DR900 Procedures

Copper DOC316.53.
01039
USEPA1 Bicinchoninate Method2 Method 8506 (CuVer 1) and Method 8026 (CuVer 2)
0.04 to 5.00 mg/L Cu Powder Pillows or AccuVac® Ampuls
Scope and application: For water, wastewater and seawater3; Method 8506 USEPA approved for reporting
wastewater analysis (digestion required)4
1 Approved, USEPA and Standard Method 3500 Cu C or E.
2 Adapted from Nakano, S., Yakugaku Zasshi, 82 486-491 (1962) [Chemical Abstracts, 58 3390e (1963)].
3 Pretreatment required for the powder pillow method- refer to the Interference section.
4 Federal Register, 45 (105) 36166 (May 29, 1980).
Test preparation
Instrument-specific information
The tables in this section show all of the instruments that have the program for this test.
Table 1 shows sample cell and orientation requirements for reagent addition tests, such
as powder pillow or bulk reagent tests. Table 2 shows sample cell and adapter
requirements for AccuVac Ampul tests.
To use either table, select an instrument, then read across to find the corresponding
information for this test.
Table 1 Instrument-specific information for powder pillows
Instrument Sample cell orientation Sample cell
DR 6000 The fill line is to the right. 2495402
DR 3800
DR 2800
DR 2700
DR 5000 The fill line is toward the user.
DR 3900
DR 900 The orientation mark is toward the user. 2401906
Table 2 Instrument-specific information for AccuVac Ampuls

Instrument Adapter Sample cell
DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
1
Before starting
Install the instrument cap on the DR 900 cell holder before ZERO or READ is pushed.
The sample must be digested with heat and acid to make sure that all forms of the metal are measured. Use the mild or
vigorous digestion. Refer to the Water Analysis Guide for more information.
For best results, measure the reagent blank value for each new lot of reagent. Replace the sample with deionized water in
the test procedure to get the reagent blank value. Subtract the reagent blank value from the sample results automatically
with the reagent blank adjust option.
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used and use any recommended personal protective
equipment.
Dispose of reacted solutions according to local, state and federal regulations. Use the Safety Data Sheets for disposal
information for unused reagents. Consult the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.
Items to collect
Powder pillows
Description Quantity
CuVer® 1 Copper Reagent Powder Pillow, 10-mL 1

Sample cells. (For information about sample cells, adapters or light shields, refer to Instrument-
2
specific information on page 1.)
Refer to Consumables and replacements items on page 7 for reorder information.
AccuVac Ampuls
CuVer® 2 Reagent AccuVac® Ampul 1

Beaker, 50-mL 1
Sample cells (For information about sample cells, adapters or light shields, refer to Instrument-
1
Stopper for 18-mm tubes and AccuVac Ampuls 1
Refer to Consumables and replacements items on page 7 for reorder information.
Sample collection and storage

• Collect samples in clean glass or plastic bottles.
• To preserve samples for later analysis, adjust the sample pH to less than 2 with
concentrated nitric acid (about 2 mL per liter). No acid addition is necessary if the
sample is tested immediately.
• If only dissolved copper is to be determined, filter the sample before the acid addition.
• Keep preserved samples at room temperature for a maximum of 6 months.
• Before analysis, adjust the pH to 4–6 with 8.0 N potassium hydroxide standard
solution (do not exceed pH 6, as copper may precipitate).
• Correct the test result for the dilution from the volume additions.
2 Copper, Bicinchoninate Method (5.00 mg/L)

Powder pillow procedure (Method 8506)
Start
1. Start program 2. Prepare the sample: Fill 3. Add the contents of one 4. Swirl to mix.
135 Copper, Bicin. For a sample cell with 10 mL of CuVer 1 Copper reagent
information about sample sample. powder pillow.
cells, adapters or light
shields, refer to Instrument-
specific information
on page 1.
Note: Although the program
name may vary between
instruments, the program
number does not change.
5. Start the instrument 6. Prepare the blank: Fill a 7. Clean the blank. 8. Insert the blank into the
timer. A 2-minute reaction second sample cell with cell holder.
time starts. 10 mL of sample.
The sample shows a purple
color when copper in the
sample mixes with the
reagent powder.
Undissolved powder does
not affect accuracy.
Zero Read
9. Push ZERO. The display 10. Clean the prepared 11. Within 30 minutes after 12. Push READ. Results
shows 0.00 mg/L Cu. sample. the timer expires, insert the show in mg/L Cu.
prepared sample into the
cell holder.
Copper, Bicinchoninate Method (5.00 mg/L) 3

AccuVac Ampul procedure (Method 8026)
Start
1. Start program 2. Prepare the blank: Fill 3. Prepare the sample: 4. Quickly invert the Ampul
140 Copper, Bicin. AV. For the sample cell with 10 mL Collect at least 40 mL of several times to mix.
information about sample of sample. sample in a 50-mL beaker.
cells, adapters or light Fill the AccuVac Ampul with
shields, refer to Instrument- sample. Keep the tip
specific information immersed while the Ampul
on page 1. fills completely.
Zero
5. Start the instrument 6. When the timer expires, 7. Insert the blank into the 8. Push ZERO. The display
timer. A 2-minute reaction clean the blank. cell holder. shows 0.00 mg/L Cu.
time starts.
The sample shows a purple
color when copper in the
sample mixes with the
reagent powder.
Read
9. Clean the AccuVac 10. Within 30 minutes of 11. Push READ. Results
Ampul. adding the reagent, insert show in mg/L Cu.
the prepared sample
AccuVac Ampul into the cell
holder.

Interferences
Table 3 gives treatments for powder pillows. Table 4 gives treatments for AccuVac
Ampuls. To differentiate free copper from that complexed to EDTA or other complexing
agents, use a 25-mL sample cell and Free Copper Reagent Powder Pillow instead of the
CuVer 1 Powder Pillow in the test procedure. Add a Hydrosulfite Reagent Powder Pillow
to the same sample and read the result again. This result is the total dissolved copper
(free and complexed). Unlike CuVer 1 Reagent, CuVer 2 Reagent Powder Pillows and
AccuVac Ampuls react directly with copper that is complexed by chelants such as EDTA.
Table 3 Interfering substances and suggested treatments for powder pillows
Interfering Interference level
substance
Acidity If the sample is extremely acidic (pH 2 or less), a precipitate may form. Add 8 N Potassium Hydroxide
Standard Solution by drops until the sample pH is above 4, then start the test.
Aluminum, Al3+ Use the powder pillow procedure, but use a CuVer 2 Copper Reagent Powder Pillow and not the
CuVer 1 Pillow. Results include total dissolved copper (free and complexed). Use a 25-mL sample
volume.
Cyanide, CN- Prevents full color development. Before the CuVer 1 Powder Pillow Reagent is added, add 0.2 mL of
formaldehyde to the 10-mL sample. Wait 4 minutes, then take the reading. Multiply the test results by
1.02 to correct for sample dilution by the formaldehyde.
Hardness Use the powder pillow procedure, but use a CuVer 2 Copper Reagent Powder Pillow and not the
volume.
Iron, Fe3+ Use the powder pillow procedure, but use a CuVer 2 Copper Reagent Powder Pillow and not the
volume.
Silver, Ag+ If a turbidity remains and turns black, silver interference is likely. Add 20 drops of 50% saturated
Potassium Chloride Solution to 75 mL of sample, then filter through a fine or highly retentive filter. Use
the filtered sample in the test procedure.
Table 4 Interfering substances and suggested treatments for AccuVac Ampuls

Interfering substance Interference level
Acidity If the sample is extremely acidic (pH 2 or less), a precipitate may form. Add 8 N Potassium
Hydroxide Standard Solution by drops until the sample pH is above 4, then start the test.
Aluminum, Al3+ Reagents accommodate high levels.
Cyanide, CN- Prevents full color development. Add 0.5 mL of formaldehyde per 25-mL of sample, then use the
CuVer 2 Reagent AccuVac Ampul. Wait 4 minutes, then take the reading. Multiply the test results by
1.02 to correct for sample dilution by the formaldehyde.
Hardness Reagents accommodate high levels.
Iron, Fe3+ Reagents accommodate high levels.
Silver, Ag+ If a turbidity remains and turns black, silver interference is likely. Add 10 drops of saturated
Potassium Chloride Solution to 75 mL of sample, then filter through a fine or highly retentive filter.
Use the filtered sample in the procedure.
Accuracy check
Standard additions method (sample spike)
Use the standard additions method (for applicable instruments) to validate the test
procedure, reagents and instrument and to find if there is an interference in the sample.
Items to collect:
• 100 mg/L Copper Standard Solution
• 5-mL volumetric pipet, Class A and pipet filler

• Mixing cylinder, 50 mL
• Deionized water
• Pipet, TenSette®, 0.1–1.0 mL and tips
1. Prepare a 12.5 mg/L copper standard solution as follows:

a. Use a pipet to add 5.00 mL of a 100 mg/L copper standard solution into a 50-mL
mixing cylinder.
b. Dilute to the 40-mL mark with deionized water. Mix well. Prepare this solution
daily.
2. Use the test procedure to measure the concentration of the sample, then keep the
(unspiked) sample in the instrument.
3. Go to the Standard Additions option in the instrument menu.
4. Select the values for standard concentration, sample volume and spike volumes.
5. Prepare three spiked samples: use the TenSette pipet to add 0.1 mL, 0.2 mL and
0.3 mL of the prepared standard solution, respectively, to three 10-mL portions of
fresh sample. Mix well.
Note: For AccuVac® Ampuls, add 0.2 mL, 0.4 mL and 0.6 mL of a 75 mg/L Copper Voluette
Ampule Standard to three 50-mL portions of fresh sample.
6. Use the test procedure to measure the concentration of each of the spiked samples.
Start with the smallest sample spike. Measure each of the spiked samples in the
instrument.
7. Select Graph to compare the expected results to the actual results.
Note: If the actual results are significantly different from the expected results, make sure that
the sample volumes and sample spikes are measured accurately. The sample volumes and
sample spikes that are used should agree with the selections in the standard additions menu. If
the results are not within acceptable limits, the sample may contain an interference.
Standard solution method

Use the standard solution method to validate the test procedure, reagents and
instrument.
Items to collect:
• Copper Standard Solution, 100-mg/L
• 100-mL volumetric flask, Class A
• Deionized water
1. Prepare a 4.00 mg/L copper standard solution as follows:

a. Use a pipet to add 4.00 mL of 100 mg/L copper standard solution into the
volumetric flask.
b. Dilute to the mark with deionized water. Mix well. Prepare this solution daily.
2. Use the test procedure to measure the concentration of the prepared standard
solution.
3. Compare the expected result to the actual result.
Note: The factory calibration can be adjusted slightly with the standard adjust option so that the
instrument shows the expected value of the standard solution. The adjusted calibration is then
used for all test results. This adjustment can increase the test accuracy when there are slight
variations in the reagents or instruments.

Method performance
The method performance data that follows was derived from laboratory tests that were
measured on a spectrophotometer during ideal test conditions. Users may get different
results under different test conditions.
Program Standard Precision (95% Confidence Interval) Sensitivity
Concentration change per 0.010 Abs change
135 1.00 mg/L Cu 0.97–1.03 mg/L Cu 0.04 mg/L Cu
140 1.00 mg/L Cu 0.97–1.03 mg/L Cu 0.03 mg/L Cu
Summary of method
Copper in the sample reacts with a salt of bicinchoninic acid contained in CuVer 1 or
CuVer 2 Copper Reagent to form a purple colored complex in proportion to the copper
concentration. The measurement wavelength is 560 nm.
Consumables and replacements items
Required reagents
Description Quantity/test Unit Item no.
CuVer® 1 Copper Reagent Powder Pillow, 10-mL 1 100/pkg 2105869

OR
CuVer® 2 Copper Reagent AccuVac® Ampul 1 25/pkg 2504025
Required apparatus
AccuVac Snapper 1 each 2405200

Beaker, 50-mL 1 each 50041H
Stoppers for 18-mm tubes and AccuVac Ampuls 2 6/pkg 173106
Recommended standards
Description Unit Item no.
Copper Standard Solution, 100-mg/L as Cu 100 mL 12842

Copper Voluette® Ampule Standard, 75-mg/L as Cu, 10-mL 16/pkg 1424710
Metals Drinking Water Standard, LR for Cu, Fe, Mn 500 mL 2833749
Metals Drinking Water Standard, HR for Cu, Fe, Mn 500 mL 2833649
Optional reagents and apparatus
Beaker, 50-mL each 50041H

CuVer 2 Copper Reagent Powder Pillow 100/pkg 2188299
Cylinder, mixing, 50-mL each 189641
100 mL
Formaldehyde, ACS 205932
MDB
Nitric Acid, concentrated 500 mL 15249
Potassium Chloride Solution, 50% saturated 25 mL 1429323

Consumables and replacements items (continued)
100 mL
Potassium Hydroxide Standard Solution, 8 N 28232H
MDB
Sample cells, 25 mL, matched, 1" square 2/pkg 2612602
AccuVac® Snapper each 2405200
Ampule Breaker, Voluette® ampules each 2196800
Sample cells, 25 mm round 6/pkg 2401906
Copper, Free and Total Reagent set, includes: each 2439200
Hydrosulfite Reagent Powder Pillows 100/pkg 2118869
Copper, Free, Reagent Powder Pillows 100/pkg 2182369
FOR TECHNICAL ASSISTANCE, PRICE INFORMATION AND ORDERING: HACH COMPANY

In the U.S.A. – Call toll-free 800-227-4224 WORLD HEADQUARTERS
Outside the U.S.A. – Contact the HACH office or distributor serving you. Telephone: (970) 669-3050
On the Worldwide Web – www.hach.com; E-mail – techhelp@hach.com FAX: (970) 669-2932
© Hach Company/Hach Lange GmbH, 1989–2013. All rights reserved. 04/2013, Edition 8

Copper DOC316.53.01038
Porphyrin Method1 Method 8143

1 to 210 µg/L Cu (LR) Powder Pillows
Scope and application: For water, wastewater and seawater.
1 Adapted from Ishii and Koh, Bunseki Kagaku, 28 (473), 1979.
Test preparation
Instrument specific information
The table in this section shows all of the instruments that have the program for this test.
as powder pillow or bulk reagent tests.
To use the table, select an instrument, then read across to find the corresponding
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
Wash all glassware with detergent. Rinse with tap water. Rinse again with 1:1 nitric acid solution. Rinse a third time with
high-quality deionized water. These steps will remove deposits that can cause slightly high results.
If samples contain high levels of metals, a slight metallic deposit or yellow buildup may form in the sample cell. Wash the cell
as described above.
equipment.
1
Items to collect
Copper Masking Reagent Powder Pillows, 10-mL 1

Porphyrin 1 Reagent Powder Pillows, 10-mL 2
Porphyrin 2 Reagent Powder Pillows, 10-mL 2
Nitric Acid Solution, 1:1 varies
Sample cells (For information about sample cells, adapters or light shields, refer to Instrument
2
Refer to Consumables and replacement items on page 5 for reorder information.

• Collect samples in acid-washed plastic bottles.
• Before analysis, adjust the pH to 2–6 with 5.0 N sodium hydroxide standard solution.
Powder pillow procedure
Start
1. Start program 2. Prepare the blank: Fill 3. Add the contents of one 4. Swirl to dissolve the
145 Copper, Porphyrin. the sample cell with 10 mL Copper Masking Reagent reagent.
For information about of sample. powder pillow to the sample
sample cells, adapters or cell to create the blank.
light shields, refer to
Instrument specific
information on page 1.
2 Copper, Porphyrin Method (210 µg/L)

5. Prepare the sample: Fill 6. Add the contents of one 7. Swirl to mix. 8. Add the contents of one
a second sample cell with Porphyrin 1 Reagent Porphyrin 2 Reagent
10 mL of sample. Powder Pillow to each Powder Pillow to each
sample cell. sample cell.
9. Swirl to mix. 10. Start the instrument 11. When the timer expires, 12. Insert the blank into the
If copper is present in the timer. A 3-minute reaction clean the blank. cell holder.
sample, the sample will time starts.
show blue, then go back to
a yellow color.
Zero Read
13. Push ZERO. The 14. Clean the prepared 15. Insert the prepared 16. Push READ. Results
display shows 0 µg/L Cu. sample. sample into the cell holder. show in µg/L Cu.
Interferences
Aluminum, Al3+ 60 mg/L
Cadmium, Cd2+ 10 mg/L
Calcium, Ca2+ 1500 mg/L
Chelating agents Interfere at all levels unless either the Digesdahl or vigorous digestion is
completed.
Chloride, Cl- 90,000 mg/L
Chromium, Cr6+ 110 mg/L
Copper, Porphyrin Method (210 µg/L) 3
Cobalt, Co2+ 100 mg/L
Fluoride, F- 30,000 mg/L
Iron, Fe2+ 6 mg/L
Lead, Pb2+ 3 mg/L
Magnesium 10,000 mg/L
Manganese 140 mg/L
Mercury, Hg2+ 3 mg/L
Molybdenum 11 mg/L
Nickel, Ni2+ 60 mg/L
Potassium, K+ 60,000 mg/L
Sodium, Na+ 90,000 mg/L
Zinc, Zn2+ 9 mg/L
Highly buffered samples or extreme sample pH Can prevent the correct pH adjustment of the sample by the reagents.
Sample pretreatment may be necessary.
Accuracy check
Items to collect:
• Copper Standard Solution, 4 mg/L (PourRite® Ampule or prepare from a dilution of a
higher concentration copper standard solution)
• Ampule breaker
4. Open the standard solution.
5. Prepare six spiked 10-mL samples: use the TenSette pipet to add 0.1 mL, 0.2 mL and
0.3 mL of the standard solution, respectively, to two 10-mL portions of fresh sample.
Mix well.
instrument.

instrument.
Items to collect:
• Copper Standard Solution, 10 mg/L
4 Copper, Porphyrin Method (210 µg/L)

• Deionized water
1. Prepare a 150 µg/L copper standard solution as follows:

a. Use a pipet to add 15.00 mL of 10-mg/L copper standard solution into the
volumetric flask.
solution.
Method performance
145 50 µg/L Cu 47–53 µg/L Cu 1 µg/L Cu
Summary of method
The porphyrin method is very sensitive to trace amounts of free copper. The method is
free from most interferences and does not require any sample extraction or concentration
before analysis. Interferences from other metals are removed with the copper masking
reagent. The porphyrin indicator forms an intense, yellow-colored complex with any free
copper present in sample. The measurement wavelength is 425 nm for
spectrophotometers or 420 nm for colorimeters.
Consumables and replacement items
Required reagents and apparatus
Nitric Acid Solution, 1:1 varies 500 mL 254049

Copper Reagent Set, Porphyrin, 10-mL 1 100/pkg 2603300
Includes:
Copper Masking Reagent Powder Pillow, 10-mL 1 100/pkg 2603449
Porphyrin 1 Reagent Powder Pillow, 10-mL 2 100/pkg 2603549
Porphyrin 2 Reagent Powder Pillow, 10-mL 2 100/pkg 2603649
Copper Standard Solution, 4 mg/L, 2 mL Pour-Rite Ampules 20/pkg 2605720

Copper Standard Solution, 10-mg/L Cu 100 mL 12932
Water, deionized 4L 27256
Copper, Porphyrin Method (210 µg/L) 5

100 mL
Sodium Hydroxide Standard Solution, 5.0 N 245032
MDB
Pipet, TenSette®, 0.1–1.0 mL each 1970001
Pipet tips for TenSette Pipet 1970001 50/pkg 2185696
Pipet, volumetric Class A, 15-mL each 1451539
Flask, volumetric, Class A, 1000-mL each 1457453
Pipet filler, safety bulb each 1465100
Sample cells, 1" square matched set 8/pkg 2495408
Paper, pH, 0–14 pH range 100/pkg 2601300
Hardness, Calcium and Magnesium DOC316.53.01043
Calmagite Colorimetric Method Method 8030

0.05 to 4.00 mg/L Ca and Mg as CaCO3
Test preparation
Table 1 Instrument-specific information for reagent solution
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
For the most accurate magnesium test results, keep the sample temperature between 21–29 °C (70–84 °F).
This test detects any calcium or magnesium contamination in the mixing cylinder, measuring droppers or sample cells. To
test cleanliness, repeat the test until the results are consistent.
Total hardness in mg/L equals mg/L Ca as CaCO3 plus mg/L Mg as CaCO3.
Traces of EDTA or EGTA that remain from previous tests will give incorrect results. Rinse sample cells thoroughly before
each use.
equipment.
1
Items to collect
Alkali Solution, for calcium and magnesium tests 1 mL

Calcium and Magnesium Indicator Solution 1 mL
EDTA Solution, 1 M 1 drop
EGTA Solution 1 drop
Cylinder, graduated mixing, 100-mL 1
Dropper, measuring, 0.5-mL and 1.0-mL 2
3

• Collect samples in acid-washed plastic bottles.
Calmagite procedure
Start
1. Start program 2. Pour 100 mL of sample 3. Use a 1.0 mL dropper to 4. Close the cylinder. Invert
225 Hardness, Mg. For into a 100-mL graduated add 1.0 mL of Calcium and the cylinder several times to
information about sample mixing cylinder. Magnesium Indicator mix.
cells, adapters or light Solution.
shields, refer to Instrument
on page 1.
2 Hardness, Ca and Mg, Calmagite Method (4.00 mg/L)

5. Use a 1.0 mL dropper to 6. Close the cylinder. Invert 7. Pour 10 mL of the 8. Blank preparation: Add
add 1.0 mL of Alkali Solution the cylinder several times to solution into each of three one drop of 1 M EDTA
for Calcium and Magnesium mix. sample cells. solution to the first sample
Test. cell.
9. Swirl to mix. 10. Magnesium sample: 11. Swirl to mix. 12. Clean the blank.
Add one drop of EGTA
Solution to the second
sample cell.
Zero
13. Insert the blank into the 14. Push ZERO. The 15. Clean the prepared 16. Magnesium sample:
cell holder. display shows 0.00 mg/L Mg sample. Insert the prepared
CaCO3. magnesium sample cell into
the cell holder.
Hardness, Ca and Mg, Calmagite Method (4.00 mg/L) 3

Read Start Zero
17. Push READ. Results 18. Do not remove the 19. Exit the magnesium 20. Push ZERO. The
show in mg/L magnesium as sample cell from the program. Start program display shows 0.00 mg/L Mg
calcium carbonate. This instrument. Record or select 220 Hardness, Ca. CaCO3.
value is the amount of STORE to save the
magnesium in the sample magnesium results before
expressed as CaCO3. the next step.
Read
21. Calcium sample: Insert 22. Push READ. Results

the third sample cell into the show in mg/L calcium as
cell holder. calcium carbonate. This
value is the amount of
calcium in the sample
expressed as CaCO3.
Interferences

Ca >1.0 mg/L; Mg >0.25 mg/L For the most accurate calcium test result, run the test again on a diluted sample if the calcium
is over 1.0 and the magnesium is over 0.25 mg/L as CaCO3. No retesting is needed if either
is below those respective concentrations.
Chromium (Cr 3+) Above 0.25 mg/L
Copper (Cu 2+) Above 0.75 mg/L
EDTA Above 0.2 mg/L as CaCO3
EDTA or EGTA Traces remaining in sample cells from previous tests will give erroneous results. Rinse cells
thoroughly before use.
Iron (Fe 2+) Above 1.4 mg/L
Iron (Fe 3+) Above 2.0 mg/L
Manganese (Mn 2+) Above 0.20 mg/L
Zinc (Zn 2+) Above 0.050 mg/L
Accuracy check
instrument.
4 Hardness, Ca and Mg, Calmagite Method (4.00 mg/L)
Items to collect:
• 2.00 mg/L (as CaCO3) Calcium Standard Solution
1. Use the test procedure to measure the concentration of the standard solution.
Method performance
220 2.00 mg/L Ca 1.90–2.10 mg/L Ca 0.05 mg/L Ca
225 2.00 mg/L Mg 1.92–2.08 mg/L Mg 0.05 mg/L Mg
Summary of method
The colorimetric method for measuring hardness supplements the conventional titrimetric
method because the colorimetric method can measure very low levels of calcium and
magnesium. Also, some metals that interfere in the titrimetric method may not interfere
when the sample is diluted to bring it within the range of this test. The indicator dye is
calmagite, which forms a purplish-blue color in a strongly alkaline solution and changes to
red when it reacts with free calcium or magnesium.
Calcium and magnesium determinations are made by chelation of calcium with EGTA to
remove the red color from calcium and then chelation of calcium and magnesium with
EDTA to remove the red color from both calcium and magnesium. The measurement of
the red color in the different states is used to measure the calcium and magnesium
concentrations. The measurement wavelength is 522 nm for spectrophotometers or
520 nm for colorimeters.
Required reagents
Hardness Reagent Set, includes: — 100 tests 2319900

100 mL
Alkali Solution, for calcium and magnesium tests 1 mL 2241732
MDB
100 mL
Calcium and Magnesium Indicator Solution 1 mL 2241832
MDB
50 mL
EDTA Solution, 1 M 2 drops 2241926
SCDB
50 mL
EGTA Solution 1 drop 2229726
SCDB
Required apparatus
Cylinder, graduated mixing, 100-mL , tall form 1 each 2088642

