Study 2212 2
Study 2212 2
Study 2212 2
Dissolved Oxygen
153
Section 4 TDEC - Fleming Training Center
Definition
Dissolved Oxygen Dissolved Oxygen
and Temperature
DO for short
measurement of the
amount of oxygen
dissolved in a unit
volume of water
indicator of
usefulness of water
for a specific
application
Applications Theory
Theory Theory
http://water.usgs.gov/software/DOTABLES/
Effect of Temperature on
Two Types of Measurement
Dissolved Oxygen
Pot of boiling water Electrode
bubbles form on
bottom & sides of
pot
number & size of
Winkler titration
bubbles increase with
temperature.
These are air bubbles
that have been
dissolved in water.
Electrodes Electrodes
Luminescent Dissolved
Common Deficiencies
Oxygen
Samples for dissolved oxygen were No membrane
collected in a bucket and then poured into
the BOD bottle No electrolyte to
The D.O. probe was immersed in the water foul or poison
during calibration
Wont affect
The D.O. probe had a water droplet on the
end during calibration readings
There was an air bubble under the Accurate & stable
membrane on the probe readings
The meter was air calibrated by placing the
probe on the counter
Winkler DO QA/QC
QC Acceptance
*QC Frequency
Control Charts
Titrate with
LFM / LFMD
ICAL / CCV
Batch Size
Corrective
Parameter
Method
Action
0.025M Sodium
DOC
MDL
LRB
LFB
Dup
Thiosulfate
Use a burette with
0.5 mL
Depends on Depends on
verify with air-saturated water
SM4500-O HachMethod
G - 2001 10360Oct. 2011
increments
Permit
Oxygen, dissolved
X X X X 20
Titrate until a pale
straw color
Permit
X X X X 20
35 TDEC - Fleming Training Center 36 TDEC - Fleming Training Center
DO SM4500-O G 2001
Membrane Electrode Method
Calibration
Calibrate daily (day of) by following
manufacturers instructions
Using barometric pressure is best
Duplicates of the sample
Run on a 5% basis, one for every 20 samples
Calculate %RPD, 20%
2014 Update For reporting purposes, all
duplicates should be reported according to
your permit limits. If your permit sets a
minimum, then the minimum value should be
reported even if falls outside your permit
limit.
Can be made at http://water.usgs.gov/software/DOTABLES/
TDEC - Fleming Training Center TDEC - Fleming Training Center
49 50
Temperature Temperature
NIST (National Institute of Standards Clean the probe end with deionized
water
& Technology) traceable thermometer
Swirl the thermometer in the sample and
Scale marked in 0.1C allow at least one minute to equilibrate
Calibrate annually by checking Suspend the thermometer away from the
sides and bottom of container
against a NIST certified thermometer Record readings to nearest 0.5C
Corrections can be made up to 4C May be measured insitu with probe if
verified against NIST traceable
thermometer
Temperature Conversion
F = (9/5 C) + 32
C = 5/9 (F - 32)
QC Acceptance
ICAL
*QC Frequency
Control Charts
LFM / LFMD
ICAL / CCV
Batch Size
Parameter
Method
Have
DOC
MDL
LRB
LFB
Dup
thermometers
verified annually
by an NIST
thermometer
NISTthemrometer
SM2550B 2000
X,verifyagainst
Corrective Action
Temperature
Annually
X QC Frequency
1.1 This method is for the measurement of dissolved oxygen (DO) in surface and ground water,
municipal and industrial wastewater, and for use in Biochemical Oxygen Demand5 (BOD5) and
carbonaceous Biochemical Oxygen Demand5 (cBOD5) determination.
1.2 The method may be used as a replacement for the modified Winkler and membrane electrode
procedures for the measurement of DO in wastewater treatment processes such as aeration and
biological nutrient basins, effluent outfalls, receiving water, and water for BOD5 and cBOD5
determination.
1.3 The method is for use in the United States Environmental Protection Agencys (EPAs) survey and
monitoring programs for the measurement of DO and for the determination of BOD5 and cBOD5
under the Clean Water Act.
1.5 This method is restricted to luminescence probe technologies where calibration is performed by
single-point water-saturated air (100% saturation).
2. Summary of Method
2.1 This luminescence-based sensor procedure measures the light emission characteristics from a
luminescence-based reaction that takes place at the sensor-water interface. A light emitting diode
(LED) provides incident light required to excite the luminophore substrate. In the presence of
dissolved oxygen the reaction is suppressed. The resulting dynamic lifetime of the excited
luminophore is evaluated and equated to DO concentration.
3. Interferences
3.1 There are no known agents that interfere with luminescence DO detection and quantification with
this method.
4. Safety
4.1 This method does not address all safety issues associated with its use. The laboratory is
responsible for maintaining a safe work environment and a current awareness file of OSHA
regulations regarding the safe handling of any chemicals specified in this method. A reference file
of material safety data sheets (MSDSs) should be available to all personnel involved in these
analyses. Additional information on laboratory safety can be found in References 17.5-17.6.
5.1 BOD bottle 300-mL with stoppers and plastic caps (Hach # 62016 and 241906, or equivalent).
5.8 Meter and LBOD Probe (Hach Catalog Number 8508500) for DO measurement in BOD bottles, or
equivalent as defined in Section 1.5 of this method)
5.9 Meter and LDO Probe (Hach Catalog Number 8505200 or 8506300) for DO measurement in open
containers and water bodies, or equivalent as defined in Section 1.5 of this method)
5.10 Dispenser Cap, for Nitrification Inhibitor (Hach Catalog Number 4590)
o
5.11 Temperature controlled environment for BOD bottle incubation, 20 1 C.
6. REAGENTS
6.1 Phosphate Buffer Solution - 1.7 g AR grade Ammonium Chloride, 8.5 g Potassium Phosphate
Monobasic, 17.7 g Sodium Phosphate Dibasic, 21.7 g Potassium Phosphate Dibasic diluted to
1000 mL with deionized water, APHA, pH 7.2, (Hach Catalog Number 43149, or equivalent)
6.2 Calcium Chloride Solution - 27.5 g AR grade Calcium Chloride diluted to 1000 mL with deionized
water, APHA, for BOD (Hach Catalog Number 42849, or equivalent)
6.3 Ferric Chloride Solution - 0.25 g Ferric Chloride diluted to 1000 mL with deionized water, APHA, for
BOD (Hach Catalog Number 42953, or equivalent)
6.4 Glucose-glutamic Acid - Standard Solution, Voluette Ampoule, 300-mg/L, 150 mg glucose and
150 mg glutamic Acid to 1000 mL in deionized water, 10 mL (Hach Catalog Number 1486510, or
equivalent) or, ezGGA Ampoules, 450 mg/L, 225 mg/L Glucose and 225 mg/L Glutamic Acid,
(Hach Catalog Number 25144-20, or equivalent)
6.5 Magnesium Sulfate Solution - 22.5 g Magnesium Sulfate diluted to 1000 mL with deionized water,
APHA, (Hach Catalog Number 43094, or equivalent)
6.7 Potassium Iodide Solution (100 g AR grade Potassium Iodide diluted to 1000 mL with deionized
water) (Hach Catalog Number 1228949, or equivalent)
6.8 Sodium Thiosulfate Solution 0.025 N (Hach Catalog Number 35253, or equivalent)
6.10 Sodium Hydroxide Pellets - ACS (Hach Catalog Number 18734, or equivalent)
6.11 Starch Indicator - 5.5 g AR grade Starch, and 1.25 g AR grade Salicylic Acid diluted to 1000 mL
with deionized water (Hach Catalog Number 34932, or equivalent)
6.12 Sulfuric Acid Solution - 0.020 N (Hach Catalog Number 104532, or equivalent)
6.13 Sulfuric Acid Solution - 1.000 N (Hach Catalog Number 127053, or equivalent)
Note: The Phosphate Buffer Solution should be refrigerated to decrease the rate of biological growth.
7.1.1 Add approximately 1 inch (2.54 cm of reagent water to a clean BOD bottle and stopper.
7.1.3 Allow for the BOD bottle and its contents to equilibrate to room temperature. Room
o
temperature should be approximately 20 3 C.
7.1.4 The stopper may now be removed from the BOD bottle and the LBOD probe inserted for
calibration purposes.
7.1.5 The luminescence technology for measuring dissolved oxygen is a superior technique from
that of Winkler titration and membrane potentiometric measurement and has no interferences
associated with the oxygen detection process. Therefore, for calibration and measurement
purposes, do not adjust the calibration luminescence measurement to that of Winker or
membrane measurement readings.
Note: Section 7.1 is a suggested procedure for the preparation of water-saturated air. Other procedures
for the preparation of water-saturated air may be used that are equally effective.
7.2 Calibration Verification, Initial Precision and Recovery, and On-going Precision and Recovery
7.2.1 Add approximately 1500 mL of organic-free water or BOD dilution water to a 2-L beaker or
PET bottle.
7.2.2 Allow the water to equilibrate to room temperature. Room temperature should be
o
approximately 20 3 C.
7.2.3 With a steady gentle stream of filtered air ( 10 40 mL per minute), aerate the water for a
minimum of 30 minutes. Alternatively, vigorously shake the reagent water or BOD dilution
water for several minutes.
7.2.4 At the completion of aeration, let water re-equilibrate to room temperature (20 3 C) for 30
o
minutes and note the barometric pressure of the laboratory during preparation. The
barometric temperature reading is used in the calculation and determination of the
theoretical DO concentration for the preparation of air-saturated water.
