to full OECD method test with GLP

Study result type Reliablility Rationale for reliability

Data source
Reference
Reference type
Author Title Bibliographic
source
Testing laboratory Owner company
Company study no.

study report
Atochem
Year

1991

Dimethyl Disulfide (DMDS): Inhalation range-finding study in the pregnant rat

Hazleton UK Atochem

Report no.

6142-514/8

Report date

1991-05-31

Data access data submitter is data owner

Materials and methods


Test guideline
Qualifier Guideline

other guideline: range-finding study

Deviations

Test materials
Test material equivalent to submission substance identity yes Test material identity
Identifier Identity

CAS number

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624-92-0

Test animals
Species rat Strain Sprague-Dawley

Administration / exposure
Route of administration inhalation Type of inhalation exposure (if applicable) whole body No. of animals per sex per dose 7 rats per group, 10, 50 and 250 ppm

Results and discussions


Effect levels
Endpoint Effect type Effect level
Basis for effect level / Remarks

NOAEL maternal toxicity < 10 ppm

Overall remarks, attachments


Overall remarks

Treatment-related reductions in body weight gain and food consumption were observed in all treatment groups, but pregnancy incidence, intrauterine death incidence, pre implantation loss, litter size, sex ratio, and the incidence of malformations were all within the expected range. Mean fetal weights showed an exposure-related reduction in all treatment groups that was considered to be an equivocal finding. The maternal NOAEL was determined to be less than 10 ppm (0.04 mg/L).

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13 Assessment Reports
Assessment Reports - DSO analysis.002
UUID Dossier UUID Author Date Remarks

IUC5-1b6fc839-f3c9-4ec8-b766-b66697dd2a13 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-05 15:24:47 CET Attachment: Appendix I.doc / 674.5 KB added

Administrative Data
Type of report other: DSO analysis Document Appendix I.doc / 674.5 KB (application/msword)

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Assessment Reports - DMDS Test Plan.003


UUID Dossier UUID Author Date Remarks

IUC5-7c43da23-377c-4572-877e-e40158ee07bf 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-05 15:25:22 CET Attachment: Appendix II.pdf / 136.52 KB added

Administrative Data
Type of report other: DMDS Test Plan Remarks Dimethyl disulfide test plan Document Appendix II.pdf / 136.52 KB (application/pdf)

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Assessment Reports - DMDS Robust Summary.004


UUID Dossier UUID Author Date Remarks

IUC5-2d034750-1f1f-4f43-a413-c6d4f992b4f1 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-05 15:25:55 CET Attachment: Appendix III.pdf / 902.07 KB added

Administrative Data
Type of report other: DMDS Robust Summaries Remarks Dimethyl disulfide robust summaries Document Appendix III.pdf / 902.07 KB (application/pdf)

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Reference substance: Disulfides, dialkyl and di-Ph, naphtha sweetening


UUID Dossier UUID Author Date Remarks

IUC5-74e0f525-9301-4ce3-a770-0cd33add3353 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-03 15:31:41 CET

General information
Reference substance name

Disulfides, dialkyl and di-Ph, naphtha sweetening

Reference substance information


CAS information
CAS number CAS name

68955-96-4 Disulfides, dialkyl and di-Ph, naphtha sweetening

Synonyms
Name Name

DSO Disulfide oil

Molecular and structural information

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Reference substance: dimethyl disulphide


UUID Dossier UUID Author Date Remarks

ECB5-76de1127-bba9-474f-a452-a6c8114a0675 0 European Commision/Joint Research Centre/European Chemicals Bureau 2007-05-10 11:00:00 CEST Created

General information
Reference substance name

dimethyl disulphide

EC inventory
EC number EC name Molecular formula

210-871-0 C2H6S2

CAS number

624-92-0

dimethyl disulphide

Reference substance information


CAS information
CAS number

624-92-0

IUPAC name
IUPAC name

dimethyl disulfide

Synonyms
Name Name

Disulfide, dimethyl Disulfide, dimethyl

DSL Category: Organics

Molecular and structural information


Molecular formula Molecular weight range SMILES notation InChI Structural formula

C2H6S2 94.199 CSSC InChI=1/C2H6S2/c1-3-4-2/h1-2H3

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Reference substance: methyl ethyl disulphide