Dropper, measuring, 0.5 and 1.0 mL plastic 2 20/pkg 2124720
Hardness, Ca and Mg, Calmagite Method (4.00 mg/L) 5


100 mL
MDB
Hydrazine DOC316.53.01046
p-Dimethylaminobenzaldehyde Method1 Method 8141

4 to 600 µg/L N2H4 (spectrophotometers) Reagent Solution or AccuVac® Ampuls
10 to 500 µg/L N2H4 (colorimeters)
Scope and application: For boiler water/feedwater.
1 Adapted from ASTM Manual of Industrial Water, D1385-78, 376 (1979).
Test preparation
Instrument-specific table
DR 3800
DR 2800
DR 2700
DR 3900
DR 900 The fill line is toward the user. 2401906

Instrument Adapter
DR 6000 —
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C)
DR 2800
DR 2700
Before starting
Samples must be analyzed immediately after collection and cannot be preserved for later analysis.
The sample temperature must be between 21 ± 4 °C (70 ± 7 °F) for accurate results.
1
The reagent that is used in this test is corrosive. Use protection for eyes and skin and be prepared to flush any spills with
running water.
equipment.
Items to collect
Reagent solution
HydraVer 2 Reagent Solution 1 mL

Deionized water 10 mL
Graduated cylinder, 25-mL 1
2
specific table PPAV.)
AccuVac Ampuls
HydraVer 2 Reagent AccuVac Ampuls 2

Deionized water 40 mL
Beaker, 50-mL 1
Stoppers for 18 mm tubes and AccuVac Ampuls 2
Sample collection
• Samples must be analyzed immediately after collection and cannot be preserved for
later analysis.
• Collect samples in clean glass or plastic bottles with tight-fitting caps. Fill the bottle
completely and immediately tighten the cap.
• Prevent agitation of the sample or exposure to air.
2 Hydrazine, p-Dimethylaminobenzaldehyde Method (600 µg/L)

Reagent solution procedure
Start
1. Start program 2. Prepare the blank: Use 3. Prepare the sample: 4. Add 0.5 mL of HydraVer
231 Hydrazine. For a graduated cylinder to pour Use a graduated cylinder to 2 Hydrazine Reagent to
information about sample 10 mL of deionized water pour 10 mL of sample into a each sample cell.
cells, adapters or light into a sample cell. second sample cell. A yellow color shows if
shields, refer to Instrument- hydrazine is present in the
specific table PPAV. sample. The blank may also
Note: Although the program show a light yellow color.
5. Swirl to mix. 6. Start the instrument 7. Clean the blank. 8. Insert the blank into the
timer. A 12-minute reaction cell holder.
time starts.
Complete the blank zero
steps and insert the
prepared sample during the
reaction period.
Zero Read
9. Push ZERO. The display 10. Clean the prepared 11. Insert the prepared 12. Immediately after the
shows 0 µg/L N2H4. sample. sample into the cell holder. timer expires, push READ.
Results show in µg/L N2H4.
Hydrazine, p-Dimethylaminobenzaldehyde Method (600 µg/L) 3

AccuVac Ampul procedure
Start
1. Start program 2. Prepare the sample: 3. Quickly invert the Ampul 4. Start the instrument
232 Hydrazine AV. For Collect at least 40 mL of several times to mix. timer. A 12-minute reaction
information about sample sample in a 50-mL beaker. time starts.
cells, adapters or light Fill one HydraVer Hydrazine Complete the blank zero
shields, refer to Instrument- AccuVac Ampul with the steps and insert the
specific table PPAV. sample. Keep the tip prepared sample during the
Note: Although the program immersed while the Ampul reaction period.
name may vary between fills completely.
5. Prepare the blank: Pour 6. Quickly invert the Ampul 7. Clean the blank. 8. Insert the blank into the
at least 40 mL of deionized several times to mix. cell holder.
water into a second 50-mL
beaker. Fill the second
HydraVer Hydrazine
AccuVac Ampul with
deionized water. Keep the
tip immersed while the
Ampul fills completely.
Zero
9. Insert the blank AccuVac 10. Push ZERO. The 11. Clean the AccuVac 12. Insert the prepared
Ampul into the cell holder. display shows 0 µg/L N2H4. Ampul. sample AccuVac Ampul into
the cell holder.

Read
13. Immediately after the

timer expires, push READ.
Results show in µg/L N2H4.
Interferences
Ammonia No interference up to 10 mg/L. May cause a positive interference of up to 20% at 20 mg/L.
Highly colored or turbid Prepare a 1:1 mixture of deionized water and household bleach. Add one drop of this mixture to
samples 25 mL of sample in a graduated mixing cylinder and invert to mix. Use this solution, instead of
deionized water, to prepare the blank in the test procedure.
Morpholine No interference up to 10 mg/L.
Accuracy check
instrument.
Items to collect:
• Hydrazine sulfate, reagent grade
• 1-L volumetric flask, Class A (2)
• Deionized water, oxygen-free
1. Prepare a 25-mg/L hydrazine stock solution as follows:

a. Add 0.1016 g of hydrazine sulfate into a 1-L volumetric flask.
b. Dilute to the mark with oxygen-free deionized water. Mix well. Prepare the stock
solution each day.
2. Prepare a 0.25 mg/L hydrazine standard solution as follows:
a. Use a pipet to add 10.00 mL of the 25-mg/L hydrazine stock solution into a 1-L
volumetric flask.
b. Dilute to the mark with oxygen-free deionized water. Mix well. Prepare the
standard solution immediately before use.
solution.
Hydrazine, p-Dimethylaminobenzaldehyde Method (600 µg/L) 5

Method performance
231 250 µg/L N2H4 247–253 µg/L N2H4 4 µg/L N2H4
232 250 µg/L N2H4 246–254 µg/L N2H4 4 µg/L N2H4
Summary of method
Hydrazine in the sample reacts with the p-dimethylaminobenzaldehyde from the
HydraVer 2 Reagent to form a yellow color which is proportional to the hydrazine
concentration. The measurement wavelength is 455 nm for spectrophotometers or
Required reagents
Water, deionized varies 4L 27256

100 mL
HydraVer® 2 Hydrazine Reagent 1 mL 179032
MDB
OR
HydraVer 2 Hydrazine Reagent AccuVac® Ampul 2 25/pkg 2524025
Required apparatus for reagent solution
Cylinder, graduated, 25-mL. 1 each 50840
Required apparatus for AccuVac Ampuls

Hydrazine Sulfate, ACS 100 g 74226

Pipet, volumetric, Class A, 10-mL each 1451538

Iron, Ferrous DOC316.53.01049
1,10-Phenanthroline Method1 Method 8146

0.02 to 3.00 mg/L Fe2+ Powder Pillows or AccuVac® Ampuls
1 Adapted from Standard Methods for the Examination of Water and Wastewater, 15th ed. 201 (1980).
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900

DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
Before starting
1
equipment.
Items to collect
Powder pillows
Ferrous Iron Reagent Powder Pillows, 25-mL 1

2
AccuVac Ampuls
Ferrous Iron Reagent AccuVac Ampuls 1

Beaker, 50-mL 1
1
Sample collection
later analysis.
• Collect samples in clean glass or plastic bottles with tight-fitting caps. Fill the bottle
completely and immediately tighten the cap.
• Prevent agitation of the sample or exposure to air.
2 Iron, Ferrous, 1,10-Phenanthroline Method (3.00 mg/L)

Start
1. Start program 255 Iron, 2. Prepare the blank: Fill 3. Prepare the sample: Fill 4. Add the contents of one
Ferrous. For information the sample cell with 10 mL a mixing cylinder to the 25- Ferrous Iron Reagent
about sample cells, of sample. mL line with sample. Powder Pillow to the mixing
adapters or light shields, cylinder.
refer to Instrument-specific An orange color shows if
information on page 1. ferrous iron is present in the
Note: Although the program sample
5. Close the cylinder. Invert 6. Start the instrument 7. When the timer expires, 8. Insert the blank into the
the cylinder several times to timer. A 3-minute reaction clean the blank. cell holder.
mix. time starts.
Zero
9. Push ZERO. The display 10. Fill a second sample 11. Clean the prepared 12. Insert the prepared
shows 0.00 mg/L Fe2+. cell with 10 mL of the sample. sample into the cell holder.
reacted prepared sample.
Iron, Ferrous, 1,10-Phenanthroline Method (3.00 mg/L) 3

Read
13. Push READ. Results

show in mg/L Fe2+.
Start
1. Start program 257 Iron, 2. Prepare the blank: Fill 3. Prepare the sample: 4. Quickly invert the Ampul
Ferrous AV. For information the sample cell with 10 mL Collect at least 40 mL of several times to mix.
about sample cells, of sample. sample in a 50-mL beaker.
adapters or light shields, Fill the AccuVac Ampul with
refer to Instrument-specific sample. Keep the tip
information on page 1. immersed while the Ampul
Note: Although the program fills completely.
Zero
timer. A 3-minute reaction clean the blank. cell holder. shows 0.00 mg/L Fe2+.
time starts.
4 Iron, Ferrous, 1,10-Phenanthroline Method (3.00 mg/L)

Read
9. Clean the AccuVac 10. Insert the prepared 11. Push READ. Results
Ampul. sample AccuVac Ampul into show in mg/L Fe2+.
the cell holder.
Accuracy check
instrument.
Items to collect:
• Ferrous Ammonium Sulfate, hexahydrate
• 1-L volumetric flask, Class A
• Deionized water
1. Prepare a 100-mg/L Fe2+ ferrous iron stock solution as follows:

a. Add 0.7022 g of ferrous ammonium sulfate, hexahydrate into a 1-L volumetric
flask.
b. Dilute to the mark with deionized water. Mix well.
2. Prepare a 2 mg/L ferrous iron standard solution as follows:
a. Use a pipet to add 2.00 mL of the 100-mg/L Fe2+ ferrous iron stock solution into a
100-mL volumetric flask.
b. Dilute to the mark with deionized water. Mix well. Prepare the standard solution
immediately before use.
solution.
Method performance
255 2.00 mg/L Fe2+ 1.99–2.01 mg/L Fe2+ 0.021 mg/L Fe2+
257 2.00 mg/L Fe2+ 1.98–2.02 mg/L Fe2+ 0.023 mg/L Fe2+
Summary of method
Iron, Ferrous, 1,10-Phenanthroline Method (3.00 mg/L) 5
The 1,10-phenanthroline indicator in the Ferrous Iron Reagent reacts with ferrous iron
(Fe2+) in the sample to form an orange color in proportion to the iron concentration. Ferric
iron (Fe3+) does not react. The ferric iron concentration can be determined by subtracting
the ferrous iron concentration from the results of a total iron test. The measurement
wavelength is 510 nm for spectrophotometers or 520 nm for colorimeters.
Required reagents
Ferrous Iron Reagent Powder Pillow, 25-mL 1 100/pkg 103769

OR
Ferrous Iron Reagent AccuVac® Ampul 1 25/pkg 2514025
Required apparatus

Stoppers for 18 mm-tubes and AccuVac Ampuls 1 6/pkg 1448000
Recommended standards and apparatus
Balance, analytical, 80 g x 0.1 mg 100-240 VAC each 2936701

Ferrous Ammonium Sulfate, hexahydrate, ACS 113 g 1125614
Pipet, volumetric, Class A, 1.00-mL each 1451535
Wipes, disposable 280/pkg 2097000
Iron DOC316.53.01048
FerroZine® Method1 Method 8147

0.09 to 1.400 mg/L Fe Reagent Solution Pillows
Scope and application: For water and seawater.
1 Adapted from Stookey, L.L., Anal. Chem., 42(7), 779 (1970).
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
Clean all glassware with 6.0 N (50%) hydrochloric acid, then rinse thoroughly with deionized water to remove contaminants.
If the sample contains rust, refer to Interferences on page 3.
Use clean clippers to open the solution pillows that are free of rust. Wipe with a dry towel. Do not allow the clippers to
contact the contents of the pillow.
As an alternative to the solution pillows, use 0.5 mL of FerroZine ® Iron Reagent Solution in the test procedure.
The FerroZine Iron Reagent can crystallize or precipitate if kept at cold temperatures during shipment. The reagent quality is
not affected. Put the reagent in warm water to dissolve the precipitate.
1
equipment.
Items to collect
FerroZine Iron Reagent Solution Pillows, OR 1

FerroZine Iron Reagent Solution 0.5 mL
Cylinder, 25-mL graduated mixing, with stopper 1
Clippers for solution pillows 1
2

• To measure only dissolved iron, filter the sample immediately after collection and
before acidification.
• Before analysis, adjust the pH to 3–5 with 10% ammonium hydroxide solution. Do not
exceed pH 5 to prevent precipitation of the iron.
Solution pillow or bulk solution procedure
Start
1. Start program 260 Iron, 2. Prepare the blank: Fill 3. Prepare the sample: Fill 4. Add the contents of one
FerroZine. For information the sample cell with 10 mL a mixing cylinder to the 25- FerroZine Iron Reagent
about sample cells, of sample. mL line with sample. Solution Pillow to the mixing
adapters or light shields, cylinder.
refer to Instrument specific
2 Iron, FerroZine Method (1.400 mg/L)

5. Close the cylinder. Invert 6. Start the instrument 7. When the timer expires, 8. Clean the blank.
the cylinder several times to timer. A 5-minute reaction pour 10 mL of the prepared
mix. time starts. sample from the mixing
A purple color shows if iron cylinder into the second
is present in the sample. sample cell.
Zero
9. Insert the blank into the 10. Push ZERO. The 11. Clean the prepared 12. Insert the prepared
cell holder. display shows 0.000 mg/L sample. sample into the cell holder.
Fe.
Read

show in mg/L Fe.
Interferences
substance
Strong chelants Interfere at all levels. Use the FerroVer® or TPTZ methods for these samples. Use the TPTZ method
(EDTA) for low iron concentrations.
Cobalt May give slightly high results
Copper May give slightly high results
Hydroxides Add the FerroZine® Iron Reagent to 25 mL of sample, then boil the sample for 1 minute in a boiling
water bath. Let the sample cool to 24 °C (75 °F), then start the instrument timer. Return the sample
volume to 25 mL with deionized water.
Iron, FerroZine Method (1.400 mg/L) 3

substance
Magnetite (black Add the FerroZine® Iron Reagent to 25 mL of sample, then gently boil the sample for 20 to 30 minutes
iron oxide) or in a boiling water bath.
Ferrites Note: Do not let the sample boil dry. A purple color will show if iron is present.
Let the sample cool to 24 °C (75 °F). Return the sample volume to 25 mL with deionized water.
Continue with the test procedure after the timer step.
Rust Add the FerroZine® Iron Reagent to 25 mL of sample, then boil the sample for 1 minute in a boiling
water bath. Cool to 24 °C (75 °F), then start the instrument timer. Return the sample volume to 25 mL
with deionized water.
Accuracy check
Items to collect:
• Iron Voluette® Ampule Standard, 10 mg/L Fe
• Ampule breaker
0.3 mL of the standard solution, respectively, to three 25-mL portions of fresh sample.
Mix well.
instrument.

instrument.
Items to collect:
• Iron Standard Solution, 100-mg/L
• Deionized water
1. Prepare a 1.0 mg/L iron standard solution as follows:

a. Use a pipet to add 5.00 mL of 100 mg/L iron standard solution into the volumetric
flask.
solution.
4 Iron, FerroZine Method (1.400 mg/L)

Method performance
260 1.000 mg/L Fe 0.985–1.015 mg/L Fe 0.009 mg/L Fe
Summary of method
The FerroZine® Iron Reagent forms a purple-colored complex with trace amounts of iron
in samples that are buffered to a pH of 3.5. This method is applicable for determining
trace levels of iron in chemical reagents and glycols and with digestion can be used to
analyze samples containing magnetite (black iron oxide) or ferrites. The measurement
Required reagents
FerroZine® Iron Reagent Solution 0.5 mL 500 mL 230149

OR
FerroZine® Iron Reagent Solution Pillows 1 50/pkg 230166
Required apparatus
Clippers for solution pillows 1 each 96800

Cylinder, graduated mixing, 25 mL with stopper 1 each 2088640
Sample cell, 10 mL square, matched pair 2 2/pkg 2495402
Iron Standard Solution, 100 mg/L Fe 100 mL 1417542

Iron Standard Solution, 10 mL Voluette® ampule, 25 mg/L Fe 16/pkg 1425310
Pipet, volumetric 5.00-mL each 1451537

Iron, FerroZine Method (1.400 mg/L) 5
100 mL
Ammonium Hydroxide, 10% 1473632
MDB
Hydrochloric Acid, 1:1, 6N 500 mL 88449
Iron, Total DOC316.53.01052
FerroMo Method1 Method 8365

0.01 to 1.80 mg/L Fe Powder Pillows
Scope and application: For cooling water that contains molybdate-based treatment.
1 Adapted from G. Frederick Smith Chemical Co., The Iron Reagents, 3rd ed. (1980).
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
Wash all glassware with detergent. Rinse with tap water. Rinse again with 1:1 hydrochloric acid solution. Rinse a third time
with high-quality deionized water. These steps will remove deposits that can cause slightly high results.
If the sample contains 100 mg/L or more molybdate (MoO42–), read the sample immediately after the instrument zero.
equipment.
1
Items to collect
FerroMo® Reagent 1 Powder Pillow 1

FerroMo® Reagent 2 Powder Pillow 1
Cylinder, graduated mixing, 25-mL with stopper 1
Cylinder, graduated mixing, 50-mL with stopper 1
2

• Collect samples in clean acid-washed glass or plastic bottles.
concentrated hydrochloric acid (about 2 mL per liter). No acid addition is necessary if
the sample is tested immediately.
• To measure only dissolved iron, filter the sample through a 0.45 micron filter or
equivalent medium immediately after collection and before acidification.
Do not exceed pH 5 to prevent precipitation of the iron.
Start
1. Start program 275 Iron, 2. Prepare the sample: Fill 3. Add the contents of one 4. Close the cylinder. Invert
FerroMo. For information a 50-mL mixing cylinder with FerroMo Iron Reagent several times to mix.
about sample cells, 50 mL of sample. 1 Powder Pillow to the
adapters or light shields, mixing cylinder.
refer to Instrument specific
2 Iron, Total, FerroMo Method (1.80 mg/L)

5. Fill a clean 25-mL mixing 6. Prepare the blank: Fill a 7. Develop the sample: 8. Close the cylinder. Invert
cylinder to the 25-mL mark second sample cell with Add the contents of one the cylinder several times to
with the prepared sample. 10 mL of the prepared FerroMo Iron Reagent mix.
Use the rest of the sample sample. 2 Powder Pillow to the A blue color will show if iron
to prepare the blank. prepared sample in the 25- is present in the sample. A
mL mixing cylinder. small amount of undissolved
reagent will not affect the
results of the test.
9. Start the instrument 10. When the timer expires, 11. Clean the blank. 12. Insert the blank into the
timer. A 3-minute reaction pour 10 mL of the cell holder.
time starts. developed sample into a
sample cell. This is the
prepared sample for the
test.
Zero Read
13. Push ZERO. The 14. Clean the developed 15. Insert the developed 16. Push READ. Results
display shows 0.00 mg/L Fe. sample. sample into the cell holder. show in mg/L Fe.
Iron, Total, FerroMo Method (1.80 mg/L) 3

Interferences
substance
pH After the addition of reagent, a sample pH of less than 3 or more than 4 may inhibit color formation, cause
the developed color to fade quickly or result in turbidity. Adjust the sample pH to between 3 and 8 in the
graduated cylinder before the addition of reagent:
1. Add by drops an applicable amount of iron-free acid or base such as 1.0 N Sulfuric Acid Standard
Solution or 1.0 N Sodium Hydroxide Standard Solution.
2. Make a volume correction if significant volumes of acid or base are used.
Accuracy check
Items to collect:
• Iron Voluette® Ampule Standard, 50 mg/L Fe
• Ampule breaker
Mix well.
instrument.

instrument.
Items to collect:
• 100 mg/L iron standard solution
• Deionized water

flask.
solution.
4 Iron, Total, FerroMo Method (1.80 mg/L)

Method performance
Summary of method
FerroMo Iron Reagent 1 contains a reducing agent combined with a masking agent. The
masking agent removes interference from high levels of molybdate. The reducing agent
converts precipitated or suspended iron, such as rust, to the ferrous state. FerroMo Iron
Reagent 2 contains the indicator combined with a buffering agent. The indicator reacts
with ferrous iron in the sample, buffered between pH 3 and 5, which results in a deep
blue-purple color. The measurement wavelength is 590 nm for spectrophotometers or
Required reagents
FerroMo® Iron Reagent Set 1 100/pkg 2544800

Includes:
FerroMo® Reagent 1 Powder Pillow 1 25/pkg 2543768
FerroMo® Reagent 2 Powder Pillow 1 50/pkg 2543866
Required apparatus
Cylinder, graduated mixing, 25 mL with stopper 1 each 2088640

Cylinder, graduated mixing, 50 mL, with stopper 1 each 2088641

Iron Standard Solution, 10 mL Voluette® ampule, 50 mg/L Fe. 16/pkg 1425410

Iron, Total, FerroMo Method (1.80 mg/L) 5

Consumables and replacement items (continued)
100 mL
MDB
100 mL
MDB
100 mL
Sulfuric Acid Standard Solution, 1 N 127032
MDB
Hydrochloric Acid, 6.0 N 1:1, 50% 500 mL 88449
dition 8
USEPA1 FerroVer® Method2 Method 8008

0.02 to 3.00 mg/L Fe Powder Pillows or AccuVac® Ampuls
Scope and application: For water, wastewater and seawater; digestion is required for determining total iron.
1 USEPA approved for reporting wastewater analysis, Federal Register, June 27, 1980; 45 (126:43459).
2 Adapted from Standard Methods for the Examination of Water and Wastewater.
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900

DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
1
Before starting
The reagent in this test procedure converts all soluble iron and most insoluble forms of iron in the sample to soluble ferrous
iron for measurement. For regulatory reporting, however, the sample must be digested with heat and acid to make sure that
all forms of the metal are measured.
For turbid samples, treat the blank with one 0.1-g scoop of RoVer Rust Remover. Swirl to dissolve.
equipment.
Items to collect
Powder pillows
FerroVer® Iron Reagent Powder Pillows, 10-mL1 1

2
1 FerroVer is a registered trademark of Hach Company.
AccuVac Ampuls
FerroVer® Iron Reagent AccuVac® Ampul 1

Beaker, 50-mL 1
1

2 Iron, Total, FerroVer Method (3.00 mg/L)

Start
1. Start program 265 Iron, 2. Prepare the sample: Fill 3. Add FerroVer Iron 4. Swirl the sample cell to
FerroVer. For information a sample cell with 10 mL of Reagent Powder Pillow to mix. Undissolved powder
about sample cells, sample. the sample cell. will not affect accuracy.
adapters or light shields,
refer to Instrument-specific
5. Start the instrument 6. Prepare the blank: Fill a 7. Clean the blank. 8. When the timer expires,
timer. A 3-minute reaction second sample cell with insert the blank into the cell
time starts. 10 mL of sample. holder.
An orange color will show if
iron is present. Let samples
that contain rust react for
5 minutes or more.
Zero Read
9. Push ZERO. The display 10. Clean the prepared 11. Insert the prepared 12. Push READ. Results
shows 0.00 mg/L Fe. sample. sample into the cell holder. show in mg/L Fe.
Iron, Total, FerroVer Method (3.00 mg/L) 3

AccuVac procedure
Start
FerroVer AV. For the sample cell with 10 mL Collect at least 40 mL of several times to mix.
information about sample of sample. sample in a 50-mL beaker. Undissolved powder will not
cells, adapters or light Fill the AccuVac Ampul with affect accuracy.
Zero
5. Start the instrument 6. Clean the blank. 7. When the timer expires, 8. Push ZERO. The display
timer. A 3-minute reaction insert the blank into the cell shows 0.00 mg/L Fe.
time starts. holder.
An orange color will show if
iron is present. Let samples
that contain rust react for
5 minutes or more.
Read
Ampul. sample AccuVac Ampul into show in mg/L Fe.
the cell holder.

