ASTA

CLEANLINESS SPECIFICATIONS
FOR
SPICES, SEEDS, AND HERBS
(FOREIGN and DOMESTICALLY PRODUCED)

REVISED APRIL 28, 1999

American Spice Trade Association, Incorporated

Web Site: www.astaspice.org

560 Sylvan Avenue, Post Office Box 1267

Englewood Cliffs, NJ 07632, U.S.A.
E-Mail: [email protected]
Phone: (201) 568-2163
Fax: (201) 568-7318
 PREFACE
In 1969 ASTA adopted its first Cleanliness Specifications for Unprocessed Spices, Seeds, and Herbs (Foreign and
Domestically Produced). Subsequent revisions were approved by the membership in 1971, 1975, 1976, 1982,
1989, 1991, 1992, 1994, 1995, 1996 and 1997. Each of these revisions focused on providing for a credible set of
Cleanliness Specifications or improving the Sampling and Analytical Procedures which are an integral part of the
Specifications.

The following is a history of the establishment of these Specifications since their inception as well as details
regarding the recent revisions.

In September 1968, the Association called a meeting of its members to consider a proposal of the New York
Regional Office of the Food and Drug Administration (FDA). At that meeting, the FDA Regional Director pointed
out that approximately 75% of all spices, seeds and herbs imported into the United States arrived through the Port
of New York. He noted that with spice imports increasing at the rate of 5% annually, the New York District Office
was forced to assign more and more inspectors to the job, with the result that by the end of 1967, nearly half of the
available manpower for import inspection was examining spices. Further, the District's laboratory was being
swamped with samples of spices - nearly 4,000 a year being drawn from shipments at the docks. This was
happening at a time when the District was being pressed to step up inspections in other areas - areas involving food
impurities that presented more significant health hazards.

According to the Regional Director, the object of the meeting was "to reach a self-regulation understanding with
spice importers so as to shift most of the work of spice sampling and analysis from the District over to industry
members themselves." The agreement sought by the FDA would give the industry the "privilege" of importing
spices under conditional release without formal FDA inspection. In exchange for this "privilege," importers would
guarantee that spice shipments which they found, by sampling and macroscopic analysis by an ASTA Approved
Laboratory, to be adulterated, would be properly cleaned or reconditioned before being put into consumer channels
or would be returned to the exporting country . At that time, the Regional Director indicated that there was a strong
probability that the program would become one of FDA national policy.

The first ASTA Specifications published in 1969 resulted from a series of meetings, first with members of the
Brokers and Agents Section, then the Importers and Dealers Section, and, finally, the Grinders and Processors
Section.

The original Specifications set in 1969 were based on the trade's experience to date. In 1982, the ASTA Standards
Committee began an annual review of all Certificates of Analysis in order to determine the percent compliance with
ASTA Specifications. This data, compiled from more than 17,000 Certificates of Analysis between 1982 and
1986, was sorted by commodity, origin, and macroscopic defect, and statistically analyzed.

The results of this four-year study indicated that lots imported and analyzed by ASTA Approved Laboratories
during this period showed relatively high compliance with the original Specifications set in 1969. No changes were
warranted at that time, according to the initial criteria applied by the trade.

In 1988 ASTA's Board of Directors charged the Standards Committee to revise ASTA's Cleanliness Specifications
so that they would be at least as stringent as FDA's Defect Action Levels (DALs). This was an issue for several
reasons. First, there were significant quantitative and qualitative differences between ASTA and FDA on
macroscopic specifications for raw, unprocessed spices, primarily in the excreta category. Second, there was an
obvious disparity in the ASTA contract, which stated that merchandise traded on that contract must comply with
both FDA regulations and ASTA Specifications. Our Approved Laboratories were only charged with analyzing
raw material samples for compliance with ASTA Specifications, not with those of FDA. Third, ASTA had just
formed its first Government Relations Committee with the firm intention of proactively influencing the impact of all
U.S. Laws and regulations, consistent with the public interest. Aligning the standards served as an important
foundation for this long term effort.

In 1989, our trade unanimously voted in favor of changes which created a unified set of macroscopic standards. In
addition, macroscopic analytical procedures were updated accordingly. In 1991, the membership unanimously
adopted a set of definitions of extraneous matter. This change in definitions also provided that in the case of Sage
and Bay Leaves, a separate column will be used on the Certificate of Analysis to report "Stems." This information
will be for economic purposes only and will not represent a pass/fail criteria.
1
A few years ago, ASTA's By-Laws were amended to permit the Standards Committee to propose standards other
than macroscopic for adoption by the Active membership. In 1992, the trade unanimously voted in favor of the
first non-macroscopic specification, providing for all oregano imported under ASTA contracts to be "Sumac
negative." In 1994 the membership voted that if the samples of Oregano are marked "Product of Mexico", analysis
for the presence of Sumac is not required.

In 1994, the membership also voted in several additional amendments. Perhaps most importantly, the name of
ASTA's Cleanliness Specifications was changed to delete the term "unprocessed". This was done to reflect the fact
that with respect to macroscopic specifications, compliance with ASTA Cleanliness Specifications would also
signify compliance with FDA's macroscopic DALs, whether or not the spice was to be further processed.
Additionally, since the Standards Committee may now propose standards other than macroscopic, the term
"unprocessed" was no longer appropriate in the title.

Other amendments passed in 1994 included the correction of an incongruity in the allowance for reconditioning
when a limited number of live insects are found, the deletion of requirements for total ash and acid insoluble ash
testing of Cumin Seed, and the requirement to test White Pepper for the presence of Black Pepper. In the case of
Cumin Seed, data was obtained which demonstrates that lots meeting ASTA's cleanliness specifications should, at
all times, meet FDA's total ash and acid insoluble ash requirements. The requirement to test White Pepper for the
percentage of Black Pepper was instituted so that buyers could determine whether or not the lots coming in meet
their requirements. This information will be for economic purposes only and will not represent a pass/fail criteria.

In 1997 the booklet was reformatted in order to make the specifications easier to use and follow.

ASTA Cleanliness Specifications for Spices, Seeds, and Herbs, and their related requirements such as sampling,
analysis, etc., are intended to insure that spices, seeds and herbs as raw unprocessed agricultural commodities have
been properly handled and stored before being further processed into acceptable finished product for consumption
at the Consumer, Food Service and Industrial levels.

In applying these Specifications to the everyday conduct of business, it is important to recognize that they are an
integral part of the American Spice Trade Association's contracts.

All products listed in these Specifications, both foreign and domestic, must comply before they can be introduced
into commerce or further processed. Imported items must be shown to comply at port of entry. Domestically
grown products must be shown to comply before they can be processed into a consumable product.