7.2.5 Transfer the aerated water to a BOD bottle until overflowing and stopper.
7.2.6 Calculate the theoretical dissolved oxygen concentration using a dissolved oxygen table
such as Hitchman referenced in Section 17.2 of this method.
Note: Section 7.2 is a suggested procedure for the preparation of air-saturated water. Other procedures
for the preparation of air-saturated water may be used that are equally effective.
8.4 See Title 40 of the Code of Federal Regulations Part 136.3, Table II for further information
regarding required sample collection containers, preservation techniques and holding times for
collection of water for measurement of DO and for the determination of BOD5 and cBOD5.
9. Quality Control
9.1 It is recommended that each laboratory that uses this method be required to operate a formal
quality assurance program (Reference 17.1). The minimum requirements of this program consist of
an initial demonstration of laboratory capability and ongoing analyses of laboratory prepared water
standards as a test of continued performance to assess accuracy and precision. Laboratory
performance is compared to established performance criteria to determine if the results of analyses
meet the performance characteristics of the method.
9.1.1 The analyst shall make an initial demonstration of the ability to generate acceptable accuracy
and precision with this method. This ability is established as described in Section 9.2.
9.1.2 The laboratory shall, on an ongoing basis, demonstrate through calibration verification and
analysis of the ongoing precision and recovery sample that the analysis system is in control.
These procedures are described in Sections 9.3 and 9.4, respectively.
9.1.3 Accompanying QC for the determination of DO is required per analytical batch. An analytical
batch is a set of samples processed during a contiguous 8-hour period, not to exceed 20
samples. Each analytical batch should be accompanied by a calibration verification and
ongoing precision and recovery sample, resulting in a minimum of three analyses (1 CV, 1
sample, and 1 OPR). Perform additional CV and OPR for each batch that exceeds 20
samples.
9.2.1 Initial precision and recovery (IPR) - To establish the ability to generate acceptable precision
and accuracy for the measurement of DO in water, the analyst shall perform the following
operations:
9.2.1.1 Prepare and measure four samples of the IPR standard (Section 7.2) according to
the procedure beginning in Section 11.
9.2.1.2 Using the results of the set of four analyses, compute the average percent recovery
(X) and the standard deviation of the percent recovery (s) for DO. Use the following
equation for calculation of the standard deviation of the percent recovery:
( x ) 2
s=
x 2
n
n 1
where:
n = Number of samples
x = Concentration in each sample
s = standard deviation of the percent recovery
9.2.1.3 Compare s and X with the corresponding limits for initial precision and recovery in
Section 18, Table 4. If s and X meet the acceptance criteria, system performance is
acceptable and analysis of samples may begin. If, however, s exceeds the precision
limit or X falls outside the range for recovery, system performance is unacceptable. In
this event correct the problem, and repeat the test.
9.3.1 Upon air calibration, prepare a calibration verification standard (Section 7.2) with each
analytical batch of 20 samples or less in an 8 hour period. Analyze according to the
procedure beginning in Section 11 and compare the recovery results to those in Section 18,
Table 4.
9.4.1 To demonstrate that the analysis system is in control, and acceptable precision and
accuracy is being maintained with each analytical batch, the analyst shall perform the
following operations
9.4.2 Prepare a precision and recovery standard (Section 7.2) with each analytical batch according
to the procedure beginning in Section 11.
9.4.3 Initially, and at the end of each analytical batch of samples, analyze a precision and recovery
standard and compare the concentration recovery with the limits for ongoing precision and
recovery in Tables 4 and 5. If the recovery is in the range specified, measurement process is
in control and analysis of samples may proceed. If, however, the recovery is not in the
specified range, the analytical process is not in control. In this event, correct the problem,
recalibrate and verify the calibration and reanalyze analytical batch, repeating the ongoing
precision and recovery test.
9.4.4 The laboratory should add results that pass the specification in Tables 4 and 5 to IPR and
previous OPR data and update QC charts to form a graphic representation of continued
laboratory performance. The laboratory should also develop a statement of laboratory data
quality for each analyte by calculating the average percent recovery (R) and the standard
deviation of the percent recovery (sr). Express the accuracy as a recovery interval from R 2sr
to R + 2sr. For example, if R = 95% and sr = 5%, the accuracy is 85% to 105%.
9.5 Depending upon specific program requirements, field replicates may be required to assess the
precision and accuracy of the sampling and sample transporting techniques.
9.6.1 Many factors can influence the BOD analysis (toxicity from sample matrix, contaminated
dilution water, poor quality seed, etc. In order to insure sufficient seeding in the BOD test, a
glucose-glutamic acid check is performed in parallel with each days BOD5 and cBOD5 test
samples. Well prepared dilution water and an active seed will produce a BOD5 of 198 30
mg/L BOD (Reference 17.7).
9.6.1.1 Prepare in triplicate a 300-mL BOD bottle with 3.0 mL of the 300 mg/L Standard
Solution of GGA (Section 6.4) or 1 ampoule (2.0 mL) of ezGGA (Section 6.4) with
each day of samples prepared for BOD determination.
9.6.1.2 When using ezGGA ampoules, place the ezGGA ampoule in the ampoule
breaker (provided with ezGGA ampoules) and rinse the assembly with reagent
water. Hold the ampoule and breaker over the rim of the BOD bottle, break and
allow the ampoule to fall into the BOD bottle. Leave the ampoule in the BOD
bottle during the incubation period and reading of DO.
3.0 mL x 0.300 mg/mL GGA x 1000 mL/L = 3.0 mg/L GGA per bottle
300 mL final volume
ezGGA Ampoule
1 ampoule (2.0 mL) x 0.450 mg/mL GGA x 1000 mL/L = 3.0 mg/L GGA per bottle
300 mL final volume
9.6.1.2 Add seed at three different volumes (typically 4 mL, 6 mL, and 8 mL,) to the GGA
bottles. Other volumes may be required, depending on the strength of the seed
being used.
9.6.1.3 Bring to volume with dilution water and analyze as described in Sections 12.9
and 12.10.
Note: GGA BOD5 recovery results outside of 198 30 mg/L should be investigated as to causation. If
toxicity of dilution water has been ruled out as a probable cause for low recovery, it is likely that the
seed is of low activity or poor quality. Either increase the seed amount or use a seed of higher
quality. High GGA recoveries are generally due to incorrect amount of GGA Standard Solution.
10.1 Because of the possible diversity of future LDO instrument hardware and, no detailed operating
conditions are provided. The analyst is advised to follow the recommended operating conditions
provided by the manufacturer. It is the responsibility of the analyst to verify that the instrument
configuration and operating conditions satisfy the analytical requirements of this method and to
maintain quality control data verifying instrument performance and analytical results.
10.3 Calibration verification (Section 7.2) is performed with air-saturated water prior to any DO sample
measurements to the method specifications in Section 14.
11. Procedure for Measuring DO in Grab Samples, Outfalls, and Open Water
Bodies
11.1.1 Follow the instrument manufacturers instructions for instrument setup (Hach Document
DOC022.53.80021 for Hach LDO IntelliCal Rugged and Standard Probes, Hach
Document Number DOC022.53.80116 for LBOD probes, and Hach Catalog Number
5790018 for Hach LDO process probes.
Note: Manufacturers instructions are only for instrument set and use. These instructions do not preclude
the calibration and performance requirements of this method.
11.2 Measurement of DO
11.2.1 For samples in an open vessel, container, or water body, place the LDO probe into the
water sample to be measured and stir gently with probe or add a stir bar. Do not put the
probe on the bottom or sides of the container. Stir the sample at a moderate rate or put the
probe in flowing conditions. Read sample. The display will show "Stabilizing" and a
progress bar as the probe stabilizes in the sample. The display will show the lock icon
when the reading stabilizes.
11.3.1 For BOD5 and CBOD5 prepared samples, insert the LBOD probe into the BOD bottle for
DO determination. Insure that there are no air bubbles that may have collected around the
probe or sensor. Turn on the stir paddle and read sample. The display will show
"Stabilizing" and a progress bar as the probe stabilizes in the sample. The display will show
the lock icon when the reading stabilizes.
12. Procedure for the Preparation and Determination of BOD5 and cBOD5
Samples
12.1 The BOD test is a 5-day test. Follow all steps carefully to make sure that the test does not have to
be repeated.
12.2 The dilution water for this test must be fully air-saturated immediately before use and determined to
not have an oxygen demand or any toxins. When incubated for 5 days at 20 1 C, the dissolved
oxygen concentration in the dilution water must not change by more than 0.2 mg/L. Air-saturation
is a function of water temperature and laboratory barometric pressure. Use an oxygen saturation
table such as in Section 17.2 of this method to insure full air-saturation of dilution water.
12.3.1 The distilled water must be prepared very carefully to make sure that no source of oxygen
demand or toxins are added. The water that is used to prepare the dilution water must be
of very high quality. The water must not have any organic compounds or any toxic
compounds such as chlorine, copper, and mercury at a concentration level that would
interfere would the BOD seed and inhibit microbiological growth of organisms.
12.3.2 For best results, use an alkaline permanganate distillation for preparing dilution water.
Resin in ionization cartridges will occasionally release organic materials that have an
oxygen demand.
o
12.3.3 Store the distilled water in clean jugs at a temperature of 20 3 C. Fill the containers to
about full and shake the jugs to saturate the water with air. Alternatively, saturate the
water with air as described in Section 7.2.1. A small aquarium pump or air compressor can
be used to saturate the water with air. Insure that the air is filtered and that the air filter
does not grow bacteria.