Interferences
Calcium, Ca2+ No effect at less than 10,000 mg/L as CaCO3.
Chloride, Cl– No effect at less than 185,000 mg/L.
Copper, Cu2+ No effect. Masking agent is contained in FerroVer Reagent.
High iron levels Inhibit color development. Dilute sample and re-test to verify results.
Iron oxide A mild, vigorous or Digesdahl digestion is necessary. After digestion, adjust the sample pH to 3–
5 with sodium hydroxide, then analyze.
Magnesium No effect at 100,000 mg/L as CaCO3.
Molybdate molybdenum No effect at 50 mg/L as Mo.
High sulfide levels, S2– Pre-treat the sample in a fume hood or well-ventilated area before analysis:
1. Add 5 mL of hydrochloric acid, ACS to 100 mL of sample in a 250-mL Erlenmeyer flask.
2. Boil for 20 minutes.
3. Let the solution cool to room temperature.
4. Adjust the pH to 3–5 with sodium hydroxide.
5. Add deionized water until the volume is 100 mL.
6. Use the treated sample in the test procedure.
Turbidity Pre-treat the sample before analysis:

1. Add 0.1 g scoop of RoVer® Rust Remover to the blank. Swirl to mix.
2. If sample remains turbid, add three 0.2 g scoops of RoVer Rust Remover to a 75 mL sample.
Let stand 5 minutes.
3. Filter through a glass membrane filter and filter holder.
Highly buffered samples Can prevent the correct pH adjustment of the sample by the reagents. Sample pretreatment may
or extreme sample pH be necessary. Adjust the pH to 3–5.
Accuracy check
Items to collect:
• Iron Voluette® Ampule Standard, 25 mg/L
• Ampule Breaker
Mix well.
Note: For AccuVac® Ampuls, add 0.2 mL, 0.4 mL and 0.6 mL of the standard solution to three
50-mL portions of fresh sample.

instrument.

instrument.
Items to collect:
• Iron standard solution, 100 mg/L
• Deionized water

a. Use a pipet to add 2 mL of the 100 mg/L iron standard solution into the volumetric
flask.
solution.
Method performance
Summary of method
FerroVer Iron Reagent converts all soluble iron and most insoluble forms of iron in the
sample to soluble ferrous iron. The ferrous iron reacts with the 1-10 phenanthroline
indicator in the reagent to form an orange color in proportion to the iron concentration.
The measurement wavelength is 510 nm for spectrophotometers or 520 nm for
colorimeters.
Required reagents
Description Quantity/Test Unit Item no.
FerroVer® Iron Reagent Powder Pillow, 10-mL 1 100/pkg 2105769

OR
FerroVer® Iron Reagent AccuVac® Ampul 1 25/pkg 2507025

Required apparatus



Filter, glass membrane, 47-mm 100/pkg 253000
Filter holder for glass membrane filter each 234000
Hydrochloric Acid, concentrated 500 mL 13449
RoVer Rust Remover 454 g 30001
100 mL
MDB
Spoon, measuring, 0.1-g each 51100

FerroVer® Method1 Method 10249

0.1 to 3.0, 1.0 to 30.0 and 10.0 to 300.0 mg/L Fe Powder Pillows
Scope and application: For oil and gas field waters; digestion is required for total iron determinations.2
1 USEPA approved for reporting wastewater analysis, Federal Register, June 27, 1980; 45 (126:43459).
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
equipment.
1
Items to collect
FerroVer® Iron Reagent Powder Pillow, 10-mL 1

EDTA solution, 1M 2 drops
1

Start
1. Start program 265 Iron, 2. Fill a clean sample cell 3. If the sample volume is 4. Swirl to mix.
FerroVer. For information with sample: less than 10 mL, add
about sample cells, deionized water to the 10-
adapters or light shields, • Use 10 mL of sample mL line.
refer to Instrument specific for the 0.02 to 3.0 mg/L
information on page 1. range.
• Use 1.0 mL of sample
name may vary between for the 0.2 to 30.0 mg/L
instruments, the program range with a dilution
number does not change. factor of 10.
• Use 0.1 mL of sample
for the 2.0 to
300.0 range with a
dilution factor of 100.
Note: Refer to Set the
dilution factor on page 4.
2 Iron, Total, FerroVer Method (multi-range: 3.0, 30.0, 300.0 mg/L)

5. Add 2 drops of EDTA 6. Swirl to mix. 7. Clean the sample cell. 8. Insert the sample cell
Solution 1 M to the sample. into the cell holder.
Zero
9. Push ZERO. The display 10. Remove the sample 11. Add the contents of one 12. Swirl to mix.
shows 0.0 mg/L Fe. from the cell holder. FerroVer Iron Reagent Accuracy is not affected by
Powder Pillow to the sample undissolved powder.
cell.
Read
13. Start the instrument 14. When the timer expires, 15. Insert the sample cell 16. Push READ. Results
timer. A 3-minute reaction clean the sample cell. into the cell holder. show in mg/L Fe.
time starts.
If iron is present in the
sample, an orange color will
show.
Iron, Total, FerroVer Method (multi-range: 3.0, 30.0, 300.0 mg/L) 3

Interferences
substance
Barium, Ba2+ The dilution of samples lowers most barium concentrations below interference levels. No effects are
seen on analyzed samples that contain less than 50 mg/L of Ba. No effects are seen when a 1.0 or
0.1 mL sample volume is used in the test procedure. A turbidity may show at higher levels. Use
5 drops of EDTA Solution in the test procedure and allow the sample to react for 5 minutes.
Calcium, Ca2+ No effect at less than 10,000 mg/L as CaCO3.
Chloride, Cl– No effect at less than 185,000 mg/L.

substance
Copper, Cu2+ No effect. Masking agent is contained in FerroVer Reagent.
High iron levels Inhibit color development. Dilute sample and re-test to verify results.
Magnesium No effect at 100,000 mg/L as CaCO3.
Molybdate No effect at 50 mg/L as Mo.
molybdenum
High sulfide levels, Pre-treat the sample in a fume hood or well-ventilated area before analysis:
S2–
1. Add 5 mL of hydrochloric acid, ACS to 100 mL of sample in a 250-mL Erlenmeyer flask.
2. Boil for 20 minutes.
3. Let the solution cool to room temperature.
4. Adjust the pH to 3–5 with sodium hydroxide.
5. Add deionized water until the volume is 100 mL.
Strontium, Sr2+ Strontium by itself does not interfere. Strontium in combination with Barium will cause a precipitate to
form. The dilution of samples lowers most strontium concentrations below interference levels. No
effects are seen on analyzed samples that contain less than 50 mg/L of combined Ba and Sr. No
effects are seen when a 1.0 or 0.1 mL sample volume is used in the test procedure. A turbidity may
show at higher levels. Use 5 drops of EDTA Solution in the test procedure and allow the sample to
react for 5 minutes.
Highly buffered Can prevent the correct pH adjustment of the sample by the reagents. Sample pretreatment may be
samples or extreme necessary. Adjust the sample pH to 3–5 before the test is started. Correct the test result for the
sample pH dilution from the volume addition.
Set the dilution factor

Instruments that have a dilution factor option can include the dilution factor in the result
and show the concentration of the original, undiluted sample. For example, if the sample
is diluted by a factor of 10, the instrument multiplies the result by 10 and shows the
calculated result in the instrument display.
1. Select Options>More>Dilution factor from the instrument menu.

Note: Colorimeters include a dilution factor when the chemical form is set. Go to
Options>Advanced Options>Chemical Form and select LR, MR or HR.
2. Enter the dilution factor:
• 1 mL sample diluted to 10 mL: dilution factor is 10.
• 0.1 mL sample diluted to 10 mL: dilution factor is 100.
3. Push OK to confirm. Push OK again.
4. Push RETURN to go back to the measurement screen.
Accuracy check
Items to collect:
• Iron Voluette® Ampule Standard, 25 mg/L
• Ampule breaker

Mix well.
instrument.

instrument.
Items to collect:
• Iron Standard Solution, 100 mg/L
• Deionized water

flask.
solution.
Method performance
Summary of method
FerroVer Iron Reagent converts all soluble iron and most insoluble forms of iron in the
sample to soluble ferrous iron. The ferrous iron reacts with the 1-10 phenanthroline
indicator in the reagent to form an orange color in proportion to the iron concentration.
colorimeters.

Required reagents
FerroVer® Iron Reagent Powder Pillow, 10-mL 1 100/pkg 2105769

50 mL
EDTA Solution, 1 M 2 drops 2241926
SCDB


100 mL
MDB
Filter, glass membrane, 47-mm 100/pkg 253000
Filter holder for glass membrane filter each 234000
RoVer Rust Remover 454 g 30001
Spoon, measuring, 0.1-g each 51100

TPTZ Method1 Method 8112

0.012 to 1.800 mg/L Fe (spectrophotometers) Powder Pillows or AccuVac® Ampuls
0.04 to 1.80 mg/L Fe (colorimeters)
1 Adapted from G. Frederic Smith Chemical Co., The Iron Reagents, 3rd ed. (1980).
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900

DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
Before starting
1
Wash all glassware with detergent. Rinse with tap water. Rinse again with 1:1 hydrochlorioc acid solution. Rinse a third time
with high-quality deionized water. These steps will remove deposits that can cause slightly high results.
equipment.
Items to collect
Powder pillows
TPTZ Iron Reagent Powder Pillows, 10-mLS 2

Sample cells For information about sample cells, adapters or light shields, refer to Instrument-
2
specific information on page 1.
AccuVac Ampuls
TPTZ Low Range Iron Reactent AccuVac Ampul 1

Beaker, 50-mL 1
1

Do not exceed pH 5 or iron may precipitate.
2 Iron, Total, TPTZ Method (1.800 mg/L)

Start
1. Start program 270 Iron, 2. Prepare the sample: Fill 3. Add the contents of one 4. Swirl the sample cell for
TPTZ. For information about a sample cell with 10 mL of 10-mL TPTZ Iron Reagent at least 30 seconds to mix.
sample cells, adapters or sample. Powder Pillow to the
light shields, refer to prepared sample .
Instrument-specific
5. Start the instrument 6. Prepare the blank: Fill a 7. Add the contents of the 8. Swirl the sample cell for
timer. A 3-minute reaction sample cell with 10 mL of contents of one 10-mL at least 30 seconds to mix.
time starts. deionized water. TPTZ Iron Reagent Powder
Prepare the blank during the Pillow to the blank. .
reaction time.
Zero
9. When the timer expires, 10. Insert the blank into the 11. Push ZERO. The 12. Clean the prepared
clean the blank. cell holder. display shows 0.000 mg/L sample.
Fe.
Iron, Total, TPTZ Method (1.800 mg/L) 3

Read
13. Insert the prepared 14. Push READ. Results

sample into the cell holder. show in mg/L Fe.
Start
TPTZ AV. For information the sample cell with 10 mL Collect at least 40 mL of several times to mix.
about sample cells, of sample. sample in a 50-mL beaker.
adapters or light shields, Fill the AccuVac Ampul with
refer to Instrument-specific sample. Keep the tip
information on page 1. immersed while the Ampul
Note: Although the program fills completely.
Zero
timer. A 3-minute reaction clean the blank. cell holder. shows 0.000 mg/L Fe.
time starts.

Read
Ampul. sample AccuVac Ampul into show in mg/L Fe.
the cell holder.
Interferences
Interferences were tested with an iron concentration of 0.5 mg/L Fe. The following do not
interfere with this method when present up to the levels shown.
Cadmium 4.0 mg/L
Chromium3+ 0.25 mg/L
Chromium6+ 1.2 mg/L
Cobalt 0.05 mg/L
Copper 0.6 mg/L
Cyanide 2.8 mg/L
Manganese 50.0 mg/L
Mercury 0.4 mg/L
Molybdenum 4.0 mg/L
Nickel 1.0 mg/L
Nitrite Ion 0.8 mg/L
Color or turbidity If the sample, without a TPTZ Iron Reagent Powder Pillow, has a color or turbidity more than the
blank (deionized water plus TPTZ Iron Reagent), then use the sample as the blank. Refer to the
powder pillow procedure.
pH After the addition of reagent, a sample pH of less than 3 or more than 4 may inhibit color formation.
The developed color fades quickly or causes turbidity. Adjust the sample pH in the sample cell
before the addition of reagent:
1. Use a pH meter or pH paper to measure the current pH.

2. Add an applicable amount of iron-free acid or base such as 1.0 N Sulfuric Acid Standard
Solution or 1.0 N Sodium Hydroxide Standard Solution to adjust the sample pH to between
3 and 4.1
3. Make a volume correction if significant volumes of acid or base are used.
1 Refer to Consumables and replacement items on page 7 for reorder information.
Accuracy check
Items to collect:
• Iron Standard Solution, 10 mg/L Fe

• Pipet, TenSette®, 0.1–1.0 mL and pipet tips
• Mixing cylinders, 50-mL (3) (for AccuVac Ampuls)
Mix well.
instrument.

instrument.
Items to collect:
• 100 mg/L iron standard solution
• Deionized water

flask.
solution.
Method performance

Summary of method
The TPTZ Iron Reagent forms a deep blue-purple color with ferrous iron (Fe2+). The
indicator is combined with a reducing agent that converts precipitated or suspended iron,
such as rust, to the ferrous state. The amount of ferric iron (Fe3+) can be determined as
the difference between the results of a ferrous iron test and the concentration of total iron.
colorimeters.
Required reagents
TPTZ Iron Reagent Powder Pillow, 10-mL 1 100/pkg 2608799

OR
TPTZ Low Range Iron Reagent AccuVac® Ampul 1 25/pkg 2510025
Required apparatus

Sample cell, 10 mL round, 25 x 54 mm 1 each 2122800
Sample cell, 10 mL round, 25 x 60 mm 1 6/pkg 2427606


50 mL
SCDB
100 mL
MDB
100 mL
MDB
Stoppers for 18-mm tubes and AccuVac Ampuls 6/pkg 173106


Pipet, volumetric 5.00-mL each 1451537
1
Nitrite DOC316.53.01074
USEPA Diazotization Method1 Method 8507

0.01 to 0.300 mg/L NO 2––N (LR, spectrophotometers) Powder Pillows or AccuVac®
0.05 to 0.350 mg/L NO 2––N (LR, colorimeters) Ampuls

1 USEPA approved for wastewater analysis, Federal Register, 44(85), 25505 (May 1, 1979).
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900

DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
2 Nitrite, Diazotization Method (0.300 mg/L)

Before starting
equipment.
Items to collect
Powder pillows
NitriVer® 3 Reagent Powder Pillows, 10-mL 1

2
AccuVac Ampuls
NitriVer® 3 Reagent AccuVac® Ampuls 1

Beaker, 50-mL 1
1

• To preserve samples for later analysis, keep the samples at or below 6 °C (43 °F) for
up to 48 hours.
• Let the sample temperature increase to room temperature before analysis.
3
Start
1. Start program 371 N, 2. Prepare the sample: Fill 3. Add the contents of one 4. Swirl to mix.
Nitrite LR PP. For a sample cell with 10 mL of NitriVer 3 Reagent Powder A pink color shows if nitrite
information about sample sample. Pillow. is present in the sample.
shields, refer to Instrument-
on page 1.
5. Start the instrument 6. Prepare the blank: 7. Clean the blank. 8. Insert the blank into the
timer. A 20-minute reaction When the timer expires, fill a cell holder.
time starts. second sample cell with
10 mL of sample.
Zero Read
2
shows 0.000 mg/L NO2––N. sample. sample into the cell holder. show in mg/L NO ––N.
Nitrite, Diazotization Method (0.300 mg/L) 3

Start
1. Start program 375 N, 2. Prepare the sample: 3. Quickly invert the Ampul 4. Start the instrument
Nitrite LR AV. For Collect at least 40 mL of several times to mix. timer. A 20-minute reaction
information about sample sample in a 50-mL beaker. A pink color shows if nitrite time starts.
cells, adapters or light Fill the AccuVac Ampul with is present in the sample.
Zero
5. Prepare the blank: 6. Clean the blank. 7. Insert the blank into the 8. Push ZERO. The display
2
When the timer expires, fill a cell holder. shows 0.000 mg/L NO ––N.
sample cell with 10 mL of
sample.
Read
Ampul. sample AccuVac Ampul into show in mg/L NO ––N.
the cell holder. 2
Interferences
Antimonous ions Interfere by causing precipitation

Auric ions Interfere by causing precipitation

Bismuth ions Interfere by causing precipitation
Chloroplatinate ions Interfere by causing precipitation
Cupric ions Cause low results
Ferric ions Interfere by causing precipitation
Ferrous ions Cause low results
Lead ions Interfere by causing precipitation
Mercurous ions Interfere by causing precipitation
Metavanadate ions Interfere by causing precipitation
Nitrate Very high levels of nitrate (>100 mg/L nitrate as N) appear to undergo a slight
amount of reduction to nitrite, either spontaneously or during the course of the
test. A small amount of nitrite will be found at these levels.
Silver ions Interfere by causing precipitation
Strong oxidizing and reducing substances Interfere at all levels
Accuracy check
instrument.
Items to collect:
• 0.150 mg/L NO2––N standard solution (Nitrite standard solutions are difficult to
prepare. Use the instructions in Standard Methods for the Examination of Water and
Wastewater, Method 4500—NO2-B)
Method performance
– – –
371 0.150 mg/L NO2 –N 0.147–0.153 mg/L NO2 –N 0.002 mg/L NO2 –N
– – –
Summary of method
Nitrite in the sample reacts with sulfanilic acid to form an intermediate diazonium salt.
This couples with chromotropic acid to produce a pink colored complex directly
proportional to the amount of nitrite present. The measurement wavelength is 507 nm for

Required reagents

NitriVer® 3 Nitrite Reagent Powder Pillow, 10-mL 1 100/pkg 2107169
OR
NitriVer® 3 Nitrite Reagent AccuVac® Ampul 1 25/pkg 2512025
Required apparatus
Recommended standards, reagents and apparatus

AccuVac® vials for sample blanks 25/pkg 2677925
AccuVac® Drainer each 4103600
Standard Methods Book, most current edition each 2270800
Sodium Nitrite, ACS 454 g 245201

Diazotization Method Method 10019

0.03 to 0.500 mg/L NO2––N (LR) Test ‘N Tube™ Vials
Test preparation
Table 1 shows adapter and light shield requirements for the instruments that use them.
Table 1 Instrument-specific information for test tubes
Instrument Adapters Light shield
DR 6000 — —
DR 5000 — —
DR 900 4846400 Cover supplied with the instrument
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
DR 3900, DR 3800, DR 2800 and DR 2700: Install the light shield in Cell Compartment #2 before this test is started.
equipment.
Items to collect
Light shield (For information about sample cells, adapters or light shields, refer to Instrument
1
Test 'N Tube™ NitriVer® 3 Nitrite Reagent Set 1
Pipet, TenSette® 1.0 to 10.0 mL with tips varies

1
up to 48 hours.
Test 'N Tube procedure
Start
1. Start program 345 N, 2. Prepare the sample: Fill 3. Put the cap on the vial. 4. Start the instrument
Nitrite LR TNT. For a Test 'N Tube NitriVer Shake to dissolve the timer. A 20-minute reaction
information about sample 3 Nitrite vial with 5 mL of powder. A pink color shows time starts.
cells, adapters or light sample. if nitrite-nitrogen is present
shields, refer to Instrument in the sample .
on page 1.
Zero
5. Prepare the blank: 6. Clean the blank vial. 7. Insert the blank vial into 8. Push ZERO. The display
2
When the timer expires, fill the 16-mm cell holder. shows 0.000 mg/L NO ––N.
and empty Test 'N Tube vial
with 5 mL of sample.
Read
9. Clean the sample vial. 10. Insert the sample vial 11. Push READ. Results
into the 16-mm cell holder. show in mg/L NO ––N.
2 Nitrite, Diazotization TNT Method (0.500 mg/L)
Interferences
Antimonous ions Interfere by causing precipitation
Auric ions Interfere by causing precipitation
Bismuth ions Interfere by causing precipitation
Chloroplatinate ions Interfere by causing precipitation
Cupric ions Cause low results
Ferric ions Interfere by causing precipitation
Ferrous ions Cause low results
Lead ions Interfere by causing precipitation
Mercurous ions Interfere by causing precipitation
Metavanadate ions Interfere by causing precipitation
Nitrate Very high levels of nitrate (>100 mg/L nitrate as N) appear to undergo a slight
amount of reduction to nitrite, either spontaneously or during the course of the
test. A small amount of nitrite will be found at these levels.
Silver ions Interfere by causing precipitation
Strong oxidizing and reducing substances Interfere at all levels
Accuracy check
instrument.
Items to collect:
• 0.300 mg/L NO2––N standard solution (Nitrite standard solutions are difficult to
prepare. Use the instructions in Standard Methods for the Examination of Water and
Method performance
– – –
Summary of method
Nitrite in the sample reacts with sulfanilic acid to form an intermediate diazonium salt.
This couples with chromotropic acid to produce a pink colored complex directly
proportional to the amount of nitrite present. The measurement wavelength is 507 nm for
Nitrite, Diazotization TNT Method (0.500 mg/L) 3

Required reagents
NitriVer® 3 Nitrite Reagent Set, Test 'N Tube™ 1 50/pkg 2608345
Required apparatus
Pipet, TenSette®, 1.0- to 10.0-mL 1 each 1970010

Pipet Tips, for TenSette Pipet 1970010 varies 50/pkg 2199796
Recommended standards, reagents and apparatus


Ferrous Sulfate Method1 Method 8153

2 to 250 mg/L NO2– (HR, spectrophotometers) Powder Pillows
2 to 150 mg/L NO2– (HR, colorimeters)™
Scope and application: For cooling systems.
1 Adapted from McAlpine, R. and Soule, B., Qualitative Chemical Analysis, New York, 476, 575 (1933).
Test preparation
Table 1 Instrument-specific information for powder pillow
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
equipment.
Items to collect
NitriVer® 2 Nitrite Reagent Powder Pillows, 10-mL 1

Deionized water varies
Items to collect (continued)
Stopper, Neoprene, solid #1 2

2

up to 48 hours.
Start
1. Start program 373 N, 2. Prepare the sample: Fill 3. Add the contents of one 4. Close the sample cell.
Nitrite HR PP. For a sample cell with 10 mL of NitriVer 2 Nitrite Reagent Shake to dissolve the
information about sample sample. Powder Pillow. A greenish- reagent.
cells, adapters or light brown color starts to show if
shields, refer to Instrument nitrite is present in the
specific information sample.
on page 1.
5. Start the instrument 6. Prepare the blank: Fill a 7. Clean the blank. 8. Insert the blank into the
timer. A 10-minute reaction second sample cell with cell holder.
To prevent low results,
leave the sample cell on a
flat surface. Do not move
or disturb the sample cell
during the reaction period.
1
Zero
9. Push ZERO. The display 10. After the timer expires, 11. Clean the prepared 12. Insert the prepared
shows 0 mg/L NO2–. gently invert the prepared sample. sample into the cell holder.
sample two times.
Excessive mixing causes
low results..
Read

show in mg/L NO2–.
Interferences
This test does not measure nitrates nor is it applicable to glycol-based samples. Dilute
glycol-based samples and follow the Low Range Nitrite procedure, Method 8507.
Accuracy check

instrument.
Items to collect:
• 200 mg/L NO2– standard solution (Nitrite standard solutions are difficult to prepare.
Use the instructions in Standard Methods for the Examination of Water and
Method performance
– – –
373 200 mg/L NO2 191–209 mg/L NO2 1.4 mg/L NO2
Summary of method
The method uses ferrous sulfate in an acidic medium to reduce nitrite to nitrous oxide.
Ferrous ions combine with the nitrous oxide to form a greenish-brown complex in direct
proportion to the nitrite present. The measurement wavelength is 585 nm for
Required reagents
NitriVer® 2 Nitrite Reagent Powder Pillow, 10-mL 1 100/ pkg 2107569
Required apparatus
Stopper, Neoprene, solid, size #1 2 12/pkg 1480801


Nitrite, Ferrous Sulfate Method (250 mg/L) 3
Nitrogen, Ammonia DOC316.53.01077
Salicylate Method1 Method 8155

0.01 to 0.50 mg/L NH3–N Powder Pillows
1 Adapted from Clin. Chim. Acta., 14, 403 (1966).
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
The reagents that are used in this test contain sodium nitroferricyanide. Keep cyanide solutions at pH > 11 to prevent
exposure to hydrogen cyanide gas. Collect the reacted samples for proper disposal.
Keep the samples sealed at all times to prevent ammonia contamination from the air.
equipment.
Items to collect
Ammonia Cyanurate Reagent Powder Pillow, 10-mL 2

Ammonia Salicylate Reagent Powder Pillow, 10-mL 2
1
Stoppers for 18-mm tubes and AccuVac Ampuls 2

2

• If the sample contains chlorine, add one drop of 0.1 N sodium thiosulfate to 1 liter of
sample to remove each 0.3 mg/L of chlorine.
concentrated sulfuric acid (about 2 mL per liter). No acid addition is necessary if the
• Keep the preserved samples at or below 6 °C (43 °F) for up to 28 days.
• Before analysis, adjust the pH to 7 with 5.0 N sodium hydroxide standard solution.
Start
1. Start program 385 N, 2. Prepare the blank: Fill a 3. Prepare the sample: Fill 4. Add the contents of one
Ammonia, Salic. For sample cell with 10 mL of a second sample cell with Ammonia Salicylate powder
information about sample deionized water. 10 mL of sample. pillow to each sample cell.
on page 1.
2 Nitrogen-Ammonia, Salicylate Method (0.50 mg/L)

5. Close the sample cell. 6. Start the instrument 7. After the timer expires, 8. Close the sample cell.
Shake to dissolve the timer. A 3-minute reaction add the contents of one Shake to dissolve the
reagent. time starts. Ammonia Cyanurate powder reagent.
pillow to each sample cell.
Zero
9. Start the instrument 10. When the timer expires, 11. Insert the blank into the 12. Push ZERO. The
timer. A 15-minute reaction clean the blank. cell holder. display shows 0.00 mg/L
time starts. NH3–N.
A green color shows when
ammonia-nitrogen is
present.
Read
13. Clean the prepared 14. Insert the prepared 15. Push READ. Results
sample. sample into the cell holder. show in mg/L NH3–N.
Interferences

Calcium 1000 mg/L as CaCO3
Iron All levels. Correct for iron interference as follows:
1. Use one of the Iron, Total procedures to measure the iron concentration of the sample.
2. Use an iron standard solution to add iron to the deionized water blank so that the blank has the
same iron concentration as the sample. The iron interference will be zeroed out from the test
result.
Nitrogen-Ammonia, Salicylate Method (0.50 mg/L) 3