The Specifications contained in the table establish limits for macroscopic extraneous matter which is removable by
further processing under good manufacturing practices to place the product in condition for consumption. They
also cover other areas covered by FDA's Defect Action Levels, such as microscopic filth in some processed spices
(namely insect fragments and rodent hairs).

FY99-59/500

2
 COMMODITY TRACKING PROGRAM

At the 1987 Annual Meeting and Convention the membership adopted the following program which had been
proposed by the Association's Standards Compliance Committee. The Commodity Tracking Program became
effective on April 1, 1989, and is a part of the ASTA Contract, unless the parties otherwise agree.

The following steps are to be followed when drawing samples in accordance with the standards paragraph of the
ASTA contract:

Phase I

l. ASTA will assign an Importer Identification Number to each importing member of the program.

2. The Sampling Order must contain the Date, Importer's Identification Number, U.S. Custom's Entry
Number, Commodity, Vessel Name, Bill of Lading Number, Container Number (where applicable),
Marks (must include, but not limited to; name of product and country of origin), Number of Bags (the
number of bags is optional on the ASTA copy), Name of Sampler, and Laboratory.

3. A copy of the Sampling Order must be received by the Sampler and the ASTA office prior to
sampling and by the ASTA Approved Laboratory prior to analysis.

4. The sampler must give all of the above information to the Approved Laboratory with the samples,
including the Sample Number, Entry Number, and Marks as observed - notifying the Importer and
Laboratory of any discrepancies on the Sampling Order.

5. Promptly after analysis, the Approved Laboratory must fax the ASTA office with the pass/fail results
of each lot tested.

In the case of a lot passing the specifications:

1. After verification that the lot has not been previously sampled, ASTA is to issue a Code Number
which then becomes the Laboratory Reporting Number.

2. The Approved Laboratory will then issue the Certificate, noting the corrected Marks by the Sampler,
with the ASTA Code Number for passing lots only noted within the ASTA stamp.

Phase II

In the case of a lot failing to pass Specifications:

1. Failed merchandise does not receive the ASTA Code Number.

2. Lots may only be resampled after reconditioning. Resampled lots must have identical information as
given under Phase I.

3. The copy of the second Sampling Order sent to ASTA must be accompanied by a letter or certificate
of reconditioning (Includes fumigation).

4. Re-do Phase I, Items 2-5.

5. After verification, ASTA issues a code number bearing an "R" for each resampling done on the
individual lot.

6. The Approved Laboratory will then issue the Certificate with the ASTA Code Number, bearing "R"
for passing lots only noted within the ASTA stamp.
3
7. If material fails to pass on resampling, return to Phase II, Items 1-6.

The ASTA Code Number will be prominently displayed on the Certificate. The "R" classification will alert the
buyer that either the merchandise has been reconditioned, or for some valid reason resampled. For example,
fumigation is an acceptable treatment in some cases where live insects are found. (Note Cleanliness Specifications,
Paragraph C. Insects). In this case, a fumigation certificate will be sent to ASTA so that when the laboratory
reports its new results, ASTA will be in a position to issue a valid "R" number.

CLEANLINESS SPECIFICATIONS

For purposes of these Specifications, extraneous matter is defined as everything foreign to the product itself and
includes, but is not restricted to: stones, dirt, wire, string, stems, sticks, nontoxic foreign seeds, excreta, manure
and animal contamination.
The level of contaminants permitted under these Specifications must fall below those shown on the following table,
except for the column ∆ "Whole Insects, Dead" which cannot exceed the limits shown.

CLEANLINESS ∆ WHOLE INSECT EXTRANEOUS/

SPECIFICATIONS INSECTS, EXCRETA, EXCRETA, DEFILED/ FOREIGN
DEAD MAMMALIAN OTHER MOLD INFESTED MATTER
Name of By By By % By % By % By
Spice, Seed or Herb Count Mg./Lb. Mg./Lb. Wgt. Wgt. Wgt.
Allspice 2 5 5.0 2.00 1.00 0.50
Anise 4 3 5.0 1.00 1.00 1.00
Sweet Basil 2 1 2.0 1.00 1.00 0.50
Caraway 4 3 10.0 1.00 1.00 0.50
Cardamom 4 3 1.0 1.00 1.00 0.50
Cassia 2 1 1.0 5.00 2.50 0.50
Cinnamon 2 1 2.0 1.00 1.00 0.50
Celery Seed 4 3 3.0 1.00 1.00 0.50
Chillies 4 1 8.0 3.00 2.50 0.50
Cloves* 4 5 8.0 1.00 1.00 1.00 *
Coriander 4 3 10.0 1.00 1.00 0.50
Cumin Seed 4 3 5.0 1.00 1.00 0.50
Dill Seed 4 3 2.0 1.00 1.00 0.50
Fennel Seed SF(2) SF(2) SF(2) 1.00 1.00 0.50
Ginger 4 3 3.0 SF(3) SF(3) 1.00
Laurel Leaves** 2 1 10.0 2.00 2.50 0.50
Mace 4 3 1.0 2.00 1.00 0.50
Marjoram 3 1 10.0 1.00 1.00 1.00
Nutmeg (Broken) 4 5 1.0 SF(4) SF(4) 0.50
Nutmeg (Whole) 4 0 0.0 SF(5) SF(5) 0.00
Oregano*** 3 1 10.0 1.00 1.00 1.00
Black Pepper 2 1 5.0 SF(6) SF(6) 1.00
White Pepper**** 2 1 1.0 SF(7) SF(7) 0.50
Poppy Seed 2 3 3.0 1.00 1.00 0.50
Rosemary Leaves 2 1 4.0 1.00 1.00 0.50
Sage** 2 1 4.0 1.00 1.00 0.50
Savory 2 1 10.0 1.00 1.00 0.50
Sesame Seed 4 5 10.0 1.00 1.00 0.50
Sesame Seed, Hulled 4 5 1.0 1.00 1.00 0.50
Tarragon 2 1 1.0 1.00 1.00 0.50
Thyme 4 1 5.0 1.00 1.00 0.50
Turmeric 3 5 5.0 3.00 2.50 0.50

4
Cleanliness Specifications - Footnotes:

* Clove Stems: Less than (<) 5% allowance by weight for unattached clove stems over and above the tolerance
for Other Extraneous Matter is permitted.

** Laurel Leaves: "Stems" will be reported separately for economic purposes and will not represent a

Sage: pass/fail criteria.

*** Oregano: Analysis for presence of Sumac shall not be mandatory if samples are marked "Product of Mexico."

**** White Pepper: "Percent Black Pepper" will be reported separately for economic purposes and will not
represent a pass/fail criteria.

(2) Fennel Seed: In the case of Fennel Seed, if 20% or more of the subsamples contain any rodent, other excreta
or whole insects, or an average of 3 mg/lb or more of mammalian excreta, the lot must be
reconditioned.