12.4.1 Using the distilled water prepared above in Section 12.3, select a BOD nutrient buffer pillow
from the BOD nutrient buffer pillows in Table 1.
12.4.2 Add the contents of the BOD Nutrient Buffer Pillow to the distilled water in a jug with ample
headspace. Cap the jug and shake vigorously for one minute to dissolve the nutrients and
to saturate the water with air.
12.4.3 Alternatively, prepare the dilution water by adding 1 mL each of the following solutions per
liter of distilled water prepared in Section 12.4:
Calcium Chloride Solution - 27.5 g AR grade Calcium Chloride to 1000 mL with deionized
water, APHA, for BOD (Hach Catalog Number 42849, or equivalent)
Ferric Chloride Solution - 0.25 g Ferric Chloride to 1000 mL deionized water, APHA, for
BOD (Hach Catalog Number (42953, or equivalent)
12.5.4 Cap the jug and shake vigorously for one minute to dissolve the nutrients and to saturate
the water with air.
Note: Dilution water should be prepared immediately before use unless it can be demonstrated that the
dilution water blank has no DO depletion greater than 0.2 mg/L.
12.6.1 Use raw sewage or other reliable sources for the bacterial seed that will yield 198 30
mg/L BOD with the GGA check sample in Section 9.6. Potential seed sources include
wastewater influent, primary effluent, soil, and domestic sewage.
o
12.6.2 Allow raw sewage to stand undisturbed at 20 3 C for 24 to 36 hours before use.
12.6.3 When seeding samples with raw sewage, always pipette from the upper portion of the
sewage.
12.7.1 Make an estimate of the sample volumes that are necessary for the test. At least 2.0 mg/L
of DO should be consumed during the test and at least 1.0 mg/L of un-depleted DO should
remain in the bottle.
12.7.2 Samples such as raw sewage will have a high BOD. Small sample volumes must be used
because large samples will deplete all of the oxygen in the sample. Samples with a low
BOD must use larger sample volumes to insure that adequate oxygen is depleted to give
accurate results.
12.7.3 Refer to the Minimum Sample Volume in Table 2 to select the minimum sample volume.
For example, if a sewage sample is estimated to contain 300 mg/L BOD, the minimum
sample volume is 2 mL. For sewage effluent with an estimated BOD of 40 mg/L, the
minimum sample volume is 15 mL.
12.7.4 Refer to the Maximum Sample Volume in Table 3 to select the maximum sample volume.
At 1000 in elevation, with an estimated BOD5 of 300 mg/L, the largest sample volume is 8
mL. For a BOD of 40 mg/L, the maximum volume of sample is 60 mL.
12.8.2 For sample matrices that contain residual chlorine, de-chlorinate with a solution of
Sodium Thiosulfate (Na2S2O3).
12.8.2.1 Measure 100 mL of sample into a 250 mL Erlenmeyer flask. Using a 10-mL
serological pipette and pipette filler, add 10 mL of 0.020 N Sulfuric Acid
Standard Solution and 10 mL of Potassium Iodide Solution, 100-g/L, to the
flask.
12.8.2.2 Add three full droppers of Starch Indicator Solution and swirl to mix.
12.8.2.3 Fill a 25-mL burette with 0.025 N Sodium Thiosulfate Standard Solution and
titrate the sample from dark blue to colorless.
12.8.2.4 Calculate the amount of 0.025 N Sodium Thiosulfate Solution to add to the
sample:
12.8.2.5 Add the required amount of 0.025 N Sodium Sulfate Standard Solution to the
sample. Mix thoroughly and wait 10 to 20 minutes before performing the BOD
test.
o
Note: Samples should be brought to a temperature of 20 3 C. prior to making dilutions.
12.9.1 Select the sample volume as described in Section 12.7. Select a minimum of three
different volumes for each sample.
12.9.2 Stir the sample gently with a pipette. Use the pipette to add the determined sample
volumes to the BOD bottles.
12.9.3 Add the appropriate seed to the individual BOD bottles as described in Section 12.6.
12.9.3.1 Separately, with each batch of BOD samples, prepare a seed sample with
dilution water. Measure the BOD of the seed for subtraction from the sample
BOD.
12.9.3.2 A seed that has a BOD5 of 200 mg/L (a typical range for domestic sewage) will
typically deplete at least 0.6 mg/L DO when added at a rate of 3 mL/L of
dilution water.
12.9.4 If the test is for cBOD5, add two potions of Nitrification Inhibitor (approximately 0.16 g) to
each bottle. The oxidation of nitrogen-based compounds will be prevented.
12.9.5 Fill each bottle to just below the lip with dilution water.
12.9.5.1 Allow the dilution water to flow down the sides of the bottle to prevent air
bottles from becoming entrapped in the bottle.
12.9.6 Fill an additional BOD bottle with only dilution water. This will be the dilution water blank.
12.9.7 Stopper the bottles carefully to prevent air bubbles from becoming entrapped.
12.9.7.1 Tightly twist the stopper and invert the bottles several times to mix.
Note: The sample preparation procedures in Section 12.9 are designed for a BOD sample analysis
volume of 300 mL. A 60-mL sample preparation volume may also be used.
12.10.1 Measure the initial dissolved oxygen concentration in each bottle with the LBOD probe
within 30 minutes of sample preparation.
12.10.2 After the initial DO measurement, stopper the bottles carefully to prevent air bubbles from
becoming entrapped.
12.10.2.1 Add dilution water to the lip of each BOD bottle to make a water seal.
o
12.10.3 Place a plastic cap over the lip of each bottle and incubate at 20 1 C for five days.
12.10.4 After 5 days, measure the remaining dissolved oxygen concentration in each bottle with
the LBOD probe.
12.10.4.2 Discard results of samples where the DO is depleted below 1.0 mg/L.
13.1 When Dilution Water Not Seed (generally influent and primary treated influent to treatment)
13.3.1 Averaged results from different dilutions are acceptable if more than one sample dilution
meets all of the following criteria:
13.3.1.2 The final DO value is at least 2 mg/L lower than the initial prepared sample DO
14. Method Performance for Dissolved Oxygen in Reference Water and GGA
BOD5 Recovery
15.1 There are no standards or reagents used in this method that when properly disposed of, pose any
threat to the environment.
16.1 It is the laboratorys responsibility to comply with all federal, state, and local regulations governing
waste management, particularly the hazardous waste identification rules and land disposal
restrictions, and to protect air, water, and land by minimizing and control all releases from fume
hoods and bench operations. Compliance with all sewage discharge permits and regulations is
also required.
16.2 For further information on waste management, consult The Waste Management manual for
Laboratory Personnel, and Less is Better: Laboratory Chemical Management for Waste
Reduction, both available from the American Societys Department of Government relations and
th
Science Policy, 1155 16 Street N.W., Washington, D.C. 20036.
17. References
17.1 Handbook of Analytical Quality Control in Water and Wastewater Laboratories, USEPA, EMSL-CI,
Cincinnati, OH 45268, EPA-600-4-79-019, March 1979.
17.2 Hitchmen, M.L. (1978) Chemical analysis. Vol. 49. Measurement of Dissolved Oxygen. Wiley and
sons, New York.
17.3 Title 40, Code of Federal Regulations (40 CFR), Part 136.
17.4 Protocol for EPA Approval of New Methods for Organic and Inorganic Analytes in Wastewater and
Drinking Water (EPA-821-B-98-003, March 1999).
17.5 OSHA Safety and Health Standards, General Industry, (29 CFR 1910), Occupational Safety and
Health Administration, OSHA 2206 (Revised, January 1976)
17.6 Safety in Academic Chemistry Laboratories, American Chemical Society, Committee on Chemical
rd
Safety, 3 Edition, 1979.
th
17.7 Standard Methods for the Examination of Water and Wastewater, 20 Edition; American Public
health Association: 1015 Fifteenth Street, NW, Washington, D.C. 2005, 1998; Method 5210B.
18. Tables
Note: Hach BOD Nutrient Pillows are formulated with the same reagents adjusted for volume preparation
as in Sections 6.1 through 6.3.
The definitions and purposes are specified to this method but have been conformed to common
usage as much as possible.
19.1.1 Symbols
o
C degrees Celsius
Test preparation
Instrument-specific information
This procedure is applicable to the meters and probes that are shown in Table 1.
Procedures for other meters and probes can be different.
Table 1 Instrument-specific information
Meter Probe
HQ30d portable single input, multi-parameter IntelliCAL LDO101 LDO
HQ40d portable dual input, multi-parameter
HQ430d benchtop single input, multi-parameter
HQ440d benchtop dual input, multi-parameter
Before starting
Refer to the meter documentation for meter settings and operation. Refer to probe documentation for probe preparation,
maintenance and storage information.
Prepare the probe before initial use. Refer to probe documentation.
When an IntelliCAL probe is connected to an HQd meter, the meter automatically identifies the measurement parameter
and is prepared for use.
The IntelliCAL LDO101 probes automatically adjust for barometric pressure, elevation and temperature.
Do not touch the probe cap with a hand, fingers or any surface that can scratch the cap.
Prepare the probe before initial use. Refer to probe documentation.
Condition the probe before use. To condition the probe, put the probe in 100 mL of tap water for 30 minutes before use.