Magnesium 6000 mg/L as CaCO3

Monochloramine Monochloramine that is in chloraminated drinking water interferes directly at all levels and gives high
results. Use a Free Ammonia and Monochloramine method to determine free ammonia in these
sample matrices.
Nitrate 100 mg/L as NO3––N
–
Nitrite 12 mg/L as NO2 –N
pH Adjust acidic or basic samples to approximately pH 7. Use 1 N sodium hydroxide standard solution
for acidic samples and 1 N hydrochloric acid standard solution for basic samples.
3–
Phosphate 100 mg/L as PO4 –P
2–
Sulfate 300 mg/L as SO4
Sulfide Sulfide will intensify the color. Remove sulfide interference as follows:
1. Measure approximately 350 mL of sample in a 500-mL Erlenmeyer flask.
2. Add the contents of one Sulfide Inhibitor Reagent Powder Pillow. Swirl to mix.
3. Filter the sample through a folded filter paper and filter funnel.
4. Use the filtered sample in the test procedure.
Other Substances Less common interferences such as hydrazine and glycine cause intensified colors in the prepared
sample. Turbidity and color will give incorrect high values. Samples with severe interferences require
distillation. Use the distillation procedure that is supplied with the distillation set.
Accuracy check
Items to collect:
• Ammonia Nitrogen Standard Solution, 10 mg/L as NH3–N
• 25-mL mixing cylinders (3)
Mix well.
instrument.

instrument.
Items to collect:
• Ammonia Nitrogen Standard Solution, 10 mg/L as NH3–N

• Deionized water
1. Prepare a 0.40 mg/L ammonia nitrogen standard solution as follows:

a. Use a pipet to add 4 mL of the 10 mg/L ammonia nitrogen standard solution into
the volumetric flask. (Alternate preparation: pipet 0.8 mL of a 50-mg/L ammonia
nitrogen standard solution into a 100-mL volumetric flask.)
solution.
Method performance
385 0.40 mg/L NH3–N 0.38–0.42 mg/L NH3–N 0.004 mg/L NH3–N
Summary of method
Ammonia compounds combine with chlorine to form monochloramine. Monochloramine
reacts with salicylate to form 5-aminosalicylate. The 5-aminosalicylate is oxidized in the
presence of a sodium nitroprusside catalyst to form a blue-colored compound. The blue
color is masked by the yellow color from the excess reagent to give a final green-colored
solution. The measurement wavelength is 655 nm for spectrophotometers or 610 nm for
colorimeters.
Pollution prevention and waste management
The ammonia salicylate reagent contains sodium nitroferricyanide and must be disposed
of as a hazardous waste. Dispose of reacted solutions according to local, state and
federal regulations.
Required reagents
Nitrogen Ammonia, reagent set, 10-mL, includes: — 100 tests 2668000

Ammonia Cyanurate Reagent Powder Pillow, 10-mL 2 100/pkg 2653199
Ammonia Salicylate Reagent Powder Pillow, 10-mL 2 100/pkg 2653299

Nitrogen Ammonia Standard Solution, 10-mg/L NH3–N 500 mL 15349
Nitrogen Ammonia Standard Solution, 10-mL Voluette® Ampule, 50-mg/L NH3–N 16/pkg 1479110


Wastewater, Effluent Inorganics, for NH3-N, NO3-N, PO4, COD, SO4, TOC 500 mL 2833249

Distillation heater and support for 2265300, 115 VAC, 60 Hz each 2274400
Distillation apparatus set, general purpose each 2265300
Flask, Erlenmeyer, 500-mL each 50549
Funnel, poly, 65-mm each 108367
Paper, filter, folded, 12.5-cm 100/pkg 189457
Pipet, serological, 2-mL each 53236
Sodium Hydroxide, 5 N 50 mL 245026
Sulfide Inhibitor Reagent Powder Pillow 100/pkg 241899
Sulfuric Acid, concentrated, ACS 500 mL 97949

Salicylate Method Method 10031

0.2 to 50.0 mg/L NH3–N (HR) Test ‘N Tube™ Vials
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
Small sample sizes (such as 0.1 mL) may not be representative of the entire sample. Mix the sample well before the test or
use a different portion of the sample to repeat the test.
To prevent airborne cross-contamination of the blank, complete the preparation of the blank before samples and standards
are opened. If the sample or standard containers are open, move to a separate area of the lab to prepare the blank.
equipment.
Items to collect
1
High Range Test 'N Tube™ AmVer® 3 Nitrogen Ammonia Reagent Set 2
1
Funnel, micro (for reagent addition) 1

Pipet, TenSette® 0.1 to 1.0 mL, with tips varies

• Analyze the samples as soon as possible for best results.
• If chlorine is known to be present, add one drop of 0.1 N Sodium Thiosulfate to 1 L of
sample for each 0.3 mg/L Cl2.
Start
1. Start program 343 N, 2. Prepare the blank: Add 3. Prepare the sample: 4. Add the contents of one
Ammonia HR TNT. For 0.1 mL of ammonia-free Add 0.1 mL of sample to Ammonia Salicylate
information about sample water to one AmVer™ one AmVer™ Diluent Reagent Powder Pillow for
cells, adapters or light Diluent Reagent Test 'N Reagent Test 'N Tube for 5-mL samples to each vial.
shields, refer to Instrument Tube for High Range High Range Ammonia
specific information Ammonia Nitrogen. Nitrogen.
on page 1.
2 Nitrogen-Ammonia, Salicylate TNT Method (50.0 mg/L)

5. Add the contents of one 6. Put the caps on both 7. Start the instrument 8. Clean the blank vial.
Ammonia Cyanurate vials. Shake thoroughly to timer. A 20-minute reaction
Reagent Powder Pillow to dissolve the powder. time starts.
each vial.
Zero
9. Insert the blank vial into 10. Push ZERO. The 11. Clean the sample vial. 12. Insert the sample vial
the 16-mm cell holder. display shows 0.0 mg/L into the 16-mm cell holder.
NH3–N.
Read

show in mg/L NH3–N.
Interferences
Calcium 50,000 mg/L as CaCO3
result.
Magnesium 300,000 mg/L as CaCO3

sample matrices.
Nitrogen-Ammonia, Salicylate TNT Method (50.0 mg/L) 3

–
Nitrate 5000 mg/L as NO3 –N
Nitrite 600 mg/L as NO2––N
Phosphate 5000 mg/L as PO43––P
Sulfate 6000 mg/L as SO42–
Accuracy check
Items to collect:
• Nitrogen, Ammonia Ampule Standard, 150-mg/L NH3–N
• Ampule breaker
Mix well.
instrument.

instrument.
Items to collect:
• 100-mg/L Ammonia Nitrogen standard

• Deionized water
1. Prepare a 40.0 mg/L ammonia nitrogen standard solution as follows:

a. Use a pipet to add 20.0 mL of 100 mg/L ammonia nitrogen standard solution into
the volumetric flask.
solution.
Method performance
Summary of method
presence of a sodium nitroprusside catalyst to form a blue colored compound. The blue
color is masked by the yellow color from the excess reagent present to give a green-
colored solution. The measurement wavelength is 655 nm for spectrophotometers or
Required reagents
Nitrogen Ammonia, Reagent Set, High Range Test 'N Tube™ AmVer™ 2 25 tests 2606945
Required apparatus
Funnel, micro, poly 1 each 2584335

Pipet Tips, for TenSette Pipet 1970001 2 50/pkg 2185696
Test tube rack 1 each 1864100

Nitrogen, Ammonia Standard Solution, 10-mg/L NH3-N 500 mL 15349

Nitrogen, Ammonia Standard Solution, 10-mL Voluette® Ampules, 150 mg/L 16/pkg 2128410

Filter paper, folded, 12.5-cm 100/pkg 69257
Hydrochloric Acid Standard Solution, 1 N 1000 mL 2321353
100 mL
MDB
50 mL
SCDB
Sodium Thiosulfate, 0.1 N 100 mL 32332


Salicylate Method1 Method 10023

0.02 to 2.50 mg/L NH3–N (LR) Test ‘N Tube™ Vials
1 Adapted from Clin. Chim. Acta, 14, 403 (1966).
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
equipment.
Items to collect
1
Nitrogen Ammonia, Reagent Set, Low Range Test 'N Tube™ AmVer™ 2
Funnel, micro, poly 1
Pipet, TenSette®, 1.0- to 10.0-mL 1
Pipet Tips, for TenSette Pipet 1970010 2
1

Start
1. Start program 342, 2. Prepare the blank: Add 3. Prepare the sample: 4. Use a funnel to add the
Ammonia LR TNT. For 2.0 mL of ammonia-free Add 2.0 mL of sample to contents of one Ammonia
information about sample water to one AmVer™ one AmVer™ Diluent Salicylate Reagent Powder
cells, adapters or light Diluent Reagent Test ‘N Reagent Test ‘N Tube for Pillow to each vial.
shields, refer to Instrument Tube for Low Range Low Range Ammonia
specific information Ammonia Nitrogen. Nitrogen.
on page 1.
5. Use a funnel to add the 6. Close the vials tightly. 7. Start the instrument 8. After the timer expires,
contents of one Ammonia Shake thoroughly to timer. A 20-minute reaction clean the blank vial.
Cyanurate Reagent Powder dissolve the powder. time starts.
Pillow to each vial.

Zero
the cell holder. display shows 0.00 mg/L into the cell holder.
NH3–N.
Read

show in mg/L NH3–N.
Interferences
result.
Magnesium 15,000 mg/L as CaCO3

sample matrices.
Nitrate 250 mg/L as NO3––N
–
Nitrite 30 mg/L as NO2 –N
3–
Phosphate 250 mg/L as PO4 –P
2–
Sulfate 300 mg/L as SO4

Accuracy check
Items to collect:
• 50 mg/L Nitrogen-Ammonia Standard Solution, 10-mL Voluette® Ampule
• Ampule breaker
Mix well.
instrument.

instrument.
Items to collect:
• 1.0 mg/L Nitrogen-Ammonia Standard Solution

Method performance
Summary of method
presence of a sodium nitroprusside catalyst to form a blue colored compound. The blue
color is masked by the yellow color from the excess reagent present to give a green-
colored solution. The measurement wavelength is 655 nm for spectrophotometers or
Required reagents
Nitrogen Ammonia, Reagent Set, Low Range Test 'N Tube ™ AmVer™ 2 25 tests 2604545
Required apparatus

Nitrogen Ammonia Standard Solution, 1.0-mg/L NH3–N 500 mL 189149


Paper, filter, folded, 12.5-cm 100/pkg 189457


Hydrochloric Acid Standard Solution, 1 N 1000 mL 2321353
100 mL
MDB
50 mL
SCDB


Nitrogen, Total Inorganic DOC316.53.01090
Titanium Trichloride Reduction Method Method 10021

0.2 to 25.0 mg/L N Test ‘N Tube™ Vials
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
For safety, wear gloves to break open the reagent ampules.
equipment.
Items to collect
Total Inorganic Nitrogen Pretreatment Reagent Set (TiCl 3 Reduction Method) 1

Test 'N Tube AmVer™ Nitrogen-Ammonia Reagent Set 1
Centrifuge 1
Funnel, micro 1
Light shield or adapter (For information about sample cells, adapters or light shields, refer to
1
Instrument specific information on page 1.)
Pipet, TenSette®, 0.1- to 1.0-mL, with pipet tips 1
Pipet, volumetric, Class A, 1.00-mL 1
1
Test tube rack 1

Water, deionized 1 mL

Start
1. Start program 346 N 2. Add 1 mL of Total 3. Prepare the sample: 4. Prepare the blank: Add
Inorganic TNT. For Inorganic Nitrogen Add 1 mL of sample to one 1 mL of deionized water to
information about sample Pretreatment Base of the vials. the second vial.
cells, adapters or light concentrate into two Total
shields, refer to Instrument Inorganic Nitrogen
specific information Pretreatment Diluent vials.
on page 1.
2 Nitrogen, Total Inorganic, Titanium Trichloride Reduction TNT Method (25.0 mg/L)
5. Put the caps on both 6. Add the contents of one 7. Put the caps on both 8. Let the vials stand for at
vials. Shake for 30 seconds Total Inorganic Nitrogen vials. Shake gently for at least one minute.
to mix. Reductant ampule to the 30 seconds to mix.
prepared sample vial. Add The precipitate should be
the contents of another black after shaking.
Total Inorganic Nitrogen Excessive shaking will
Reductant ampule to the cause the precipitate to
blank vial. change to a white color and
A black precipitate will form give low test results.
immediately.
9. Put the vials in a 10. Start the instrument 11. After the timer expires, 12. Add 2 mL of blank from
centrifuge. timer. A 3-minute reaction add 2 mL of prepared the centrifuge to a second
time starts. sample from the centrifuge AmVer Diluent Reagent
Put the vials in a centrifuge to an AmVer Diluent Test 'N Tube for Low Range
and run the centrifuge to for Reagent Test 'N Tube for Ammonia Nitrogen.
three minutes. If no Low Range Ammonia
centrifuge is available, let Nitrogen.
the vials sit for 30 minutes Do not disturb the sediment
so the solids settle at the in the bottom of the vials.
bottom of the vials.
13. Add the contents of one 14. Add the contents of one 15. Put the caps on both 16. Start the instrument
Ammonia Salicylate Ammonia Cyanurate vials. Shake to dissolve the timer. A 20-minute reaction
Reagent Powder Pillow (for Reagent Powder Pillow )for powder completely. A green time starts.
5-mL samples) to each vial. 5-mL samples) to each vial. color shows if nitrogen is
present.
Nitrogen, Total Inorganic, Titanium Trichloride Reduction TNT Method (25.0 mg/L) 3
Zero
17. When the timer expires, 18. Insert the blank vial into 19. Push ZERO. The 20. Clean the sample vial.
clean the blank vial. the 16-mm cell holder. display shows 0.0 mg/L N.
Read
21. Insert the sample vial 22. Push READ. Results

into the 16-mm cell holder. show in mg/L N.
Interferences
The substances in Table 2 may interfere when present. The substances in Table 3 do not
interfere below the levels listed.
Table 2 Interfering substances
Calcium Causes a positive interference at 1000 mg/L as CaCO3
Manganese (IV) Causes a negative interference at 3 mg/L
Magnesium Causes a positive interference at 1000 mg/L as CaCO3
Sulfide Causes a negative interference at 3 mg/L
Sulfate Causes a negative interference at 250 mg/L
Table 3 Non-interfering substances

Al3+ 8 mg/L
Ba2+ 40 mg/L
Cu2+ 40 mg/L
Fe3+ 8 mg/L
Zn2+ 80 mg/L
F– 40 mg/L
3--
PO4 P 8 mg/L
SiO2 80 mg/L
EDTA 80 mg/L
Accuracy check
Items to collect:
• Nitrate Nitrogen PourRite Ampule Standard, 500-mg/L NO3––N
• Ampule breaker
Mix well.
instrument.

instrument.
Items to collect:
• 10.0-mg/L Nitrate Nitrogen Standard Solution
Method performance
– – –
Species recovery
The total inorganic nitrogen test is designed to provide an estimate of the total nitrite,
nitrate and ammonia nitrogen load in a water or wastewater sample. This test is most
applicable to the monitoring of samples taken from an industrial process stream or a
wastewater treatment stream where it is important to track the inorganic nitrogen load as
it passes through the treatment process. The test does show different recoveries of each
Nitrogen, Total Inorganic, Titanium Trichloride Reduction TNT Method (25.0 mg/L) 5
of the three nitrogen species, as shown in Table 4. The test is not recommended for use
when quantifying only one of the three species. In that case, specific procedures for each
particular analyte would be more appropriate.
Table 4 Species recovery
Nitrogen form Percent recovery
NH3–N 112%
NO3 ––N 100%
NO2––N 77%
Summary of method
Titanium (III) ions reduce nitrate and nitrite to ammonia in a basic environment. After
centrifugation to remove solids, the ammonia is combined with chlorine to form
monochloramine. Monochloramine reacts with salicylate to form 5-aminosalicylate. The 5-
aminosalicylate is oxidized in the presence of a sodium nitroprusside catalyst to form a
blue colored compound. The blue color is masked by the yellow color from the excess
reagent present to give a final green colored solution. The measurement wavelength is
655 nm for spectrophotometers or 610 nm for colorimeters.
Required reagents
Nitrogen, Total Inorganic, Pretreatment Reagent Set (TiCl3 Reduction

— 25 tests 2604945
Method)
Nitrogen Ammonia, Reagent Set, Low Range Test 'N Tube ™ AmVer™ 2 25 tests 2604545
Water, deionized varies 100 mL 27242
Required apparatus
Centrifuge, 115 VAC, 6 x 15 mL 1 each 2676500

OR
Centrifuge, 220 VAC, 6 x 15 mL 1 each 2676502
Gloves, nitrile, large (other sizes available) 1 pair 100/pkg 2550503
Flask, volumetric, 50-mL each 1457441

Nitrate Nitrogen Standard Solution, 10.0-mg/L NO3--N 500 mL 30749
Nitrate Nitrogen Standard Solution, 2-mL PourRite® Ampule, 500 mg/L 20/pkg 1426020


50 mL
SCDB
Sodium Thiosulfate, 0.1 N 100 mL 32332
®
PourRite Ampule Breaker, 2-mL each 2484600
Nitrate Nitrogen Standard Solution, 1-mg/L NH3-N 500 mL 204649
1
Nitrogen, Total Kjeldahl DOC316.53.01091
Nessler Method1 Method 8075

1 to 150 mg/L TKN Reagent Solution
Scope and application: For water, wastewater and sludge; digestion is required.
1 Adapted from Hach, et. al., Journal of Association of Official Analytical Chemists, 70(5) 783-787 (1987); Hach, et. al., Journal of
Agricultural and Food Chemistry, 33(6) 1117-1123 (1985); Standard Methods for the Examination of Water and Wastewater.
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
If the Pour-Thru Cell is used (for applicable instruments), clean the cell periodically. Pour a few sodium thiosulfate
pentahydrate crystals into the cell funnel or rinse the cell with a solution of sodium thiosulfate. Flush the crystals through the
funnel and cell with enough deionized water to dissolve. Rinse the cell with deionized water.
The Nessler reagent contains mercuric iodide. Both the reacted sample and blank will contain mercury. Do not pour these
solutions down the drain. Collect the reacted samples and the blank for proper disposal.
Hold the reagent droppers and dropper bottles vertically, not at an angle, when the reagent is added.
Use the Standard Adjust option with each new lot of reagent for the best results.
equipment.
2 Nitrogen, Total Kjeldahl, Nessler Method (150 mg/L)

Items to collect
Boiling chips, silicon carbide 2-3

Cylinder, graduated mixing, 25-mL 2
Finger cots 2
Digesdahl Digestion Apparatus 1
Hydrogen Peroxide, 50% 20 mL
Mineral Stabilizer 6 drops
Nessler Reagent 2 mL
Polyvinyl Alcohol Dispersing Agent 6 drops
Potassium Hydroxide (KOH) Standard Solution, 1.0 N varies
Potassium Hydroxide (KOH) Standard Solution, 8.0 N varies
Sulfuric Acid, ACS, concentrated 6 mL
TKN Indicator Solution 2 drops
Pipet, TenSette, 0.1–1.0 mL, plus tips 1
Safety shield 1
2

3
Nessler method
Start
1. Start program 2. Prepare the sample: 3. Prepare the blank: 4. Use a pipet to move an
399 Nitrogen, TKN. For Use the Digesdahl Digestion Digest an equal amount of analysis volume of the
information about sample Apparatus Instruction deionized water for use as digested sample to a
cells, adapters or light Manual to digest the sample the blank . graduated mixing cylinder.
shields, refer to Instrument amount. Refer to Digested Refer to Digested sample
specific information sample volumes volumes on page 5.
on page 1. on page 5.
5. Use a pipet to transfer an 6. Add one drop of TKN 7. If the aliquot is less that 8. Add 1.0 N KOH to each
equal amount of digested Indicator to eacy cylinder. 1 mL, go to step 8. If the cylinder, one drop at a time.
deionized water to a second aliquot is greater than 1 mL, Mix after each addition.
graduated mixing cylinder. add drops of 8.0 KOH to Continue until the first
each cylinder until the first permanent blue color
flash of blue color shows. shows.
Put the stopper in the
cylinder and invert after
each addition.
9. Fill both cylinders to the 10. Add three drops of 11. Put the stoppers in the 12. Add three drops of
20-mL mark with deionized Mineral Stabilizer to each cylinders and invert to mix. Polyvinyl Alcohol Dispersing
water. cylinder. Agent to each cylinder.
Nitrogen, Total Kjeldahl, Nessler Method (150 mg/L) 3

13. Put the stoppers in the 14. Fill both cylinders to the 15. Put the stoppers in the 16. Use a pipet to add
cylinders and invert to mix. 25-mL mark with deionized cylinders and invert several 1.00 mL of Nessler Reagent
water. times to mix. to each cylinder.
17. Put the stoppers in the 18. Start the instrument 19. When the timer expires, 20. Clean the blank.
cylinders and invert to mix. timer. A 2-minute reaction pour the contents of each
The solution should not be time starts. cylinder into separate
hazy. Any turbidity (haze) sample cells.
will cause incorrect results.
Zero
21. Insert the blank into the 22. Push ZERO. The 23. Clean the prepared 24. Insert the prepared
cell holder. display shows 0 mg/L TKN. sample. sample into the cell holder.

Read
25. Push READ. Results 26. Calculate the sample

show in mg/L TKN. TKN in ppm:
TKN = (75 x A) ÷ (B x C)
Where:
• A = mg/L read from the

display
• B = g (or mL of water)
sample taken for the
digestion
• C = mL analysis volume
of the digested sample
Digested sample volumes

Table 2 Aqueous samples (solutions or suspensions in water—less than 1% solids)
Expected nitrogen concentration (mg/L) Analysis volume (mL)
0.5–28 10
2–112 5
11–560 2
45–2250 1
425–22500 0.5
Table 3 Dry samples

42–2200 10
106–5600 5
350–18,000 2
1000–56,000 1
4200–220,000 0.5
Table 4 Oils and fats

85–4500 10
210–11,000 5
2100–110,000 1

Accuracy check
Digestion method
To validate the digestion method, use the Primary Standards for Kjeldahl Nitrogen that
are given in the Accuracy Check section of the Digesdahl® Digestion Apparatus
Instruction Manual. Use the accuracy check procedure to find the digestion efficiency and
the amount of bound nitrogen that is released during digestion.
Use the digested Kjeldahl standard in the Nessler test procedure to measure the TKN of
the primary standard. The TKN value should be within ± 3% of the value of the prepared
Kjeldahl standard.

Items to collect:
• 1.0-mg/L NH3–N standard solution
• TKN indicator
• Dropper
• 25-mL graduated mixing cylinders (2)
• Deionized water
• Mineral Stabilizer
• Polyvinyl Alcohol Dispersing agent
1. Add one drop of TKN Indicator to each 25-mL graduated mixing cylinder.
2. Fill one cylinder to the 20-mL mark with deionized water. Fill the other cylinder to the
20-mL mark with a 1.0-mg/L NH3–N standard solution.
3. Add 3 drops of Mineral Stabilizer to each cylinder. Invert several times to mix.
4. Add 3 drops of Polyvinyl Alcohol Dispersing agent to each cylinder. Invert several
times to mix.
5. Continue with the TKN procedure to measure the concentration of the standard
solution. Accurate calibrations will show 26–27 mg/L TKN.
Method performance
399 76 mg/L NH3–N 70–82 mg/L NH3–N 1 mg/L NH3–N
Summary of method
The term Total Kjeldahl Nitrogen refers to the combination of ammonia and organic
nitrogen. However, only the organic nitrogen compounds that are present as organically
bound nitrogen in the trinegative state are determined in this test. Nitrogen in this form is
converted into ammonium salts by the action of sulfuric acid and hydrogen peroxide. The
ammonia is then analyzed by a modified Nessler method test. The measurement
The Nessler reagent contains mercuric iodide. The reacted samples and blanks will
contain mercury and must be disposed of as a hazardous waste. Dispose of reacted
solutions according to local, state and federal regulations.

