FACULTY OF CIVIL AND ENVIRONMENTAL ENGINEERING

(FKAAS)

BFC 32501
TRANSPORTATION AND ENVIRONMENTAL ENGINEERING
LABORATORY

SECTION 1

GROUP WORK
PROPOSAL

GROUP MEMBERS:
1) ALIS SYAKINA BINTI AHMAD AF160057
2) MUHAMMAD MAHATHIR BIN ABDUL SHUKUR AF160143
3) NORMADIANA BINTI NOH DF170056
4) NURUL ASHIKEN BINTI ALI AF160047
5) SITI NUR EZIANIE BINTI MOHAMAD A’ASHRI DF170121

LECTURER: Ts. Dr. ROSLINDA BINTI SESWOYA

DATE: 26th FEBRUARY 2019
OBJECTIVE
To determine whether the lake water of Kemajuan Lake, UTHM is suitable as an alternative water
resource for irrigation purposes according to its water quality index.

SOURCE OF WATER
The source of water for sampling are from an artificial lake/reservoir (Kemajuan Lake), located
near G3 building in UTHM as shown in Figure 1 below. Figure 2 and Figure 3 shows the map of
the Kemajuan Lake that can be obtained from Google Maps.

Figure 1: Kemajuan Lake, UTHM

Figure 2: Map (earth view) of Kemajuan Lake, UTHM

Figure 3: Map of Kemajuan Lake, UTHM

Figure 4 shows the general flow chart of sampling process. Sampling process for each method
were shown in attachment of established methods at the last part of this proposal.

Figure 4: Sampling Flow Chart

PARAMETERS

According to standard methods for examination of water and wastewater, the parameters that will
be measured are:

Parameters Indicates

Biochemical Oxygen Demand (BOD)
Chemical Oxygen Demand (COD)
Determination of pH value
Total Suspended Solid (TSS)
Dissolved Oxygen (DO)
Ammonia Nitrogen
For determining how fast biological organism use
oxygen in body of water
Indicates the amount of organic pollution in water
Contamination and acidification
Small solid particles which remain in suspension in
water as a colloid or due to motion in water
Measures amount of oxygen dissolved in water.
Measure the amount of ammonia, a toxic pollutant
CALCULATION OF WATER QUALITY INDEX (WQI)

Water Quality Index is calculated using Equation 1 below:

𝑾𝑸𝑰 = (𝟎. 𝟐𝟐∗ 𝑺𝑰𝑫𝑶) + (𝟎. 𝟏𝟗∗ 𝑺𝑰𝑩𝑶𝑫) + (𝟎. 𝟏𝟔∗ 𝑺𝑰𝑪𝑶𝑫) + (𝟎. 𝟏𝟓∗ 𝑺𝑰𝑨𝑵) +
(𝟎. 𝟏𝟔∗ 𝑺𝑰𝑺𝑺) + (𝟎. 𝟏𝟐∗ 𝑺𝑰𝒑𝑯) --------- Equation 1

where;

SIDO = Subindex Dissolved Oxygen (% saturation)

SIBOD = Subindex Biochemical Oxygen Demand

SICOD = Subindex Chemical Oxygen Demand

SIAN = Subindex Ammoniacal Nitrogen

SISS = Subindex Suspended Solid

SIpH = Subindex pH value

CLASSIFICATION

From calculation using equation 1, the result will be classified according to few classifications
which are:
 Class Based
 Pollution Status Based

Table 1: DOE Water Quality Index Classification

Table 2: Water Classes and Uses

Table 3: DOE Quality Classification Based on Water Quality Index

ANALYTICAL METHOD OF PARAMETERS

No. Parameters Method References

1 Biochemical Oxygen Demand
(BOD)
2 Chemical Oxygen Demand
(COD)
3 Determination of pH value
4 Total Suspended Solid (TSS)
5 Dissolved Oxygen (DO)
6 Ammonia Nitrogen
ASTM WK28466 Ernieza Hazlin binti Mohd Hata. (June
Method 8043 2017). “Groundwater Quality
Improvement by Using Aeration and
Filtration Methods”. UTHM.
ASTM D1252 – Muhammad Syazwan bin Sapiren.
06(2012)1 (June 2017). “Lake Water Quality
Method 8000 Improvement by Using Waste Clam
Shells as an Adsorbent”. UTHM.
ASTM D1293 Ayu Zulaikha binti Yahya. (July 2017).
Method 8156 “Domestic Greywater System
Treatment (DGST) by Using Used
Ceramic Filtration in Village Houses”.
UTHM.
ASTM D5907 – 18 Muhammad Syazwan bin Sapiren.
Method 8006 (June 2017). “Lake Water Quality
Improvement by Using Waste Clam
Shells as an Adsorbent”. UTHM.
ASTM D888 – 18 Syafik Akmal bin Mohd Tajuddin.
Method 8229 (January 2017). “Water Quality and
Trophic Status Study in Sembrong
Reservoir during Monsoon Season”.
UTHM.
ASTM D1426-15 Syafik Akmal bin Mohd Tajuddin.
Method 8038 (January 2017). “Water Quality and
Trophic Status Study in Sembrong
Reservoir during Monsoon Season”.
UTHM.
ATTACHMENT
Oxygen Demand, Biochemical DOC316.53.01200

Dilution Method1 Method 8043

Scope and application: For water and wastewater.

1 Adapted from Standard Methods for the Examination of Water and Wastewater and from Klein, R.L.; Gibbs, C. Journal of Water Pollution
Control Federation, 1979, 51(9), 2257.

Test preparation

Before starting
This test is a 5-day test. Complete all the steps carefully to make sure that the test does not have to be done again.
Prepare the dilution water with a BOD Nutrient Buffer Pillow. Make sure that the dilution water for this test does not contain
an oxygen demand or toxins. When incubated for 5 days at 20 °C, the dissolved oxygen concentration in the dilution water
must not change by more than 0.2 mg/L.
Carbonaceous BOD (CBOD) can be determined by the addition of nitrification inhibitor. A test for CBOD is recommended for
biologically-treated effluents, samples with bacterial seed, samples with biologically treated effluents and river water.
As an alternative, use the method BOD calculation-Graphical Method on page 6 to calculate the test results. The
graphical calculation method is also a tool for troubleshooting problems in BOD measurements. The graphical calculation
method is not approved for regulatory reporting.
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used. Use the recommended personal protective
equipment.
Dispose of reacted solutions according to local, state and federal regulations. Refer to the Safety Data Sheets for disposal
information for unused reagents. Refer to the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.

Items to collect
Description Quantity

BOD bottle, 300 mL, glass, with glass stoppers and plastic caps 6
BOD bottle cap, plastic 6
Dilution water (refer to Prepare the dilution water on page 3) varies
Pipet, seriological, 1 mL, 5 mL and 10 mL 1
Incubator 1
Nitrification Inhibitor (for the CBOD test only) 1 bottle

Refer to Consumables and replacement items on page 10 for order information.

Sample collection
The main consideration with sample collection is to prevent contamination of the sample
with atmospheric oxygen.
• Analyze the samples immediately. The samples cannot be preserved for later
analysis.
• Collect samples in 300 mL glass BOD bottles. Completely fill the bottles.

1
Test procedure
1. Identify five sample
volumes to use for this test.
Refer to Select the sample
volumes on page 4.
Note: If the minimum
sample volume is 3 mL or
more, determine the
dissolved oxygen in the
undiluted sample. Ignore
this determination if sewage
and settled effluents with a
dissolved oxygen content
near 0 mg/L are analyzed. If
industrial effluents and
disinfected samples are
analyzed, refer to
Interferences on page 8.
5. Fill each bottle with
prepared dilution water.
Refer to Prepare the dilution
water on page 3. To
prevent air bubbles, pour
the water down the inner
surface of the bottle.
2 Oxygen Demand, Biochemical, dilution method
2. Prepare the samples:
Gently stir the sample.
6. Carefully insert a stopper
in each bottle to prevent
trapped air bubbles. Push
down on the stopper. Invert
the bottles several times to
mix.
3. Use a pipet to add the 4. If the test is for CBOD,
sample volumes to five add two portions of
300‑mL BOD bottles. Nitrification Inhibitor
(approximately 0.16 g) to
each bottle to prevent the
oxidation of nitrogen
compounds. Record the
results as CBOD.
7. Prepare the blank: Fill 8. Use a probe or use a
another 300‑mL BOD bottle titration procedure to
with the prepared dilution measure the dissolved
water. To prevent air oxygen concentration in
bubbles, pour the water each bottle. If titration is
down the inner surface of used for the measurement,
the bottle. prepare two sets of BOD
bottles. Make sure to
measure the dissolved
oxygen of the blank.
9. Carefully insert a stopper
in each of the prepared
sample bottles to prevent
trapped air bubbles. Add
dilution water above the
stopper of the BOD bottles
to make a water seal.
10. Add a cap to each bottle
to prevent evaporation.
11. Keep the prepared 12. After 5 days, measure
sample bottles in an the remaining dissolved
incubator at 20 °C (68°F). oxygen in each of the
Do not move the prepared prepared samples.
sample bottles for 5 days. For accurate results, the
prepared samples must
contain a minimum 1.0 mg/L
DO concentration after
incubation.