For probes that are continuously in aqueous solutions, condition the sensor cap for 72 hours.
Calibrate the probe before initial use. Refer to Calibration on page 3.
Salinity affects the concentration of dissolved oxygen in the sample. To correct for salinity effects, refer to the probe
documentation.
Analyze the samples immediately. The samples cannot be preserved for later analysis.
Stir the samples at a slow and constant rate to prevent the formation of a vortex.
Air bubbles under the sensor tip can cause slow response or measurement errors. To remove the bubbles, carefully shake
the probe.
For rugged electrodes, it may be necessary to remove the shroud before measurement and calibration.
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used. Use the recommended personal protective
equipment.
Dispose of reacted solutions according to local, state and federal regulations. Refer to the Safety Data Sheets for disposal
information for unused reagents. Refer to the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.
Sample collection
The main consideration with sample collection is to prevent contamination of the sample
with atmospheric oxygen.
Analyze the samples immediately. The samples cannot be preserved for later
analysis.
Analyze the samples at the collection site if possible.
Do not introduce air into the sample.
Test procedure
1. Rinse the probe with 2. Laboratory test: Put the 3. Push Read. A progress 4. When the value is stable,
deionized water. Dry the probe in a beaker that bar is shown. When the store or record the mV value
probe with a lint-free cloth. contains the solution. Do not measurement is stable, the and the temperature value.
let the probe touch the stir lock icon is shown.
bar, bottom or sides of the
container. Remove the air
bubbles from under the
probe tip. Stir the sample at
a slow to moderate rate.
Field test: Put the probe in
the sample. Move the probe
up and down to remove
bubbles from the probe tip.
1. Add a small amount of 2. Insert a stopper and 3. If the probe cap is wet, 4. Remove the stopper. Put
water (approximately 1 cm) shake the bottle vigorously carefully dry the probe cap the probe in the bottle.
to the bottom of narrow- for several minutes. with a soft cloth.
neck bottle, such as a BOD
bottle.
5. Push Calibrate. 6. Push Read. A progress 7. Push Done. The 8. Push Store to accept the
bar is shown. When the calibration summary shows. calibration.
measurement is stable, the The slope value is the
lock icon is shown. comparison between the
latest calibration and the
factory calibration shown as
a percentage.
1. Rinse the probe with deionized water. Blot dry with a lint-free cloth.
2. If harsh contaminants are attached to the probe, polish the probe tip with a soft cloth
or cotton swab to remove the contaminants.
3. Soak the probe in deionized water for 1 minute.
Summary of method
The oxygen sensor is a clear, oxygen impermeable hard substrate. An oxygen-sensitive
luminescent dye and a scattering agent is on the substrate. A final overlay of dark
pigment is added to keep stray light out of the measurement cell. The luminescent dye
gives a red light when exposed to blue light. The scattering agent distributes the light in
the sensor matrix and contributes to the opacity of the sensor. Pulses from a red LED are
used as an internal reference. The duration of the luminescence is proportional to the
concentration of dissolved oxygen in the sample.
Accessories
181
Section 5 TDEC - Fleming Training Center
OUTLINE
Introduction to BOD
What is BOD?
3 4
5 6
Source: http://en.wikipedia.org/wiki/Biochemical_oxygen_demand
7 8
Source: http://en.wikipedia.org/wiki/Biochemical_oxygen_demand Source: http://en.wikipedia.org/wiki/Biochemical_oxygen_demand
9 10
+ Food + Oxygen (O2) Carbon Dioxide (CO2) + Ammonia + Oxygen (O2) Nitrate(NO3-)
(Bacteria) or Nitrite (NO2 )
-
(Bacteria)
Food - Organic material (carbon), exerts Food Reduced forms of nitrogen, exert nitrogenous
carbonaceous oxygen demand (cBOD) oxygen demand
11 12
13 14
15 16
oxygen concentration of
Influent Effluent the receiving water body.
BOD may be regulated by
WWTP
permit requirements.
300 mg/L 30 mg/L
90% Removal
17 18
Duplicates
19 20
21 22
Running Standards
23 24
25 26
27 28
Dilution water blanks must meet quality control limits, Tubing constructed of oxygen-demand leaching material
< 0.2 mg/L DO (preferably <0.1 mg/L) (consider medical grades)
Otherwise discard and prepare fresh solution
No seed or nitrification inhibitor is added for dilution
water blank
Run one nitrification inhibitor blank
31 32
33 34
35 36
37 38
39 40
41 42
DETERMINING RANGE AND SAMPLE VOLUME DETERMINING RANGE AND SAMPLE VOLUME
Fill bottles to top without adding bubbles
If > 67% (200 mL) sample after dilution, add nutrients, First Last Dilution
Dilution 1mg/L
TDEC - Fleming Training Center
45 46
DETERMINING RANGE AND SAMPLE VOLUME DETERMINING RANGE AND SAMPLE VOLUME
Selection of sample volumes used in the test depends on
two factors:
TDEC - Fleming Training Center
Type of sample
Elevation
47 48
DETERMINING RANGE AND SAMPLE VOLUME DETERMINING RANGE AND SAMPLE VOLUME
If our sample is approximately 300mg/L BOD, what
should the minimum and maximum sample volumes be?
DETERMINING RANGE AND SAMPLE VOLUME DETERMINING RANGE AND SAMPLE VOLUME
If our sample is approximately 300mg/L BOD, what
should the minimum and maximum sample volumes be?
TDEC - Fleming Training Center
51 52
DETERMINING RANGE AND SAMPLE VOLUME DETERMINING RANGE AND SAMPLE VOLUME
TDEC - Fleming Training Center
53 54
DETERMINING RANGE AND SAMPLE VOLUME DETERMINING RANGE AND SAMPLE VOLUME
Nitrification:
Can add nitrification Nitrosomonas + NH3 + O2 NO2-
55 56
DETERMINING RANGE AND SAMPLE VOLUME DETERMINING RANGE AND SAMPLE VOLUME
Nitrification inhibitor Fill bottles past the neck with dilution water and invert
Prevents Nitrosomonas from oxidizing ammonia to nitrite, to mix (no air bubbles).
preventing nitrogenous oxygen demand in the sample
TDEC - Fleming Training Center
0 mL 2 mL 4 mL 6 mL 8 mL
57 58
59 60
CALIBRATION CALIBRATION
Winkler titration - best; most accurate Probe: Water-saturated air (most common)
Relies on chemistry Air-calibration chamber calibrate at sample temperature.
61 62
sension156
TDEC - Fleming Training Center
0.00
DO mg/L
20.0oC
2 mL
63 64
mg/L
20.0oC
Read all 5
bottles
65 66
2 mL
69 70
P
Read all 5
bottles
71 72
75 76
77 78
79 80
81 82
Acceptance
ICAL / CCV
Corrective
Batch Size
Control
Method
Method
Charts
Action
Blank
MDL
DOC
LFB
Dup
daily
X X X X X X X 20
83 84
Dup Batch Size Dilution water quality check (nutrient, mineral, buffer)
QC Frequency
must not be more than 0.2 mg/l (0.1 is preferred)
ICAL/CCV
Seed control of three dilutions. Smallest to give at least
2.0 mg/l depletion and the largest to at least 1.0 mg/l
residual.
85 86
87 88
89 90
93
Initial DO (D1)
Oxygen Demand
(D1 D2)
(-) Depletion due to seed
(B1 B2)f
Dilution Factor (1/P)
Vol/300mL= P
Avg. mg/L
For unseeded samples: (D1 - D2) (D1 - D2) - (B1 - B2) f
BOD5, mg/L = For seeded samples: BOD5, mg/L =
P P
Add 4mL of settled primary clarifier effluent or settled raw sewage for each liter of seeded dilution water. Manufactured seed material is also available.
Seed Control
mL. used Seed Dilution Water
OR % Concentration (P) (Oxygen Demand Calculation)
EXAMPLE WORKSHEET
207
Section 6 TDEC - Fleming Training Center
pH
One of the most important and frequently used tests in
water chemistry
A measure of the intensity of the acidic or alkaline
character of a solution
Logarithmic scale of ionic activity
0 to 14 s.u.
pH pH values cannot be averaged
pH Measurement pH Theory
pH is typically measured Acid
with a meter and probe increases the hydrogen ion (H+) concentration in a solution
This is an Base
electrochemical method increases the hydroxide ion (OH-) concentration in a solution
of analysis
pH Theory pH Scale
pH is defined as the negative log of the molar hydrogen
ion concentration in aqueous solution
Acid Base
0 7 14
Neutral
Vinegar Ammonia
pH 3 pH 11.5
208 pH
TDEC - Fleming Training Center Section 6
10X
0 7 14
4 5 6
100X
Internal Filling
Solution
Reference
Electrolyte
Salt Bridge
9 TDEC - Fleming Training Center Junction 10 TDEC - Fleming Training Center
pH 7 Solution pH 7 Solution
H+ conc. the Hydrogen ion H+ conc. the same
same both inside concentration fixed both inside and 0.0mV
and outside glass at pH 7 outside glass bulb
bulb
*No potential
*No potential develops
H+ H+
develops H+ H+ H+ H+
H+ H+
H+ H+ + H+
H H+
H+ H+ H+
pH 209
Section 6 TDEC - Fleming Training Center
pH 4 Solution pH 4 Solution
H+ conc. 1000x Hydrogen ion H+ conc. 1000x
greater outside concentration fixed greater outside
glass bulb glass bulb 180mV
at pH 7
*Potential develops *Potential develops
1000H+ H+ 1000H+
H+ H+ H+ H+
1000H+ 1000H+
H+ + H+
1000H+ H
1000H + 1000H +
pH 10 Solution pH 10 Solution
H+ conc. 1000x Hydrogen ion H+ conc. 1000x
greater inside glass concentration fixed greater inside glass
bulb bulb -180mV
at pH 7
*Potential develops *Potential develops
H+ H+
H+ H+ H+ H+
H+ + H+ H+
H
Calibration Calibration
A calibration curve allows The optimal slope for pH is -58 3 mV/decade.
the meter to convert a
measured millivolt
potential into a pH
reading.
mV
pH
210 pH
TDEC - Fleming Training Center Section 6
Calibration Calibration
+180 -180mV difference measured between pH4 and pH7
pH4 to pH7 (3 pH units) is 1000x concentration change
Decade = 10-fold concentration change
mV 0 = 1pH unit
-180/3 = -60 -58mV/decade
-180
4 7 10
pH
The mV values from the previous demos (4, 7, and 10 buffers) are plotted on a
graph of mV versus pH. A line is drawn between the points and the slope
determined. Since slope is rise over run, it will be in units of mV per pH unit, or
mV per decade (since the difference between pH units is 10X).