Required reagents
Nitrogen Reagent Set, 0-150 mg/L, Nessler Method — 250 tests 2495300
Includes:
Hydrogen Peroxide, 50% 20 mL 490 mL 2119649
50 mL
Mineral Stabilizer 6 drops 2376626
SCDB
Nessler Reagent 2 mL 500 mL 2119449
50 mL
Polyvinyl Alcohol Dispersing Agent 6 drops 2376526
SCDB
50 mL
Potassium Hydroxide Standard Solution, 1.0 N varies 2314426
SCDB
100 mL
Potassium Hydroxide Standard Solution, 8.0 N varies 28232H
MDB
Sulfuric Acid, concentrated, ACS varies 500 mL 97949
50 mL
TKN Indicator Solution 2 drops 2251926
SCDB
Required apparatus
Boiling chips, silicon carbide 2-3 500 g 2055734

Cylinder, graduated mixing, 25-mL 2 each 2636240
Digesdahl® Digestion Apparatus, 115 VAC 1 each 2313020
OR
Digesdahl® Digestion Apparatus, 220 VAC 1 each 2313021
Finger cots 2 2/pkg 1464702
Safety shield 1 each 5003000
Kjeldahl Nitrogen Primary Standard Set set of 3 2277800

Wastewater Influent Standard, Mixed Parameter, for NH3-N, NO3-N, PO4, COD, SO4,
500 mL 2833149
TOC
Sodium Thiosulfate, Pentahydrate 454 g 46001

Pour-Thru Cell Kit (DR 2700, DR 2800) each 5940400

Pour-Thru Cell Kit (DR 5000) each LZV479

PourRite® Ampule Breaker, 2-mL each 2484600
Paper, for weighing, 100 x 100 mm 500/pkg 1473885
Pipet, TenSette®, 1.0 to 10.0 mL each 1970010
Nitrogen Ammonia Standard Solution, 100-mg/L as NH3–N 500 mL 2406549
Nitrogen, Ammonia Standard Solution, 1000-mg/L NH3-N 1L 2354153
Nitrogen Ammonia Standard Solution, 2-mL PourRite® Ampules, 50 mg/L 20/pkg 1479120


Nitrogen, Total DOC316.53.01085
Persulfate Digestion Method Method 10072

2 to 150 mg/L N (HR) Test ‘N Tube™ Vials
Scope and application: For water and wastewater.
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
Digestion is required for total nitrogen determinations.
The vials must be mixed carefully for accurate results. Start each vial inversion with the vial in the vertical position, with the
cap on the top. Turn the vial upside-down and wait for all of the solution to flow down to the cap. Return the vial to the
vertical position and wait for all of the solution to flow down to the bottom of the vial. This mixing method equals one
inversion.
If the test result is over-range, dilute a fresh portion of sample and repeat the complete test procedure. The digestion must
be repeated for accurate results.
Use the deionized water that is supplied in the reagent set or organic-free water for the blank vial and for the preparation of
standard solutions.
equipment.
Items to collect
Test 'N Tube HR Total Nitrogen Reagent Set 1

DRB200 Reactor 1
Finger cots 2
1
Funnel, micro 1
1
Test tube rack 1

Persulfate digestion for Test 'N Tubes
1. Start the 2. Use a funnel to add the 3. Prepare the sample: 4. Prepare the blank: Add
DRB200 Reactor. Set the contents of one Total Add 0.5 mL of sample to 0.5 mL of deionized water
temperature to 105 °C. Nitrogen Persulfate Reagent one of the vials. (included in the kit) to the
Powder Pillow to each of second vial.
two HR Total Nitrogen Use only water that is free of
Hydroxide Digestion all nitrogen-containing
Reagent vials. species as a substitute for
Make sure to clean any the provided deionized
reagent that gets on the lip water.
of the vials or on the vial
threads.
2 Nitrogen, Total, Persulfate Digestion TNT Method (150 mg/L)

Start
5. Put the caps on both 6. Put the vials in the 7. At 30 minutes, use finger 8. Start program 394 N,
vials. Shake vigorously for reactor and close the lid. cots to immediately remove Total HR TNT. For
at least 30 seconds to mix. Leave the vials in the the vials from the reactor. information about sample
Undissolved powder will not reactor for exactly Let the vials cool to room cells, adapters or light
affect the accuracy of the 30 minutes. temperature. shields, refer to Instrument
test. specific information
on page 1.
9. Add the contents of one 10. Put the caps on both 11. Start the instrument 12. After the timer expires,
Total Nitrogen (TN) Reagent vials. Shake for 30 seconds. timer. A 3-minute reaction remove the caps from the
A Powder Pillow to each time starts. vials. Add one TN Reagent
vial. B Powder Pillow to each
vial.
13. Put the caps on both 14. Start the instrument 15. Prepared sample: 16. Blank: When the timer
vials. Shake vigorously for timer. A 2-minute reaction When the timer expires, use expires, use a pipet to put
15 seconds to mix. The time starts. a pipet to put 2 mL of the 2 mL of the digested,
reagent will not dissolve digested, treated prepared treated blank into the
completely. Undissolved sample into one TN second TN Reagent C vial.
powder will not affect the Reagent C vial.
accuracy of the test.
The solution will start to turn
yellow.
Nitrogen, Total, Persulfate Digestion TNT Method (150 mg/L) 3

17. Put the caps on both 18. Start the instrument 19. When the timer expires, 20. Insert the blank vial into
vials. Invert 10 times to mix. timer. A 5-minute. reaction clean the blank vial. the 16-mm cell holder.
Use slow, deliberation time starts.
inversions for complete The yellow color will
recovery. The vials will be intensify.
warm to the touch.
Zero Read
21. Push ZERO. The 22. Clean the sample vial. 23. Insert the sample vial 24. Push READ. Results
display shows 0 mg/L N. into the 16-mm cell holder. show in mg/L N.
Blanks for colorimetric measurement

The reagent blank can be used for up to 7 days for measurements that use the same lot
of reagents. Keep the reagent blank in the dark at room temperature (18–25 °C). If a
small amount of white floc appears within a week, discard the reagent blank and prepare
a new one.
Interferences
The substances in the Table 2 have been tested and found not to interfere up to the
indicated levels (in mg/L). Interfering substances that resulted in a concentration change
of ±10% appear in the Table 3.
Barium 10.4 mg/L
Calcium 1200 mg/L
Chromium (3+) 2 mg/L
Iron 8 mg/L
Lead 26.4 µg/L
Magnesium 2000 mg/L
Organic Carbon 600 mg/L
Phosphorus 400 mg/L
Silica 600 mg/L

Table 2 Non-interfering substances (continued)
Silver 3.6 mg/L
Tin 6 mg/L

Bromide > 240 mg/L; positive interference
Chloride > 3000 mg/L; positive interference
This test performed with standard nitrogen solutions prepared from the following
compounds obtained 95% recovery:
• Ammonium chloride
• Ammonium sulfate
• Ammonium acetate
• Glycine
• Urea
Ammonium chloride or nicotinic-PTSA spikes in domestic influent, effluent and the ASTM
standard specification for substitute wastewater (D 5905-96) also resulted in ≥ 95%
recovery.
The large amounts of nitrogen-free organic compounds in some samples may decrease
digestion efficiency by consuming some of the persulfate reagent. Samples known to
contain high levels of organics should be diluted and re-run to verify digestion efficiency.
Accuracy check
Digestion method
For proof of accuracy use Primary Standards for Kjeldahl Nitrogen. This method generally
gives 95–100% recovery on organic nitrogen standards. Analysts have found Ammonia-
PTSA (p-Toluenesulfonate) to be the most difficult to digest. Other compounds may yield
different percent recoveries.
Items to collect:
• Primary Standard for Kjeldahl Nitrogen (Ammonia-PTSA, Glycine-PTSA or Nicotinic-
PTSA)
• Deionized water (use the deionized water supplied in the reagent set or water that is
free of all organic and nitrogen-containing species)
1. Prepare a 120-mg/L N equivalent standard.

a. Weigh the applicable standard:
• Ammonia-PTSA: 1.6208 g
• Glycine-PTSA: 2.1179 g
• Nicotinic-PTSA: 2.5295 g
b. Use a funnel to add the standard to the volumetric flask.
c. Add deionized water to the flask and mix to dissolve the standard.
d. Dilute to the mark with deionized water. Mix well.
2. Use the test procedure to measure the concentration of the nitrogen standard.
Calculate the percent recovery as follows:
% recovery = [(measured concentration)/120] x 100
Note: The minimum expected % recovery for each standard is 95%.

Items to collect:
• Ammonia Nitrogen Standard Solution, 1000-mg/L as NH3–N
• Ampule breaker
Mix well.
instrument.

instrument.
Items to collect:
• 100-mg/L ammonia nitrogen standard solution
Method performance
394 100 mg/L NH3–N 98–102 mg/L N 0.5 mg/L N
Summary of method
An alkaline persulfate digestion converts all forms of nitrogen to nitrate. Sodium
metabisulfite is added after the digestion to eliminate halogen oxide interferences. Nitrate
then reacts with chromotropic acid under strongly acidic conditions to form a yellow
complex. The measurement wavelength is 410 nm for spectrophotometers or 420 nm for
colorimeters.

Required reagents
Nitrogen, Total, Test 'N Tube™ Reagent Set 50 vials 2714100
Required apparatus
DRB200 Reactor, 110 V, 15 x 16 mm 1 each LTV082.53.40001

OR
Pipet, TenSette 1.0–10.0 mL 1 each 1970010
Nitrogen Ammonia Standard Solution, 1000-mg/L as NH3–N 1L 2354153

Nitrogen Ammonia Standard Solution, 100-mg/L as NH3–N 500 mL 2406549
Sulfuric Acid, ACS 500 mL 97949
500 mL 2833149
TOC
Water, deionized 500 mL 27249
Water, organic-free 500 mL 2641549
PourRite® Ampule Breaker, 2-mL each 2484600
Nitrogen Ammonia Standard Solution, 150-mg/L NH3–N, 10-mL Voulette® Ampules 16/pkg 2128410
Nitrogen Ammonia Standard Solution, 2-mL PourRite® Ampules, 50 mg/L 20/pkg 1479120

04/2013, Edition 8
Nitrogen, Total DOC316.53.01086
Persulfate Digestion Method Method 10071

0.5 to 25.0 mg/L N (LR) Test ‘N Tube™ Vials
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
Digestion is required for total nitrogen determinations.
The vials must be mixed carefully for accurate results. Start each vial inversion with the vial in the vertical position, with the
cap on the top. Turn the vial upside-down and wait for all of the solution to flow down to the cap. Return the vial to the
vertical position and wait for all of the solution to flow down to the bottom of the vial. This mixing method equals one
inversion.
If the test result is over-range, dilute a fresh portion of sample and repeat the complete test procedure. The digestion must
be repeated for accurate results.
Use the deionized water that is supplied in the reagent set or organic-free water for the blank vial and for the preparation of
standard solutions.
equipment.
Items to collect
Test 'N Tube LR Total Nigtrogen Reagent Set 1

DRB200 Reactor 1
Finger cots 2
1
Funnel, micro 1
1
Test tube rack 1 to 3

Persulfate digestion for Test 'N Tubes
1. Start the 2. Use a funnel to add the 3. Prepare the sample: 4. Prepare the blank: Add
DRB200 Reactor. Set the contents of one Total Add 2 mL of sample to one 2 mL of deionized water
temperature to 105 °C. Nitrogen Persulfate Reagent of the vials. (included in the kit) to the
Powder Pillow to each of second vial.
two Total Nitrogen Use only water that is free of
Hydroxide Digestion all nitrogen-containing
Reagent vials. species as a substitute for
Make sure to clean any the provided deionized
reagent that gets on the lip water.
of the vials or on the vial
threads.
2 Nitrogen, Total, Persulfate Digestion TNT Method (25.0 mg/L)

Start
5. Put the caps on both 6. Put the vials in the 7. At 30 minutes, use finger 8. Start program 350 N,
vials. Shake vigorously for reactor and close the lid. cots to immediately remove Total LR TNT. For
at least 30 seconds to mix. Leave the vials in the the vials from the reactor. information about sample
Undissolved powder will not reactor for exactly Let the vials cool to room cells, adapters or light
affect the accuracy of the 30 minutes. temperature. shields, refer to Instrument
test. specific information
on page 1.
9. Add the contents of one 10. Put the caps on both 11. Start the instrument 12. After the timer expires,
Total Nitrogen (TN) Reagent vials. Shake for 15 seconds. timer. A 3-minute reaction remove the caps from the
A Powder Pillow to each time starts. vials. Add one TN Reagent
vial. B Powder Pillow to each
vial.
13. Put the caps on both 14. Start the instrument 15. Prepared sample: 16. Blank: When the timer
vials. Shake for 15 seconds timer. A 2-minute reaction When the timer expires, use expires, use a pipet to put
to mix. The reagent will not time starts. a pipet to put 2 mL of the 2 mL of the digested,
dissolve completely. digested, treated prepared treated blank into the
Undissolved powder will not sample into one TN second TN Reagent C vial.
affect the accuracy of the Reagent C vial.
test.
The solution will start to turn
yellow.
Nitrogen, Total, Persulfate Digestion TNT Method (25.0 mg/L) 3

17. Put the caps on both 18. Start the instrument 19. When the timer expires, 20. Insert the blank vial into
vials. Invert 10 times to mix. timer. A 5-minute reaction clean the blank vial. the 16-mm cell holder.
Use slow, deliberation time starts.
inversions for complete The yellow color will
recovery. The vials will be intensify.
warm to the touch.
Zero Read
21. Push ZERO. The 22. Clean the sample vial. 23. Insert the sample vial 24. Push READ. Results
display shows 0.0 mg/L N. into the 16-mm cell holder. show in mg/L N.
Multiple samples can be
measured after "zero" is set
with the blank.
Blanks for colorimetric measurement

The reagent blank can be used for up to 7 days for measurements that use the same lot
of reagents. Keep the reagent blank in the dark at room temperature (18–25 °C). If a
small amount of white floc appears within a week, discard the reagent blank and prepare
a new one.
Interferences
The substances in the Table 2 have been tested and found not to interfere up to the
indicated levels (in mg/L). Interfering substances that resulted in a concentration change
of ±10% appear in the Table 3.
Barium 2.6 mg/L
Calcium 300 mg/L
Chromium (3+) 0.5 mg/L
Iron 2 mg/L
Lead 6.6 µg/L
Magnesium 500 mg/L
Organic Carbon 150 mg/L
Phosphorus 100 mg/L
Table 2 Non-interfering substances (continued)
Silica 150 mg/L
Silver 0.9 mg/L
Tin 1.5 mg/L

Bromide > 60 mg/L; positive interference
Chloride > 1000 mg/L; positive interference
This test performed with standard nitrogen solutions prepared from the following
compounds obtained 95% recovery:
• Ammonium chloride
• Ammonium sulfate
• Ammonium acetate
• Glycine
• Urea
Ammonium chloride or nicotinic-PTSA spikes in domestic influent, effluent and the ASTM
standard specification for substitute wastewater (D 5905-96) also resulted in ≥ 95%
recovery.
The large amounts of nitrogen-free organic compounds in some samples may decrease
digestion efficiency by consuming some of the persulfate reagent. Samples known to
contain high levels of organics should be diluted and re-run to verify digestion efficiency.
Accuracy check
Digestion method
For proof of accuracy use Primary Standards for Kjeldahl Nitrogen. This method generally
gives 95–100% recovery on organic nitrogen standards. Analysts have found Ammonia-
PTSA (p-Toluenesulfonate) to be the most difficult to digest. Other compounds may yield
different percent recoveries.
Items to collect:
• Primary Standard for Kjeldahl Nitrogen (Ammonia-PTSA, Glycine-PTSA or Nicotinic-
PTSA)
• Deionized water (use the deionized water supplied in the reagent set or water that is
free of all organic and nitrogen-containing species)
1. Prepare a 25-mg/L N equivalent standard.

a. Weigh the applicable standard:
• Ammonia-PTSA: 0.3379 g
• Glycine-PTSA: 0.4416 g
• Nicotinic-PTSA: 0.5274 g
b. Use a funnel to add the standard to the volumetric flask.
c. Add deionized water to the flask and mix to dissolve the standard.
d. Dilute to the mark with deionized water. Mix well.
2. Use the test procedure to measure the concentration of the nitrogen standard.
Calculate the percent recovery as follows:
% recovery = [(measured concentration)/25] x 100
Note: The minimum expected % recovery for each standard is 95%.

Items to collect:
• Ammonia Nitrogen Standard Solution, 1000-mg/L as NH3–N
• Ampule breaker
Mix well.
instrument.

instrument.
Items to collect:
• 10-mg/L ammonia nitrogen standard solution
Method performance
350 10 mg/L NH3–N 9.6–10.4 mg/L N 0.5 mg/L N
Summary of method
An alkaline persulfate digestion converts all forms of nitrogen to nitrate. Sodium
metabisulfite is added after the digestion to eliminate halogen oxide interferences. Nitrate
then reacts with chromotropic acid under strongly acidic conditions to form a yellow
complex. The measurement wavelength is 410 nm for spectrophotometers or 420 nm for
colorimeters.

Required reagents
Nitrogen, Total, LR, Test 'N Tube™ Reagent Set 50 vials 2672245
Required apparatus

OR
Nitrogen, Ammonia Standard Solution, 1000-mg/L NH3-N 1L 2354153

500 mL 2833149
TOC
Water, deionized 500 mL 27249
Water, organic-free 500 mL 2641549

Ampule Breaker, PourRite® ampules each 2484600
Nitrogen, Ammonia Standard Solution, 2-mL PourRite Ampule, 50 mg/L 20/pkg 1479120
Nitrogen, Ammonia Standard Solution, 10-mL Voluette Ampules, 50 mg/L 16/pkg 1479110




Organic Carbon, Total DOC316.53.01093
Direct Method1 Method 10129

0.3 to 20.0 mg/L C (LR) Test ‘N Tube™ Vials
Scope and application: For water, drinking water and wastewater
1 U.S. Patent 6,368,870
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
A reagent blank is required for each series of samples.
To test for higher concentrations of TOC, use Method 10173 or method 10128.
equipment.
Items to Collect
Total Organic Carbon Direct Method Low Range Test 'N Tube Reagent Set 1
Cylinder, graduated, 10-mL 1
DRB200 Reactor 1
Flask, Erlenmeyer, 50-mL 1
Light shield and adapter (For information about sample cells, adapters or light shields, refer to
1
Magnetic stirrer 1
1
Items to Collect (continued)
Paper, pH 1
Stir bar, magnetic 1
Test tube rack 1
Water, organic-free 3.0 mL
Wipes, disposable 1
Sample collection
• Collect samples in clean glass bottles.
• Homogenize samples that contain solids to get a representative sample.
• Rinse the sample bottle several times with the sample to be collected.
• Fill the bottle completely full, then tighten the cap on the bottle.
• Acid preservation is not recommended.
1. Start the 2. Add 10 mL of sample to 3. Add 0.4 mL of Buffer 4. Put the flask on a stir
DRB200 Reactor. Select the a 50-mL Erlenmeyer flask. Solution to the Erlenmeyer plate. Stir at a moderate
TOC program. Put the stir bar in the flask, pH 2.0. Use pH paper speed for 10 minutes.
Erlenmeyer flask. to make sure that the
sample pH is 2.
2 Organic Carbon, Total, Direct TNT Method (20.0 mg/L)

5. Put a label that says 6. Prepare the blank: Add 7. Prepare the sample: 8. Use deionized water to
"Reagent Blank" on one 3.0 mL of organic-free water Add 3.0 mL of sample from rinse two blue Low Range
Low Range Acid Digestion to the blank vial. the Erlenmeyer flask to the Indicator Ampules. Clean
vial. Put a lable that says sample vial. the ampules with a wipe. Do
"Sample" on a second Low not touch the sides of the
Range Acid Digestion vial. ampules after they are
Add the contents of one clean. Hold the ampules by
TOC Persulfate Powder the top.
Pillow to each Acid
Digestion Vial.
9. Put one unopened 10. Close the vials tightly. 11. Close the reactor. Let 12. After two hours, remove
ampule into each Acid Insert them into the reactor. the vials react for 2 hours at the vials from the reactor.
Digestion Vial. Snap the top 103 to 105 °C. Put them in a test tube rack
off of the ampule when the to cool for one hour. Make
score aligns with the top of sure that the vials stay in an
the vial. Let the ampules upright position at all times.
drop into the vials. The liquid in the blank
Do not invert or tilt the should show a dark blue
vials after the ampule is color.
inside.
Organic Carbon, Total, Direct TNT Method (20.0 mg/L) 3

Start Zero
13. Start program 14. Clean the blank vial. 15. Insert the blank vial into 16. Push ZERO. The
427 Organic Carbon LR. the 16-mm cell holder. display shows 0.0 mg/L C.
For information about
sample cells, adapters or
Instrument specific
Read
into the 16-mm cell holder. show in mg/L C.
Interferences
If the sample contains more than 600 mg/L CaCO3 alkalinity, add sulfuric acid to lower
the sample pH to less than 7, then start the test procedure.
Most sample turbidity is either dissolved during the digestion stage or settled during the
cooling period. Sample turbidities up to 50 NTU have been tested without interference.
The table that follows shows the substances that were tested for interference and did not
interfere up to the levels shown.
Aluminum 10 mg/L Al
Ammonia Nitrogen 1000 mg/L as N
ASTM Wastewater No effect
Bromide 500 mg/L Br–
Bromine 25 mg/L Br2
Chloride 500 mg/L Cl–
Chlorine 10 mg/L Cl2
Chlorine Dioxide 6 mg/L ClO2

Copper 10 mg/L Cu
Cyanide 10 mg/L CN–
Iodide 50 mg/L I–
Iron (II) 10 mg/L Fe2+
Iron (III) 10 mg/L Fe3+
Manganese (VII) 1 mg/L Mn
Monochloramine 14 mg/L NH2Cl as Cl2
Nitrite 500 mg/L NO2-
Ozone 2 mg/L O3
Phosphate 3390 mg/L PO43-
Silica 100 mg/L SiO2
2-
Sulfate 5000 mg/L SO4
Sulfide 20 mg/L S2-
2-
Sulfite 50 mg/L SO3
Zinc 5 mg/L Zn
Reagent blank water

Water that is used for the reagent blank must contain less than 0.05 mg/L carbon. If the
organic-free water container is left open for extended periods, the water can absorb
carbon dioxide (CO2) from the atmosphere and contaminate the blank. To remove the
dissolved CO2 from the organic-free water, acidify the water and stir for 10 minutes as in
the test procedure.
Generally, water that is stored in plastic containers is not suitable for low-range TOC
blanks. Water that is stored in plastic can become contaminated with organic compounds
from the container walls. These leached organic compounds usually cannot be removed
by the acid-sparge process.
Accuracy check
Items to collect:
• TOC Standard Solution Ampule, 1000 mg/L C
• 15-mL volumetric pipet and pipet filler
• Organic-free water
1. Prepare a 150-mg/L total organic carbon standard solution as follows:

a. Use a pipet to add 15.00 mL of a 1000 mg/L TOC standard solution into the
volumetric flask.
b. Dilute to the mark with organic-free water. Mix well. Prepare this solution daily.

0.3 mL of the prepared standard solution, respectively, to three Acid Digestion Vials.
6. Add the contents of one TOC Persulfate Powder Pillow to each vial.
7. Add 3.0 mL of sample to each vial. Swirl to mix.
instrument.

instrument.
Items to collect:
• TOC Standard Solution Ampule 1000 mg/L C
• Organic-free reagent water
1. Prepare a 10.0 mg/L C standard solution as follows:

a. Use a pipet to add 10.00 mL of 1000 mg/L total organic carbon standard solution
into the volumetric flask.
b. Dilute to the mark with organic-free reagent water. Mix well. Prepare this solution
daily.
solution.
Method performance
427 10.0 mg/L C 9.1–10.9 mg/L C 0.2 mg/L C
Summary of method
The total organic carbon (TOC) concentration is determined by first sparging the sample
under slightly acidic conditions to remove the inorganic carbon. In the outside vial,
organic carbon in the sample is digested by persulfate and acid to form carbon dioxide.
During digestion, the carbon dioxide diffuses into a pH indicator reagent in the inner
ampule. The absorption of carbon dioxide into the indicator forms carbonic acid. Carbonic
acid changes the pH and thus the color of the indicator solution. The amount of color
change is related to the original amount of carbon present in the sample. The
measurement wavelengths are 598 and 430 nm for spectrophotometers or 610 nm for
colorimeters.