(3) Ginger: More than 3% moldy pieces and/or insect infested pieces by weight.

(4) Broken Nutmeg: More than 5% mold/insect defiled combined by weight.

(5) Whole Nutmeg: More than 10% insect infested and/or moldy pieces, with a maximum of 5% insect defiled
pieces by count.

(6) Black Pepper: 1% moldy and/or infested pieces by weight.

(7) White Pepper: 1% moldy and/or infested pieces by weight.

∆ Whole Insects, Dead: Cannot exceed the limits shown.

Extraneous Matter: Includes other plant material, e. g. foreign leaves

GROUND PROCESSED SPICE *

(Cannot exceed limit shown)

Whole Insect Other Rats/ Animal

Spices Equivalent Fragments Mites Insects Mouse Hairs
Insects Hairs

Ground Average of Average of

Paprika more than 75 more than
fragments 11 rodent
/25g hairs/25g

* Microanalytical Methods for Paprika and Ground Capsicums can be found in the “Analytical Procedures” section
of this book.

5
IN ADDITION TO THE PRECEDING SPECIFICATIONS, A LOT MUST ALSO BE
RECONDITIONED:

A. MAMMALIAN EXCRETA -
if the average of the total number of subsamples exceeds the listed milligram per pound Specification.
Exception: In the case of fennel seed, see footnote (2).

B. OTHER EXCRETA -

if the average weight expressed as milligrams per pound for all subdivisions of the sample exceeds the
specified values shown in the table.
Exception: In the case of fennel, see footnote (2).

NOTE: The Food and Drug Administration specifies only mammalian excreta, ASTA will count all types of non-
mammalian excreta as Other Excreta. FDA does retain the right to detain merchandise containing excessive
amounts of excreta which is of non-mammalian origin on the basis that the merchandise was exposed to Insanitary
Conditions.

C. INSECTS -

if the total number of whole dead insects found in the total number of the subsamples exceeds the specified
value shown in the table.

if live insects are found in the original sample reconditioning is to include fumigating. If the number of live
and dead insects exceed the cleanliness specification for whole dead insects for that spice, then reconditioning
must also include sifting and blowing. Thereafter, in accordance with Procedures A. Sampling, new samples
shall be drawn, analyzed and a new Certificate of Analysis issued.
Exceptions:

a. In the case of fennel seed, if 20% or more of the subsamples contain any whole insects, the lot
must be reconditioned. (For example, if two or more subsamples of a ten-unit sample each
contains one whole insect, the lot must be reconditioned.)

b. If the number of live insects found in the total number of subsamples are less than the cleanliness
specifications for whole insects for that spice, the lot must be fumigated and then resampled.

c. If it appears to the unaided eye that 50 or more mites and psocids are present, the lot must be
fumigated, sifted and blown. Mites and/or psocids are not to be counted as insects.

D. MOLD -

if mold is present, as expressed by percent by weight of the total quantity of spice in all subsamples, in excess
of the specified values shown in the table. A product is classified as moldy if it contains mold, visible to the
naked eye, exceeding 1/4 of its surface area and confirmed by the presence of mycelial filaments and spores
when examined with the aid of a microscope (40 X magnification or less).

E. INSECT DEFILED/INFESTED -
6
 if the total sample quantity exceeds the specified values shown in the table expressed as percent by weight of
insect infested, bored, or otherwise defiled seeds, leaves or roots. Before reconditioning a lot is considered
defiled whenever a sample shows visible evidence of webbing or definite insect feeding.

F. RECONDITIONING AND RECONDITIONED ITEMS -

Reconditioning may include, but shall not be limited to, techniques such as fumigating, washing, cutting,
sifting, aspirating and blowing.

Insect channels or insect-bored holes in reconditioned spices will not be counted as insect defiled, as they
would in the examination of non-reconditioned spices, provided there are no insects, webbing or excreta in
those channels or holes.

G. LIGHT BERRIES - Black Pepper -

if the light berries, though not considered extraneous matter, exceed 4% by weight.

7
 SAMPLING AND ANALYTICAL PROCEDURES

A. SAMPLING -

i. Each lot distinguished by specific chop marks and/or numbers must be sampled/analyzed
separately. For the purpose of an ASTA certificate, a lot shall be defined as not to exceed one
container load, where applicable. No commingling is permitted.

ii. In sampling merchandise for analysis under these specifications, the number of samples drawn
must be equal to the square root of the packages, bags or containers in the lot with a maximum of
ten samples drawn. In the event that this is for ground/crushed processed spices the number of
samples drawn must be six.

iii. The sample size shall be 3/4 to 1 pound for the high density items. These include: Black and
White Pepper, Cassia, Cinnamon (Seychelle), Nutmeg (Whole and Broken), Ginger, Cloves,
Allspice/Pimento, Turmeric, Celery Seed, Poppy Seed, Sesame Seed, Caraway Seed, Cardamom
Seed, Anise Seed, Coriander Seed, Cumin Seed, Dill Seed, and Fennel Seed.

The sample size shall be 1/2 to 3/4 of a pound for the low density items. These include: Chillies,
Capsicums, Mace, Sage, Oregano Leaves, Basil Leaves, Laurel Leaves, Thyme Leaves,
Rosemary Leaves, Tarragon Leaves, Marjoram Leaves and Savory Leaves.

iv. The sampling size shall be 4-6 ounces (113-170 grams) for ground/crushed processed spices.

All samples shall be drawn by an independent and impartial public sampler and shall be
forwarded by him directly to an ASTA Approved Laboratory that is to make the analysis.

v. In all cases, each subsample must be analyzed. In the case of high and low density items the
entire sample must be analyzed, in the samples of ground and crushed processed spices the
amount to be analyzed is determined by the method used.

B. PROCEDURES -

Utilize the appropriate methods as given under the “Analytical Procedures” section of this manual.

C. RESAMPLING/REANALYSIS -

i. No resampling or reanalysis is permitted except in those instances where fumigation and/or

reconditioning has taken place.

D. LABORATORIES -

The list of laboratories approved by the Association to be qualified to make the required macroscopic analyses
has been published. The names and addresses of these firms and others that are subsequently added to the list
can be secured through the Association.

8
ANALYTICAL PROCEDURES

9
 ASTA ANALYTICAL METHODS

Method 14.0
Light Berries and Extraneous Matter in Black and White Pepper

Purpose: To determine the amount of light berries and extraneous matter in black or white pepper.

A. Apparatus:

1. A standard pepper sieve, (No. 9 1/2 round screen with a frame 18 to 22 inches in diameter and 2 3/4
inches in height. The bottom is a metal sheet perforated with round holes of 7/64 inch in diameter,
with an average of 5 1/2 holes per linear inch. Screen only with standard pepper sieve obtainable
from: McNichols Company, 5501 Gray Street, Tampa, Florida 33609 (813) 876-4100 or (800) 237-
3820. U.S. Standard No. 8 sieve (0.0937 in. or 2.38 square mm opening) provide equivalent sieve
opening.