19 TDEC - Fleming Training Center 20 TDEC - Fleming Training Center
pH 211
Section 6 TDEC - Fleming Training Center
Measurement Troubleshooting
Place probe into sample, stir, and wait for readings to mV reading in pH 7 buffer
stabilize Should read 0 30 mV in pH 7 buffer
Rinse and blot dry between measurements Response time
Storage between measurements May require cleaning if slow in buffered solution
As suggested by probe manufacturer Slope
pH electrode storage solution (best) Optimal slope is -58 3 mV/decade
212 pH
TDEC - Fleming Training Center Section 6
Cleaning
Slow response may indicate need for cleaning
Alternate soaking in dilute hydrochloric acid and dilute
sodium hydroxide
Rinse with deionized water
Condition in pH 7 buffer before use
Filler hole button was not removed
pH 213
Section 6 Fleming Training Center TDEC - Fleming Training Center
pH Calibration Record
214
Month___________ pH pH Meter _______________
Section 7
Solids
215
Section 7 TDEC - Fleming Training Center
Serve as refuge for harmful bacteria Control of Biological and Physical treatment
process
Poor data = Poor decisions
Unsightly appearance
216 Solids
TDEC - Fleming Training Center Section 7
Solids 217
Section 7 TDEC - Fleming Training Center
218 Solids
TDEC - Fleming Training Center Section 7
Solids 219
Section 7 TDEC - Fleming Training Center
220 Solids
TDEC - Fleming Training Center Section 7
Solids 221
Section 7 TDEC - Fleming Training Center
222 Solids
TDEC - Fleming Training Center Section 7
Solids 223
Section 7 TDEC - Fleming Training Center
TSS Exercise
24. Stir sample with magnetic stirrer at a speed to shear larger particles, __________, to obtain
a more uniform (_________________) particle size.
25. ____________ force may separate particles by ______________, resulting in
_____________ when point of sample withdrawal is varied.
26. While _________, pipet a measured volume onto the seated glass-fiber filter.
27. For homogeneous samples, pipet from approximate _________ of container but ___ in
______.
28. Choose a point both ________ and ______ between wall and vortex.
29. Wash filter with _____ successive _______ volumes of reagent-grade water, allow
________ drainage between washings, and ________ suction for about ____ after filtration
is ___________.
30. Samples with _____ dissolved solids may require additional __________.
31. Carefully remove filter from filtration apparatus and ___________ to an aluminum weighing
dish as a _______.
32. Dry for _____________ at 103 to 105oC in an oven, cool in a desiccator to balance
temperature, and weight.
33. _______ the cycle of drying, cooling, desiccating, and weight until a constant weight is
obtained or until the weight change is less than ____ of the previous weight or _________
whichever is less.
34. Analyze __________ of all samples in ______________.
35. ___________ determinations should agree within ___ of their ________ weight.
Solids 225
Section 7 TDEC - Fleming Training Center
Blank 100
Effluent 100
Raw 25
Mixed Liquor 5
Dup Raw 25
Blank 100
226 Solids
Section 8
WET Testing
227
Section 8 TDEC - Fleming Training Center
Approaches to Evaluating
Environmental Samples
Test sample for specific chemical substances
dangerous to environment (ex: ammonia,
Whole Effluent Toxicity metals, etc.)
Laboratory knows what to look for in sample
Identification techniques specific to analytes
WET Testing
May overlook a compound harmful to
environment
Approaches to Evaluating
WET Test
Environmental Samples
Treat a living organism with a portion of Whole Effluent Toxicity (WET) refers to the
aggregate toxic effect to aquatic organisms from
sample and see if it exhibits effects: all pollutants contained in a facility's wastewater
Many samples screened quickly (effluent).
Results somewhat dependent on species used in It is one way we implement the Clean Water
tests Act's prohibition of the discharge of toxic
pollutants in toxic amounts.
If toxicity found, no indication of exact cause of WET tests measure wastewater's effects on
toxicity specific test organisms' ability to survive, grow
and reproduce.
Biomonitoring Requirements,
Effluent Sampling
Chronic
Collected as composite over 24 hrs NPDES Permit 3.4
Chilled to 4C during and after sampling The permittee shall conduct a 3-Brood Ceriodaphnia
dubia Survival and Reproduction Test and a 7-day
Hold time 36 hrs from last aliquot collection Fathead Minnow (Pimephales promelas) Larval Survival
until beginning of testing and Growth Test on samples on final effluent
Sample size: 4L or 1gal cubitainer The measured endpoint for toxicity will be the
inhibition concentration causing 25% reduction in
survival, reproduction and growth (IC25) of the test
organisms.
Biomonitoring Requirements,
Acute Bioassay
Acute
NPDES Permit 3.5 Conducted with Pimephelas promelas and
The permittee shall conduct a 48-hour static acute toxicity Ceriodaphnia dubia
test on two test species on samples of final effluent
The test species to be used are Water Fleas (Ceriodaphnia Determine end of pipe conditions
dubia) and Fathead Minnows (Pimephales promelas). Effects in 100% effluent
The measured endpoint for toxicity will be the Last 48-96 hrs
concentration causing 50% lethality (LC50) of the test
organisms. Objective: determine effluent concentration that
The LC50 shall be determined based on a 50% lethality as causes 50% lethality during short term exposure
compared to controls, and as derived from linear
interpolation.
233
Section 9 TDEC - Fleming Training Center
Introduction
Importance in Wastewater
Alkalinity
Treatment
Defined as the measurement of a waters Chemical and biological treatment systems
capacity to neutralize an acid
An acid releases H+ Biological nutrient removal
The alkalinity in the water will absorb H+
Most common ions that add alkalinity are Anaerobic digestion control
OH , CO3 ,HCO3
Ammonia removal by air stripping
234 Alkalinity
TDEC - Fleming Training Center Section 9
Alkalinity Apparatus
Alkalinity caused by OH is called Buret and stand
hydroxyl alkalinity
Alkalinity caused by CO3 is called
carbonate alkalinity Beaker, 250 mL
Alkalinity caused by HCO3 is called
bicarbonate alkalinity Stir plate
The combined effect of all three types is called
total alkalinity
Stir bar
Collect samples in clean plastic or glass Highly colored or turbid samples may mask
bottles the color change at the end point.
Avoid excessive agitation or prolonged Use a pH meter for these samples.
exposure to air
Analyze as soon as possible Chlorine may interfere with indicators.
May be stored for 24 hrs at 4C Add one drop 0.1N sodium thiosulfate to
Warm to room temperature before analysis. eliminate this interference.
Alkalinity 235
Alkalinity, BT, 8221
Section 9 TDEC - Fleming Training Center
Alkalinity DOC316.53.01151
Test preparation
A pH meter is required for NPDES reporting and is recommended for best results.
Substitute six drops of Phenolphthalein Indicator Solution for the Phenolphthalein Indicator Powder Pillow if necessary
Substitute six drops of Bromcresol Green-Methyl Red Indicator Solution for the Bromcresol Green-Methyl Red Powder Pillow
if necessary.
Description Quantity
Funnel, Micro 1
Support Stand 1
See
Table 1
1. Select a sample 2. Use a graduated 3. Transfer the sample 4. Add the contents of
volume from the Sample cylinder or pipet to into a 250-mL Erlenmeyer one Phenolphthalein
volume selection for measure the sample flask. Dilute to about Indicator Powder Pillow.
expected concentration volume. 50-mL with deionized Swirl to mix. (Omit this
table that corresponds to water if necessary. step when using a
the expected alkalinity pH meter.)
concentration in mg/L as
calcium carbonate
(CaCO3).
5. Fill a 25-mL buret to 6. While swirling the 7. Calculate: 8. Add the contents of
the zero mark with 0.020 N flask, titrate the sample mL Titrant multiplier one Bromcresol Green-
Sulfuric Acid standard until the solution color used = mg/L Methyl Red Indicator
solution. changes from pink to phenolphthalein alkalinity Powder Pillow to the
colorless (pH 8.3). as CaCO3. titrated sample. Swirl to
If the solution is colorless mix.
before titrating with sulfuric Do not add indicator if a
acid, the phenolphthalein pH meter is used.
alkalinity is zero. Specific sample
composition may require
titration to a specific pH
(see the
Alkalinity relationship
table).