13. Calculate the BOD

value. Refer to BOD
calculation—Standard
Methods on page 6 or
BOD calculation-Graphical
Method on page 6.

Prepare the dilution water

Make sure that no source of oxygen demand or toxins are added when the dilution water
is prepared.
Items to collect:
• Dilution water (refer to the dilution water guidelines)
• BOD Nutrient Buffer Pillow1
• Raw sewage2 for the bacterial seed, 3 mL (if the sample is low in bacteria)

1 Different sizes are available for different quantities of water (e.g., 3 L and 6 L).
2 Keep the raw sewage at 20 °C (68°F) and do not move for 24–36 hours before use. Pipet from the upper
portion of the sewage.

Oxygen Demand, Biochemical, dilution method 3

 Dilution water guidelines
• For the best results, use distilled water from an alkaline permanganate distillation.
• Use high-quality water that does not contain organic compounds or toxic compounds
(e.g., chlorine, copper and mercury).
• Do not use deionized water from ion exchange columns. The resins in the cartridges
(mostly new cartridges) occasionally release organic materials that have an oxygen
demand. In addition, bacteria can grow on the columns, which adds contamination to
the dilution water.
• The dissolved oxygen concentration of the dilution water must not change by more
than 0.2 mg/L when incubated for 5 days at 20 °C (68 °F).
Prepare the dilution water as follows:

1. Keep the dilution water in clean jugs in an incubator at 20 °C (68°F). Shake the jugs
to saturate the water with air. As an alternative, loosely put the cap on the jugs and
wait a minimum of 24 hours before use.
2. (Optional) Use a small aquarium pump or an air compressor to saturate the water
with air. Make sure to use filtered air. Make sure that the filter does not grow bacteria.
3. Shake the BOD Nutrient Buffer Pillow to mix the contents.
4. Add the contents of the BOD Nutrient Buffer Pillow to the distilled water.
5. Put the cap on the jug. Shake the jug vigorously for 1 minute to dissolve the nutrients
and to saturate the water with air.
6. If the sample is known to be low in bacteria (e.g., industrial waste or sewage that has
been disinfected), immediately before the test, add 3 mL of raw sewage to each liter
of the dilution water.
Measure the BOD of the raw sewage collected. The BOD of the raw sewage will be
subtracted from the BOD of the sample.

Conventional method (optional)

As an alternative, prepare the dilution water with the conventional method as follows:
1. Pipet 1 mL of each of the solutions that follow per liter of distilled water at 20 °C:
Note: Be careful to prevent contamination of the solutions.

• Calcium Chloride Solution

• Ferric Chloride Solution
• Magnesium Sulfate Solution
• Phosphate Buffer Solution3
2. Put the cap on the bottle. Shake the bottle vigorously for 1 minute.

Select the sample volumes

Select the five sample volumes to use for this test. The sample volumes change based on
the sample BOD and the altitude of the laboratory.
• High BOD samples (e.g., raw sewage)—Use small sample volumes so that all the
oxygen is not consumed.
• Low BOD samples—Use large sample volumes so that sufficient oxygen is
consumed to give an accurate result.
At higher altitudes, the amount of oxygen that dissolves in water decreases, so less
oxygen is available to microorganisms. Refer to Table 1.
Note: At least 2.0 mg/L of dissolved oxygen (DO) must be consumed during the test and at least
1.0 mg/L DO must be in the BOD bottle after the test.

1. Identify the minimum sample volume. Refer to Table 2.

3 Keep the phosphate buffer solution in a refrigerator to decrease the rate of biological growth.

4 Oxygen Demand, Biochemical, dilution method

 For example, if a sewage sample contains approximately 300 mg/L BOD, the
minimum sample volume is 2 mL. If the sewage effluent contains approximately
40 mg/L BOD, the minimum sample volume is 15 mL.
2. Identify the maximum sample volume. Refer to Table 3.
For example, if the sample contains approximately 300 mg/L BOD and the laboratory
altitude is 305 m (1000 ft), the maximum sample volume is 8 mL. If the sample
contains approximately 40 mg/L BOD and the laboratory altitude is 305 m (1000 ft),
the maximum sample volume is 60 mL.
3. Select three other sample volumes between the minimum and maximum volumes so
that there are five sample volumes total.
Table 1 Oxygen values at different altitudes—20 °C (68 °F)
Altitude Oxygen value in water saturated with air Altitude Oxygen value in water saturated with air
Sea level 9.2 mg/L 1219 m (4000 ft) 7.9 mg/L
305 m (1000 ft) 8.9 mg/L 1524 m (5000 ft) 7.6 mg/L
610 m (2000 ft) 8.6 mg/L 1829 m (6000 ft) 7.4 mg/L
914 m (3000 ft) 8.2 mg/L

Table 2 Minimum sample volume

Sample type BOD (mg/L) Volume (mL) Sample type BOD (mg/L) Volume (mL)
Strong trade waste 600 1 Oxidized effluents 50 12
Raw and settled sewage 300 2 40 15
200 3 30 20
150 4 20 30
120 5 10 60
100 6 Polluted river waters 6 100
75 8 4 200
60 10 2 300

Table 3 Maximum sample volume

mg/L BOD—Sea level mg/L BOD—305 m (1000 ft) BOD—1524 m (5000 ft) Volume (mL)
2460 2380 2032 1
1230 1189 1016 2
820 793 677 3
615 595 508 4
492 476 406 5
410 397 339 6
304 294 251 8
246 238 203 10
205 198 169 12
164 158 135 15
123 119 101 20
82 79 68 30
41 40 34 60
25 24 21 100

Oxygen Demand, Biochemical, dilution method 5

 Table 3 Maximum sample volume (continued)
mg/L BOD—Sea level mg/L BOD—305 m (1000 ft) BOD—1524 m (5000 ft) Volume (mL)
12 12 10 200
8 8 7 300

BOD calculation—Standard Methods

Use the Standard Methods calculation when the results will be given to a regulatory
agency. Give the results as CBOD5 if nitrification inhibitor was added in the test.
When a bacterial seed is not added to the dilution water, calculate the BOD as follows:
BOD5 mg/L = (D1 – D2) ÷ P
When a bacterial seed is added to the dilution water, calculate the BOD as follows:
BOD5 mg/L = ((D1 – D2) – (B1 – B2)× f) ÷ P
Where:
BOD5 = BOD value from the 5-day test (mg/L)
D1 = DO of the prepared sample immediately after preparation (mg/L)
D2 = DO of the prepared sample after incubation in mg/L
P = Decimal volumetric fraction of sample used
B1 = DO of the bacterial seed control immediately after preparation (mg/L)
B2 = DO of bacterial seed control after 5-days at 20 °C (68 °F) in mg/L
f = ratio of the bacterial seed in the diluted sample to the bacterial seed in the bacterial
seed control. f = (% seed in diluted sample) ÷ (% seed in seed control)
OR
If the bacterial seed material is added directly to sample or to the bacterial seed control
bottles:
f = (volume of the bacterial seed in the diluted sample) ÷ (volume of the bacterial seed in
the bacterial seed control)
Averaged results are satisfactory if all the criteria that follows is true for more than one of
the sample dilutions:
• The remaining DO is at least 1 mg/L.
• The final DO value is at least 2 mg/L less than the initial DO value.
• Toxicity at higher sample concentrations is not seen.
• There are no obvious anomalies.

BOD calculation-Graphical Method

NOTICE
Do not use the Graphical Method when the results must be given to a regulatory agency.

1. On a graph, record on the y-axis the remaining dissolved oxygen (DO) (mg/L) in each
of the prepared samples after 5 days. Record the sample volume (mL) on the x-axis.
Draw the best straight line through the plotted points. Refer to Figure 1.
Note: At least three points should be on the graph line or very near to the graph line. Ignore an
error point if seen at this time. For unseeded dilution water, the graph line should cross the
mg/L oxygen remaining scale near or at less than the oxygen saturation value for the altitude of
the laboratory as shown in Prepare the dilution water on page 3.
2. To calculate the BOD, use the equation that follows, which is mathematically the
same as the BOD equation. Refer to BOD calculation—Standard Methods on page 6.
mg/L BOD = (A × 300) – B + C
Where:
A = the slope of the graph line. The slope of the graph line is equal to the mg/L DO
consumed for each mL of sample. Select a point on the line and subtract the mg/L