Required reagents
pH Paper 1 5/pkg 39133

Water, Organic-free 3.0 mL 500 mL 2641549
Reagent Set, Total Organic Carbon Direct Method Low Range Test 'N
— 50 vials 2760345
Tube™
Includes:
Acid Digestion Solution Vials, Low Range TOC (not sold separately) 1 50/pkg —
Buffer Solution, Sulfate (not sold separately; see alternate size below) 0.4 mL 25 mL 45233
Indicator Ampule, Low Range TOC (not sold separately) 1 10/pkg —
TOC Persulfate Powder Pillows (not sold separately) 1 50/pkg —
Required apparatus
Cylinder, graduated, 10-mL 1 each 50838

OR
Flask, Erlenmeyer, 50-mL 1 each 50541
Magnetic Stirrer 1 each 2881200
Stir bar, magnetic 1 each 4531500
Wipes, Disposable 1 280/pkg 2097000
TOC Standard Solution Ampule (KHP Standard, 1000-mg/L C) 5/pkg 2791505

Buffer Solution, Sulfate pH 2.0 500 mL 45249


Potassium Acid Phthalate, ACS 500 g 31534
Sulfuric Acid Standard Solution, 5.25 N 100 mL 244932



15 to 150 mg/L C (MR) Test ‘N Tube™ Vials
Scope and application: For water, drinking water and wastewater
1 U.S. Patent 6,368,870
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
To test for higher concentrations of TOC, use Method 10128. To test for lower concentrations of TOC, use Method 10129.
equipment.
Items to collect
Total Organic Carbon Direct Method Mid Range Test 'N Tube Reagent Set 1
DRB200 Reactor 1
1
Magnetic stirrer 1
1
Paper, pH 1
Test tube rack 1
Wipes, disposable 1
Sample collection
sample pH is 2.
2 Organic Carbon, Total, Direct TNT Method (150 mg/L)

5. Add the contents of one 6. Prepare the blank: Add 7. Prepare the sample: 8. Use deionized water to
TOC Persulfate Powder 1.0 mL of organic-free water Add 1.0 mL of sample from rinse two blue Mid/High
Pillow to each Acid to the blank vial. the Erlenmeyer flask to the Range Indicator Ampules.
Digestion Vial. Put a label sample vial. Clean the ampules with a
that says "Reagent Blank" wipe. Do not touch the sides
on one Mid Range Acid of the ampules after they
Digestion vial. Put a lable are clean. Hold the ampules
that says "Sample" on a by the top.
second Mid Range Acid
Digestion vial.
Digestion Vial. Snap the top 103 to 105 °C. Put them in a test tube rack
off of the ampule when the to cool for one hour. The
score aligns with the top of liquid in the blank should
the vial. Let the ampules show a dark blue color.
drop into the vials.
Do not invert or tilt the vials
after the ampule is inside.
Organic Carbon, Total, Direct TNT Method (150 mg/L) 3

Start Zero
425 Organic Carbon MR. the 16-mm cell holder. display shows 0 mg/L C.
Instrument specific
Read
Interferences
Aluminum 10 mg/L Al
Bromine 25 mg/L Br2

Copper 10 mg/L Cu
Iodide 50 mg/L I–
Ozone 2 mg/L O3
2-
Sulfide 20 mg/L S2-
2-
Sulfite 50 mg/L SO3
Zinc 5 mg/L Zn
Reagent blank water

the test procedure.
Accuracy check
Items to collect:

volumetric flask.

instrument.

instrument.
Items to collect:
1. Prepare a 100 mg/L C standard solution as follows:

daily.
solution.
Method performance
425 70 mg/L C 68–72 mg/L C 1.2 mg/L C
Summary of method
change is related to the original amount of carbon in the sample. The measurement
wavelengths are 598 and 430 nm for spectrophotometers or 610 nm for colorimeters.

Required reagents

Reagent Set, Total Organic Carbon Direct Method Mid Range Test 'N
— 50 vials 2815945
Tube™
Includes:
Acid Digestion Solution Vials, High Range TOC (not sold separately) 1 50/pkg —
Buffer Solution (not sold separately; see alternate size below) 0.4 mL 25 mL —
Indicator Ampule, MR/HR TOC (not sold separately) 1 10/pkg —
Required apparatus

OR




100 to 700 mg/L C (HR, spectrophotometers) Test ‘N Tube™ Vials
20 to 700 mg/L C (HR, colorimeters)
Scope and application: For wastewater and industrial water
1 U.S. Patent 6,368,870
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
To test for lower concentrations of TOC, use Method 10128 or Method 10129.
equipment.
Items to collect
Total Organic Carbon Direct Method High Range Test 'N Tube Reagent Set 1
DRB200 Reactor 1
1
Magnetic stirrer 1
1
Paper, pH 1
Test tube rack 1
Wipes, disposable 1
Sample collection
Organic Carbon Total HR
sample pH is 2.

5. Add the contents of one 6. Prepare the blank: Add 7. Prepare the sample: 8. Use deionized water to
TOC Persulfate Powder 0.3 mL of organic-free water Add 0.3 mL of sample from rinse two blue Mid/High
Pillow to each Acid to the blank vial. the Erlenmeyer flask to the Range Indicator Ampules.
Digestion Vial. Put a label sample vial. Clean the ampules with a
that says "Reagent Blank" wipe. Do not touch the sides
on one High Range Acid of the ampules after they
Digestion vial. Put a label are clean. Hold the ampules
that says "Sample" on a by the top.
second High Range Acid
Digestion vial.
Digestion Vial. Snap the top 103 to 105 °C. Keep the vials in an upright
off of the ampule when the position at all times. Put
score aligns with the top of them in a test tube rack to
the vial. Let the ampules cool for one hour. The liquid
drop into the vials. in the blank should show a
Do not invert or tilt the vials dark blue color.
after the ampule is inside.

Start Zero
426 Organic Carbon HR. the 16-mm cell holder. display shows 0 mg/L C.
Instrument specific
Read
Interferences
Aluminum 10 mg/L Al
Bromine 25 mg/L Br2

Copper 10 mg/L Cu
Iodide 50 mg/L I–
Ozone 2 mg/L O3
2-
Sulfide 20 mg/L S2-
2-
Sulfite 50 mg/L SO3
Zinc 5 mg/L Zn
Reagent blank water

the test procedure.
Accuracy check
Items to collect:

volumetric flask.

instrument.

instrument.
Items to collect:
1. Prepare a 300 mg/L C standard solution as follows:

daily.
solution.
Method performance
426 350 mg/L C 337–363 mg/L C 4 mg/L C
Summary of method
change is related to the original amount of carbon in the sample. The measurement
wavelengths are 598 and 430 nm for spectrophotometers or 610 nm for colorimeters.

Required reagents

Total Organic Carbon Direct Method High Range Test ’N Tube™
— 50 vials 2760445
Reagent Set
Includes:
Acid Digestion Solution Vials, High Range TOC (not sold separately) 1 50/pkg —
Buffer Solution, Sulfate (not sold separately; see alternate size below) 0.4 mL 25 mL —
Indicator Ampules, MR/HR TOC (not sold separately) 1 10/pkg —
Required apparatus

OR





Oxygen Demand, Chemical DOC316.53.01099
USEPA1 Reactor Digestion Method2 Method 8000

0.7 to 40.03 mg/L COD (ULR); 3 to 150 mg/L COD (LR); 20 to 1500 mg/L
COD (HR); 200 to 15,000 mg/L COD (HR Plus)
Scope and application: For water and wastewater. Digestion is required.
1 Ranges 3 to 150 mg/L COD and 20 to 1500 mg/L COD are USEPA approved for wastewater analyses (Standard Method 5220 D), Federal
Register, April 21, 1980, 45(78), 26811-26812.
2 Jirka, A.M.; Carter, M.J., Analytical Chemistry, 1975, 47(8), 1397.
3 The ULR is only available with spectrophotometers that can measure at a wavelength of 350 nm.
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
The reagent that is used in this test is corrosive and toxic. Use protection for eyes and skin and be prepared to flush any
spills with running water.
The reagents that are used in this test contain mercury. Collect the reacted samples for proper disposal.
equipment.
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
is on the container label. Refer to Blanks for colorimetric determination on page 4.
Store unused (light sensitive) vials in a closed box.
If the samples contain high concentrations of chloride, refer to the Alternate reagents section.
1
Items to collect
Beaker, 250-mL 1
Blender 1
COD Digestion Reagent vials varies
DRB200 Reactor 1
1
Magnetic stirrer and stir bar 1
Opaque shipping container for storage of unused, light-sensitive reagent vials varies
Pipet, TenSette, 0.1- to 1.0-mL, with pipet tips (for use with the 200–15,000 mg/L range) 1
Pipet, volumetric, 2.00-mL 2
Pipet filler safety bulb 1
Test tube rack 2

• Collect samples in clean glass bottles. Use plastic bottles only if they are known to be
free of organic contamination.
• Test biologically active samples as soon as possible.
Reactor digestion procedure
1. Put 100 mL of sample in 2. For the 200–15,000 mg/L 3. Set the DRB200 Reactor 4. Prepare the sample:
a blender. Blend for range or to improve power to on. Preheat to Remove the cap from a vial
30 seconds or until accuracy and reproducibility 150 °C. for the selected range. Hold
homogenized. of the other ranges, pour the Refer to the DRB200 User the vial at an angle of
For samples with large homogenized sample into a Manual for selecting pre- 45 degrees. Use a clean
amounts of solids, increase 250-mL beaker and gently programmed temperature pipet to add 2.00 mL of
the homogenization time. If stir with a magnetic stir applications. sample to the vial.
the sample does not contain plate. For 250–15,000 mg/L vials:
suspended solids, omit Use a TenSette Pipet to add
steps 1 and 2. 0.20 mL of sample to the
vial.
2 Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L)
5. Prepare the blank: 6. Close the vials tightly. 7. Hold the vials by the cap, 8. Put the vials in the
Remove the cap from a Rinse the vials with water over a sink. Invert gently preheated DRB200 reactor.
second vial for the selected and wipe with a clean paper several times to mix. Close the lid.
range. Hold the vial at an towel. The vials get very hot
angle of 45 degrees. Use a during mixing.
clean pipet to add 2.00 mL
of deionized water to the
vial.
For 250–15,000 mg/L vials:
Use a TenSette Pipet to add
0.20 mL of deionized water
to the vial.
9. Heat the vials for 10. Set the reactor power to 11. Invert each vial several 12. Put the vials in a tube
2 hours. off. Let the vials cool in the times while it is still warm. rack to cool to room
reactor for about temperature.
20 minutes. The vials should
cool to 120 °C or less.
Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L) 3
Colorimetric procedure
Start Zero
1. Start program 431 COD 2. Clean the blank. 3. Insert the blank into the 4. Push ZERO. The display
ULR, 430 COD LR or cell holder. shows 0 or 0.0 mg/L COD.
435 COD HR. For
information about sample
on page 1.
Read
5. Clean the prepared 6. Insert the prepared 7. Push READ. Results 8. If using High Range Plus
sample. sample into the cell holder. show in mg/L COD. COD digestion reagent
vials, multiply the result by
10. For the most accurate
results with samples near
1500 or 15,000 mg/L COD,
repeat the analysis with a
diluted sample.
Blanks for colorimetric determination

The blank vial can be used again and again for measurements that use the same lot of
reagent vials. Measure the absorbance of the blank vial over time and prepare a new
blank vial when the absorbance changes.
1. Put the instrument in the absorbance mode at the applicable wavelength. Refer to
Table 3 on page 7.
2. Add 5 mL of deionized water into an empty vial.
3. Put the vial in the instrument and zero the instrument.
4. Put the blank vial that is used in the test procedure into the instrument and record the
absorbance value.
5. Keep the blank vial in the dark.
6. Prepare a new blank when the absorbance has changed by approximately
0.01 absorbance units.
Interferences
Chloride is the primary interference in this test procedure. Each COD vial contains
mercuric sulfate that removes chloride interference to the level specified in Column 1 of
Table 2. Dilute samples that have higher chloride concentrations to the level given in
Column 2.
Note: For best results, use the low range and ultra-low range vials for samples that have high
chloride concentrations (near maximum concentration) and low COD concentrations.
If sample dilution causes the COD concentration to be too low for accurate
measurements, add 0.50 g of mercuric sulfate (HgSO4) to each COD vial before the
sample is added. The additional mercuric sulfate will increase the maximum chloride
concentration to the level given in Column 3.
Note: Bromide interference is not removed with mercuric sulfate.
Table 2 Chloride concentration limits in the sample
Vial range Column 1 (maximum mg/L Column 2 (mg/L Cl– for Column 3 (maximum mg/L
Cl–) diluted samples) Cl– with mercuric sulfate)
ULR1 (0.7–40.0 mg/L) 2000 1000 N/A
LR (3–150 mg/L) 2000 1000 8000
HR (20–1500 mg/L) 2000 1000 4000
HR Plus (200–15,000 mg/L) 20,000 10,000 40,000
1 The ULR is only available for spectrophotometers that can measure at a wavelength of 350 nm.
Accuracy check
Items to collect:
• 1000 mg/L COD standard solution
• Volumetric pipets, Class A and pipet filler
• Deionized water
• Potassium acid phthalate (KHP), dried overnight at 120 °C (HR Plus only)
0.7 to 40.0 mg/L ULR
1. Prepare a 30-mg/L COD standard solution as follows:
a. Use a pipet to add 3.00 mL of the 1000 mg/L standard solution into a 100-mL
volumetric flask.
3 to 150 mg/L LR
1. Prepare a 100-mg/L COD standard solution as follows:
a. Use a pipet to add 10 mL of the 1000 mg/L standard solution into a 100-mL
volumetric flask.
20 to 1500 mg/L HR
1. Use the test procedure with a 300-mg/L, 800 mg/L or 1000 mg/L COD standard
solution to measure the concentration of the standard solution.
200 to 15,000 mg/L HR Plus

1. Prepare a 10,000 mg/L COD standard solution as follows:
a. Dissolve 8.500 g of dried KHP in 1000-mL of organic-free deionized water.
Alternate reagents Mercury-free COD2 Reagents are available as a mercury-free alternative. These
reagents are fully compatible with test procedures and stored programs in the
instruments. Chloride and ammonia determinations are recommended for accurate
results.
NOTICE
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. More
information is available for use with specific applications.
Method performance
431 (ULR) 30 mg/L COD 28.8–31.2 mg/L COD 0.5 mg/L COD
430 (LR) 80 mg/L COD 77–83 mg/L COD 3 mg/L COD
435 (HR) 800 mg/L COD 785–815 mg/L COD 23 mg/L COD
435 (HR Plus) 8000 mg/L COD 7850–8150 mg/L COD 230 mg/L COD
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 2 hours with
sulfuric acid and a strong oxidizing agent, potassium dichromate. Oxidizable organic
compounds react, reducing the dichromate ion (Cr 2O7 2–) to green chromic ion (Cr3+).
When the 0.7–40.0 or the 3–150 mg/L colorimetric method is used, the amount of Cr6+
that remains is measured. When the 20–1500 mg/L or 200–15,000 mg/L colorimetric
method is used, the amount of Cr3+ that is produced is measured. 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 that are specified in Table 3.
Table 3 Range-specific test wavelengths
Range in mg/L COD Wavelength
0.7 to 40.0 mg/L 350 nm (for applicable instruments)
3 to 150 mg/L 420 nm
20 to 1500 620 nm (610 nm for colorimeters)
2000 to 15,000 mg/L 620 nm (610 nm for colorimeters)

Reacted samples contain mercury, silver and chromium and must be disposed of as a
hazardous waste. Dispose of reacted solutions according to local, state and federal
regulations. Users in the United States can use the ez COD Recycling Service for
disposal of COD vials. Refer to Consumables and replacement items on page 7.
Required reagents
COD, Ultra Low Range, 0.7 to 40 mg/L 1-2 vials 25/pkg 2415825
COD, Low Range, 3 to 150 mg/L 1-2 vials 25/pkg 2125825
COD, High Range, 20 to 1500 mg/L 1-2 vials 25/pkg 2125925
COD, High Range Plus, 200 to 15,000 mg/L 1-2 vials 25/pkg 2415925
Alternate reagents and package sizes
COD2, Low Range, 0 to 150 mg/L COD 1-2 vials 25/pkg 2565025
COD2, High Range, 0 to 1500 mg/L COD 1-2 vials 25/pkg 2565125
COD2, High Range, 0 to 1500 mg/L COD 1-2 vials 150/pkg 2565115
COD2, High Range Plus, 0 to 15,000 mg/L COD 1-2 vials 25/pkg 2834325
COD Digestion Reagent Vials, 3 to 150 mg/L COD 1-2 vials 150/pkg 2125815
COD Digestion Reagent Vials, 200 to 1500 mg/L COD 1-2 vials 150/pkg 2125915
COD Digestion Reagent Vials, ULR 0.7-40.0 mg/L 1-2 vials 150/pkg 2415815
COD Digestion Reagent Vials, HR plus,200-25,000 mg/L 1-2 vials 150/pkg 2415915
Required apparatus
Blender, 2-speed, 120 VAC 1 each 2616100

OR
OR
Pipet filler, safety bulb 1 each 1465100
Pipet, volumetric, Class A, 2.00-mL 1 each 1451536

COD Standard Solution, 300-mg/L 200 mL 1218629
COD Standard Solution, 300-mg/L 500mL 1218649
Oxygen Demand Standard (BOD, COD, TOC), 10-mL ampules 16/pkg 2833510
Stir bar, octagonal each 2095352
Stirrer, electromagnetic, 120 VAC, with electrode stand each 4530001
Stirrer, electromagnetic, 230 VAC, with electrode stand each 4530002
Test tube rack each 1864100
Wipes, disposable 70/pkg 2096900

Mercuric Sulfate 28 g 191520
500 mL 2833149
TOC
EZ COD™ Recycling Service with 5-gal bucket-mail back option (For US customers only.
each 2895405
20 and 55 gallon sizes are also available. )
EZ COD™ Recycling Service with 5-gal bucket- pick up option. (For US customers only.
each 2895405P
20 and 55 gallon sizes are also available. )
Finger cots 2/pkg 1464702
Gloves, chemical resistant, size 9-9.5 pair 24101041
Safety goggles, vented each 2550700
1 Other sizes available
Oxygen Demand, Chemical DOC316.53.01101
Manganese III Reactor Digestion Method (with chloride removal) Method 10067
30 to 1000 mg/L COD Mn Test ‘N Tube™ Vials
Scope and application: For water and wastewater
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
To find if the sample contains chloride, use Quantab ® Titrator Strips for low range chloride.
If the sample COD is expected to exceed 1000 mg/L, dilute the sample. Refer to Multiplication factors for sample dilutions
on page 6.
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
is on the container label.
The reagent blank vial can be used for multiple tests. Fill a clean COD vial with deionized water and use this vial to zero the
instrument, then measure the absorbance of the reagent blank vial. The absorbance value should be approximately 1.41–
1.47. Prepare a new reagent blank vial when the absorbance is outside of this range.
To oxidize resistant organics, samples can be digested for up to four hours. Digest the blank for the same time period as the
samples.
Make sure that the filter disc is not in the center of the vial during the Zero and Read steps. Make sure that the filter disc is
more than 20 mm (0.8 in.) or less than 10 mm (0.4 in.) from the bottom of the vial. If necessary, move the filter disk by gently
swirling or by lightly tapping the vial on a table top.
The Chloride Removal Cartridge can be used only once.
If the sample boils during the digestion, the vial is not properly sealed. Test results will be invalid.
Spilled reagent will affect test accuracy and is hazardous. Do not run tests with spilled vials.
The maximum range of the VPD gauge is 40 inches of water; it will not indicate the full vacuum level obtained. Full vacuum
is 20–25 inches of mercury; this can be measured at the vacuum pump with a gauge calibrated for inches of mercury.
1
equipment.
Items to collect
Blender 1
DRB200 Reactor 1
Forceps, extra fine 1
1
Manganese III COD Reagent Vials, 20 to 1000 mg/L COD 1
Pipet, TenSette, 0.1- to 1.0-mL, with pipet tips 1
Pipet, TenSette, 10.0 to 10.0-mL, with pipet tips 1
Sulfuric Acid, concentrated ACS 1 mL
Test tube rack 1
Vacuum pretreatment device 1
Vacuum pump 1
Vial, glass, for sample and acid 2
Water, deionized varies

• Collect samples in clean glass bottles. Use plastic bottles only if they are known to be
free of organic contamination.
• Test biologically active samples as soon as possible.
2 Oxygen Demand, Chemical, Manganese III Method with chloride removal (1000 mg/L)
Acidified sample preparation
1. Set the DRB200 Reactor 2. Put 100 mL of sample in 3. Prepare the blank: Use 4. Prepare the sample:
power to on. Preheat to a blender. Blend for a pipet to add 9.0 mL of Use a pipet to add 9.0 mL of
150 °C or set to the COD 30 seconds or until deionized water to a glass homogenized sample to a
program. homogenized. mixing cell. second glass mixing cell.
If suspended solids are
present, continue to mix the
sample while the sample is
moved to the mixing cell for
the prepared sample.
5. Use a pipet or dispenser 6. Close the mixing cells

to add 1.0 mL of tightly. Invert several times.
concentrated Sulfuric Acid The cells get hot during
to both mixing cells. mixing. Let the mixing cells
cool to room temperature.
Go to the Vacuum
pretreatment procedure
on page 4.
Oxygen Demand, Chemical, Manganese III Method with chloride removal (1000 mg/L) 3
Vacuum pretreatment procedure
1. Attach the vacuum 2. Write a sample identifier 3. Put the VPD top on the 4. Start the vacuum pump.
pretreatment device (VPD) on each vial. Insert the Mn base. Insert a fresh Chloride Adjust the vacuum regulator
to a vacuum pump that can III COD vials in the Removal Cartridge (CRC) valve on top of the VPD until
create a vacuum of 20– numbered holes in the VPD directly above each Mn III the internal gauge reads
25 inches of mercury. base. Remove the caps COD Reagent Vial. Close 20 inches of water.
Do not use an aspirator type from the vials. any open holes in the VPD
vacuum. with the supplied stoppers.
5. Prepare the blank: Use 6. Prepare the sample: 7. Close the vacuum 8. Open the VPD regulator
a pipet to add 0.60 mL of Use a pipet to add 0.60 mL regulator valve completely to release the vacuum. Turn
acidified blank into the CRC. of each acidified sample into to achieve full vacuum. After the pump off. Remove the
It should take 30– the CRC. one minute of full vacuum, VPD top and set it aside.
45 seconds to pull the liquid move the VPD back and Dispose of the used
through the CRC into the forth several times to Chloride Removal
vial. remove drops that cling to Cartridges.
Note: If the liquid does not the CRC. Go to Sample preparation
flow through the CRC, and measurement
increase the vacuum until on page 5.
the flow starts, then reduce
the vacuum back to
20 inches of water.
Sample preparation and measurement
2. Insert each filter into the 3. Remove the Mn III COD 4. Insert the vials in the
1. Use forceps to remove corresponding Mn III COD vial from the vacuum DRB200 Reactor at 150 °C.
the filter from the top of the vial. Use the numbers on chamber. Put the original Close the cover. Digest the
CRC. the VPD as a guide. Use a caps on and close tightly. samples for one hour.
If the sample does not clean towel or deionized Invert the vials several times
contain suspended solids, it water to clean the forceps to mix.
is not necessary to transfer between samples.
the filter to the digestion vial.
Start
6. Let the vials cool to room 7. Start program 432 COD 8. Clean the blank vial.
5. After an hour, remove temperature in a cool water Mn III. For information about
the vials from the DRB200. bath, or hold under running sample cells, adapters or
Let the vials cool in a rack tap water for several light shields, refer to
for two minutes. minutes. Instrument specific
If the solution develops a information on page 1.
colorless upper layer and a
purple lower layer, invert the name may vary between
vials several times to mix. instruments, the program
Zero
the 16-mm cell holder. Make display shows 0 mg/L COD into the 16-mm cell holder.
sure that the filter disc does Mn. Make sure that the filter disc
not block the instrument does not block the
light beam. Refer to Before instrument light beam. Refer
starting on page 1. to Before starting on page 1.
Read

show in mg/L COD Mn.
Multiplication factors for sample dilutions

If the sample COD is expected to exceed 1000 mg/L, dilute the sample as shown in
Table 2. For other dilutions not shown, add the sample volume and the deionized water
and divide by the sample volume to obtain the multiplication factor. All dilutions require
that the ratio of sample to sulfuric acid remain at 9:1.
Note: Mixing concentrated sulfuric acid and water is not additive. Adding 1.0 mL of concentrated
sulfuric acid to 9.0 mL of sample does not result in a final volume of 10.0 mL. This factor is built into
the calibration curve.
Table 2 Multiplication factors
Sample (mL) Deionized water (mL) Range (mg/L COD) Multiplication factor
6.0 3.0 30–1500 1.5
3.0 6.0 60–3000 3
1.0 8.0 180–9000 9
0.5 8.5 360–18,000 18
For best results, use a minimum of 0.5 mL sample for the dilution. If the sample values
exceed 18,000 mg/L COD, use a separate sample dilution, then start the sample chloride
removal procedure.
Example: Dilute the sample to a range of 90–4500 mg/L COD.
Sample Volume (2.0 mL) + Deionized water (7.0 mL) = Total Volume (9.0 mL)
Multiplication factor = (total volume)/(sample volume) = 9.0 mL/2.0 mL = 4.5
Standard test range is 50 to 1000 mg/L COD.
Example test range = 4.5(50) to 4.5(1000) = 225 to 4500 mg/L COD
Interferences
Inorganic materials may also be oxidized by trivalent manganese and constitute a positive
interference when present in significant amounts. Chloride is the most common
interference and is removed by sample pretreatment with the Chloride Removal
Cartridge. If chloride is known to be absent or present in insignificant levels, the
pretreatment can be omitted. A simple way to examine if chloride will affect test results is
to run routine samples with and without the chloride removal, then compare results. Other
inorganic interferences (i.e., nitrite, ferrous iron, sulfide) are not usually present in
significant amounts. If necessary, these interferences can be corrected after finding their
concentrations with separate methods and adjusting the final COD test results
accordingly.
Ammonia nitrogen is known to interfere in the presence of chloride; it does not interfere if
chloride is absent.
Accuracy check
instrument.
Items to collect:
• COD standard solution, 800 mg/L (use 0.60 mL in place of the sample)
Method performance
432 600 mg/L COD 576–624 mg/L COD 8 mg/L COD
Estimated detection limit