0500 50 20353 20
4001000 25 20353 40
10002500 10 20353 100
20005000 5 20353 200
The end points in the Alkalinity endpoints table are recommended for determining total alkalinity in
water samples of various compositions and alkalinity concentrations.
Total alkalinity primarily includes hydroxide, carbonate, and bicarbonate alkalinities. The
concentration of these types in a sample may be determined when the phenolphthalein and total
alkalinities are known ( Alkalinity relationship table).
Table 3 Alkalinity relationship
Hydroxide Alkalinity Carbonate Alkalinity Bicarbonate
Row Result of Titration
Equals: Equals: Alkalinity Equals:
1 Phenolphthalein Alkalinity equal to 0 0 0 Total Alkalinity
2 Phenolphthalein Alkalinity equal to Total Total Alkalinity 0 0
Alkalinity
3 0 Phenolphthalein Alkalinity Total Alkalinity minus
Phenolphthalein Alkalinity less than times 2 two times
one-half of Total Alkalinity Phenolphthalein
Alkalinity
4 Phenolphthalein Alkalinity equal to one- 0 Total Alkalinity 0
half of Total Alkalinity
5 2 times 2 times the difference 0
Phenolphthalein Alkalinity greater than Phenolphthalein between Total and
one-half of Total Alkalinity Alkalinity minus Total Phenolphthalein Alkalinity
Alkalinity
2. Does the phenolphthalein alkalinity equal total alkalinity? If yes, use Row 2.
4. Select Row 3, 4 or 5 based on comparing the result of step c (one-half total alkalinity) with the
phenolphthalein alkalinity.
6. Check your results. The sum of the three alkalinity types will equal the total alkalinity.
Example:
A sample has 170 mg/L as CaCO3 phenolphthalein alkalinity and 250 mg/L as CaCO3 total
alkalinity. What is the concentration of hydroxide, carbonate, and bicarbonate alkalinities?
a. The phenolphthalein alkalinity does not equal zero but 170 mg/L.
b. The phenolphthalein alkalinity does not equal total alkalinity (170 mg/L vs. 250 mg/L).
c. One-half of the total alkalinity equals 125 mg/L.
d. Because the phenolphthalein alkalinity of 170 mg/L is greater than one-half the total
alkalinity of 125 mg/L, select Row 5.
The hydroxide alkalinity is equal to:
2 170 = 340
250 170 = 80
Interferences
Chlorine at levels above 3.5 mg/L cause a yellow-brown color upon the addition of the Bromcresol
Green-Methyl Red Indicator Powder Pillow. Residual chlorine interference with the indicator may
be removed by adding a drop of 0.1 N Sodium Thiosulfate Standard Solution* before adding the
indicator.
Highly colored or turbid samples may mask the color change at the end point. Use a pH meter for
these samples, titrating to pH 8.3 for phenolphthalein alkalinity and the appropriate pH (see the
Alkalinity endpoints table) for total alkalinity.
Sampling and storage
Collect samples in plastic or glass bottles. Fill completely and cap tightly. Avoid excessive agitation
and prolonged exposure to air. Samples should be analyzed as soon as possible after collection
but can be stored at least 24 hours by cooling to 4 C (39 F) or below. Warm to room temperature
before analyzing.
Accuracy check
End point confirmation
To accurately determine the phenolphthalein alkalinity end point, mix the contents of one
Phenolphthalein Indicator Powder Pillow and the contents of one pH 8.3 Buffer Powder Pillow
with 50 mL of deionized water in a 250-mL Erlenmeyer flask. The resulting color is the end
point.
To accurately determine the total alkalinity end point, mix the contents of one pH 4.5 Buffer
Powder Pillow and the contents of one Bromcresol Green-Methyl Red Indicator Powder Pillow
with 50 mL of deionized water in a 250-mL Erlenmeyer flask. Titrate to a light pink color
change.
Standard additions method (Sample Spike)
Perform the standard additions method check as follows:
1. Break the top off an Alkalinity Voluette Ampule Standard Solution, 0.500 N.
2. Use the TenSette Pipet* to add 0.1 mL of standard to the sample titrated in step 6 or step 9.
Resume titration back to the same end point. Record the volume of titrant needed.
3. Repeat, using two more additions of 0.1 mL. Titrate to the end point after each addition.
4. The mL of titrant required should increase by 2.5 mL for each 0.1 mL increment of standard
added.
Summary of method
Alkalinity is expressed as P (phenolphthalein) alkalinity or as T (total) alkalinity. Both types are
determined by titration with a Sulfuric Acid Standard Solution to an end point evidenced by the
color change of an indicator solution or determined with a pH meter. The P alkalinity is determined
by titration to a pH of 8.3 and registers the total hydroxide and one half the carbonate present. The
T alkalinity is determined by titration to a pH of 4.5. The total alkalinity includes all carbonate,
bicarbonate and hydroxide alkalinity. Alternatively, total alkalinity end points may be determined by
using a pH meter and titrating to the specific pH required for the sample composition.
Required apparatus
Description Quantity/test Unit Catalog number
Buret Clamp, double 1 each 32800
Buret, Class A, 25-mL 1 each 2636540
Select one or more based on sample volume:
Cylinder, graduated, 5-mL each 50837
Cylinder, graduated, 10-mL each 50838
Cylinder, graduated, 25-mL each 50840
Cylinder, graduated, 50-mL each 50841
Flask, Erlenmeyer, 250-mL 1 each 50546
Pipet, volumetric, Class A, 5-mL, each 1451537
Pipet, volumetric, Class A, 10-mL each 1451537
Pipet Filler, Safety Bulb each 1465100
Ampule Breaker each 2196800
Funnel, Micro 1 each 2584335
Support Stand 1 each 56300
Required standards
Description Unit Catalog number
Alkalinity Standard Solution, Voluette Ampules, 0.500 N, 10-mL 16/pkg 1427810
Buffer Powder Pillows, pH 4.5 25/pkg 89568
Buffer Powder Pillows, pH 8.3 25/pkg 89868
Water, deionized 4L 27256
Optional items
Description Unit Catalog number
Sodium Thiosulfate Standard Solution, 0.1 N 32332
TenSette Pipet, 0.11.0 mL 1970001
Tips for Tensette Pipet 50/pkg 2185696
Alkalinity
Hach Company, 2007. All rights reserved. Printed in the U.S.A. 241
Updated February 2008, Edition 5
Section 9 TDEC - Fleming Training Center
Influent Effluent
mL sample used = mL sample used =
242 Alkalinity
Section 10
Chemical Oxygen Demand
243
Section 10 TDEC - Fleming Training Center
1 2
3 4
5 6
Watertown, TN
7 8
9 10
Titrimetric Colorimetric:
COD, mg/L = (A-B)N x 8000 Direct reading in mg/L from instrument
S
A = mL of titrant for blank If different sample volumes used then,
B = mL of titrant for sample COD, mg/L = (mg O2 in final vol.)(1000)
N = Normality of titrant mL sample
S = mL of sample used
11 12
13 14
Test preparation
DR 5000
DR 2800 LZV646
DR 2700 LZV646
DR/2500
DR/2400 5945700
DR 2800 and DR 2700: Install the light shield in Cell Compartment #2 before performing this test.
DR 2700 and DR/2400: Ultra low range (0.7 to 40.0 mg/L) is not available.
Some of the chemicals and apparatus used in this procedure may be hazardous to the health and safety of the user if
inappropriately handled or accidentally misused. Please read all warnings and associated MSDS sheets.
Run one blank with each set of samples. Run all tests (the samples and the blank) with the same lot of vials. The lot number
appears on the container label. See Blanks for colorimetric determination.
Spilled reagent will affect test accuracy and is hazardous to skin and other materials. Be prepared to wash spills with
running water.
Wear appropriate eye protection and clothing for adequate user protection. If contact occurs, flush the affected area with
running water. Review and follow reagent MSDS safety instructions carefully.
If high chloride samples are being tested, refer to the Alternate reagents section.
Description Quantity
Beaker, 250-mL 1
Blender 1
DRB200 Reactor 1
Opaque shipping container for storage of unused, light-sensitive reagent vials varies
Pipet, TenSette, 0.1 to 1.0 mL, with tips (for 20015,000 mg/L range) 1
1. Homogenize 100 mL 2. For the 200 3. Turn on the DRB200 4. Remove the caps from
of sample for 30 seconds 15,000 mg/L range or to Reactor. Preheat to two COD Digestion
in a blender. For samples improve accuracy and 150 C. Reagent Vials. (Be sure to
containing large amounts reproducibility of the other See the DRB200 User use vials for the
of solids, increase the ranges, pour the Manual for selecting pre- appropriate range.)
homogenization time. homogenized sample into programmed temperature
If the sample does not a 250-mL beaker and applications.
contain suspended solids, gently stir with a magnetic
omit steps 1 and 2. stir plate.
Oxygen
248 Demand, Chemical Chemical Oxygen Demand
Page 2 of 12
Oxygen Demand, Chemical
TDEC - Fleming Training Center Section 10
5. Prepared Sample: 6. Blank Preparation: 7. Cap the vials tightly. 8. Hold the vials by the
Hold one vial at a Hold a second vial at a Rinse them with water and cap over a sink. Invert
45-degree angle. Use a 45-degree angle. Use a wipe with a clean paper gently several times to
clean volumetric pipet to clean volumetric pipet to towel. mix. The sample vials
add 2.00 mL of sample to add 2.00 mL of deionized become very hot during
the vial. water to the vial. mixing.