6 Oxygen Demand, Biochemical, dilution method

 DO remaining at that point from the mg/L DO where the line crosses with the DO
scale (Y-intercept, mg/L DO remaining). Divide the difference by the mL of sample at
the point selected.
300 = the volume of the BOD bottle (300 mL)
B = the Y-intercept. The Y-intercept is the DO value where the line crosses the “DO
remaining” scale. (The Y-intercept should be very near to the actual dilution water
blank value.)
C = the sample DO. The sample DO is the DO of the undiluted sample.
A different way to write this equation is:
mg/L BOD = (slope × 300) – Y-intercept + sample DO
Note: If the best straight line is supplied by linear regression through the use of a calculator,
change the sign (–) of the slope (+ slope) before the slope is multiplied by 300.
For example:
The mg/L DO remaining was determined for a series of four dilutions of domestic
sewage after 5 days of incubation. The results are given in Table 4.
If a set of BOD dilutions is done correctly with a homogeneous sample, a graph of the
mg/L DO remaining versus the sample volume results in a straight line. The Y-
intercept value is equal to the DO content of the dilution water after the 5-day
incubation. But the Y-intercept value is not actually measured.
In this example, the Y-intercept is equal to 9.0 mg/L and the DO of the domestic
sewage sample is thought to be zero. Refer to Table 4. If another type of sample is
used, measure the DO of an undiluted sample by the Winkler titration or
potentiometrically. The slope in the example is calculated as follows:
(DO value at Y-intercept – DO value at Point A) ÷ (sample volume at Point A –
sample volume at Y-intercept)
(9.0 mg/L –3.0 mg/L) ÷ 8 mL – 0 mL = 0.75 mg/L per mL = slope
American Public Health Association formula—The calculation for BOD can be
written as follows (not approved for reporting purposes):
Slope = (mg/L DO remaining with a smaller sample volume – mg/L DO remaining
with a larger sample volume) ÷
(mL of a larger sample volume – mL of a smaller sample volume)
slope × 300 – DOD + S = mg/L BOD
Where:
DOD = mg/L DO of dilution water
S = mg/L DO of sample

Oxygen Demand, Biochemical, dilution method 7

Figure 1 DO per mL of sample

1 mL of sample 2 mg/L DO remaining 3 Y-intercept

Table 4 Sample volume versus DO remaining

Sample volume DO remaining
2.0 mL 7.50 mg/L
3.0 mL 6.75 mg/L
6.0 mL 4.50 mg/L
9.0 mL 2.25 mg/L

Interferences
To get good BOD results, special handling is necessary to analyze chlorinated and
industrial effluents. Usually, careful experimentation with the sample shows the changes
that should be made to the test procedure. Toxins in the sample have an adverse effect
on any microorganisms in the sample, which results in lower BODs.
The substances in Table 5 interfere in the determination of oxygen demand at the given
concentrations.
Table 5 Interfering substances
Interfering substance Interference level
Chlorine Small quantities of residual chlorine—Let the sample sit for 1 to 2 hours at room temperature.
Larger quantities of chlorine—Refer to Remove chlorine from the sample on page 9.
Phenols Dilute the sample with high quality distilled water. As an alternative, acclimatize the bacterial seed
used in the dilution water to tolerate such materials. Refer to Acclimatize the bacterial seed
Heavy metals on page 9.
Cyanide
Highly buffered Less than pH 6.5 or more than pH 7.5 interfere. Adjust to pH 7.2 with acid (Sulfuric Acid, 1 N or
samples or extreme Phosphate Buffer Solution) or base (Sodium Hydroxide, 1 N).
sample pH
Cold temperature Cold samples can be supersaturated with oxygen and will have low BOD results. Fill a 1-liter (1-
quart) bottle ½ full with cold sample. Shake the bottle vigorously for 2 minutes. Let the sample
temperature increase to 20 °C (68 °F).

8 Oxygen Demand, Biochemical, dilution method

Remove chlorine from the sample
Items to collect:
• 250-mL Erlenmeyer flask
• 10-mL serological pipet and a pipet filler
• 25-mL buret
• 0.020 N Sulfuric Acid Standard Solution, 10 mL
• 100-g/L Potassium Iodide Solution, 10 mL
• 0.025 N Sodium Thiosulfate Standard Solution, 25 mL
• Starch Indicator Solution, 3 full droppers

1. Add 100 mL of sample to a 250-mL Erlenmeyer flask.

2. Use a 10-mL serological pipet and a pipet filler to add 10 mL of 0.020 N Sulfuric Acid
Standard Solution to the flask.
3. Use a 10-mL serological pipet and a pipet filler to add 10 mL of 100-g/L Potassium
Iodide Solution to the flask.
4. Add 3 full droppers of Starch Indicator Solution. Swirl to mix.
5. Fill a 25-mL buret with 0.025 N Sodium Thiosulfate Standard Solution.
6. Titrate the sample from dark blue to colorless.
7. Calculate the amount of 0.025 N Sodium Thiosulfate Standard Solution to add to the
sample.
mL 0.025 N Sodium Thiosulfate Standard Solution = (mL titrant used × volume of
remaining sample) ÷ 100
8. Add the calculated quantity of 0.025 N Sodium Thiosulfate Standard Solution to the
sample.
9. Mix fully. Wait 10–20 minutes before the test is done.

Acclimatize the bacterial seed

1. Fill a 4-liter (1-gallon) stainless steel or plastic container with domestic sewage.
2. Aerate the sewage for 24 hours.
3. Let the heavier material collect on the bottom for 1 hour.
4. Use a siphon to remove 3 liters (3 quarts) of the material from the top and discard.
5. Fill the container with a mixture of 90% sewage and 10% wastes that contain the
toxic material.
6. Aerate for 24 hours.
7. Do steps 3–5 again with more and more quantities of waste until the container holds
100% toxic waste material.

Accuracy check
Standard solution method
Use the standard solution method to validate the test procedure and the instrument.
Items to collect:
• 300-mg/L BOD Standard Solution4, Voluette Ampule, 10 mL
• Seeded dilution water
• Four 300-mL BOD bottles
• Pipet, volumetric, Class A, 1.0–4.0 mL
• TenSette Pipet

4 300-mg/L of glucose and 300-mg/L of glutamic acid

Oxygen Demand, Biochemical, dilution method 9

 1. Use the TenSette pipet to add 1.00, 2.00, 3.00 and 4.00 mL of the standard solution
to four 300-mL BOD bottles.
2. Fill each bottle with the seeded dilution water. Refer to Prepare the dilution water
on page 3. To prevent air bubbles, let the water flow over and down the outer surface
of the bottle.
3. Use the test procedure to measure the DO concentration of the diluted standard
solutions and then again after 5 days. Do not add Nitrification Inhibitor. Record the
values.
4. Calculate the BOD value.
5. Divide the calculated BOD value by 2. The expected result when compared to
Standard Methods is 198 (± 30.5) mg/L.
Note: The ampule includes 2 mL of 450 mg/L Glucose plus Glutamic Acid (GGA). Pour all the
ampule is equivalent to add 6 mL of 150 mg/L solution with the Standard Methods. Calculate
the BOD concentration as though there were 6 mL added to the bottle instead of 2 mL. The
dilution factor for this standard is 50x.

Summary of method
Biochemical oxygen demand (BOD) is a measurement of the oxygen requirements of
municipal and industrial wastewaters and sewage. The test results are used to calculate
the effect of waste discharges on the oxygen resources of the receiving waters. The BOD
test gives a limited value in the measurement of the actual oxygen demand because the
environmental factors (e.g., temperature change, biological population, water movement,
sunlight, oxygen concentration and other factors) cannot be simulated accurately in the
laboratory. The BOD test is a very important value after patterns of oxygen uptake for a
specified effluent and receiving water are identified.
For this test, a sealed wastewater sample (or a prepared dilution) is incubated for the
standard 5-day period. Then, the change in dissolved oxygen content is identified. The
BOD value is calculated from the results of the dissolved oxygen tests.
Consumables and replacement items
Required reagents

Description Quantity/Test Unit Item no.

BOD Nutrient Buffer Pillow, 3 mL (for 3 L of dilution water) 1 pillow 50/pkg 1486166

Required apparatus

Description Quantity/test Unit Item no.

BOD bottle with glass stopper, 300 mL 6 each 62100

BOD bottle cap, plastic 6 6/pkg 241906
Pipet, serological, 1 mL, glass 1 50/pkg 2093135
Pipet, serological, 5 mL 1 each 53237
Pipet, serological, graduated, 10 mL 1 each 53238
Pipet filler, safety bulb 1 each 1465100

Recommended standards

Description Unit Item no.

BOD Standard Solution, Voluette® Ampule, 300 mg/L, 10 mL 16/pkg 1486510

ez GGA BOD Standard Solution, 300 mg/L, 2-mL ampules 20/pkg 2514420

10 Oxygen Demand, Biochemical, dilution method

Optional reagents and apparatus

Description Unit Item no.