UUID Dossier UUID Author Date Remarks

IUC5-2ead9865-02d4-4a38-b146-4c980bc95a1f 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-09-25 12:58:10 CEST

General information
Reference substance name

methyl ethyl disulphide

Reference substance information


CAS information
CAS number

20333-39-5

IUPAC name
IUPAC name

methyldisulfanylethane

Molecular and structural information

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Reference substance: isopropyl methyl disulphide


UUID Dossier UUID Author Date Remarks

ECB5-df9aa4e7-5225-4652-a16b-b2a2bf927bd1 0 European Commision/Joint Research Centre/European Chemicals Bureau 2007-05-10 11:00:00 CEST Created

General information
Reference substance name

isopropyl methyl disulphide

EC inventory
EC number EC name Molecular formula

254-808-5 C4H10S2

CAS number

40136-65-0

isopropyl methyl disulphide

Reference substance information


CAS information
CAS number

40136-65-0

IUPAC name
IUPAC name

2-(methyldisulfanyl)propane

Synonyms
Name

Disulfide, methyl 1-methylethyl

Molecular and structural information


Molecular formula Molecular weight range SMILES notation InChI Structural formula

C4H10S2 122.2522 CSSC(C)C InChI=1/C4H10S2/c1-4(2)6-5-3/h4H,1-3H3

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Reference substance: diethyl disulphide


UUID Dossier UUID Author Date Remarks

ECB5-6a0faffe-0e34-4d54-9b66-e3d3719f8c5d 0 European Commision/Joint Research Centre/European Chemicals Bureau 2007-05-10 11:00:00 CEST Created

General information
Reference substance name

diethyl disulphide

EC inventory
EC number EC name Molecular formula

203-805-7 C4H10S2

CAS number

110-81-6

diethyl disulphide

Reference substance information


CAS information
CAS number

110-81-6

IUPAC name
IUPAC name

1,1'-disulfanediyldiethane

Synonyms
Name Name

Disulfide, diethyl Disulfide, diethyl

DSL Category: Organics

Molecular and structural information


Molecular formula Molecular weight range SMILES notation InChI Structural formula

C4H10S2 122.2522 CCSSCC InChI=1/C4H10S2/c1-3-5-6-4-2/h3-4H2,1-2H3

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and d... Page 50 of 58

Reference substance: methyl propyl disulphide


UUID Dossier UUID Author Date Remarks

ECB5-7a7513ce-2d09-4236-bc86-997d2e91325a 0 European Commision/Joint Research Centre/European Chemicals Bureau 2007-05-10 11:00:00 CEST Created

General information
Reference substance name

methyl propyl disulphide

EC inventory
EC number EC name Molecular formula

218-551-2 C4H10S2

CAS number

2179-60-4

methyl propyl disulphide

Reference substance information


CAS information
CAS number

2179-60-4

IUPAC name
IUPAC name

1-(methyldisulfanyl)propane

Synonyms
Name Name Name

Disulfide, methyl propyl


Disulfide, methyl propyl
Disulfide, methyl propyl

DSL Category: Organics

Molecular and structural information


Molecular formula Molecular weight range SMILES notation InChI Structural formula

C4H10S2 122.2522 CCCSSC InChI=1/C4H10S2/c1-3-4-6-5-2/h3-4H2,1-2H3

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and d... Page 51 of 58

Reference substance: ethyl isopropyl disulphide


UUID Dossier UUID Author Date Remarks

IUC5-e0b3a767-308b-48ea-90c3-59268fe4df22 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-09-25 13:41:24 CEST

General information
Reference substance name

ethyl isopropyl disulphide

Reference substance information


CAS information
CAS number

53966-36-2

IUPAC name
IUPAC name

2-(ethyldisulfanyl)-propane

Molecular and structural information

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and d... Page 52 of 58

Reference substance: ethyl n-propyl disulphide


UUID Dossier UUID Author Date Remarks

IUC5-4a2af2ef-9e2b-4acf-a8ff-72e6cfc98594 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-11-03 15:29:50 CET

General information
Reference substance name

ethyl n-propyl disulphide

Reference substance information


CAS information
CAS number

30453-31-7

IUPAC name
IUPAC name

1-(ethylsulfanyl)-propane

Molecular and structural information

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and d... Page 53 of 58

Reference substance: diisopropyl disulphide


UUID Dossier UUID Author Date Remarks

IUC5-6768b0b0-0c19-4e74-8a5e-c8c2727d6811 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-09-25 13:38:39 CEST