2. Balance -- sensitivity 0.01 g.

3. Beaker, 600 ml. Griffin, Low form, pyrex approximately 85 mm. in diameter and 120 mm. in height
is recommended. (Note 1).

4. Blotting paper or other similar absorbent material.

5. Tweezers.

6. Stereoscopic, Binocular, wide-field microscope (40-50x).

B. Reagents:

1. Alcohol-water solution of a specific gravity 0.80-0.82 at 25°/25°. The alcohol may be ethanol,
denatured ethanol (Note 2) or isopropanol.

C. Preparation of Sample:

1. The number of samples drawn must be equal to the square root of the packages, bags, or containers
in the lot, with maximum of 10 samples drawn.

2. The sample size shall be 3/4 to l pound (340 g to 454 g).

3. Each entire subsample must be analyzed.

10
 ASTA ANALYTICAL METHODS

Method 14.0
Light Berries and Extraneous Matter in Black and White Pepper

D. Procedure:

1. To Determine Excreta, Insects, Mites, Psocids, Mold, and Percent Black Pepper in White
Pepper:
a. Weigh each sub-sample to the nearest gram. Sprinkle and examine a small portion at a time,
with a good light and against a white background into a standard pepper sieve. Pick out any
bird, rodent, or other animal excreta. Separate mammalian from non-mammalian excreta.
Weigh to the nearest 0.1 mg and record. Do not remove other extraneous/foreign material at
this time.
b. Shake pepper sieve moderately back and forth, examine siftings collected on white
background for live and dead insects and for excreta.
c. Accumulate the siftings.
d. Mix sub-sample of pepper on sieve and weight 50 g of aliquot into a pan. Hand-pick moldy
peppercorns and weigh. In case of white pepper, additionally hand pick for black peppercorns
(with skin coat attached) and record. Each sub-sample is examined in sequence in a similar
manner and the results are averaged.

2. Extraneous/Foreign Matter by Sifting:

a. Weigh to nearest 0.1 g the cumulated siftings and calculate the percentage by weight. Percent
siftings must be determined after the removal of small berries that pass through the pepper
sieve. (See Calculations)

3. Light Berry Determination for Black Pepper:

a. Combine sufficient material from each sub-sample to give a composite sample of
approximately 5 lbs. Mix composite well.
b. Form the composite into a pile shaped like a cone. Quarter the cone designating each quarter
as A, B, C, or D in a clockwise sequence.
c. Set aside two opposite quarters such as A and C.
d. Mix and reduce each quarter separately.
e. Remove a 50.0 g sample from each reduced quarter.
f. Place the weighed sample in the 600 mL Griffin, low-form pyrex beaker and add 300 mL of
the alcohol-water solution.
g. Stir the material in the beaker with a spoon and allow to settle two minutes; then spoon off the
berries which float.
h. Repeat the stirring, settling and removal of the floating berries until two successive additional
stirrings raise no more berries to the surface. Remove only the berries that actually float
(Note 3).
i. Blot the removed berries to free them from excess liquid and spread them out to dry on a piece
of paper towel or other absorbent material.

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 ASTA ANALYTICAL METHODS

Method 14.0
Light Berries and Extraneous Matter in Pepper

j. Air dry for one hour and weigh the air dried light berries to the nearest 0.01 g and calculate
and report the percent of light berries to the nearest 0.1%.
(See Calculations)
k. If the range of two determinations is not over 0.8%, the two determinations shall be averaged
and reported as percent light berries. If the difference is greater than 0.8%, determine the light
berries in a third sample obtained from either quarter B or D. Average all three values and
report as percent light berries.
4. Extraneous/Foreign Matter by Hand Picking
a. From the two opposite quarters set aside for light berries determination such as A & C, weigh
100 grams from each and hand pick for any sticks, stones, stems, foreign seeds, other
extraneous matter and make note of its nature.
b. Weigh the pickings and calculate. (See Calculations)
E. Calculations:
Wt. of combined sifting (g)
% Extraneous/Foreign Matter by Sifting = ____________________________________________________ x 100
Combined wt. of sub-samples (g)

weight of light berries (g)

% Light Berries = ____________________________________________________________ x 100
weight of samples (50 g)

A (in grams) + C (in grams)

% Extraneous/Foreign Matter by Hand Picking = __________________________________________________
2
F. Statistics:
TBD
G. Notes:

1. Other transparent beakers may be used, but they should be between 75 and 100 mm. in diameter and
between 100 and 140 mm in height.

2. Specially denatured alcohols no. 3A, 23A, or 30 are recommended.

a. SDA no. 3A: 5 gallons of methyl alcohol plus 100 gallons 95% ethyl alcohol.
b. SDA no. 23A: 10 gallons USP acetone plus 100 gallons 95% ethyl alcohol.
c. SDA no. 30: 10 gallons methyl alcohol plus 100 gallons 95% ethyl alcohol.

3. Some berries may remain suspended some distance below the surface of the liquid. These are not
considered as floaters.
H. References:
Macroanalytical Procedures Manual 1984, Chapter 5.

12
 Revised January, 1997

ASTA ANALYTICAL METHODS

Method 14.1
Extraneous Matter in Spices (Excluding Black and White Pepper)

Purpose: To determine the amount of extraneous matter in spices (excluding black and white
pepper).

A. Apparatus:

1. A standard pepper sieve, (No. 9 1/2 round screen with a frame 18 to 22 inches in diameter and 2 3/4
inches in height. The bottom is a metal sheet perforated with round holes of 7/64 inch in diameter,
with an average of 5 1/2 holes per linear inch. Screen only with standard pepper sieve obtainable
from: McNichols Company, 5501 Gray Street, Tampa, Florida 33609 (813) 876-4100 or (800) 237-
3820. U.S. Standard No. 8 sieve (0.0937 in. or 2.38 square mm opening) provide equivalent sieve
opening.

2. Balance -- sensitivity 0.01 g.

3. Tweezers

4. Binocular, wide-field microscope (40-50x).

B. Reagents:

None required.

C. Preparation of Sample:

1. The number of samples drawn must be equal to the square root of the package, bags or containers in
the lot with a maximum of ten samples drawn.

2. The sample size shall be 3/4 to 1 pound for high density items. These include: Cassia, Cinnamon
(Seychelle), Nutmeg (Whole and Broken), Ginger, Cloves, Allspice/Pimento, Turmeric, Celery Seed,
Poppy Seed, Sesame Seed, Caraway Seed, Cardamom Seed, Anise Seed, Coriander Seed, Cumin
Seed, Dill Seed, and Fennel Seed.