BOD bottle with glass stopper, 300 mL 6/pkg 62106

BOD bottle with glass stopper, 300 mL 24 pkg 62124
BOD Nutrient Buffer Pillow, 0.5 mL (for 300 mL of dilution water) 50/pkg 1416066
BOD Nutrient Buffer Pillow, 3 mL (for 3 L of dilution water) 50/pkg 1486166
BOD Nutrient Buffer Pillow, 4 mL (for 4 L of dilution water) 50/pkg 2436466
BOD Nutrient Buffer Pillow, 6 mL (for 6 L of dilution water) 50/pkg 1486266
BOD Nutrient Buffer Pillow, 19 mL (for 19 L of dilution water) 25/pkg 1486398
Calcium Chloride Solution, APHA, for BOD 500 mL 42849
Clippers each 96800
Ferric Chloride Solution, APHA, for BOD 1L 42953
Flask, Erlenmeyer, 250 mL each 50546
Magnesium Sulfate Solution, APHA, for BOD 500 mL 43049
Nitrification Inhibitor 35 g 253335
Nitrification Inhibitor 500 g 253334
Nitrification Inhibitor, dispenser cap each 45901
Phosphate Buffer Solution, APHA, for BOD, pH 7.2 500 mL 43149
Potassium Iodide Solution, 100 g/L 500 mL 1228949
Probe clips, color-coded, for IntelliCAL probes 50/pkg 5818400
Sodium Hydroxide, pellets, ACS 500 g 18734
Sodium Hydroxide Standard Solution, 1.00 N 100 mL MDB 104532
Sodium Thiosulfate Standard Solution, 0.025 N 1L 35253
Sodium Thiosulfate Standard Solution, 0.1 N 100 mL 32332
Starch Indicator Solution 100 mL MDB 34932
Sulfuric Acid Standard Solution, 0.020 N 1L 20353
Sulfuric Acid Solution, 1.00 N 1000 mL 127053

Oxygen Demand, Biochemical, dilution method 11

 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
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© Hach Company/Hach Lange GmbH, 2007–2015, 2017. All rights reserved. 08/2017, Edition 10
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

Instrument-specific information
Table 1 shows all of the instruments that have the program for this test. The table also
shows adapter and light shield requirements for the instruments that use them.
To use the table, select an instrument, then read across to find the applicable information
for this test.
Table 1 Instrument-specific information for test tubes
Instrument Adapters Light shield
DR 6000, DR 5000 — —
DR 3900 — LZV849
DR 3800, DR 2800, DR 2700 — LZV646
DR 1900 9609900 (D1) —
DR 900 4846400 Cover supplied with the instrument
1 The D adapter is not available with all instrument versions.

Before starting
Install the instrument cap on the DR 900 cell holder before ZERO or READ is pushed.
DR 3900, DR 3800, DR 2800 and DR 2700: Install the light shield in Cell Compartment #2 before this test is started.
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.
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used. Use the recommended personal protective
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.
Dispose of reacted solutions according to local, state and federal regulations. Refer to the Safety Data Sheets for disposal
information for unused reagents. Refer to the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.

1
Items to collect
Description Quantity

Beaker, 250-mL 1
Blender 1
COD Digestion Reagent vials varies
DRB200 Reactor 1
Light shield or adapter (For information about sample cells, adapters or light shields, refer to
1
Instrument-specific information on page 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

Refer to Consumables and replacement items on page 7 for order information.

Sample collection and storage

• 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.
• Homogenize samples that contain solids to get a representative sample.
• To preserve samples for later analysis, adjust the sample pH to less than 2 with
concentrated sulfuric acid (approximately 2 mL per liter). No acid addition is
necessary if the sample is tested immediately.
• Keep the preserved samples at 2–6 °C (36–43 °F) for a maximum of 28 days.
• Correct the test result for the dilution caused by the volume additions.

Reactor digestion procedure

1. Put 100 mL of sample in
a blender. Blend for
30 seconds or until
homogenized.
For samples with large
amounts of solids, increase
the homogenization time. If
the sample does not contain
suspended solids, go to step
3.
2. For the 200–15,000 mg/L
range or to improve
accuracy and reproducibility
of the other ranges, pour the
homogenized sample into a
250‑mL beaker and gently
stir with a magnetic stir
plate.
3. Set the DRB200 Reactor
power to on. Preheat to
150 °C.
Refer to the DRB200 User
Manual for selecting pre-
programmed temperature
applications.
4. Prepare the sample:
Remove the cap from a vial
for the selected range. Hold
the vial at an angle of
45 degrees. Use a clean
pipet to add 2.00 mL of
sample to the vial.
For 250–15,000 mg/L vials:
Use a TenSette Pipet to add
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:
Remove the cap from a
second vial for the selected
range. Hold the vial at an
angle of 45 degrees. Use a
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
2 hours.
Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L)
6. Close the vials tightly.
Rinse the vials with water
and wipe with a clean paper
towel.
10. Set the reactor power to
off. Let the vials cool in the
reactor for approximately
20 minutes to 120 °C or
less.
7. Hold the vials by the cap,
over a sink. Invert gently
several times to mix.
The vials get very hot
during mixing.
11. Invert each vial several
times while it is still warm.
8. Put the vials in the
preheated DRB200 reactor.
Close the lid.
12. Put the vials in a tube
rack to cool to room
temperature.
3
Colorimetric procedure

Start Zero

1. Start program 431 COD 2. Clean the blank sample 3. Insert the blank into the 4. Push ZERO. The display
ULR, 430 COD LR or 435 cell. cell holder. shows 0 or 0.0 mg/L COD.
COD HR. For information
about sample cells,
adapters or light shields,
refer to Instrument-specific
information on page 1.
Note: Although the program
name can be different
between instruments, the
program number does not
change.

Read

5. Clean the prepared
sample cell.
6. Insert the prepared
sample into the cell holder.
7. Push READ. Results
show in mg/L COD.
8. If using High Range Plus
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.

4 Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L)
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
Standard solution method
Items to collect:
• 1000 mg/L COD standard solution
• 100-mL volumetric flask, Class A
• 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.
b. Dilute to the mark with deionized water. Mix well.
2. Use the test procedure to measure the concentration of the 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.

Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L) 5
 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.
b. Dilute to the mark with deionized water. Mix well.
2. Use the test procedure to measure the concentration of the 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.

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.
2. 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.

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.
2. Use the test procedure to measure the concentration of the 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.

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
The method performance data that follows was derived from laboratory tests that were
measured on a spectrophotometer during ideal test conditions. Users can get different
results under different test conditions.
Program Standard Precision (95% Confidence Interval) Sensitivity
Concentration change per 0.010 Abs change
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

6 Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L)
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 (Cr2O72–) 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
0.7–40.0 mg/L 350 nm (for applicable instruments)
3–150 mg/L 420 nm
20–1500 620 nm (610 nm for colorimeters)
200–15,000 mg/L 620 nm (610 nm for colorimeters)

Pollution prevention and waste management

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.
Consumables and replacement items
Required reagents

Description Quantity/test Unit Item no.

COD, Ultra Low Range, 0.7–40 mg/L 1–2 vials 25/pkg 2415825
COD, Low Range, 3–150 mg/L 1–2 vials 25/pkg 2125825
COD, High Range, 20–1500 mg/L 1–2 vials 25/pkg 2125925
COD, High Range Plus, 200–15,000 mg/L 1–2 vials 25/pkg 2415925
Water, deionized varies 4L 27256

Alternate reagents and package sizes

Description Quantity/test Unit Item no.

COD2, Low Range, 0–150 mg/L COD 1–2 vials 25/pkg 2565025
COD2, High Range, 0–1500 mg/L COD 1–2 vials 25/pkg 2565125
COD2, High Range, 0–1500 mg/L COD 1–2 vials 150/pkg 2565115
COD2, High Range Plus, 0–15,000 mg/L COD 1–2 vials 25/pkg 2834325
COD Digestion Reagent Vials, 3–150 mg/L COD 1–2 vials 150/pkg 2125815
COD Digestion Reagent Vials, 200–1500 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

Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L) 7
Required apparatus

Description Quantity/test Unit Item no.

Blender, 2-speed, 120 VAC option 1 each 2616100

OR
Blender, 2-speed, 240 VAC option 1 each 2616102
DRB 200 Reactor, 110 VAC option, 15 x 16-mm wells 1 each LTV082.53.40001
OR
DRB 200 Reactor, 220 VAC option, 15 x 16-mm wells 1 each LTV082.52.40001
Pipet filler, safety bulb 1 each 1465100
Pipet, volumetric, Class A, 2.00-mL 1 each 1451536

Recommended standards and apparatus

Description Unit Item no.

Beaker, 250-mL each 50046H

COD Standard Solution, 300-mg/L 200 mL 1218629
COD Standard Solution, 300-mg/L 500mL 1218649
COD Standard Solution, 800-mg/L 200 mL 2672629
COD Standard Solution, 1000-mg/L 200 mL 2253929
Oxygen Demand Standard (BOD, COD, TOC), 10-mL ampules 16/pkg 2833510
®
Pipet, TenSette , 0.1–1.0 mL each 1970001
®
Pipet tips for TenSette Pipet, 0.1–1.0 mL 50/pkg 2185696
®
Pipet tips for TenSette Pipet, 0.1–1.0 mL 1000/pkg 2185628
Potassium Acid Phthalate (KHP), ACS 500 g 31534
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, stainless steel each 1864100
Wipes, disposable 70/pkg 2096900

Optional reagents and apparatus

Description Unit Item no.