The EDL for program 432 is 4 mg/L COD. The EDL is the calculated lowest average
concentration in a deionized water matrix that is different from zero with a 99% level of
confidence.
Summary of method
Chemical Oxygen Demand (COD) is defined as a measure of the oxygen equivalent of
the organic matter content of a sample that is susceptible to oxidation by a strong
chemical oxidant (APHA Standard Methods, 19th ed., 1995). Trivalent manganese is a
strong, non-carcinogenic chemical oxidant that changes quantitatively from purple to
colorless when it reacts with organic matter. It typically oxidizes about 80% of the organic
compounds. Studies have shown that the reactions are highly reproducible and test
results correlate closely to Biochemical Oxygen Demand (BOD) values and hexavalent
chromium COD tests. None of the oxygen demand tests provide 100% oxidation of all
organic compounds.
A calibration is provided which is based on the oxidation of Potassium Acid Phthalate
(KHP). A different response may be seen in analyzing various wastewaters. The KHP
calibration is adequate for most applications. The highest degree of accuracy is obtained
when test results are correlated to a standard reference method such as BOD or one of
the chromium COD methods. Special waste streams or classes will require a separate
calibration to obtain a direct mg/L COD reading or to generate a correction factor for the
precalibrated KHP response. The sample digestion time can be extended up to four hours
for samples that are difficult to oxidize. Test results are measured at 510 nm in
spectrophotometers and 520 nm in colorimeters.
Required reagents
Manganese III COD Reagent Vials, 20–1000 mg/L COD 1 25/pkg 2623425
Chloride Removal Cartridge (CRC) 1 25/pkg 2661825
Sulfuric Acid, concentrated, ACS 75 mL 2.5 L 97909

Required apparatus

Cap, with inert Teflon liner, for mixing bottle varies 12/pkg 2401812
Cell, mixing 2 each 2427700
Forceps, extra fine point 1 each 2669600
Vacuum Pretreatment Device (VPD) 1 each 4900000
Vacuum Pump, 1.2 CFM 115 V 1 each 2824800

Oxygen Demand Standard (BOD, COD, TOC), 10-mL ampules 16/pkg 2833510
500 mL 2833149
TOC
Dispenser, automatic, 1.0–5.0 mL each 2563137

Titrator Strips, Quantab®, for low range chloride 40 tests 2744940
COD Standard Solution, 300-mg/L 500mL 1218649
Oxygen, Dissolved DOC316.53.01096
HRDO Method Method 8166

0.3 to 15.0 mg/L O2 (HR) AccuVac® Ampuls
Scope and application: For water and wastewater
Test preparation
Table 2 shows sample cell and adapter requirements for AccuVac Ampul tests.
DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
Before starting
equipment.
Items to collect
High Range Dissolved Oxygen AccuVac® Ampuls 1

Polypropylene beaker, 50-mL 1
Stoppers, for 18-mm tubes and AccuVac Ampuls 1
1
specific table on page 1.)
1
Sample collection
Good sample collection and handling technique are important to get meaningful results.
The dissolved oxygen content of the water that is tested can change with depth,
turbulence, temperature, sludge deposits, light, microbial action, mixing, travel time and
other factors. A single dissolved oxygen test rarely reflects the accurate overall condition
of a body of water. Several samples taken at different times, locations and depths are
recommended for most reliable results.
The main consideration with sample collection is to prevent contamination of the sample
with atmospheric oxygen.
• Samples must be analyzed immediately after collection, although only a small error
results if the reading on a capped ampule is taken several hours later. The
absorbance will decrease by approximately 3% during the first hour and will not
change significantly afterwards.
• Make sure to put the cap on the ampule before the ampule is removed from the
sample.
AccuVac® Ampul procedure
Start
1. Start program 2. Prepare the blank: Fill 3. Fill a blue Ampul cap 4. Prepare the sample:
445 Oxygen, Dis HR AV. the sample cell with 10 mL with sample. Collect at least 40 mL of
For information about of sample. sample in a 50-mL beaker.
sample cells, adapters or Fill the AccuVac Ampul with
light shields, refer to sample. Keep the tip
Instrument-specific table immersed while the Ampul
5. Hold the Ampul with the 6. Shake the Ampul for 7. Start the instrument 8. When the timer expires,
tip down. Immediately put 30 seconds. timer. A 2-minute reaction shake the Ampul for
the Ampul into the Ampul A small amount of time starts. 30 seconds.
cap. undissolved reagent will not The oxygen that has Let all of the bubbles
The cap prevents affect results. degassed during aspiration dissipate before the next
contamination from dissolves again and reacts. step.
atmospheric oxygen.
2 Oxygen, Dissolved, HRDO Method (15.0 mg/L)

Zero
9. Clean the blank. 10. Insert the blank into the 11. Push ZERO. The 12. Clean the AccuVac
cell holder. display shows 0.0 mg/L O2. Ampul.
Read

sample AccuVac Ampul into show in mg/L O2.
the cell holder.
Interferences
Cr3+ More than 10 mg/L
Cu2+ More than 10 mg/L
Fe2+ More than 10 mg/L
Mg2+ Magnesium is commonly present in seawater and causes a negative interference. If the sample
contains more than 50% seawater, the oxygen concentration obtained by this method will be 25%
less than the true oxygen concentration. If the sample contains less than 50% seawater, the
interference will be less than 5%.
Mn2+ More than 10 mg/L
Ni2+ More than 10 mg/L
NO2- More than 10 mg/L
Accuracy check
Comparison method
To validate the test results, measure the concentration of the same sample with a
dissolved oxygen meter or with a titrimetric method.
Method performance
445 6.7 mg/L O2 6.2–7.3 mg/L O2 0.09 mg/L O2
Oxygen, Dissolved, HRDO Method (15.0 mg/L) 3
Summary of method
The High Range Dissolved Oxygen AccuVac Ampul contains reagent vacuum-sealed in a
glass Ampul. When the AccuVac Ampul is opened in a sample that contains dissolved
oxygen, the solution forms a yellow color which turns purple. The purple color
development is proportional to the concentration of dissolved oxygen. The measurement
Required reagents
High Range Dissolved Oxygen AccuVac® Ampuls 1 25/pkg 2515025
Required apparatus
Beaker, polypropylene, 50-mL, low form 1 each 108041
Optional reagents, apparatus and meters

AccuVac® Sampler each 2405100
HQ30d Meter with Standard LDO Dissolved Oxygen Probe (1 meter cable; additional
each HQ30d53301000
probes and cable lengths are available)
HQ40d Meter with Standard LDO Probe (1 meter cable; additional probes and cable
each HQ40d53301000
lengths are available)

Oxygen, Dissolved DOC316.53.01098
Indigo Carmine Method Method 8316

6 to 800 µg/L O2 (LR, spectrophotometers) AccuVac® Ampuls
10 to 1000 µg/L O2 (LR, colorimeters)
Scope and application: For boiler feedwater
Test preparation
Table 2 shows sample cell and adapter requirements for AccuVac Ampul tests.
DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
Before starting
The dissolved oxygen reading is only stable for 30 seconds. After 30 seconds, the Ampul solution will absorb oxygen from
the air.
equipment.
Items to collect
Low Range Dissolved Oxygen AccuVac® Ampuls 1

Polypropylene beaker, 50-mL 1
Stoppers, for 18-mm tubes and AccuVac Ampuls 1
1
specific table on page 1.)
1
Sample collection
The main consideration with sample collection is to prevent contamination of the sample
with atmospheric oxygen.
later analysis.
• For best results, collect the sample from a stream of water that is hard-plumbed to the
sample source.
• Use a funnel to maintain a continual flow of sample and yet collect enough sample to
immerse the Ampul.
• Do not introduce air in place of the sample.
• Rubber tubing, if used, will introduce unacceptable amounts of oxygen into the
sample unless the length of tubing is minimized and the flow rate is maximized.
• Flush the sampling system with sample for at least 5 minutes.
AccuVac® Ampul procedure
Start
1. Start program 2. Prepare the blank: Fill 3. Clean the blank. 4. Insert the blank into the
446 Oxygen, Dis LR AV. the sample cell with 10 mL cell holder.
For information about of sample.
on page 1.
Zero Read
5. Push ZERO. The display 6. Prepare the sample: 7. Immediately clean and 8. Push READ. Results
shows 0 µg/L O2. Open the AccuVac Ampul. then insert the Ampul into show in µg/L O2.
Fill the Ampul with sample. the cell holder.
For best results, collect the
sample from a stream of
water that is hard-plumbed
to the sample source. Refer
to Sample collection
on page 2.
Interferences
Excess amounts of thioglycolate, ascorbate, ascorbate + sulfite, ascorbate + cupric
sulfate, nitrite, sulfite, thiosulfate and hydroquinone will not reduce the oxidized form of
the indicator and do not cause significant interference.
Hydrazine 100,000 fold excess will begin to reduce the oxidized form of the indicator solution.
Sodium hydrosulfite Reduces the oxidized form of the indicator solution and will cause a significant interference.
Accuracy check
Reagent blank measurement
A reagent blank for this test can be measured as follows:
1. Fill a 50-mL beaker with sample.

2. Add one sodium hydrosulfite powder pillow and mix.
3. Fill a Low Range Dissolved Oxygen AccuVac Ampul with this sample.
4. Measure the dissolved oxygen concentration as shown in the test procedure. The
result should be 0 ± 6 µg/L O2.
Method performance
446 N/A not determined 6 µg/L O2
Summary of method
The Low Range Dissolved Oxygen AccuVac Ampul contains reagent vacuum-sealed in
an Ampul. When the AccuVac Ampul is broken open in a sample containing dissolved
oxygen, the yellow solution will turn blue. The blue color development is proportional to
the concentration of dissolved oxygen. Test results are measured at 610 nm.
Required reagents
Low Range Dissolved Oxygen AccuVac® Ampuls 1 25/pkg 2501025
Required apparatus
Beaker, polypropylene, 50-mL, low form 1 each 108041
Hydrosulfite Reagent Powder Pillows 100/pkg 2118869
Oxygen, Dissolved, Indigo Carmine Method (800 µg/L) 3


AccuVac® Sampler each 2405100

Oxygen, Dissolved, Indigo Carmine Method (800 µg/L) 3

Oxygen Scavengers DOC316.53.01105
Iron Reduction Method Method 8140

3 to 450 µg/L DEHA; 5 to 600 µg/L carbohydrazide; 9 to 1000 µg/L Powder Pillows
hydroquinone; 13 to 1500 µg/L iso-ascorbic acid (ISA); 15 to 1000 µg/L
methylethyl ketoxime (MEKO)
Scope and application: For testing residual corrosion inhibitors (oxygen scavengers) in boiler feed water or
condensate
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
The sample temperature should be 25 ± 3 °C (77 ± 5 °F).
Clean all glassware with 6.0 N (50%) hydrochloric acid, then rinse thoroughly with deionized water to remove iron
contaminants.
To measure the ferrous iron concentration, repeat the test procedure but do not add the DEHA Reagent 2. To automatically
subtract the ferrous iron concentration from the test results, use the reagent blank adjust option. Use the ferrous iron
concentration as the reagent blank value.
equipment.
1
Items to collect
Bottle, glass mixing, with 25-mL mark 2

DEHA Reagent 1 Powder Pillows 2
DEHA Reagent 2 Solution 1 mL
Dropper, 0.5 and 1.0 mL marks 1
Hydrochloric acid, 1:1, 6.0 N varies
2
Sample collection
later analysis.
• Collect samples in clean, dry glass or plastic bottles with tight-fitting caps.
• Rinse the container several times with the sample before collection.
• Prevent agitation of the sample or exposure to sunlight or air.
• Fill the bottle completely and let the sample overflow. Immediately tighten the cap so
that there is no headspace above the sample.
Start
1. Start program 180 O 2. Prepare the blank: Fill a 3. Prepare the sample: Fill 4. Add the contents of one
Scav-Carbohy, 181 O mixing bottle with 25 mL of a second mixing bottle with DEHA Reagent 1 Powder
Scav-DEHA, 182 O Scav- deionized water. 25 mL of sample. Pillow to each mixing bottle.
Hydro, 183 O Scav-ISA or To measure oxygen
184 O Scav-MEKO. For scavenders that react
information about sample quickly with oxygen at room
cells, adapters or light temperature, close the
shields, refer to Instrument bottle.
on page 1.
2 Oxygen Scavengers, Iron Reduction Method (450 µg/L DEHA)

5. Swirl to mix. 6. Use a pipet to add 7. Swirl to mix. 8. Start the instrument
0.5 mL of DEHA Reagent Put both mixing bottles in a timer. A 10-minute (2-minute
2 Solution to each bottle. dark location. A purple color for hydroquinone) reaction
shows if an oxygen time starts.
scavender is present in the Keep the mixing bottles in
sample. the dark during the reaction
period.
Zero
9. When the timer expires, 10. Clean the blank. 11. Insert the blank into the 12. Push ZERO. For
transfer the blank and cell holder. greater accuracy, read the
prepared samples to the result immediately after the
sample cells. timer expires.
Read
13. Clean the prepared 14. Insert the prepared 15. Push READ. Results
sample. sample into the cell holder. show in µg/L.
Interferences
Substances which reduce ferric iron will interfere. Substances which complex iron
strongly may also interfere.
Borate (as Na2B4O7) More than 500 mg/L
Cobalt More than 0.025 mg/L
Copper More than 8.0 mg/L
Ferrous Iron All levels. Measure and subtract (refer to Before starting on page 1)
Oxygen Scavengers, Iron Reduction Method (450 µg/L DEHA) 3

Hardness (as CaCO3) More than 1000 mg/L
Light Light may interfere. Keep sample cells in the dark during color development.
Lignosulfonates More than 0.05 mg/L
Manganese More than 0.8 mg/L
Molybdenum More than 80 mg/L
Nickel More than 0.8 mg/L
Phosphate More than 10 mg/L
Phosphonates More than 10 mg/L
Sulfate More than 1000 mg/L
Temperature Sample temperatures below 22 °C or above 28 °C (72 °F or 82 °F) may affect test accuracy.
Zinc More than 50 mg/L
Method performance
180 299 µg/L 295–303 µg/L 4 µg/L
181 226 µg/L 223–229 µg/L 3 µg/L
182 600 µg/L 591–609 µg/L 8 µg/L
183 886 µg/L 873–899 µg/L 12 µg/L
184 976 µg/L 962–990 µg/L 14 µg/L
Summary of method
Diethylhydroxylamine (DEHA) or other oxygen scavengers in the sample react with ferric
iron in DEHA Reagent 2 Solution to produce ferrous ion in an amount that is equivalent to
the DEHA concentration. This solution then reacts with DEHA 1 Reagent, which forms a
purple color with ferrous iron that is proportional to the concentration of the oxygen
scavenger. This method reacts with all oxygen scavengers and does not differentiate
when the sample contains more than one type of oxygen scavenger. The measurement
Required reagents
Hydrochloric Acid, 6.0 N varies 500 mL 88449

Oxygen Scavenger Reagent Set — — 2446600
Includes:
DEHA Reagent 1 Powder Pillows 2 100/pkg 2167969
DEHA Reagent 2 Solution 1 mL 100 mL 2168042
Required apparatus
4 Oxygen Scavengers, Iron Reduction Method (450 µg/L DEHA)

Bottle, square, with 25 mL mark 1 each 1704200

Dropper, measuring, 0.5 and 1.0 mL plastic 2 20/pkg 2124720
Optional apparatus
Oxygen Scavengers, Iron Reduction Method (450 µg/L DEHA) 5

DOC316.53.01113
Phosphorus, Reactive
(Orthophosphate)
Amino Acid Method1 Method 8178
0.23 to 30.00 mg/L PO43– Reagent Solution
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
The contents of one Amino Acid Reagent Powder Pillow can be used as an alternative to the 1 mL of Amino Acid Reagent
Solution in the test procedure.
equipment.
1
Items to collect
Amino Acid Reagent 1 mL

Cylinder, 25-mL, graduated mixing 1
Molybdate Reagent 1 mL
2

• Collect samples in clean glass or plastic bottles that have been cleaned with
1:1 hydrochloric acid and rinsed with deionized water.
• Do not use a detergent that contains phosphate to clean the sample bottles. The
phosphate in the detergent will contaminate the sample.
• If prompt analysis is not possible, immediately filter and keep the samples at or below
6 °C (43 °F) for a maximum of 48 hours.
Start
1. Start program 485 P 2. Prepare the sample: Fill 3. Prepare the sample: 4. Add 1 mL of Amino Acid
React. Amino. For a mixing cylinder to the 25- Add 1 mL of Molybdate Reagent Solution.
information about sample mL line with sample. Reagent.
on page 1.
2 Phosphorus, Reactive, Amino Acid Method (30.00 mg/L)

5. Close the cylinder. Invert 6. Start the instrument 7. Prepare the blank: Fill a 8. When the timer expires,
the cylinder several times to timer. A 10-minute reaction sample cell with 10 mL of clean the blank.
mix. time starts. untreated sample.
A blue color shows if Prepare the blank while the
phosphate is present in the timer is counting down.
sample.
Zero
9. Insert the blank into the 10. Push ZERO. The 11. Fill a second sample 12. Clean the prepared
cell holder. display shows 0.00 mg/L cell with 10-mL of the sample.
PO43–. prepared sample.
Read

sample into the cell holder. show in mg/L PO 3–.
4
Interferences

substance
Calcium More than 10,000 mg/L as CaCO3
Chloride More than 150,000 mg/L Cl–
Colored samples Add 1 mL of 10 N Sulfuric Acid Standard Solution to another 25-mL sample. Use this instead of
untreated sample as the blank to zero the instrument. Use a pipet and pipet filler to measure the
sulfuric acid standard.
High salt levels (Na+) May cause low results. To eliminate this interference, dilute the sample until two successive dilutions
give about the same result.
Magnesium More than 40,000 mg/L as CaCO3
Phosphorus, Reactive, Amino Acid Method (30.00 mg/L) 3

substance
Nitrite (NO –) Bleaches the blue color. Remove nitrite interference by adding 0.05 g of sulfamic acid to 25 mL
2
sample. Swirl to mix. Use this treated sample in the test procedure.
Phosphates, high As the concentration of phosphate increases, the color changes from blue to green, then to yellow
levels (PO43–) and finally to brown. The brown color may suggest a concentration as high as 100,000 mg/L PO 43–. If
a color other than blue is formed, dilute the sample and retest.
Sulfide (S2–) Sulfide interferes. For samples with sulfide concentration less than 5 mg/L sulfide interference may
be removed by oxidation with Bromine Water as follows:
1. Measure 50 mL of sample into an Erlenmeyer flask.

2. Add Bromine Water drop-wise with constant swirling until permanent yellow color develops.
3. Add Phenol Solution by drops until the yellow color just disappears. Use this treated sample in
the test procedure.
Temperature For best results, sample temperature should be 21 ±3 °C (70 ±5 °F).

Turbidity May give inconsistent results for two reasons. Some suspended particles may dissolve because of
the acid used in the test. Also, desorption of orthophosphate from particles may occur. For highly
turbid samples, add 1 mL of 10 N Sulfuric Acid Standard Solution to another 25-mL sample. Use this
instead of untreated sample as the blank to zero the instrument. Use a pipet and pipet filler to
measure the sulfuric acid standard.
Highly buffered Can prevent the correct pH adjustment of the sample by the reagents. Sample pretreatment may be
samples or extreme necessary.
sample pH
Accuracy check
Items to collect:
• Phosphate 2-mL Ampule Standard, 500-mg/L PO43–
• Ampule breaker
• Mixing cylinders, 25-mL (3)
Mix well.
instrument.

instrument.
4 Phosphorus, Reactive, Amino Acid Method (30.00 mg/L)

Items to collect:
• 10-mg/L Phosphate Standard Solution
Method performance
3– 3–
485 10.00 mg/L PO4 9.86–10.14 mg/L PO43– 0.20 mg/L PO4
Summary of method
In a highly acidic solution, ammonium molybdate reacts with orthophosphate to form
molybdophosphoric acid. This complex is then reduced by the amino acid reagent to yield
an intensely colored molybdenum blue compound. The measurement wavelength is
Required reagents
High Range Reactive Phosphorus Reagent Set — 100 tests 2244100

Includes:
100 mL
Amino Acid Reagent 1 mL 193432
MDB
100 mL
Molybdate Reagent 1 mL 223632
MDB
Required apparatus
Cylinder, graduated mixing, 25-mL 1 each 189640
Phosphate Standard Solution, 10-mg/L as PO4 946 mL 1420416

Phosphate Standard Solution, 2-mL PourRite® Ampule, 500-mg/L PO4 3–
16/pkg 1424220
500 mL 2833149
TOC
Ampule Breaker, PourRite® ampules each 2484600

Amino Acid Reagent Powder Pillow 100/pkg 80499

Bromine Water, 30 g/L 29 mL 221120
Flask, Erlenmeyer, 125-mL each 50543
Phenol Solution, 30-g/L 29 mL 211220
Sulfamic Acid, 454 g each 234401
Sulfuric Acid Standard Solution, 10 N 1000 mL 93153
Thermometer, non-mercury, -10 to +225 °C each 2635700
Bottle, sampling, with cap, low density polyethylene, 250-mL 12/pkg 2087076
Optional standards

Phosphate Standard Solution, 50-mg/L, 10-mL Voluette® Ampules 16/pkg 17110
Phosphate Standard Solution, 10-mL Ampule, 500 mg/L as PO4 16/pkg 1424210


DOC316.53.01115
(Orthophosphate)
Molybdovanadate Method1 Method 8114
0.3 to 45.0 mg/L PO43– Reagent Solution or AccuVac® Ampuls
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900

DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
Before starting
For best results, the sample temperature should be 20–25 °C (68–77 °F).
1
equipment.
Items to collect
Reagent solution
Molybdovanadate reagent 1.0 mL

2
AccuVac Ampuls
Molybdovanadate reagent AccuVac® Ampuls 2

Beaker, 50-mL 1
Stoppers for 18 mm tubes and AccuVac Ampuls 2

2 Phosphorus, Reactive, Molybdovanadate Method (45.0 mg/L)

Reagent solution procedure
Start
1. Start program 480 P 2. Prepare the blank: Fill a 3. Prepare the sample: Fill 4. Add 0.5 mL of
React. Mo. For information sample cell with 10 mL of a second sample cell with Molybdovanadate reagent to
about sample cells, deionized water. 10 mL of sample. each cell.
adapters or light shields,
5. Swirl to mix. 6. Start the instrument 7. When the timer expires, 8. Insert the blank into the
timer. A 7-minute reaction clean the blank. cell holder.
time starts.
If the sample concentration
is greater than 30 mg/L
PO43–, read at exactly
7 minutes or make a
1:1 dilution of the sample
and repeat the test.
Zero Read
4
shows 0.0 mg/L PO43–. sample. sample into the cell holder. show in mg/L PO 3–.
Phosphorus, Reactive, Molybdovanadate Method (45.0 mg/L) 3

AccuVac Ampuls procedure
Start
1. Start program 482 P 2. Prepare the blank: Pour 3. Prepare the sample: 4. Start the instrument
React. Mo. AV. For at least 40 mL of deionized Collect at least 40 mL of timer. A 7-minute reaction
information about sample water into a 50-mL beaker. sample in a 50-mL beaker. time starts.
cells, adapters or light Fill an AccuVac Ampul with Fill the AccuVac Ampul with If the sample concentration
shields, refer to Instrument- deionized water. Keep the sample. Keep the tip is greater than 30 mg/L
specific information tip immersed while the immersed while the Ampul PO43–, read at exactly
on page 1. Ampul fills completely. fills completely. 7 minutes or make a
Note: Although the program 1:1 dilution of the sample
name may vary between and repeat the test.
Zero
5. When the timer expires, 6. Insert the blank AccuVac 7. Push ZERO. The display 8. Clean the AccuVac
clean the blank AccuVac Ampul into the cell holder. shows 0.0 mg/L PO43–. Ampul.
Ampul.
Read

sample AccuVac Ampul into show in mg/L PO 3–.
4
the cell holder.

Interferences
Table 3 shows the interferences and interference levels. Table 4 shows the substances
that do not interfere at or below the indicated levels.
Arsenate Causes a positive interference if the sample is warm when the reagent is added. The sample can
be gently warmed to room temperature without interference.
Iron, ferrous Causes a blue color which interferes at more than 100 mg/L.
Molybdate Causes a negative interference at more than 1000 mg/L.
Silica Causes a positive interference if the sample is warm when the reagent is added. The sample can
Sulfide Causes a negative interference. Correct for this interference as follows:
2. Add Bromine Water drop-wise with constant swirling until a permanent yellow color remains.
3. Add Phenol Solution drop-wise until the yellow color just disappears.
Use this sample in the test procedure.

or extreme sample pH be necessary. The pH should be approximately 7.
Fluoride, thorium, Causes a negative interference.
bismuth, thiosulfate or
thiocyanate
Temperature Temperatures below 18 °C (64 °F) cause a negative interference. Temperatures above 25 °C
(77 °F) cause a positive interference. The sample can be gently warmed to room temperature
without interference.
Table 4 Substances that do not interfere at less than 1000 mg/L

Pyrophosphate Tetraborate Selenate Benzoate
Citrate Oxalate Lactate Tartrate
Formate Salicylate Al3+ Fe3+
Mg2+ Ca2+ Ba2+ Sr2+
+
Li+ Na+ K+ NH4
Cd2+ Mn2+ NO3– NO2–
2– 2–
SO4 SO3 Pb2+ Hg+
Hg2+ Sn2+ Cu2+ Ni2+
–
Ag+ U4+ Zr4+ AsO3
–
Br– CO32– ClO4 CN–
–
IO3 SiO44– — —
Accuracy check
Items to collect:
• Phosphate standard solution, 500 mg/L PO43– ampule
• Ampule breaker

Mix well.
instrument.

instrument.
Items to collect:
• 10 mg/L phosphate standard solution
Method performance
3– 3–
480 30.0 mg/L PO4 29.6–30.4 mg/L PO4 0.3 mg/L PO43–
3– 3–
Summary of method
In the molybdovanadate method, orthophosphate reacts with molybdate in an acid
medium to produce a mixed phosphate/molybdate complex. In the presence of vanadium,
yellow molybdovanadophosphoric acid is formed. The intensity of the yellow color is
proportional to the phosphate concentration. The measurement wavelength is 430 nm for
Note: Product and Article numbers may vary for some selling regions. Contact the appropriate
distributor or refer to the company website for contact information.