For the 20015,000 mg/L For the 20015,000 mg/L Insert the vials in the
vials: Use a TenSette vials: Use a TenSette preheated DRB200
Pipet to add 0.20 mL of Pipet to add 0.20 mL of Reactor. Close the
sample to the vial. sample to the vial. protective lid.
9. Heat the vials for two 10. Turn the reactor off. 11. Invert each vial 12. Place the vials into a
hours. Before removing the vials, several times while still rack and cool to room
wait about 20 minutes for warm. temperature.
the vials to cool to 120 C Proceed to Colorimetric
or less. determination.
Colorimetric determination
Stored Programs
Start
1. Select the test. 2. Clean the outside of 3. Insert the blank into 4. ZERO the instrument.
Insert an adapter or light the vials with a damp towel the 16-mm cell holder. The display will show:
shield if required (see followed by a dry one.
0 mg/L COD
Instrument-specific or
information). 0.0 mg/L COD
Refer to the user manual
for orientation.
Read
5. Insert the sample vial 6. READ the results in 7. If using High Range
into the 16-mm cell holder. mg/L COD. Plus COD Digestion
Reagent Vials, multiply the
result by 10.
For most accurate results
with samples near 1500 or
15,000 mg/L COD, repeat
the analysis with a diluted
sample.
2. Zero the instrument in the absorbance mode. Use a vial containing 5 mL of deionized water
and measure the absorbance of the blank. Record the value.
3. Prepare a new blank when the absorbance has changed by about 0.01 absorbance units.
Oxygen
250 Demand, Chemical Chemical Oxygen Demand
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Oxygen Demand, Chemical
TDEC - Fleming Training Center Section 10
Interferences
Chloride is the primary interference when determining COD concentration. Each COD vial
contains mercuric sulfate that will eliminate chloride interference up to the level specified in
Column 1 of the Interfering substances table. Dilute samples with higher chloride concentrations.
Dilute the sample enough to reduce the chloride concentration to the level given in Column 2.
Note: For best results, use the low range and ultra-low range test for samples with high chloride
concentrations (approaching maximum concentration) and low COD concentrations.
If sample dilution will cause the COD concentration to be too low for accurate determination, add
0.50 g of mercuric sulfate (HgSO4) to each COD vial before the sample is added.
The additional mercuric sulfate will raise the maximum chloride concentration allowable to the
level given in Column 3.
Accuracy check
Standard solution method
Note: Refer to the instrument user manual for specific software navigation instructions.
2. Use 2 mL of the 30 mg/L COD solution in place of the sample. Follow the Colorimetric
determination test procedure. The result should be 30 mg/L. Refer to the Standard adjust
instructions in this procedure to adjust the curve with the reading obtained from the standard.
2. Use 2 mL of the 100-mg/L solution in place of the sample. Follow the Colorimetric
determination test procedure. The result should be 100 mg/L. Refer to the Standard adjust
instructions in this procedure to adjust the curve with the reading obtained from the standard
2. Use 2 mL of the 500 mg/L COD solution in place of the sample. Follow the Colorimetric
determination test procedure. The result should be 500 mg/L. Refer to the Standard adjust
instructions in this procedure to adjust the curve with the reading obtained from the standard.
Note: Alternately, use 2 mL of 300 mg/L, 800 or 1000 mg/L COD standards for accuracy check.
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TDEC - Fleming Training Center Section 10
2. Use 0.2 mL of the 10,000 mg/L COD solution in place of the sample. Follow the Colorimetric
determination test procedure. The result should be 10,000 mg/L (after multiplying by 10).
Refer to the Standard adjust instructions in this procedure to adjust the curve with the reading
obtained from the standard.
Standard adjust
1. To adjust the calibration curve using the reading obtained with the standard solution, navigate
to Standard Adjust in the software.
2. Turn on the Standard Adjust feature and accept the displayed concentration. If an alternate
concentration is used, enter the concentration and adjust the curve to that value.
Alternate reagents
Mercury-free COD2 Reagents can provide a mercury-free testing option for non-reporting
purposes. For process control applications, COD2 Reagents will eliminate mercury waste and
save on disposal costs. These reagents are fully compatible with test procedures and calibration
curves programmed into the spectrophotometer. Determine chloride and ammonia for
accurate results.
Important Note: COD2 reagents are not approved for USEPA reporting purposes. Because
COD2 reagents do not contain mercury as a masking agent, they exhibit a positive
interference from chloride. Request a copy of the COD Reagent Vial Information Brochure,
Lit. No. 1356, for more information about specific applications.
Method performance
Precision Sensitivity
Program Instrument Standard 95% Confidence Limits Concentration change
of Distribution per 0.010 Abs change
430 DR 5000 30 mg/L COD (Ultra Low 7783 mg/L COD 3 mg/L COD
(Range, DR 2800 Range); 7783 mg/L COD 3 mg/L COD
3150 mg/L) 80 mg/L COD (Low Range);
DR 2700 800 mg/L COD (High Range); 7783 mg/L COD 3 mg/L COD
8000 mg/L COD (High Range
Plus)
DR/2500 30 mg/L COD (Ultra Low 77.682.4 mg/L COD 3 mg/L COD
DR/2400 Range); 77.682.4 mg/L COD 3 mg/L COD
80 mg/L COD (Low Range);
800 mg/L COD (High Range);
10,000 mg/L COD (High
Range Plus)
431 DR 5000 30 mg/L COD (Ultra Low 28.831.2 mg/L COD 0.5 mg/L COD
(Range, DR 2800 Range); 28.831.2 mg/L COD 0.5 mg/L COD
0.540.0 mg/L) 80 mg/L COD (Low Range);
800 mg/L COD (High Range);
8000 mg/L COD (High Range
Plus)
431 DR/2500 30 mg/L COD (Ultra Low 29.031.0 mg/L COD 0.7 mg/L COD
(Range, Range);
0.740.0 mg/L) 80 mg/L COD (Low Range);
800 mg/L COD (High Range);
10,000 mg/L COD (High
Range Plus)
435 DR 5000 30 mg/L COD (Ultra Low 785815 mg/L COD 23 mg/L COD
(Range, DR 2800 Range); 785815 mg/L COD 23 mg/L COD
201500 mg/L) 80 mg/L COD (Low Range);
DR 2700 800 mg/L COD (High Range); 785815 mg/L COD 23 mg/L COD
8000 mg/L COD (High Range
Plus)
DR/2500 30 mg/L COD (Ultra Low 778822 mg/L COD 20 mg/L COD
DR/2400 Range); 778822 mg/L COD 20 mg/L COD
80 mg/L COD (Low Range);
800 mg/L COD (High Range);
10,000 mg/L COD (High
Range Plus)
435 DR 5000 30 mg/L COD (Ultra Low 78508150 mg/L COD 230 mg/L COD
(Range, DR 2800 Range); 78508150 mg/L COD 230 mg/L COD
20015,000 mg/L) 80 mg/L COD (Low Range);
DR 2700 800 mg/L COD (High 78508150 mg/L COD 230 mg/L COD
Range);8000 mg/L COD (High
Range Plus)
DR/2500 30 mg/L COD (Ultra Low 978010,220 mg/L COD 200 mg/L COD
DR/2400 Range); 978010,220 mg/L COD 200 mg/L COD
80 mg/L COD (Low Range);
800 mg/L COD (High Range);
10,000 mg/L COD (High
Range Plus)
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TDEC - Fleming Training Center Section 10
Summary of method
The results in mg/L COD are defined as the milligrams of O2 consumed per liter of sample under
the conditions of this procedure. The sample is heated for two hours with sulfuric acid and a strong
oxidizing agent, potassium dichromate. Oxidizable organic compounds react, reducing the
dichromate ion (Cr2O72) to green chromic ion (Cr3+).
When the 0.740.0 or the 3150 mg/L colorimetric method is used, the amount of Cr6+ remaining
is determined. When the 201500 mg/L or 20015,000 mg/L colorimetric method is used, the
amount of Cr3+ produced is determined. The COD reagent also contains silver and mercury ions.
Silver is a catalyst, and mercury is used to complex chloride interferences.
Test results are measured at the wavelengths specified in the Range-specific test
wavelengths table.
Required reagents
Description Quantity/Test Unit Catalog number
Select the appropriate COD Digestion Reagent Vial:
Ultra Low Range, 0.7 to 40 mg/L COD 12 vials 25/pkg 2415825
Low Range, 3 to 150 mg/L COD 12 vials 25/pkg 2125825
High Range, 20 to 1500 mg/L COD 12 vials 25/pkg 2125925
High Range Plus, 200 to 15,000 mg/L COD 12 vials 25/pkg 2415925
Water, deionized varies 4L 27256
Alternate reagents1
Description Quantity/Test Unit Catalog number
Select the appropriate COD Digestion Reagent Vial:
COD2, Low Range, 0 to 150 mg/L COD 12 vials 25/pkg 2565025
COD2, High Range, 0 to 1500 mg/L COD 12 vials 25/pkg 2565125
COD2, High Range, 0 to 1500 mg/L COD 12 vials 150/pkg 2565115
COD2, High Range Plus, 0 to 15,000 mg/L COD 12 vials 25/pkg 2834325
COD Digestion Reagent Vials, 3 to 150 mg/L COD 150/pkg 2125815
COD Digestion Reagent Vials, 200 to 1500 mg/L COD 150/pkg 2125915
1 These reagents are not approved for USEPA reporting purposes. Request a copy of the COD Reagent Vial Information Brochure,
Lit. No. 1356, for more information about specific applications.