Balance, analytical, 80 g x 0.1 mg 100–240 VAC each 2936701

Flask, volumetric, Class A, 1000-mL glass each 1457453
Flask, volumetric, Class A, 100-mL glass each 1457442
Mercuric Sulfate 28 g 191520
Pipet, volumetric, Class A, 3-mL each 1451503
Pipet, volumetric, Class A, 10-mL each 1451538
Sulfuric Acid, ACS 500 mL 97949
Wastewater Influent Standard Solution, Mixed Parameter, for NH3-N, NO3-N, PO4,
500 mL 2833149
COD, SO4, TOC

8 Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L)
Optional reagents and apparatus (continued)

Description Unit Item no.

EZ COD™ Recycling Service with 5-gal bucket-mail back option (For US customers
each 2895405
only. 20 and 55 gallon sizes are also available. )
EZ COD™ Recycling Service with 5-gal bucket- pick up option. (For US customers
each 2895405P
only. 20 and 55 gallon sizes are also available. )
Finger cots 2/pkg 1464702
Gloves, chemical resistant, size 9–9.5 pair 24101041
Paper, for weighing, 100 x 100 mm 500/pkg 1473885
Safety goggles, vented each 2550700
Wastewater Effluent Standard Solution, Mixed Parameter, for NH3-N, NO3-N, PO43–,
500 mL 2833249
COD, SO42–, TOC
1 Other sizes available

Oxygen Demand, Chemical, Dichromate Method (multi-range: 40.0, 150, 1500, 15,000 mg/L) 9
 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, 1989–2014. All rights reserved. 10/2014, Edition 10
pH DOC316.53.01245

USEPA Electrode Method Method 8156

pH electrode
Scope and application: For drinking water1, wastewater2 and process water.
1 Based on Standard Method 4500-H+B, ASTM Method D1293-95 and USEPA Method 150.1
2 Based on Standard Method 4500-H+B, ASTM Method D1293-84(90)/(A or B) and USEPA Method 150.1

Test preparation

Instrument specific information

This procedure is applicable to the meters and probes that are shown in Table 1.
Procedures for other meters and probes can be different.
Table 1 Instrument-specific information
Meter Probe
HQ11d portable single input, pH/ORP IntelliCAL PHC101, PHC201, PHC281 or PHC301 pH
HQ30d portable single input, multi-parameter
HQ40d portable dual input, multi-parameter
HQ411d benchtop single input, pH/mV
HQ430d benchtop single input, multi-parameter
HQ440d benchtop dual input, multi-parameter
sensION™+ MM156 portable pH/EC/DO sensION+ 5049 multi-parameter
sensION™+ pH1 portable pH sensION+ 5050T, 5051T or 5052T combination pH
sensION™+ MM110 portable pH/ORP sensION+ 5045, 5048 or 5059 multi-parameter
sensION™+ MM150 portable pH/ORP/EC
sensION™+ pH3 lab pH sensION+ 5010T, 5011T, 5014T or 5021T combination pH
sensION™+ pH31 GLP lab pH
sensION™+ MM340 lab dual input, pH/mV/ISE
sensION™+ MM374 lab dual input, pH/mV/EC/ISE
sensION™+ MM378 lab dual input, pH/ISE/EC/DO

Before starting
Refer to the meter documentation for meter settings and operation. Refer to probe documentation for probe preparation,
maintenance and storage information.
Prepare the probe before initial use. Refer to probe documentation.
When an IntelliCAL™ probe is connected to an HQd meter, the meter automatically identifies the measurement parameter
and is prepared for use.
Condition the electrode for the best response time. To condition the electrode, soak the electrode for several minutes in a
solution that has almost the same pH and ionic strength as the sample.
Calibrate the probe before initial use. Refer to Calibration procedure on page 3.
For rugged electrodes, it may be necessary to remove the shroud before measurement and calibration.
Air bubbles under the sensor tip can cause slow response or measurement errors. To remove the bubbles, carefully shake
the probe.
To save data automatically, set the measurement mode to Press to Read or Interval. When the measurement mode is
Continuous, select Store to save data manually.
Rinse the electrode between measurements to prevent contamination.

1
 Keep the electrode in a pH storage solution when not in use. Refer to the probe documentation.
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used. Use the recommended personal protective
equipment.
Dispose of reacted solutions according to local, state and federal regulations. Refer to the Safety Data Sheets for disposal
information for unused reagents. Refer to the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.
This procedure is specified for the HQd meters. The sensION+ meters can be used, but the menus and navigation will be
different.

Items to collect
Description Quantity

Beaker or sample containers 3

Wash bottle with deionized water 1
pH buffers (4.0, 7.0, 10.0) 3

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

Sample collection
• Analyze the samples immediately. The samples cannot be preserved for later
analysis.
• Collect samples in clean glass or plastic bottles.

Test procedure

1. Rinse the probe with
deionized water. Dry the
probe with a lint-free cloth.
2. Laboratory test: Put the 3. Push Read. A progress 4. Rinse the probe with
probe in a beaker that bar is shown. When the deionized water. Dry the
contains the solution. Do not measurement is stable, the probe with a lint-free cloth.
let the probe touch the stir lock icon is shown.
bar, bottom or sides of the
container. Remove the air
bubbles from under the
probe tip. Stir the sample at
a slow to moderate rate.
Field test: Put the probe in
the sample. Move the probe
up and down to remove
bubbles from the electrode.
Make sure to put the
temperature sensor fully in
the sample.

2 pH, USEPA electrode method

Calibration procedure

1. Prepare two or three
fresh buffer solutions in
separate beakers. If two
buffers are used, use a 7.0
and a 4.0 or a 7.0 and a
10.0 pH buffer solution.
2. Add a stir bar and put the
beaker on a magnetic
stirrer. Stir at a moderate
rate.
3. Rinse the probe with
deionized water. Dry the
probe with a lint-free cloth.
4. Put the probe in the
solution. Do not let the
probe touch the stir bar,
bottom or sides of the
container. Remove the air
bubbles from under the
probe tip.

5. Push Calibrate. The
standard solution value is
shown.
6. Push Read. A progress
bar is shown. When the
measurement is stable, the
lock icon is shown.
7. Measure the remaining
buffer solutions.
8. Push Done. A calibration
summary is shown when the
minimum number of
calibration standards are
measured.

9. Push Store to accept the

calibration.

Low ionic strength or high-purity water measurements

NOTICE
Do not keep the probe in LIS samples for a long period of time because this can decrease the
probe life. Put the probe in electrode storage solution or 3 M KCl when LIS measurements are
complete.

Low ionic strength (LIS) solutions have very low buffering capacity and absorb carbon
dioxide from the air. When a sample absorbs carbon dioxide from the atmosphere,
carbonic acid forms. Carbonic acid decreases the sample pH, which causes inaccurate

pH, USEPA electrode method 3

 readings. One solution to this problem is to measure the sample in a low volume, airtight
sample chamber such as a low ionic strength chamber.
Use refillable or platinum series electrodes for measurement of pH in LIS or high purity
waters.
Before an LIS sample is measured, condition the probe as follows:
1. Soak the probe in a solution equivalent to the sample in ionic strength and pH for
10 to 15 minutes.
2. Rinse the probe with deionized water.
3. Dry the probe with a soft paper towel.
Between measurements, keep the probe in the sample or a neutral LIS solution (e.g., tap
water) for a maximum of 2 hours.
Interferences
The sodium error is low but increases at pH values that are higher than pH 11. The acid
error is negligible. Refer to the electrode or the meter documentation.
Accuracy check
Slope test
The electrode operation is satisfactory when the calibration slope is within the specified
range (typically –58 mV (±3) at 25 °C).

Calibration accuracy
Measure the pH of a fresh buffer solution. A calibration is satisfactory when the measured
pH value agrees with the known pH value of the buffer solution.
Clean the probe
Clean the probe when:
• Drifting/inaccurate readings occur as a result of contamination on the sensing
element or incorrect storage conditions.
• Slow response time occurs as a result of contamination on the sensing element.
• The slope is out of range as a result of contamination on the sensing element.
For general contamination, complete the steps that follow.

1. Rinse the probe with deionized water. Blot dry with a lint-free cloth.
2. Soak the glass bulb for 12 to 16 hours in Hach Electrode Cleaning Solution.
3. Rinse or soak the probe for 1 minute in deionized water.
4. Soak the probe in pH 4 buffer for up to 20 minutes, then rinse with deionized water.
5. Blot dry with a lint-free cloth.
6. If harsh contaminants are attached to the probe, polish the probe tip with a soft cloth
or cotton swab to remove the contaminants.
7. Soak for up to 20 minutes in pH 4 buffer, then rinse with deionized water.

Method performance
The accuracy of the measurements is dependent on many factors that are related with
the overall system, which includes the meter, the probe and calibration solutions. Refer to
the meter or probe documentation for more information.
Summary of method
A combination pH electrode develops a potential at the glass/liquid interface. At a
constant temperature, this potential varies linearly with the pH of the solution.
The pH is the hydrogen ion activity in a solution and is defined as –log10a(H+), where
a(H+) is the activity of the hydrogen ion. The sample pH can change when carbon dioxide
is absorbed from the atmosphere. In water that has a high conductivity, the buffer
capacity is typically high and the pH does not change much.