Required reagents
100 mL
Molybdovanadate Reagent 1.0 mL 2076032
MDB
OR
Molybdovanadate Reagent AccuVac® Ampul 2 25/pkg 2525025
Required apparatus
AccuVac Snapper 1 each 2405200


500 mL 2833149
TOC

Bromine Water, 30-g/L 29 mL 221120


DOC316.53.01116
(Orthophosphate)
Molybdovanadate Method1 Method 8114
1.0 to 100.0 mg/L PO43– (HR) Test ‘N Tube™ Vials
Test preparation
DR 6000 — —
DR 5000 — —
DR 3900 — LZV849
DR 3800 — LZV646
DR 2800 —
Before starting
The blank vial that is prepared in the test procedure can be used more than once. At room temperature, the reagent blank is
stable for a maximum of three weeks. Prepare a new blank vial when a new lot of reagent is used.
The 7-minute reaction time in the test procedure is for samples that are at 23 °C (73 °F). If the sample temperature is 13 °C
(55 °F), wait 15 minutes. If the sample temperature is 33 °C (91 °F), wait 2 minutes.
The reagent that is used in this test is corrosive. Use protection for eyes and skin and be prepared to flush any spills with
running water.
equipment.
Items to collect
High Range Reactve Phosphorus Test 'N Tube Vials 1

1
1

Test tube rack 1

Molybdovanadate method for Test 'N Tubes
Start
1. Start program 540 P 2. Prepare the blank: Add 3. Put the cap on the vial. 4. Prepare the sample:
React. HR TNT. For 5.0 mL of deionized water to Invert to mix. Add 5.0 mL of sample to a
information about sample a Reactive High Range second Reactive High
cells, adapters or light Phosphorus Test 'N Tube Range Phosphorus Test 'N
shields, refer to Instrument Vial. Tube Vial.
on page 1.
2 Phosphorus, Reactive, Molybdovanadate TNT Method (100.0 mg/L)

5. Put the cap on the vial. 6. Start the instrument 7. When the timer expires, 8. Insert the blank into the
Invert to mix. timer. A 7-minute reaction clean the blank. cell holder.
time starts.
Measure the prepared
sample within two minutes
after the timer expires.
Zero Read
9. Push ZERO. The display 10. Clean the sample vial. 11. Insert the sample vial 12. Push READ. Results
shows 0.0 mg/L PO43–. into the 16-mm cell holder. show in mg/L PO 3–.
4
Interferences
Table 2 shows the interferences and interference levels. Table 3 shows the substances
that do not interfere at or below the indicated levels.
Arsenate Causes a positive interference if the sample is warm when the reagent is added. The sample can
Iron, ferrous Causes a blue color which interferes at more than 100 mg/L.
Molybdate Causes a negative interference at more than 1000 mg/L.
Silica Causes a positive interference if the sample is warm when the reagent is added. The sample can
Sulfide Causes a negative interference. Correct for this interference as follows:
2. Add Bromine Water drop-wise with constant swirling until a permanent yellow color remains.
3. Add Phenol Solution drop-wise until the yellow color just disappears.
Use this sample in the test procedure.

or extreme sample pH be necessary. The pH should be approximately 7.
Phosphorus, Reactive, Molybdovanadate TNT Method (100.0 mg/L) 3

Table 2 Interfering substances (continued)
Fluoride, thorium, Causes a negative interference.
bismuth, thiosulfate or
thiocyanate
Temperature Temperatures below 18 °C (64 °F) cause a negative interference. Temperatures above 25 °C
(77 °F) cause a positive interference. The sample can be gently warmed to room temperature
without interference.
Table 3 Substances that do not interfere at less than 1000 mg/L

Pyrophosphate Tetraborate Selenate Benzoate
Citrate Oxalate Lactate Tartrate
Formate Salicylate Al3+ Fe3+
Mg2+ Ca2+ Ba2+ Sr2+
+
Li+ Na+ K+ NH4
Cd2+ Mn2+ NO3– NO2–
2– 2–
SO4 SO3 Pb2+ Hg+
Hg2+ Sn2+ Cu2+ Ni2+
–
Ag+ U4+ Zr4+ AsO3
–
Br– CO32– ClO4 CN–
–
IO3 SiO44– — —
Accuracy check
Items to collect:
• 10-mL Voluette® Ampule of Phosphate Standard Solution, 500-mg/L PO43–
• Ampule breaker
Mix well.
instrument.
4 Phosphorus, Reactive, Molybdovanadate TNT Method (100.0 mg/L)

instrument.
Items to collect:
• 50-mg/L PO43– standard solution
Method performance
3– 3–
Summary of method
Orthophosphate reacts with molybdate in an acid medium to produce a mixed
phosphate/molybdate complex. In the presence of vanadium, yellow
molybdovanadophosphoric acid forms. The intensity of the yellow color is proportional to
the phosphate concentration. Test results are measured at 420 nm.
Reacted samples contain molybdenum and must be disposed of as a hazardous waste.
Dispose of reacted solutions according to local, state and federal regulations.
Required reagents
High Range Reactive Phosphorus Test ’N Tube™ Reagent Set — 50 vials 2767345
Includes:
Reactive High Range Phosphorus Test ’N Tube Vials (not sold
1 50/pkg —
separately)
Water, deionized varies 100 mL 27242
Required apparatus



500 mL 2833149
TOC
Silica Standard Solution, 1-mg/L SiO2 500 mL 110649

Bromine Water, 30 g/L 29 mL 221120
Dropper, LDPE, 0.5 –1.0 mL 20/pkg 2124720
Optional standards



DOC316.53.01119
(Orthophosphate)
USEPA1 PhosVer 3® (Ascorbic Acid) Method2 Method 8048
0.02 to 2.50 mg/L PO43– Powder Pillows or AccuVac® Ampuls
1 USEPA Accepted for reporting for wastewater analyses. Procedure is equivalent to USEPA and Standard Method 4500-P-E for
wastewater.
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900

DR 6000 — 2427606
DR 5000
DR 900
DR 3900 LZV846 (A)
DR 3800 LZV584 (C) 2122800
DR 2800
DR 2700
1
Before starting
equipment.
Items to collect
Powder pillows
PhosVer® 3 Phosphate Reagent powder pillow, 10-mL 1

2
AccuVac Ampuls
PhosVer® 3 Phosphate Reagent AccuVac® Ampul 1

Beaker, 50-mL 1
1

2 Phosphorus, Reactive, PhosVer 3 Method (2.50 mg/L)

Start
1. Start program 490 P 2. Prepare the sample: Fill 3. Add the contents of one 4. Immediately close the
React. PP. For information a sample cell with 10 mL of PhosVer 3 Phosphate sample cell. Shake
about sample cells, sample. Reagent Powder Pillow to vigorously for 30 seconds.
adapters or light shields, the cell.
5. Start the instrument 6. Prepare the blank: Fill a 7. When the timer expires, 8. Insert the blank into the
timer. A 2-minute reaction second sample cell with clean the blank. cell holder.
If the sample was digested
using the Acid Persulfate
digestion, a 10-minute
reaction period is
necessary.
Zero Read
shows 0.00 mg/L PO43–. sample. sample into the cell holder. show in mg/L PO 3–.
4
Phosphorus, Reactive, PhosVer 3 Method (2.50 mg/L) 3

Start
1. Start program 492 P 2. Prepare the blank: Fill 3. Prepare the sample: 4. Close the AccuVac
React. PV AV. For the sample cell with 10 mL Collect at least 40 mL of Ampul. Shake for
information about sample of sample. sample in a 50-mL beaker. approximately 30 seconds.
cells, adapters or light Fill the AccuVac Ampul with Accuracy is not affected by
shields, refer to Instrument- sample. Keep the tip undissolved powder.
Zero
timer. A 2-minute reaction clean the blank. cell holder. shows 0.00 mg/L PO 3–.
4
time starts.
If the sample was digested
using the Acid Persulfate
digestion, a 10-minute
reaction period is
necessary.
Read
Ampul. sample AccuVac Ampul into show in mg/L PO 3–.
4
the cell holder.

Interferences
Aluminum More than 200 mg/L
Arsenate Interferes at any level.
Chromium More than 100 mg/L
Copper More than 10 mg/L
Hydrogen Sulfide Interferes at any level.
Iron More than 100 mg/L
Nickel More than 300 mg/L
Highly buffered samples or Can prevent the correct pH adjustment of the sample by the reagents. Sample pretreatment
extreme sample pH may be necessary. A pH range of 2–10 is recommended.
Silica More than 50 mg/L
Silicate More than 10 mg/L
Turbidity or color May cause inconsistent results. The acid in the powder pillow can dissolve some of the
suspended particles and the desorption of orthophosphate from the particles can vary. For
highly turbid or colored samples, add the contents of one Phosphate Pretreatment Powder
Pillow to 25 mL of sample. Mix well. Use this solution to zero the instrument.
Zinc More than 80 mg/L
Accuracy check
Items to collect:
• Phosphate standard solution, 50 mg/L PO43– ampule
• Ampule breaker
Mix well.
instrument.

instrument.
Items to collect:
• 50 mg/L phosphate standard solution
• Deionized water
1. Prepare a 2.00 mg/L phosphate standard solution as follows:

a. Use a pipet to add 4.00 mL of 50 mg/L phosphate standard solution into the
volumetric flask. (Alternately, use one of the available mixed parameter
standards. These standards contain 2.0 mg/L phosphate.)
solution.
Method performance
3– 3– 3–
490 2.00 mg/L PO4 1.98–2.02 mg/L PO4 0.02 mg/L PO4
3– 3– 3–
492 2.00 mg/L PO4 1.98–2.02 mg/L PO4 0.02 mg/L PO4
Summary of method
Orthophosphate reacts with molybdate in an acid medium to produce a mixed
phosphate/molybdate complex. Ascorbic acid then reduces the complex, giving an
intense molybdenum blue color. The measurement wavelength is 880 nm for
Required reagents
PhosVer® 3 Phosphate Reagent Powder Pillow, 10-mL 1 100/pkg 2106069

OR
PhosVer® 3 Phosphate Reagent AccuVac® Ampul 1 25/pkg 2508025
Required apparatus


Phosphate Standard Solution, 10-mL Voluette® Ampul, 50-mg/L as PO4 16/pkg 17110
Drinking Water Standard, Mixed Parameter, Inorganic for F -, NO3, PO4, SO4 500 mL 2833049

Phosphate Treatment Powder Pillow 100/pkg 1450199

Silica DOC316.53.01132
Heteropoly Blue Method1 Method 8186

0.010 to 1.600 mg/L SiO2 (LR, spectrophotometers) Powder Pillows
0.01 to 1.60 mg/L SiO2 (LR, colorimeters)
Scope and application: For boiler and ultrapure water.
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
The reaction times in the test procedure are for samples that are at 20 °C (68 °F). If the sample temperature is 10 °C (50 °F),
wait 8 minutes for the first (4-minute) reaction time and 2 minutes for the second (1-minute) reaction time. If the sample
temperature is 30 °C (86 °F), wait 2 minutes for the first (4-minute) reaction time and 30 seconds for the second (1-minute)
reaction time.
To test for very low concentrations, use the ULR rapid liquid method for the best results.
equipment.
1
Items to collect
Amino Acid F Reagent powder pillows, 10-mL 1

Citric acid powder pillows, 10-mL 2
Molybdate 3 Reagent solution 1 mL
2
Sample collection
• Collect samples in clean plastic bottles with tight-fitting caps. Do not use glass
bottles, which will contaminate the sample.
• If prompt analysis is not possible, keep the samples at or below 6 °C (43 °F) for up to
28 days.
Start
1. Start program 651 Silica 2. Prepare the blank: Fill a 3. Prepare the sample: Fill 4. Add 14 drops of
LR. For information about sample cell with 10 mL of a second sample cell with Molybdate 3 reagent
sample cells, adapters or sample. 10 mL of sample. solution to each cell.
Instrument specific
2 Silica, Heteropoly Blue Method (1.600 mg/L)

5. Swirl to mix. 6. Start the instrument 7. After the timer expires, 8. Swirl to mix.
timer. A 4-minute reaction add the contents of one
time starts. Citric Acid Reagent powder
pillow to each sample cell.
9. Start the instrument 10. After the timer expires, 11. Swirl to mix. 12. Start the instrument
timer. A 1-minute reaction add the contents of one timer. A 2-minute reaction
time starts. Amino Acid F Reagent time starts.
The destruction of possible powder pillow to the A blue color shows if silica is
phosphate interference prepared sample cell. present.
occurs during this period. Blank: The sample without
the Amino Acid F Reagent is
the blank.
Zero
13. When the timer expires, 14. Insert the blank into the 15. Push ZERO. The 16. Clean the prepared
clean the blank. cell holder. display shows 0.000 mg/L sample.
SiO2.
Silica, Heteropoly Blue Method (1.600 mg/L) 3

Read

sample into the cell holder. show in mg/L SiO2.
Interferences

substance
Color Does not interfere when the original sample is used to zero the instrument.
Iron Large amounts of both ferrous and ferric iron interfere.
Phosphate Does not interfere at levels less than 50 mg/L PO4. At 60 mg/L PO4, an interference of –2% occurs. At
75 mg/L PO4, the interference is –11%.
Slow reacting forms Occasionally a sample contains silica which reacts very slowly with molybdate. The nature of these
of silica "molybdate-unreactive" forms is not known. A pretreatment with sodium bicarbonate, then sulfuric acid
will make these forms reactive to molybdate. The pretreatment is given in Standard Methods for the
Examination of Water and Wastewater under Silica-Digestion with Sodium Bicarbonate. A longer
reaction time with the sample and the molybdate and acid reagents (before the citric acid is added) can
help as an alternative to the bicarbonate pretreatment.
Sulfides Interfere at all levels.
Turbidity Does not interfere when the original sample is used to zero the instrument.
Accuracy check
Items to collect:
• Silica Standard Solution, 25 mg/L SiO2
• Pipet, TenSette®, 0.1–1.0 mL
• Pipet tips
Mix well.
instrument.
4 Silica, Heteropoly Blue Method (1.600 mg/L)
instrument.
Items to collect:
• Silica Standard Solution, 1.00 mg/L SiO2
Method performance
651 1.000 mg/L SiO2 0.990–1.010 mg/L SiO2 0.012 mg/L SiO2
Summary of method
Silica and phosphate in the sample react with molybdate ion under acidic conditions to
form yellow silicomolybdic acid complexes and phosphomolybdic acid complexes.
Addition of citric acid destroys the phosphate complexes. An amino acid is then added to
reduce the yellow silicomolybdic acid to an intense blue color, which is proportional to the
silica concentration. The measurement wavelength is 815 nm for spectrophotometers or
Required reagents
Silica Reagent Set, low range, includes: — 100 tests 2459300

Amino Acid F Reagent Powder Pillow, 10-mL 1 100/pkg 2254069
Citric Acid Powder Pillow, 10-mL 2 100/pkg 2106269
Molybdate 3 Reagent Solution 1 mL 50 mL 199526
Silica Standard Solution, 1-mg/L SiO2 500 mL 110649

Silica Standard Solution, 25-mg/L as SiO2 236 mL 2122531
Sodium Bicarbonate 454 g 77601
Silica, Heteropoly Blue Method (1.600 mg/L) 5

Sulfuric Acid, 1.00 N 1000 mL 127053
Silica DOC316.53.01133
Silicomolybdate Method Method 8185

1 to 100 mg/L SiO2 (HR, spectrophotometers) Powder Pillows
1 to 75 mg/L SiO2 (HR, colorimeters)
Scope and application: For water and seawater.
Test preparation
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
The sample temperature must be between 15–25 °C (59–77 °F) for accurate results.
Use the Standard Adjust option with each new lot of reagent for the best results.
equipment.
Items to collect
High Range SIlica Reagent Set 1

2
1
Sample collection
• Collect samples in clean plastic bottles with tight-fitting caps. Do not use glass
bottles, which will contaminate the sample.
• If prompt analysis is not possible, keep the samples at or below 6 °C (43 °F) for up to
28 days.
Start
1. Start program 656 Silica 2. Prepare the sample: Fill 3. Add the contents of one 4. Swirl until the reagent is
HR. For information about a sample cell with 10 mL of Molybdate Reagent Powder completely dissolved.
sample cells, adapters or sample. Pillow for High Range Silica
light shields, refer to to the sample cell.
Instrument specific
5. Add the contents of one 6. Swirl to mix. 7. Start the instrument 8. When the timer expires,
Acid Reagent Powder Pillow timer. A 10-minute reaction add the contents of one
for High Range Silica. time starts. Citric Acid Powder Pillow to
A yellow color shows if silica the sample cell.
or phosphorus is present in Any yellow color caused by
the sample. phosphorous is removed
during this step.
2 Silica, Silicomolybdate Method (100 mg/L)

9. Start the instrument 10. Prepare the blank: Fill 11. When the timer expires, 12. Insert the blank into the
timer. A 2-minute reaction a second sample cell with clean the blank. cell holder.
time starts. 10 mL of the original
Complete the rest of the sample.
procedure within three
minutes after the timer
expires.
Zero Read
13. Push ZERO. The 14. Clean the prepared 15. Insert the prepared 16. Push READ. Results
display shows 0 mg/L SiO2. sample. sample into the cell holder. show in mg/L SiO2.
Interferences

substance
Color Does not interfere when the original sample is used to zero the instrument.
Iron Large amounts of both ferrous and ferric iron interfere.
Phosphate Does not interfere at levels less than 50 mg/L PO4. At 60 mg/L PO4, an interference of –2% occurs. At
75 mg/L PO4, the interference is –11%.
Slow reacting forms Occasionally a sample contains silica which reacts very slowly with molybdate. The nature of these
of silica "molybdate-unreactive" forms is not known. A pretreatment with sodium bicarbonate, then sulfuric acid
will make these forms reactive to molybdate. The pretreatment is given in Standard Methods for the
Examination of Water and Wastewater under Silica-Digestion with Sodium Bicarbonate. A longer
reaction time with the sample and the molybdate and acid reagents (before the citric acid is added) can
help as an alternative to the bicarbonate pretreatment.
Sulfides Interfere at all levels.
Turbidity Does not interfere when the original sample is used to zero the instrument.
Accuracy check
Items to collect:
• Silica Standard Solution, 1000 mg/L
Silica, Silicomolybdate Method (100 mg/L) 3

• Pipet, TenSette®, 0.1–1.0 mL
• Pipet tips
Mix well.
instrument.

instrument.
Items to collect:
• Silica Standard Solution, 50 mg/L
Method performance
656 50 mg/L SiO2 48–52 mg/L SiO2 1.0 mg/L SiO2
Summary of method
Silica and phosphate in the sample react with molybdate ion under acidic conditions to
form yellow silicomolybdic acid complexes and phosphomolybdic acid complexes.
Addition of citric acid destroys the phosphate complexes. Silica is then determined by
measuring the remaining yellow color. The measurement wavelength is 452 nm for
Required reagents

High Range Silica Reagent Set, 10-mL — 100 tests 2429600

Includes:
Acid Reagent Powder Pillows for High Range Silica, 10-mL 1 100/pkg 2107469
Citric Acid Powder Pillow, 10-mL 2 100/pkg 2106269
Molybdate Reagent Powder Pillows for High Range Silica, 10-mL 1 100/pkg 2107369

Sodium Bicarbonate 454 g 77601

100 mL
MDB
Silica, Silicomolybdate Method (100 mg/L) 5

Suspended Solids DOC316.53.01139
Photometric Method1 Method 8006

5 to 750 mg/L TSS
1 Adapted from Sewage and Industrial Wastes, 31, 1159 (1959).
Test preparation
Table 1 Instrument-specific information for reagent addition
DR 3800
DR 2800
DR 2700
DR 3900
Before starting
Do not use the Pour-Thru Cell or sipper module (for applicable instruments) with this test.
Items to collect
Beaker, 600-mL, polypropylene 1

Blender 1
Cylinder, 500-mL polypropylene, graduated 1
2

1
up to 7 days.
Photometric procedure
Start
1. Start program 2. Blend 500 mL of sample 3. Pour the blended sample 4. Prepare the sample:
630 Suspended Solids. For in a blender at high speed into a 600-ml beaker. Stir the sample and
information about sample for exactly two minutes. immediately pour 10 mL of
cells, adapters or light the blended sample into a
shields, refer to Instrument sample cell.
on page 1.
Zero
5. Prepare the blank: Fill a 6. Clean the blank. 7. Insert the blank into the 8. Push ZERO. The display
second sample cell with cell holder. shows 0 mg/L TSS.
10 mL of tap water or
deionized water.
Read
9. Swirl the prepared 10. Clean the prepared 11. Insert the prepared 12. Push READ. Results
sample to remove any gas sample. sample into the cell holder. show in mg/L TSS.
bubbles and uniformly
suspend any residue.
2 Suspended Solids, Photometric Method (750 mg/L)
Interferences
Samples that absorb strongly at the measurement wavelength, such as blue dyes, may
give false, high-bias readings. A user-entered calibration is advised for these samples.
Accuracy check

Calibration for this test is based on the gravimetric technique with parallel sewage
samples from a municipal sewage plant. For most samples, this calibration supplies
satisfactory results. When higher accuracy is required, run parallel spectrophotometric
and gravimetric determinations with portions of the same sample. Make the new
calibration on the particular sample using a gravimetric technique as a basis.
Summary of method
This method of determining total suspended solids (TSS) is a simple, direct measurement
which does not require the filtration or ignition/weighing steps that gravimetric procedures
do. The USEPA specifies the gravimetric method for solids determinations, while this
method is often used for checking in-plant processes. The measurement wavelength is
Required apparatus
Beaker, 600-mL, polypropylene 1 each 108052

Cylinder, graduated, 500-mL, polypropylene 1 each 108149

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Copper DOC316.53.

Table 2 Instrument-specific information for AccuVac Ampuls

CuVer® 1 Copper Reagent Powder Pillow, 10-mL 1

Refer to Consumables and replacements items on page 7 for reorder information.

CuVer® 2 Reagent AccuVac® Ampul 1

Refer to Consumables and replacements items on page 7 for reorder information.

Sample collection and storage

2 Copper, Bicinchoninate Method (5.00 mg/L)

Copper, Bicinchoninate Method (5.00 mg/L) 3

4 Copper, Bicinchoninate Method (5.00 mg/L)

Table 4 Interfering substances and suggested treatments for AccuVac Ampuls

Copper, Bicinchoninate Method (5.00 mg/L) 5

1. Prepare a 12.5 mg/L copper standard solution as follows:

Standard solution method

1. Prepare a 4.00 mg/L copper standard solution as follows:

6 Copper, Bicinchoninate Method (5.00 mg/L)

Description Quantity/test Unit Item no.

CuVer® 1 Copper Reagent Powder Pillow, 10-mL 1 100/pkg 2105869

Description Quantity/test Unit Item no.

AccuVac Snapper 1 each 2405200

Description Unit Item no.

Copper Standard Solution, 100-mg/L as Cu 100 mL 12842

Optional reagents and apparatus

Description Unit Item no.

Beaker, 50-mL each 50041H

Copper, Bicinchoninate Method (5.00 mg/L) 7

FOR TECHNICAL ASSISTANCE, PRICE INFORMATION AND ORDERING: HACH COMPANY

Copper, Bicinchoninate Method (5.00 mg/L) 9

Porphyrin Method1 Method 8143

Copper Masking Reagent Powder Pillows, 10-mL 1

Refer to Consumables and replacement items on page 5 for reorder information.

Sample collection and storage

Powder pillow procedure

2 Copper, Porphyrin Method (210 µg/L)

Standard solution method

4 Copper, Porphyrin Method (210 µg/L)

1. Prepare a 150 µg/L copper standard solution as follows:

Description Quantity/test Unit Item no.

Nitric Acid Solution, 1:1 varies 500 mL 254049

Description Unit Item no.

Copper Standard Solution, 4 mg/L, 2 mL Pour-Rite Ampules 20/pkg 2605720

Optional reagents and apparatus

Description Unit Item no.

Copper, Porphyrin Method (210 µg/L) 5

Calmagite Colorimetric Method Method 8030

Alkali Solution, for calcium and magnesium tests 1 mL

Refer to Consumables and replacement items on page 5 for reorder information.

Sample collection and storage

2 Hardness, Ca and Mg, Calmagite Method (4.00 mg/L)

Hardness, Ca and Mg, Calmagite Method (4.00 mg/L) 3

21. Calcium sample: Insert 22. Push READ. Results

Interfering substance Interference level

Description Quantity/test Unit Item no.

Hardness Reagent Set, includes: — 100 tests 2319900

Description Quantity/test Unit Item no.

Cylinder, graduated mixing, 100-mL , tall form 1 each 2088642

Hardness, Ca and Mg, Calmagite Method (4.00 mg/L) 5

Description Unit Item no.