3. The sample size shall be 1/2 to 3/4 of a pound for low density items. These include: Chillies,
Capsicums, Mace, Sage, Oregano Leaves, Basil Leaves, Laurel Leaves, Thyme Leaves, Rosemary
Leaves, Tarragon Leaves, Marjoram Leaves and Savory Leaves.

4. Regardless of sample size, the entire subsample must be analyzed.

13
 ASTA ANALYTICAL METHODS

Method 14.1
Extraneous Matter in Spices (Excluding Pepper)

D. Procedure:

1. Whole and Broken Nutmegs:

Whole Nutmegs:
a. Weigh and shake each subsample on the sieve. Examine siftings for live and dead insects,
extraneous matter and mammalian or other excreta.
b. Select at random 100 Nutmegs from each subsample. Cut the Nutmegs in half longitudinally.
Examine the cut surfaces of each Nutmeg for evidence of insects or insect damage and presence
of mold filaments. Report as rejects Nutmegs containing insects or insect parts, insect excreta,
insect channeling, and those showing mold filaments on 25% or more of the cut surface of each.
(Note 3) Check borderline or doubtful specimens using magnification. Report results for Insect
Defiled/Infested and for Mold separately for each subsample in % by count and average.

Broken Nutmegs:
a. Weigh and shake each subsample on the sieve. Examine siftings for live and dead insects,
extraneous matter. (Note 1)
b. Mix thoroughly, weigh out 50 grams of each subsample and examine the surfaces of the
Nutmegs for evidence of insects or insect damage and presence of mold filaments. Report as
rejects Nutmegs containing insects or insect parts, excreta, insect channeling, and those showing
mold filaments on 25% or more of the cut surface of each. (Note 3) Check borderline or
doubtful specimens using magnification. Report results of each subsample in % by weight and
average. Results of Insect Defiled/Infested and Mold can be combined in the case of Broken
Nutmegs. (Note 1)

2. Chillies:

a. The individual subsample is weighed, shaken from the bag, a small portion at a time, with a
good light, on the sieve with white paper beneath. As the chillies are discharged on the sieve,
they are examined for extraneous/foreign matter, mammalian, or other excreta. (Note 1)

b. When the entire sample is on the sieve, it is shaken back and forth a few times. The siftings on
the white paper are also examined for live and dead insects, mammalian, or other excreta.

14
 ASTA ANALYTICAL METHODS

Method 14.1
Extraneous Matter in Spices (Excluding Pepper)

c. The sample is mixed and a 25 gram portion for chillies up to 2 1/2 inches in length or 100 gram
portion for chillies over 2 1/2 inches in length is taken at random for examination of mold or
insect defiled/infested chillies. (Note 3) The chillies are broken and examined inside. Moldy
chillies (mold exceeding 1/4 of its area) or insect defiled/infested pods are weighed and % by
weight determined.
d. Each sample representing the lot is done in sequence in this manner.

3. Basil, Marjoram, Oregano (Note 2), Rosemary Leaves, Savory, Tarragon, and Thyme:
a. The individual subsamples are weighed. Each subsample is shaken from the bag, a small
portion at a time on white paper, with good light.
b. As the sample is discharged and spread out on the paper, examine and pick out live and dead
insects, mammalian and other excreta, mold and insect defiled/infested pieces. (Note 1 and
Note 3)
c. Report by count (Whole Insects) or by weight (Mammalian Excreta. Other Excreta, Mold and
Insect Defiled/Infested Pieces) on the Certificates of Analysis.
d. Extraneous/Foreign Matter is defined as everything foreign to the product itself and includes,
but is not restricted to: stones, dirt, wire, string, stems, sticks, nontoxic foreign seeds and hair
and other plant materials, e.g. foreign leaves. (Note 5)
e. From a composite of the subsamples weigh out and hand pick 100 grams of sample for
extraneous/foreign matter.
f. Record results in percent by weight.

4. Anise Seed, Caraway Seed, Celery Seed, Cloves, Coriander, Cumin Seed, Dill Seed, Fennel Seed,
Mace (siftings), Poppy Seed, Sesame Seed, Hulled Sesame Seed:

a. The individual subsamples are weighed. Each subsample is shaken from the bag, a small
portion at a time on white paper, with good light.
b. As the sample is discharged and spread out on the paper, examine and pick out live and dead
insects, mammalian or other excreta, mold and insect defiled/infested pieces. (Note 1, Note 3)
c. Report by count (Whole Insects) or by weight (Mammalian Excreta. Other Excreta, Mold and
Insect Defiled/Infested Pieces).
d. Extraneous/Foreign Matter is defined as everything foreign to the product itself and includes,
but is not restricted to: stones, dirt, wire, string, stems, sticks, nontoxic foreign seeds and hair.

15
 ASTA ANALYTICAL METHODS

Method 14.1
Extraneous Matter in Spices (Excluding Pepper)

e. From a composite of the subsamples weigh out and hand pick 50 grams of sample for
extraneous/foreign matter.
f. Record results in percent by weight.

5. Allspice/Pimento, Whole Cardamom, Broken Cassia, Madagascar and Seychelle Cinnamon,

Ginger, Laurel Leaves (Fig. 1), Mace (whole), Sage, Turmeric (Note 4, Note 5):
a. The individual subsample is weighed, shaken from the bag, a small portion at a time, with a
good light, on the sieve with white paper beneath. As the sample is discharged on the sieve,
examine for extraneous/foreign matter, mammalian, or other excreta.
b. When the entire sample is on the sieve, it is shaken back and forth a few times. The siftings
on the white paper are also examined for live and dead insects, mammalian, or other
excreta. (Note 1)
c. Examine entire sample for mold or insect defiled/infested pieces. (Note 3)
d. Report by count (Whole Insects) or by weight in milligrams (Mammalian Excreta, Other
Excreta).
e. Each sample representing the lot is done in sequence in this manner.

6. Cassia Sticks or Vera AA Cassia:

a. Break each stick separately from the entire subsample into pieces with a hammer or weight.
b. Examine the pieces for mold or insect defiled/infested pieces. (Note 3)
c. The entire stick is considered in the calculations where evidence of contamination is found.
d. Report results of each subsample in % by weight in milligrams.

E. Calculations:

1. Excreta mg/lb = Weight Excreta (mg) 454g

x
Weight of Product (g) 1 lb.

2. % Moldy/Insect Defiled/Infested Product = Weight Reject Product (g) x 100

Weight Product (g)

3. % Siftings = Weight Siftings (g) x 100

Weight Product (g)

4. %Extraneous Matter = Weight Extraneous Matter (g) x 100

Weight Product (g)

16
 ASTA ANALYTICAL METHODS

Method 14.1
Extraneous Matter in Spices (Excluding Pepper)

F. Statistics:

TBD

G. Notes:

1. Calculate the average milligrams of mammalian excreta and the average milligram of other excreta
across all of the subsamples analyzed and report each value separately, and as mg/lb (see
Calculations).