4 pH, USEPA electrode method

Consumables and replacement items
HQd meters and probes

Description Unit Item no.

HQ11d portable single input, pH/ORP meter each HQ11D53000000

HQ30d portable single input, multi-parameter meter each HQ30D53000000
HQ40d portable dual input, multi-parameter meter each HQ40D53000000
HQ411d benchtop single input, pH/mV meter each HQ411D
HQ430d benchtop single input, multi-parameter meter each HQ430D
HQ440d benchtop dual input, multi-parameter meter each HQ440D
IntelliCAL™ pH gel probe, standard with 1 m cable each PHC10101
IntelliCAL™ pH gel probe, standard with 3 m cable each PHC10103
IntelliCAL™ pH gel probe, rugged with 5 m cable each PHC10105
IntelliCAL™ pH gel probe, rugged with 10 m cable each PHC10110
IntelliCAL™ pH gel probe, rugged with 15 m cable each PHC10115
IntelliCAL™ pH gel probe, rugged with 30 m cable each PHC10130
IntelliCAL™ pH gel probe, standard with 1 m cable each PHC20101
IntelliCAL™ pH gel probe, standard with 3 m cable each PHC20103
IntelliCAL™ pH gel probe, ultra with 1 m cable each PHC28101
IntelliCAL™ pH gel probe, ultra with 3 m cable each PHC28103
IntelliCAL™ pH liquid probe, standard with 1 m cable each PHC30101
IntelliCAL™ pH liquid probe, standard with 3 m cable each PHC30103

sensION+ meters and probes

Description Unit Item no.

sensION™+ pH1 portable pH meter each LPV2500.97.0002

sensION™+ MM110 portable pH/ORP meter each LPV2600.97.0002
sensION™+ MM150 portable pH/ORP/EC meter each LPV4000.97.0002
sensION™+ MM156 portable pH/EC/DO meter each LPV4030.97.0002
sensION™+ pH3 lab pH meter each LPV2010T.97.002
sensION™+ pH31 GLP lab pH meter each LPV2110T.97.002
sensION™+ MM340 lab dual input, pH/mV/ISE meter each LPV2200.97.0002
sensION™+ MM374 lab dual input, pH/mV/EC/ISE meter each LPV4110.97.0002
sensION™+ MM378 lab dual input, pH/ISE/EC/DO meter each LPV4130.97.0002
sensION™+ 5010T combination pH probe each LZW5010T.97.002
sensION™+ 5011T combination pH probe each LZW5011T.97.002
sensION™+ 5014T combination pH probe each LZW5014T.97.002
sensION™+ 5021T combination pH probe each LZW5021T.97.002
sensION™+ 5050T combination pH probe each LZW5050T.97.002
sensION™+ 5051T combination pH probe each LZW5051T.97.002
sensION™+ 5052T combination pH probe each LZW5052T.97.002

pH, USEPA electrode method 5

sensION+ meters and probes (continued)

Description Unit Item no.

sensION™+ 5045 multi-parameter probe each LZW5045.97.0002

sensION™+ 5048 multi-parameter probe each LZW5048.97.0002
sensION™+ 5049 multi-parameter probe each LZW5049.97.0002
sensION™+ 5059 multi-parameter probe each LZW5059.97.0002

Recommended standards

Description Unit Item no.

pH color-coded buffer solution kit (NIST), 500 mL, includes: 1 2947600

pH 4.01 ± 0.02 pH buffer (NIST) 500 mL 2283449
pH 7.00 ± 0.02 pH buffer (NIST) 500 mL 2283549
pH 10.01 ± 0.02 pH buffer (NIST) 500 mL 2283649
Powder pillows:
pH 4.01 ± 0.02 pH buffer powder pillow (NIST) 50/pkg 2226966
pH 7.00 ± 0.02 pH buffer powder pillow (NIST) 50/pkg 2227066
pH 10.01 ± 0.02 pH buffer powder pillow (NIST) 50/pkg 2227166
Radiometer Analytical (IUPAC Series certified pH standards):
pH 1.679 ± 0.010 at 25 °C (77 °F) 500 mL S11M001
pH 4.005 ± 0.010 at 25 °C (77 °F) 500 mL S11M002
pH 7.000 ± 0.010 at 25 °C (77 °F) 500 mL S11M004
pH 10.012 ± 0.010 at 25 °C (77 °F) 500 mL S11M007
pH buffer 1.09, technical 500 mL S11M009
pH buffer 4.65, technical 500 mL S11M010
pH buffer 9.23, technical 500 mL S11M011

Accessories

Description Unit Item no.

Beaker, polypropylene, 50-mL, low form each 108041

Beaker, polypropylene, 100-mL each 108042
Bottle, wash, 500-mL each 62011
Stir bar, magnetic, 2.2 x 0.5 cm (7/8 x 3/16 in.) each 4531500
Stirrer, electromagnetic, 120 VAC, with electrode stand each 4530001
Stirrer, electromagnetic, 230 VAC, with electrode stand each 4530002
Sample bottle with screw-top cap, polypropylene, 500-mL each 2758101
Water, deionized 4L 27256

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, 2007–2015. All rights reserved. 03/2015, Edition 8
Suspended Solids DOC316.53.01139

Photometric Method1 Method 8006

5 to 750 mg/L TSS
Scope and application: For water and wastewater.
1 Adapted from Sewage and Industrial Wastes, 31, 1159 (1959).

Test preparation

Instrument-specific information
Table 1 shows all of the instruments that have the program for this test. The table also
shows sample cell and orientation requirements for reagent addition tests, such as
powder pillow or bulk reagent tests.
To use the table, select an instrument, then read across to find the applicable information
for this test.
Table 1 Instrument-specific information
Instrument Sample cell orientation Sample cell
DR 6000 The fill line is to the right. 2495402
DR 3800
DR 2800
DR 2700
DR 1900
DR 5000 The fill line is toward the user.
DR 3900
DR 900 The orientation mark is toward the user. 2401906

Before starting
For turbidimetric methods, install the instrument cap or cover on all instruments before ZERO or READ is pushed.
Do not use the Pour-Thru Cell or sipper module (for applicable instruments) with this test.

Items to collect
Description Quantity

Beaker, 600-mL, polypropylene 1

Blender 1
Cylinder, 500-mL polypropylene, graduated 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 replacement items on page 3 for order information.

1
Sample collection and storage
• Collect samples in clean glass or plastic bottles.
• To preserve samples for later analysis, keep the samples at or below 6 °C (43 °F) for
up to 7 days.
• Let the sample temperature increase to room temperature before analysis.

Photometric procedure

Start

1. Start program 630 2. Blend 500 mL of sample 3. Pour the blended sample 4. Prepare the sample:
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.
specific information
on page 1.
Note: Although the program
name can be different
between instruments, the
program number does not
change.

Zero

5. Prepare the blank: Fill a 6. Clean the blank sample 7. Insert the blank into the 8. Push ZERO. The display
second sample cell with cell. cell holder. shows 0 mg/L TSS.
10 mL of tap water or
deionized water.

2 Suspended Solids, Photometric Method (750 mg/L)

 Read

9. Swirl the prepared 10. Clean the prepared 11. Insert the prepared 12. Push READ. Results
sample to remove any gas sample cell. sample into the cell holder. show in mg/L TSS.
bubbles and uniformly
suspend any residue.

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
Standard solution method
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
810 nm (DR 1900: 800 nm) for spectrophotometers or 610 nm for colorimeters.
Consumables and replacement items
Required apparatus

Description Quantity/test Unit Item no.

Beaker, 600-mL, polypropylene 1 each 108052

Blender, 2-speed, 120 VAC option 1 each 2616100
Blender, 2-speed, 240 VAC option 1 each 2616102
Cylinder, graduated, 500-mL, polypropylene 1 each 108149

Suspended Solids, Photometric Method (750 mg/L) 3

 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, 1989–2014. All rights reserved. 01/2014, Edition 9
Oxygen, Dissolved DOC316.53.01161

Azide Modification of Winkler Method1, 2 Method 8229

1 to more than 10 mg/L Buret Titration
Scope and application: For water, wastewater and seawater.
1 USEPA approved.
2 Adapted from Standard Methods for the Examination of Water and Wastewater, Standard Method 4500 O C.

Test preparation

Before starting
As an alternative to the powder pillows, use standard APHA solutions for dissolved oxygen. Use 1 mL of Manganous Sulfate
Solution, 1 mL of Alkaline Iodide-Azide Reagent and 1 mL of Sulfuric Acid (concentrated) as an alternative to the powder
pillows. Add the solutions below the surface of the liquid.
The optional TitraStir Titration Stand can hold the buret and stir the sample.
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used. Use the recommended personal protective
equipment.
Dispose of reacted solutions according to local, state and federal regulations. Refer to the Safety Data Sheets for disposal
information for unused reagents. Refer to the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.