Required apparatus
Description Quantity/Test Unit Catalog number
Blender, 2-speed, 120 VAC 1 each 2616100
Blender, 2-speed, 240 VAC 1 each 2616102
DRB200 Reactor, 110 V, 15 x 16 mm 1 each LTV082.53.40001
DRB200 Reactor, 220 V, 15 x 16 mm 1 each LTV082.52.40001
Pipet Filler, safety bulb 1 each 1465100
Pipet, Volumetric, Class A, 2.00 mL 1 each 1451536
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Oxygen Demand, Chemical
TDEC - Fleming Training Center Section 10
COD Laboratory
261
Section 11 TDEC - Fleming Training Center
1 2
Solutions
Basically, an even mixture of two or more
chemicals
Solution Chemistry A solution consist of two parts:
Solute
Fleming Training Center TDEC - Fleming Training Center
Solvent
The solute part of the solution is dissolved in the
solvent
The most common solvent is water
3 4
5 6
262 Solutions
TDEC - Fleming Training Center Section 11
7 8
9 10
11 12
Step 2 - Calculate the equivalent weight for Na2CO3 #Equivalent weights = total weight
equivalent weights
Equivalent weight = molecular weight
#Equivalent weights = 105.99 g
# of (+) charges
53 g/equivalent
Equivalent weight = 105.99 #Equivalents = 2
2
Equivalent weight = 53 grams
Solutions 263
Section 11 TDEC - Fleming Training Center
13 14
Normality Normality
OR Step 2c - Calculate the grams required Step 3 - Calculate Normality
Normality = # of equivalents
g required = (#equivalents)(g/equivalents) volume of solvent
Normality = 2 equivalents
g required = (2 equivalent wts)(53g/equivalents) 1 L of water
Normality = 2N
g required = 106 Every 105.99 grams of Na2CO3 in 1L of water
gives a 2N solution
to make a 1N solution, cut the grams in half
15 16
17 18
264 Solutions
TDEC - Fleming Training Center Section 11
3. You are given 100 mL of 2.8N HCl. How many mL of water should be
added to make 0.4N HCl?
Solutions 265
Section 11 TDEC - Fleming Training Center
266 Solutions
TDEC - Fleming Training Center Section 11
10. You are given 8 mL of 15N H2SO4. How much water (in mL) should be
added to make 0.4N H2SO4?
Solutions 267
Section 11 TDEC - Fleming Training Center
Answers
1. 52 mg/L 8. 50 mL 13. 40 g for 1M
2. 18.2% 9. 2.25N 40 g for 1N
3. 600 mL to add 10. 292 mL to add 14. 999.78 g for 5M
4. 0.75N 11. 40 mL 166.7 g for 5N
5. 5N 12. 84.01 g for 1M
6. 525.5 mL to add 84.01 g for 1N
7. 66.67 mL
268 Solutions
TDEC - Fleming Training Center Section 11
Common Valences
1+ 2+ 3+
1- 2- 3-
Solutions 269
Section 11 TDEC - Fleming Training Center
C
Symbol Relative atomic masses are based
+4 on 12C = 12.000
Group Group
1 2 Note: Mass numbers in parentheses 13 14 15 16 17 18
6.941 +1 9.01218 +2
Atomic Number
6 are mass numbers of the most 10.81 +3 12.0111 4 14.0067 3 15.9994
2
2 18.998403 1 20.179 0
Electron Configuration 2-4 +2
2
3
Li 4
Be stable or common isotope.
5
B 6
C +4
7
N 1
+1
+2
+3
8
O 9
F 10
Ne
+4
2-1 2-2 2-3 2-4 2-5 +5 2-6 2-7 2-8
22.98977 +1 24.305 +2 26.98154 +3 28.0855 4 30.97376 3 32.06 2 35.453 1 39.948 0
Al
+2 +3 +1
Na Mg Si P Cl Ar
+4
3
11 12
Group
13 14
+4
15
+5
16
S +6
17
+3
+5
+7 18
2-8-1 2-8-2 3 4 5 6 7 8 9 10 11 12 2-8-3 2-8-4 2-8-5 2-8-6 2-8-7 2-8-8
39.0983 +1 40.08 +2 44.9559 +3 47.88 +2 50.9415 +2 51.996 +2 54.9380 +2 55.847 +2 58.9332 +2 58.69 +2 63.546 +1 65.39 +2 69.72 +3 72.59 4 74.9216 3 78.96 2 79.904 1 83.80 0
+2 +4 +2
K Ca Sc Ti
+3 +3 +3 +1
Cu Zn Ga Ge As Se Kr
+3 +3 +3 +3 +3 +2
4
19 20 21 22
+4
23
V +4
+5
24
Cr Mn Fe +6
25
+4
+7
26 27
Co 28
Ni 29 30 31 32
+4
33
+5
34
+6
35
Br +5
36
2-8-8-1 2-8-8-2 2-8-9-2 2-8-10-2 2-8-11-2 2-8-13-1 2-8-13-2 2-8-14-2 2-8-15-2 2-8-16-2 2-8-18-1 2-8-18-2 2-8-18-3 2-8-18-4 2-8-18-5 2-8-18-6 2-8-18-7 2-8-18-8
85.4678 +1 87.62 +2 88.9059 +3 91.224 +4 92.9064 +3 95.94 +3 (98) +4 101.07 +3 102.906 +3 106.42 +2 107.868 +1 112.41 +2 114.82 +3 118.71 +2 121.75 3 127.60 2 126.905 1 131.29 0
+5 +6 +4 +3 +4
Zr Nb Mo Tc Ru Rh Pd Ag Cd Te
+4
In Sn Sb
+6 +1 +2
5
37
Rb 38
Sr 39
Y 40 41 42 43
+7
44 45 46 47 48 49 50 51
+5
52
+6
53
l +5
+7
54
Xe +4
+6
2-8-18-8-1 2-8-18-8-2 2-8-18-9-2 2-8-18-10-2 2-8-18-12-1 2-8-18-13-1 2-8-18-14-1 2-8-18-15-1 2-8-18-16-1 2-8-18-18 2-8-18-18-1 2-8-18-18-2 2-8-18-18-3 2-8-18-18-4 2-8-18-18-5 2-8-18-18-6 2-8-18-18-7 2-8-18-18-8
132.905 +1 137.33 +2 138.906 +3 178.49 +4 180.948 +5 183.85 +6 186.207 +4 190.2 +3 192.22 +3 195.08 +2 196.967 +1 200.59 +1 204.383 +1 207.2 +2 208.980 +3 (209) +2 (210) (222) 0
+6 +4 +4 +4 +3 +2 +3
Cs Ba La Ta At Rn
+4 +5
Hf W Re Os Ir Pt Au Hg Tl
+4
6
55 56 57 72 73 74 75
+7
76 77 78 79 80 81 82
Pb 83
Bi 84
Po 85 86
2-8-18-18-8-1 2-8-18-18-8-2 2-8-18-18-9-2 **18-32-10-2 -18-32-11-2 -18-32-12-2 -18-32-13-2 -18-32-14-2 -18-32-15-2 -18-32-17-1 -18-32-18-1 -18-32-18-2 -18-32-18-3 -18-32-18-4 -18-32-18-5 -18-32-18-6 -18-32-18-7 -18-32-18-8
(223) +1 226.025 +2 227.028 +3 (261) (262) (263) (264) (265) (268) (269) (272) (277) (285)
7
87
Fr 88
Ra Ac 89 104
Rf Db Sg Bh Hs Mt Uun Uuu Uub
105 106 107 108 109 111 112
Uuq
114
-18-32-18-8-1 -18-32-18-8-2 -18-32-18-9-2
*The systematic names and symbols for elements of atomic numbers above 109
**Denotes the presence of (2-8-) will be used until the approval of trivial names by IUPAC.
for elements 72 and above
140.12 +3 140.908 +3 144.24 +3 (145) +3 150.36 +2 151.96 +2 157.25 +3 158.925 +3 162.50 +3 164.930 +3 167.26 +3 168.934 +3 173.04 +2 174.967 +3
+3
Er Tm Yb Lu
+3
Pr Nd Pm Sm Eu Gd Tb Dy Ho
+4 +3
58
Ce 59 60 61 62 63 64 65 66 67 68 69 70 71
232.038 +4 231.036 +4 238.029 +3 237.048 +3 (244) +3 (243) +3 (247) +3 (247) +3 (251) +3 (252) (257) (258) (259) (260)
+5 +4 +4 +4 +4
Es Fm Md No
+4
90
Th Pa 91 92
U +5
+6
93
Np Pu Am Cm Bk
+5
+6
94
+5
+6
95
+5
+6
96 97 98
Cf 99 100 101 102 103
Lr
270 Solutions 51
8 Reference Tables for Physical Setting/ CHEMISTRY Reference Tables for Physical Setting/ CHEMISTRY 9
TDEC - Fleming Training Center Section 11
Solutions 271
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272 Solutions
TDEC - Fleming Training Center Section 11
Solutions 273
Section 11 TDEC - Fleming Training Center
274 Solutions