2. In the case of Oregano, analysis for the presence of Sumac must be performed (See Method 26.0).
However, if the samples are marked "Product of Mexico", analysis for the presence of Sumac shall not
be mandatory.

3 . Classification of Mold and Insect Defiled - Insect Defiled -- Any product material exhibiting definite
evidence of insect feeding or webbing.

Mold -- any material bearing mold, visible to the naked eye, exceeding 1/4 of its area and confirmed
by the presence of mycelial filaments and spores when examined the aid of a microscope (40X
magnification or less).

4. In the case of Sage and Bay leaves only, a separate column will be used on the Certificate of Analysis
to report "Stems." This information will be for economic purposes only and will not represent a
pass/fail criteria.

17
 ASTA ANALYTICAL METHODS

Method 14.1
Extraneous Matter in Spices (Excluding Pepper)

5. The definitions of extraneous plant material/stem:

SAGE - If any pithy plant material exceeds one (1) millimeter in diameter at any point on its
contiguous body, the attached leaves are stripped and if the length exceeds twelve and a half (12.5)
millimeters, the remaining material is defined as a stem. In addition, if any pithy plant material
exceeds two (2) millimeters in diameter at any point, it is considered to be a stem regardless of its
length. That material is to be weighed and the total weight of stems reported as a percentage.

BAY (LAUREL) LEAVES - If any pithy plant material, excluding the "bud," exceeds one (1)
millimeter in diameter at any point on its contiguous body, the attached leaves are stripped and if
the length exceeds twelve and a half (12.5) millimeters, the remaining material is defined as a stem.
In addition, if any pithy plant material exceeds two (2) millimeters in diameter at any point, it is
considered to be a stem regardless of its length. That material is to be weighed and the total weight
of stems reported as a percentage.

ROSEMARY and SAVORY - The analyst establishes an average size of the leaves in a particular
lot. Any pithy plant material other than the leaves exceeding that size/dimension is defined as
extraneous matter.

THYME - If any pithy plant material exceeds the suggested length of twelve and a half (12.5)
millimeters (1/2"), it is defined as extraneous matter.

CLOVES - If a stem attached to a clove is greater in length than the clove itself, it shall be broken
off and counted as a clove stem.

OREGANO - If any pithy plant material exceeds one (1) millimeter in diameter at any point on its
contiguous body, the attached leaves are stripped and if the length exceeds twelve and a half (12.5)
millimeters (1/2"), the-remaining material is defined as extraneous matter. In addition, if any pithy
plant material exceeds two (2) millimeters in diameter at any point, it is considered to be
extraneous matter regardless of its length.

SWEET BASIL - If any pithy plant material exceeds one (1) millimeter in diameter at any point on
its contiguous body, the attached leaves are stripped and if the length exceeds twelve and a half
(12.5) millimeters (1/2"), the remaining material is defined as extraneous matter. In addition, if
any pithy plant material exceeds two (2) millimeters in diameter at any point, it is considered to be
extraneous matter regardless of its length.

H. References:

Macronalytical Procedures Manual (MPM) 1984, Chapter V.

Revised January, 1997

18
 THIS PAGE CONTAINS
FIGURE 1
CLASSIFICATION OF INSECT DAMAGE IN BAY LEAVES
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19
 ASTA ANALYTICAL METHODS

Method 22.1
Microanalytical Analysis of Paprika

Purpose: To isolate extraneous material of insect, rodent, other animal and bird origin
from ground paprika for microscopic detection and enumeration.

A. Apparatus:

1. Stereoscopic binocular microscope - wide field with following minimum specifications: 3 parfocal
objectives - 1X, 3 X and 6X or 7.5X; paired 10X wide field oculars, mounted on a base and
capable of illumination by reflected light. Ordinarily 30X magnification is used for routine
examination of filter papers. Confirmation of suspect material at higher magnification may be
required.

2. Microscope illuminator - preferably with a transformer or rheostat to vary light intensity, a

focusing adjustment to give uniformly lighted field of view, and blue-white color from a cool low-
voltage source.

3. Wildman trap flask - consists of a 2L Erlenmeyer flask into which is inserted a close-fitting rubber
stopper supported on a stiff metal rod, 3/16" diam., and about 4" longer than height of flask. Rod
is threaded at lower end and furnished with nuts and washers to hold it in place on the stopper.
Countersink lower nut and washer in the rubber stopper to prevent striking flask.

4. Filter paper: a) 32 cm folded rapid flow (S & S 588, or equivalent).

b) 9 cm high wet strength, ruled, 5mm apart (S&S #8 or equivalent).

5. Hirsch funnel, porcelain, 56 mm plate diameter.

6. Büchner funnel, porcelain, 114 mm diameter.

7. Suction flask to provide suction by means of an H2O aspirator or electric vacuum pump.

8. Sieve U.S. Standard No. 230, 8" or 12" diameter (plain-not twill weave).

9. Magnetic stirrer - hot plate.

10. Teflon coated magnetic stirring bar, 1 3/4" - 2" x 3/8" (44.4 mm - 50.8 mm x 9.5 mm).

11. Beakers, glass - 1 liter, funnels, glass or metal, 6" diameter or greater

20
 ASTA ANALYTICAL METHODS

Method 22.1
Microanalytical Analysis of Paprika

B. Reagents

1. Isopropanol and 40% isopropanol in water.

2. Premix Tween 80-40% isopropanol-Tetrasodium EDTA: mix 250mL of (a) and 250mL of (b):
a) Mix and filter 40mL Tween 80 and 2l0mL 40% isopropanol.
b) Dissolve 5g Na4 EDTA in l50mL H2O add l00mL isopropanol, mix and filter.
Mixed reagent is stable several weeks. Store in non-metal containers.

3. Mineral Oil - paraffin oil, white, light 125/135, saybolt viscosity (38°), specific gravity 0.84-0.86
(24°). Fisher Scientific Co. No. 0-119 or equivalent.

C. Preparation of Sample

1. The number of samples drawn should be six.

2. The sample size shall be 4 - 6 ounces (113 to 170 grams).

D. Procedure

I. Weigh 25.0g of paprika and place in a filter paper cup formed by fitting a 32cm filter paper around
a 400mL beaker. Place cup with ground paprika in a 1L beaker.

2. Pour 400mL 99% isopropanol into the paper cup in the beaker. Place on a pre-heated hot plate,
bring to a boil, then boil gently exactly 10 minutes (a "cold finger" should be used to condense
vapors).