Items to collect
Description Quantity

BOD bottle, 300 mL 1

Alkaline Iodide-Azide Reagent Powder Pillow 1
Manganous Sulfate Powder Pillow 1
Sulfamic Acid Powder Pillow 1
Sodium Thiosulfate Standard Solution (titrant), 0.025 N varies
Starch Indicator Solution varies
Buret, Class A, 25 mL 1
Graduated cylinder, 250 mL 1
Funnel, micro 1
Support stand with buret clamp 1
Water, deionized varies

Refer to Consumables and replacement items on page 4 for order information.

Sample collection
Good sample collection and handling techniques are important to get correct results. The
dissolved oxygen content of the sample can change with depth, turbulence, temperature,
sludge deposits, light, microbial action, mixing, travel time and other factors. A single
dissolved oxygen test frequently is not an accurate reflection of the overall condition of a
body of water. Several samples taken at different times, locations and depths are
recommended for most reliable results.
• Collect samples in clean BOD Bottles.

1
 • If prompt analysis is not possible, do steps 1 through 4 of the procedure and keep the
samples protected from light at 10 to 20 °C (50 to 68 °F).
• Pour a small quantity of water into the flared lip area of the stopper to seal the bottle.
• Push a BOD bottle cap on the flared lip.
• Keep samples for a maximum of 8 hours. For analysis start with step 5.

Test procedure

1. Collect a water sample in
a clean 300‑mL BOD bottle.
Let the sample overflow the
bottle for 2 or 3 minutes to
remove trapped air bubbles
and to make sure that a
representative sample is
collected.
2. Add the contents of one
Manganous Sulfate Powder
Pillow and one Alkaline
Iodide-Azide Reagent
Powder Pillow to the
sample.
3. Immediately put the
stopper in the bottle. Make
sure that no air is inside the
bottle.
4. Invert the bottle at least 5
times to mix.
A flocculent precipitate
forms. The floc is
orange/brown if oxygen is in
the sample or white if there
is no oxygen.
The floc forms slowly in salt
water (approximately
5 minutes more are
necessary). When the floc
formation is complete,
proceed to next step.

5. Again, invert the bottle at 6. Remove the stopper and 7. Immediately put the 8. Invert the bottle at least 5
least 5 times to mix. add the contents of one stopper in the bottle. Make times to mix.
Wait until the top half of the Sulfamic Acid Powder Pillow sure that no air is inside the The floc dissolves and a
solution is clear and the floc to the sample. bottle. yellow color develops if
collects at the bottom to oxygen is in the sample.
make sure that the reaction
of the sample and reagents
is complete. Some
suspended floc will have no
effect on accuracy.

2 Oxygen, Dissolved
9. Pour the prepared
sample into a 250‑mL
graduated cylinder to the
200‑mL mark.
10. Pour the contents of the
graduated cylinder into a
clean, 250‑mL Erlenmeyer
flask.
11. Fill a 25‑mL buret to the
zero mark with 0.025 N
Sodium Thiosulfate
Standard Solution.
12. Put the flask under the
buret. Swirl the flask. Add
titrant until the color
changes yellow to a pale
yellow.

13. Add 1 mL of Starch
Indicator Solution.
A dark blue color develops.
14. Put the flask under the
buret. Swirl the flask. Add
titrant until the color
changes from a dark blue to
a colorless end point.
15. Calculate the
concentration.
mL of titrant = mg/L DO

Interferences
Nitrite interference is removed by the azide in the reagents. Other reducing or oxidizing
substances may interfere. If these are in the sample, use an alternative method, such as
the High Range Dissolved Oxygen Method (colorimetric, Method 8166) or a dissolved
oxygen electrode.

Prepare activated sludge samples

Prepare activated sludge samples for the test as follows:

1. Add 10 mL of Copper Sulfate-Sulfamic Acid Inhibitor Solution to a clean, 1000-mL

graduated cylinder.
2. Use a tube to fill the cylinder with sample. Put the end of the tube near the bottom of
the cylinder. Let approximately 200 mL of sample flow over the cylinder.
3. Swirl the cylinder to mix the contents. Let the suspended solids go to the bottom of
the cylinder.
4. Use a siphon tube to collect the transparent top layer in a 300-mL BOD bottle. Put the
end of the tube near the bottom of the BOD bottle. When the water flows over the
BOD bottle, remove the tube. Make sure that no air bubbles are in the bottle.
5. Start at step 2 of the test procedure.

Accuracy check
Standard solution method
Use the standard solution method to validate the test procedure and the concentration of
the titrant.

Oxygen, Dissolved 3
 Items to collect:
• Iodate-Iodide Standard Solution, 0.00125 N (equivalent to 10 mg/L as O2)

1. Add 200 mL of 0.00125 N Iodate-Iodide Standard Solution to a 250-mL Erlenmeyer

flask.
2. Add one Sulfamic Acid Powder Pillow to the bottle.
3. Start at step 12 of the test procedure. Titrate the prepared standard solution to the
endpoint. Record the mL of titrant added in step 14.
4. Compare the actual result to the correct result. The correct result for this titration is
10.0 mL of titrant. If more than 10.5 mL of titrant was used, discard the titrant. Get
new titrant.

Summary of method
The Azide Modification of the Winkler Method is the standard test for dissolved oxygen. In
the analysis, manganous ion reacts with the dissolved oxygen in the alkaline solution to
form a manganese (IV) oxide hydroxide flocculent. Then, Azide is added to suppress
interference from nitrite, which would react with the iodide. Then, the solution is acidified
and the manganese (IV) floc is decreased by iodide to make free iodine as I3– in
proportion to the oxygen concentration. Then, the liberated iodine is titrated to the starch-
iodide end point.
Consumables and replacement items
Required reagents

Description Quantity/test Unit Item no.

Alkaline Iodide-Azide Powder Pillows 1 pillow 50/pkg 107266

Manganous Sulfate Powder Pillows 1 pillow 50/pkg 107166
Sodium Thiosulfate Standard Solution, 0.025 N varies 1L 2409353
Starch Indicator Solution 1 mL 100 mL MDB1 34932
Sulfamic Acid Powder Pillows 1 pillow 50/pkg 2076266

Required apparatus

Description Quantity/test Unit Item no.

Bottle, with stopper, BOD, 300-mL 1 each 62100

Buret clamp, double 1 each 32800
Buret, Class A, 25 mL 1 each 2636540
Support stand 1 each 56300
Funnel, micro 1 each 2584335
Cylinder, graduated, 250 mL 1 each 50846
Flask, Erlenmeyer, 250 mL 1 each 50546

Recommended standards

Description Unit Item no.

Iodate-Iodide Standard Solution, 0.00125 N 500 mL 40149

1 MDB is Marked Dropper Bottle

4 Oxygen, Dissolved
Optional reagents and apparatus

Description Unit Item no.

APHA reagents: — —
Alkaline Iodide-Azide Reagent Solution 500 mL 27749
Manganous Sulfate Solution 500 mL 27549
Sodium Thiosulfate Standard Solution, 0.025 N 1L 35253
Sulfuric Acid, ACS 500 mL 97949
Cap, BOD Bottle Snap-over 6/pkg 241906
BOD Bottle, serialized (#1–24), 300 mL 24/pkg 2898700
Copper Sulfate-Sulfamic Acid Inhibitor Solution 100 mL 35732
Copper Sulfate-Sulfamic Acid Inhibitor Solution 500 mL 35749
Stir bar, octagonal each 2095352
TitraStir® Titration Stand, 115 VAC each 1940000
TitraStir® Titration Stand, 230 VAC each 1940010

Oxygen, Dissolved 5
 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, 2007–2015. All rights reserved. 05/2015, Edition 8
Nitrogen, Ammonia DOC316.53.01078

USEPA1 Nessler Method2 Method 8038

0.02 to 2.50 mg/L NH3–N Reagent Solution
Scope and application: For water, wastewater and seawater. Distillation is required for wastewater and
seawater.
1 USEPA accepted for wastewater analysis (distillation required), Method 350.2.
2 Adapted from Standard Methods for the Examination of Water and Wastewater, 4500-NH3 B & C, 15th Edition.

Test preparation

Instrument-specific information
Table 1 shows all of the instruments that have the program for this test. The table also
shows sample cell and orientation requirements for specific instruments.
To use the table, select an instrument, then read across to find the applicable information
for this test.
Table 1 Instrument-specific information
Instrument Sample cell orientation Sample cell
DR 6000 The fill line is to the right. 2495402
DR 3800
DR 2800
DR 2700
DR 1900
DR 5000 The fill line is toward the user.
DR 3900

Before starting
Hold the reagent droppers and dropper bottles vertically, not at an angle, when the reagent is added.
The reagents that are used in this test contain mercury. Collect the reacted samples for safe disposal.
If the Pour-Thru Cell is used, clean the cell periodically. To clean, add several crystals of sodium thiosulfate pentahydrate
into the cell funnel. Add deionized water to dissolve the crystals. Rinse fully with deionized water.
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used. Use the recommended personal protective
equipment.
Dispose of reacted solutions according to local, state and federal regulations. Refer to the Safety Data Sheets for disposal
information for unused reagents. Refer to the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.