General information
Reference substance name

diisopropyl disulphide

EC inventory
EC number EC name Molecular formula

224-225-0 C6H14S2

CAS number

4253-89-8

diisopropyl sulphide

Reference substance information


CAS information
CAS number

4253-89-8

IUPAC name
IUPAC name

2-propan-2-yldisulfanylpropane

Molecular and structural information

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and d... Page 54 of 58

Reference substance: ethyl n-butyl sulphide


UUID Dossier UUID Author Date Remarks

IUC5-8d4a86ae-93cb-481f-b05d-fb5622842ba0 0 Chris3 / Bootman Chemical Safety Ltd 3 / Diss / United Kingdom 2008-09-25 13:43:46 CEST

General information
Reference substance name

ethyl n-butyl sulphide

Reference substance information


CAS information
CAS number

63986-03-8

IUPAC name
IUPAC name

1-(ethylsulfanyl)butane

Molecular and structural information

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and d... Page 55 of 58

Reference substance: dipropyl disulphide


UUID Dossier UUID Author Date Remarks

ECB5-a8709cff-7bc8-4ccd-9b4a-5f5ef6df0796 0 European Commision/Joint Research Centre/European Chemicals Bureau 2007-05-10 11:00:00 CEST Created

General information
Reference substance name

dipropyl disulphide

EC inventory
EC number EC name Molecular formula

211-079-8 C6H14S2

CAS number

629-19-6

dipropyl disulphide

Reference substance information


CAS information
CAS number

629-19-6

IUPAC name
IUPAC name

1,1'-disulfanediyldipropane

Synonyms
Name Name Name

Disulfide, dipropyl Disulfide, dipropyl Disulfide, dipropyl

DSL Category: Organics

Molecular and structural information


Molecular formula Molecular weight range SMILES notation InChI Structural formula

C6H14S2 150.3054 CCCSSCCC InChI=1/C6H14S2/c1-3-5-7-8-6-4-2/h3-6H2,1-2H3

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and d... Page 56 of 58

Reference substance: benzene


UUID Dossier UUID Author Date Remarks

ECB5-86bfd927-e2e9-46e6-bde9-ce721eca359b 0 European Commision/Joint Research Centre/European Chemicals Bureau 2007-05-10 11:00:00 CEST Created

General information
Reference substance name

benzene

EC inventory
EC number EC name Molecular formula

200-753-7 benzene C6H6

CAS number

71-43-2

Reference substance information


CAS information
CAS number

71-43-2

IUPAC name
IUPAC name

benzene

Synonyms
Name Name

Benzene Benzene

DSL Category: Organics

Molecular and structural information


Molecular formula Molecular weight range SMILES notation InChI Structural formula

C6H6 78.1118 c1ccccc1 InChI=1/C6H6/c1-2-4-6-5-3-1/h1-6H

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Substance: Disulfides, dialkyl and di-Ph, naphtha sweetening / Disulfides, dialkyl and d... Page 57 of 58

Legal entity: Lyondell Chemie Nederland B.V.


UUID Dossier UUID Author Date Remarks

IUC5-a6de8fc0-4d1d-4406-849b-79d230119bb8 0 jrfowles / Lyondell Chemie Nederland B.V. / Rotterdam / Netherlands 2008-11-10 14:40:13 CET

General information
Legal entity name Legal entity type

Lyondell Chemie Nederland B.V. company

Identifiers
Legal entity identifiers
Identifier type ID

DUNS 405790762

Identifier type ID

VAT NL811582929B01

Other IT system identifiers


IT system ID

LEO 8789

Contact information
Contact address
Address Postal code Town Country E-mail Web site

Weena 737 3013 AM Rotterdam Netherlands [email protected] http://www.lyondellbasell.com

Contact persons
Organisation Department Title First name Last name E-mail Address

Lyondell Chemie Nederland B.V. Health Safety and Environment Toxicology Manager Marcy Banton [email protected]

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Weena 737
Postal code Town Country

3013 AM Rotterdam Netherlands

Sites
Botlek Maasvlakte

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