3. Remove cup from beaker without delay and place in a Büchner funnel and aspirate to slow drip.
Discard liquid.

4. Replace cup in 1 liter beaker and repeat Step 2 and 3 twice using 400mL 99% isopropanol each time
to remove oil and pigment.

5. Using a gentle stream of water, quantitatively transfer the sample to a prewashed No. 230 sieve.
Avoid splashing and loss of sample.

21
 ASTA ANALYTICAL METHODS

Method 22.1

Microanalytical Analysis of Paprika

6. Wash the sample with a forceful stream of warm (55 -70°C) water using a Fisher aerator until foam
is gone and drainings are clear. Higher flow rates may cause breakage of fragments.

7. Add 400mL 40% isopropanol to wash bottle. Place 6" diameter funnel in trap flask. Wash sample
to the edge of the sieve and quantitatively transfer to the trap flask with 40% isopropanol. Wash
walls of flask and pour remainder of 400mL into flask.

8. Place on hot plate, bring to a boil, then boil gently 10 minutes, using gentle magnetic stirring to
avoid splashing. Wash sides of flask every 2 minutes to prevent material from accumulating and
drying on flask wall.

9. Remove from hot plate and immediately add 100mL premixed Tween 80-40% isopropanol - Na4
EDTA solution down stirring rod.

10. Stir mag., gently, about 1 minute. Let stand 10 minutes.

11. Dilute to 800mL with 40% isopropanol added slowly down stirring rod, positioned with stopper just
above liquid level.

12. Add 50mL mineral oil down stirring rod and stir mag. 3 minutes with stopper located above liquid
level.

13. Add 40% isopropanol slowly down stirring rod to bring oil into neck of flask. Let stand about 10
minutes.

14. Raise stopper to middle of flask and swirl gently to hasten rising of oil droplets.

15. Rinse rod with 40% isopropanol and clamp so that stopper is at mid point of flask.

16. Add 40% isopropanol down rod to bring bottom of oil layer to level 1 cm above raised stopper.

17. Let stand 10 minutes and swirl very gently again.

18. Let stand 10 minutes undisturbed and trap off into beaker, or onto ruled filter paper in Hirsch funnel.

22
 ASTA ANALYTICAL METHODS

Method 22.1
Microanalytical Analysis of Paprika

19. Add 35mL mineral oil and hand stir 1 minute at speed to sufficiently keep oil moving through trap
flask. Add about 20mL 40% isopropanol, stir gently at about 5 minute intervals for 20 - 25 minutes,
then let stand undisturbed 5 - 10 minutes.

20. Trap off into second beaker or onto ruled filter paper in Hirsch funnel and rinse neck of flask with
alcohol or undiluted isopropanol.

21. Filter onto ruled paper, rinsing beaker with isopropanol, and examine at 30X.

E. Calculation:

Report separately numbers of insect, rodent hair, animal hair and feather barbule fragments.

F. Statistics:

Repeatability Reproducibility
Rodent hairs 6.41% 8.59%
Elytra squares 5.89% 6.24%

G. Notes

1. Periodically check the 32cm filter paper used under microscope at 30X for completeness of transfer
of fragments

2. When 230 sieve draining slows, wash with detergent, then 50% sodium hydroxide (heated, if
necessary).

3. Complete analysis without overnight interruption.

H. References

AOAC Official Methods of Analysis - 16.14.22 (977.25).

JAOAC 60 114 (1977).

Revised January, 1997

23
 ASTA ANALYTICAL METHODS

Method 22.2
Microanalytical Analysis of Ground Capsicums (Excluding Paprika)

Purpose: To isolate extraneous material of insect, rodent, other animal and bird origin
from ground capsicums, excluding paprika, for microscopic detection and enumeration.

A. Apparatus:

1. Stereoscopic binocular microscope - wide field with following minimum specifications: 3 parfocal
objectives - 1X, 3X and 6X or 7.5X; paired 10X wide field oculars, mounted on a base and capable of
illumination by reflected light. Ordinarily 30X magnification is used for routine examination of filter
papers. Confirmation of suspect material at higher magnification may be required.

2. Microscope illuminator - preferably with a transformer or rheostat to vary light intensity, a focusing
adjustment to give uniformly lighted field of view, and blue-white color from a cool low-voltage
source.

3 . Wildman trap flask - consists of a 2L Erlenmeyer flask into which is inserted a close-fitting rubber
stopper supported on a stiff metal rod, 3/16" diam., and about 4" longer than height of flask. Rod is
threaded at lower end and furnished with nuts and washers to hold it in place on the stopper.
Countersink lower nut and washer in the rubber stopper to prevent striking flask.

4. Filter paper: a) 32cm folded rapid flow (S & S 588, or equivalent).

b) 9 cm high wet strength, ruled, 5mm apart (S&S #8 or equivalent).

5. Hirsch funnel - porcelain, 56 mm plate diameter.

6. Büchner funnel, porcelain, 114 mm diameter.

7. Suction flask to provide suction by means of an H2O aspirator or electric vacuum pump.

8. Sieve U.S. Standard No. 230, 8" or 12" diameter (plain-not twill weave).

9. Magnetic stirrer - hot plate.

10. Teflon covered stirring bars about 47mm x 9mm (egg-shaped, round or octagonal)

11. Beakers, glass - 1 liter, funnels, glass or metal, 6" diameter or greater.

24
 ASTA ANALYTICAL METHODS

Method 22.2
Microanalytical Analysis of Ground Capsicums (Excluding Paprika)

B. Reagents:

1. Isopropanol (IPA) - 99% and 40% by volume.

2. Ethanol 95% and 60% by volume.

3. Tween 80-ethanol-tetrasodium EDTA Premix: pour 420mL 60% ethanol in 1L graduate. Add 80mL
Tween 80 to 250mL glass stoppered graduate. Invert 250mL graduate over 3L beaker and drain
briefly. Rinse 250mL graduate with several portions of the 420mL 60% ethanol, pouring each into
beaker. Add rest of 60% ethanol to beaker and start mag. stirring. Add 10g Tetrasodium EDTA to
beaker while stirring rapidly. Add 500mL 60% ethanol and stir until uniform. Store in non-metal
containers. Mixed reagent is stable several weeks.

4. Mineral Oil - Paraffin oil, white, light 125/135, saybolt viscosity (38°), specific gravity 0.84-0.86
(24°). Fisher Scientific Co. No. 0-119 (or equivalent).

5. Heptane - commercial heptane containing less than 8% toluene.

6. Flotation liquid - mineral oil and heptane (85+15).

C. Preparation of Sample

1. The number of samples drawn should be six.

2. The sample size shall be 4 - 6 ounces (113 to 170 grams).

D. Procedure:

1. Weigh 25 g of capsicum and place in a filter paper cup formed by fitting a 32cm filter paper around a
400mL beaker. Place cup with ground capsicums in a 1L beaker.