Items to collect
Description Quantity

Ammonia Nitrogen Reagent Set 1

Water, deionized 25 mL
Pipet, serological, 1-mL 1
Mixing cylinder, graduated, 25 mL, glass stopper 2
Sample cells (For information about sample cells, adapters or light shields, refer to Instrument-
2
specific information on page 1.)

1
Refer to Consumables and replacement items on page 5 for order information.

Sample collection and storage

• Collect samples in clean glass or plastic bottles.
• If the sample contains chlorine, add one drop of 0.1 N sodium thiosulfate for each
0.3 mg/L chlorine in 1 liter of sample.
• To preserve samples for later analysis, adjust the sample pH to less than 2 with
concentrated sulfuric acid (approximately 2 mL per liter). No acid addition is
necessary if the sample is tested immediately.
• Keep the preserved samples at or below 6 °C (43 °F) for a maximum of 28 days.
• Let the sample temperature increase to room temperature before analysis.
• Before analysis, adjust the pH to ~7 with 5 N sodium hydroxide solution.
• Correct the test result for the dilution caused by the volume additions.

Test procedure

Start

1. Start program 380 N, 2. Prepare the sample: Fill 3. Prepare the blank: Fill a 4. Add 3 drops of Mineral
Ammonia, Ness. For a mixing cylinder to the 25- mixing cylinder to the 25‑mL Stabilizer to each mixing
information about sample mL line with sample. line with deionized water. cylinder.
cells, adapters or light
shields, refer to Instrument-
specific information
on page 1.

5. Put the stopper on the 6. Add 3 drops of Polyvinyl 7. Put the stopper on the 8. Use a pipet to add 1.0
mixing cylinders. Invert the Alcohol Dispersing Agent to mixing cylinders. Invert the mL of Nessler Reagent to
mixing cylinders several each mixing cylinder. mixing cylinders several each mixing cylinder.
times to mix. times to mix.

2 Nitrogen-Ammonia, Nessler Method (2.50 mg/L)

9. Put the stopper on the 10. Start the instrument 11. Pour 10 mL from the 12. When the timer expires,
mixing cylinders. Invert the timer. A 1-minute reaction blank cylinder into a sample clean the blank sample cell.
mixing cylinders several time starts. cell.
times to mix.

Zero

13. Insert the blank into the 14. Push ZERO. The 15. Pour 10 mL from the 16. Clean the prepared
cell holder. display shows 0.00 mg/L sample cylinder into a sample cell.
NH3–N. second sample cell.

Read

17. Insert the prepared 18. Push READ. Results

sample into the cell holder. show in mg/L NH3–N.

Interferences
Table 2 Interfering substances
Interfering substance Interference level
Chlorine Remove residual chlorine from a 250 mL sample by adding 1 drop of sodium thiosulfate for each
mg/L chlorine (Cl2). Sodium arsenite can be used instead of sodium thiosulfate. Refer to Sample
collection and storage on page 2.
Hardness A solution containing a mixture of 500 mg/L CaCO3 and 500 mg/L Mg as CaCO3 does not
interfere. If the hardness concentration is more than these concentrations, add extra Mineral
Stabilizer.
Iron Interferes at all levels by causing turbidity with Nessler Reagent.

Nitrogen-Ammonia, Nessler Method (2.50 mg/L) 3


Interfering substance
Seawater
Sulfide
Glycine, various aliphatic
and aromatic amines,
organic chloramines,
acetone, aldehydes and
alcohols
Table 2 Interfering substances (continued)
Interference level
Add 1.0 mL (27 drops) of Mineral Stabilizer to the sample before analysis. This complexes the
high magnesium concentrations found in sea water, but the sensitivity of the test is reduced by
30% due to the high chloride concentration. For best results, make a calibration with standards
that contain the same chloride concentration as seawater, or distill the sample.
Interferes at all levels by causing turbidity with Nessler Reagent.
May cause greenish or other off colors or turbidity. Distill the sample if these compounds are
present.

Pollution prevention and waste management

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.
Distillation
To eliminate most interferences, distill the sample, then use the distilled sample in the test
procedure.

1. Set up the distillation apparatus for general purpose distillation. Refer to the
Distillation Apparatus manual for proper assembly.
2. Measure 250 mL of sample into a 250-mL graduated cylinder.
3. Pour the sample into a 400-mL beaker. If the sample contains chlorine, add 1 drop of
0.1 N sodium thiosulfate solution for each 1 mg/L Cl2 to remove the chlorine.
4. Add 25 mL of borate buffer solution and mix. Adjust the pH to approximately 9.5 with
1 N sodium hydroxide solution. Use a pH meter to monitor the pH.
5. Pour the solution into the distillation flask.
6. Add a magnetic stir bar and 5 glass beads.
7. Use a graduated cylinder to measure 25 mL of deionized water into a 250-mL
Erlenmeyer flask. Add the contents of one Boric Acid Powder Pillow. Mix thoroughly.
8. Set the flask under the distillation apparatus drip tube. Elevate the flask so that the
end of the tube is immersed in the solution.
9. Set the stirrer power to on. Set the stir control to 5.
10. With the thermometer inserted, set the heat control to 10. The yellow pilot lamp is an
indication that the heater is on.
11. Turn on the water and adjust to maintain a steady flow through the condenser.
12. When 150 mL of distillate has been collected, turn the heater off. Immediately remove
the collection flask. Measure the distillate to make sure 150 mL was collected (total
volume = 175 mL).
13. Adjust the pH to approximately 7 with 1 N sodium hydroxide solution. Use a pH meter
to monitor the pH.
14. Quantitatively transfer the distillate into a 250-mL volumetric flask. Dilute to the mark
with deionized water. Mix well. Use the diluted distillate in the test 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:
• 50-mg/L Nitrogen-Ammonia Standard Solution

4 Nitrogen-Ammonia, Nessler Method (2.50 mg/L)

 • Mixing cylinders, 25-mL (3x)
• TenSette Pipet and pipet tips

1. Use the test procedure to measure the concentration of the sample, then keep the
(unspiked) sample in the instrument.
2. Go to the Standard Additions option in the instrument menu.
3. Select the values for standard concentration, sample volume and spike volumes.
4. Open the standard solution.
5. Prepare three spiked samples: use the TenSette pipet to add 0.1 mL, 0.2 mL and
0.3 mL of the standard solution, respectively, to three 25-mL portions of fresh sample.
Mix well.
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, the reagents and the
instrument.
Items to collect:
• 1-mg/L Nitrogen-Ammonia Standard Solution

1. Use the test procedure to measure the concentration of the standard solution.
2. 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 small
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 can get different
results under different test conditions.
Program Standard Precision (95% confidence interval) Sensitivity
Concentration change per 0.010 Abs change
380 1.00 mg/L NH3–N 0.99–1.01 mg/L NH3–N 0.02 mg/L NH3–N

Summary of Method
The Mineral Stabilizer complexes hardness in the sample. The Polyvinyl Alcohol
Dispersing Agent helps the color formation in the reaction of Nessler Reagent with
ammonia and certain other amines. A yellow color is formed that is proportional to the
ammonia concentration. The measurement wavelength is 425 nm.
Consumables and replacement items
Required reagents

Description Quantity/test Unit Item no.

Ammonia Nitrogen Reagent Set, includes: — — 2458200

Nessler Reagent 2 mL 500 mL 2119449
Mineral Stabilizer 6 drops 50 mL SCDB 2376626

Nitrogen-Ammonia, Nessler Method (2.50 mg/L) 5

Consumables and replacement items (continued)
Description Quantity/test Unit Item no.

Polyvinyl Alcohol Dispersing Agent 6 drops 50 mL SCDB 2376526

Water, deionized varies 4L 27256

Required apparatus

Description Quantity/test Unit Item no.

Mixing cylinder, graduated, 25 mL with stopper 1 each 2088640

Pipet, serological, 1 mL, glass 1 50/pkg 2093135
Pipet filler, safety bulb 1 each 1465100

Recommended standards

Description Unit Item no.

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

®
Nitrogen Ammonia Standard Solution, 10-mL Voluette Ampule, 50-mg/L NH3–N 16/pkg 1479110
Wastewater Effluent Standard Solution, Mixed Parameter, for NH3-N, NO3-N, PO43–,
500 mL 2833249
COD, SO42–, TOC

Optional reagents and apparatus

Description Unit Item no.

®
Ampule Breaker, 10-mL Voluette Ampules each 2196800
Distillation apparatus set, general purpose each 2265300
Distillation heater and support for apparatus set, 115 VAC option each 2274400
Distillation heater and support for apparatus set, 230 VAC option each 2274402
®
Pipet, TenSette , 0.1–1.0 mL each 1970001
®
Pipet tips for TenSette Pipet, 0.1–1.0 mL 50/pkg 2185696
Sodium Hydroxide Standard Solution, 5.0 N 100 mL MDB 245032
Sodium Thiosulfate, 0.1 N 100 mL 32332
Sulfuric Acid, ACS 500 mL 97949

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© Hach Company/Hach Lange GmbH, 2007, 2010, 2012, 2014, 2017. All rights reserved. 04/2017, Edition 9