2. Pour 400mL 99% isopropanol into the paper cup in the beaker. Place on a pre-heated hot plate, bring
to a boil, then boil gently 10 min.

3 . Remove cup from beaker without delay and place in a Büchner funnel and aspirate to slow drip.
Discard liquid.

4 . Replace cup in 1 liter beaker and repeat Step 2 and 3 twice using 400mL 99% isopropanol each time
to remove oil and pigment.

25
 ASTA ANALYTICAL METHODS

Method 22.2
Microanalytical Analysis of Ground Capsicums (Excluding Paprika)

5. Using a gentle stream of water, quantitatively transfer the sample to a prewashed No. 230 sieve.
Avoid splashing and loss of sample.

6. Wash the sample with a forceful stream of warm (55-70°C) water using a Fisher aerator until foam
is gone and drainings are clear. Higher flow rates may cause breakage of fragments. (Note: Longer
washing time than for paprika is needed.)

7. Add 600mL 60% ethanol to wash bottle. Place 6" diameter funnel in trap flask. Wash sample to the
edge of the sieve and quantitatively transfer to the trap flask with 60% ethanol. Wash walls of flask
and pour remainder of 600mL into flask.

8. Place on hot plate, bring to a boil, then boil gently 10 minutes, using gentle magnetic stirring to
avoid splashing. Wash sides of flask every 2 minutes to prevent material from accumulating and
drying on flask wall.

9. Remove from hot plate and cool to between 20 and 25° with cold water. Add 40mL flotation liquid
down stirring rod.

10. Dilute to 800mL with 60% ethanol and stir mag. 5 minutes.

11. Set aside, add 100 mL Tween 80-ethanol-tetrasodium EDTA premix, down stirring rod, and mix
through liquid by gently swirling (to prevent foaming) stopper about 1 minute. Let stand 3 minutes.
Wash sides of flask with 60% ethanol to keep solids down.

12. Slowly add 60% ethanol down trap rod, maintaining stopper above oil layer, until oil just reaches
neck of flask.

13. Gently swirl stopper through lower portion of flask to suspend settling.

14. Add 60% ethanol down rod to bring bottom of oil layer to a level 1 cm above raised stopper.

15. Clamp rod with stopper at mid point of flask. Let stand 15 minutes. Then gently swirl stopper
through upper half of liquid to hasten rising of oil droplets.

16. Let stand 15 minutes undisturbed and trap into beaker, rinsing neck of flask with 60% ethanol.
Filter onto ruled paper.

17. Add 30mL flotation liquid and stir manually 1 minute, with an up and down motion.

26
 ASTA ANALYTICAL METHODS

Method 22.2
Microanalytical Analysis of Ground Capsicums (Excluding Paprika)

18. Clamp rod at mid point and let stand 10 minutes. Swirl stopper gently through upper half of liquid
and adjust oil level.

19. Let stand 15 minutes undisturbed and trap off.

20. Rinse neck of flask with 95% ethanol.

21. Filter onto ruled paper, rinsing beaker with 95% ethanol, and examine at 30X.

E. Calculation:

Report numbers of insect, rodent hair, animal hair and feather barbule fragments.

F. Statistics:

Repeatability Reproducibility
Rodent hairs 13.43% 14.66%
Elytra squares 8.10% 9.18%

G. Notes:

1. Periodically check the 32cm filter paper used under microscope for completeness of transfer of
fragments.

2. When 230 sieve draining slows, wash with detergent, then 50% sodium hydroxide (heated if
necessary).

3. Complete analysis without overnight interruption.

4 Do not use any plastic equipment because fragments or hairs might adhere.

H. Reference:

AOAC Official Methods of Analysis 16.14.10 (978.22).

JAOAC 61 900 (1978).

Revised January, 1997

27
 ASTA ANALYTICAL METHODS

Method 26.0
Sumac in Oregano

Purpose: To determine the presence of sumac in oregano.

A. Apparatus:

1. Stereoscopic binocular microscope--widefield with the following minimum

specifications: 3 parfocal objectives--1X, 3X and 6X or 7.5X; paired 10X widefield oculars, mounted
on a base and capable of illumination by transmitted or reflected light. Ordinarily 30X magnification
is used for routine examination of filter papers. Confirmation of suspect material at higher
magnification may be required.

2. Compound polarizing microscope - microscope with the following minimum

specifications: 4 parfocal achromatic objectives of ca 4, 10, 20, and 40X; revolving 4-place
nosepiece; Abbe condenser with N.A. of 1.25; 10X Huygenian or widefield eyepieces; fine adjustment;
mechanical stage; fitted with polarizing prisms below and above the mechanical stage.

3. 100 mm Petri plates.

4 . 7 cm (or 9 cm) ruled filter papers.

5. Microscope slides and cover slips.

6. Eye dropper.

7. Dissecting needle and forceps.

B. Reagents:

1. Xylene - reagent grade.

C. Preparation of Sample:

1. Utilize sample procedures for original sampling (see Method 14. 1). From each sub-sample, take a
representative 10 g (approximately 75 mL) sample of oregano leaf. These samples will be composited
and gently blended.

2. Prepare a 5 to 10 g analytical sample from above using a sampling splitter or quartering procedure.

3. Maintain leaf integrity; do not break up leaves.

28
 ASTA ANALYTICAL METHODS

Method 26.0
Sumac in Oregano

D. Procedure:

1. Gently shake enough oregano into one Petri plate lined with standard 7 cm (or 9 cm) ruled filter paper
to cover the filter paper in a single layer (approximately 500 leaf fragments per plate; in no case is less
than 400 mg of oregano fragments to be viewed).

2. Examine the plate of material under the 30X to 60X magnification of the stereoscopic binocular
microscope looking for the following distinguishing characteristics:

Oregano - Oil glands, jointed hairs

Sumac - No oil glands, single cell hairs with central cavity

3. If no sumac is found in the plate, report as "Sumac Negative."

4. If sumac is found in the plate, report as Presumptive Sumac Positive (PSP) and continue to the
confirmatory steps.

5. To confirm the presence of sumac, place leaf fragments of oregano and the PSP material on a
microscope slide and place a cover slip over the material. Using an eye dropper, introduce enough
xylene to fill the cover slip.

6. Polarizing Microscope -
Observe with crossed polars at 40X to 100X magnification for the following:

Oregano - No oxalate crystals

Sumac - Oxalate crystals, birefringence of the sumac hairs and their central cavity.
(Note)

7. If the presence of sumac is confirmed, report as "sumac positive".

E. Calculation:

N/A

F. Statistics:

N/A

G. Notes:

Upon removing the polarization, the oxalate crystals in the sumac disappear.

H. References:

N/A Revised January 1997

29