2540 A.

Introduction
Solids refer to matter suspended or dissolved in water or wastewater. Solids analyses are important in
the control of biological and physical wastewater treatment processes and for assessing compliance
with regulatory agency wastewater effluent limitations.

1. Definitions ‘‘Total solids’’ is the term applied to the material residue left in the vessel after
evaporation of a sample and its subsequent drying in an oven at a defined temperature

2. Apparatus
Apparatus listed in Section 2540B.2 and Section 2540C.2 is required, except for evaporating dishes,
steam bath, and 180°C drying oven. In addition: Aluminum weighing dishes.

3. Procedure
A. Preparation of glass-fiber filter disk
If pre-prepared glass fiber filter disks are used, eliminate this step. Insert disk with
wrinkled side up in filtration apparatus. Apply vacuum and wash disk with three
successive 20-mL portions of reagent-grade water. Continue suction to remove all
traces of water, turn vacuum off, and discard washings. Remove filter from filtration
apparatus and transfer to an inert aluminum weighing dish. If a Gooch crucible is
used, remove crucible and filter combination. Dry in an oven at 103 to 105°C for 1 h.
If volatile solids are to be measured, ignite at 550°C for 15 min in a muffle furnace.
Cool in desiccator to balance temperature and weigh. Repeat cycle of drying or
igniting, cooling, desiccating, and weighing until a constant weight is obtained or until
weight change is less than 4% of the previous weighing or 0.5 mg, whichever is less.
Store in desiccator until needed.
b. Selection

B. Selection of filter and sample sizes

Choose sample volume to yield between 2.5 and 200 mg dried residue. If volume
filtered fails to meet minimum yield, increase sample volume up to 1 L. If complete
filtration takes more than 10 min, increase filter diameter or decrease sample
volume.

C. Sample analysis
 Assemble filtering apparatus and filter and begin suction. Wet filter with a small
volume of reagent-grade water to seat it. Stir sample with a magnetic stirrer at a
speed to shear larger particles, if practical, to obtain a more uniform (preferably
homogeneous) particle size. Centrifugal force may separate particles by size and
density, resulting in poor precision when point of sample withdrawal is varied. While
stirring, pipet a measured volume onto the seated glass-fiber filter.

4. Calculation
mg total suspended solids/L = (A-B)*1000
sample volume, ml
where: A = weight of filter + dried residue, mg, and B = weight of filter, mg.

5210 BIOCHEMICAL OXYGEN DEMAND (BOD)*#(1)

Introduction
1. General Discussion
The biochemical oxygen demand (BOD) determination is an empirical test in which
standardized laboratory procedures are used to determine the relative oxygen
requirements of wastewaters, effluents, and polluted waters

2. Dilution Requirements
The BOD concentration in most wastewaters exceeds the concentration of dissolved
oxygen (DO) available in an air-saturated sample.

2. Apparatus
A. Incubation bottles

B. Air incubator or water bath

3. Procedure
A. Preparation of dilution water
B. Dilution water storage
C. Glucose-glutamic acid check
D. Seeding
E. Sample pretreatment
4. Calculation
BOD5, mg/L = D1-D2
P
5210 Ultimate BOD Test (PROPOSED)
1. Apparatus
A. Incubation bottles
B. Reservoir bottle
C. Incubator or water bath
D. Oxygen-sensitive membrane electrode

3. Procedure
a. River water samples.
4. Calculations
BODt = UBOD (1 − e–kt)
where: BODt = oxygen uptake measured at time t, mg/L, and

k = first-order oxygen uptake rate.

5210 D. Respirometric Method (PROPOSED)

 1. Apparatus
a. Respirometer system
b. Incubator or water bath.
2. Calculations
C = [A − B(SA/SB)](1000/NA)

where:

C = corrected oxygen uptake of sample, mg/L,

A = measured oxygen uptake in seeded sample, mg,
B = measured oxygen uptake in seed control, mg,
SA = volume of seed in Sample A, mL,
SB = volume of seed in Sample B, mL, and
NA = volume of undiluted sample in Sample A, mL.

METHOD 9131
TOTAL COLIFORM: MULTIPLE TUBE FERMENTATION
TECHNIQUE
1.1 SUMMARY OF METHOD
1.2 The multiple-tube fermentation technique is a three-stage procedure in which the results
are statistically expressed in terms of the Most Probable Number (MPN). These stages --
the presumptive stage, confirmed stage, and completed test -- are briefly summarized
below. (For the analysis to be accurate, a five-tube test is required.)
1.2.1 Presumptive Stage: A series of lauryl tryptose broth primary fermentation
tubes are inoculated with graduated quantities of the sample to be tested. The
inoculated tubes are incubated at 35 + 0.5EC for 24 + 2 hr, at which time the tubes
are examined for gas formation. For the tubes in which no gas is formed, continue
incubation and examine for gas formation at the end of 48 + 3 hr. Formation of gas
in any amount within 48 + 3 hr is a positive presumptive test.
1.2.2 Confirmed Stage: The confirmed stage is used on all primary fermentation
tubes showing gas formation during the 24-hr and 48-hr periods. Fermentation
 tubes containing brilliant green lactose bile broth are inoculated with medium from
the tubes showing a positive result in the presumptive test. Inoculation should be
performed as soon as possible after gas formation occurs. The inoculated tubes
are incubated for 48 + 3 hr at 35 + 0.5EC. Formation of gas at any time in the tube
indicates a positive confirmed test.
1.2.3 Completed Test: The completed test is performed on all samples showing a
positive result in the confirmed test. It can also be used as a quality control
measure on 20% of all samples analyzed. One or more plates of eosin methylene
blue are streaked with sample to be analyzed. The streaked plates are incubated
for 24 + 2 hr at 35 + 0.5EC. After incubation, transfer one or more typical colonies
(nucleated, with or without metallic sheen) to a lauryl tryptose broth fermentation
tube and a nutrient agar slant. The fermentation tubes and agar slants are
incubated at 35 + 0.5EC for 24 + 2 hr, or for 48 + 3 hr if gas is not produced. From
the agar slants corresponding to the fermentation tubes in which gas
formation occurs, gram-stained samples are examined microscopically. The
formation of gas in the fermentation tube and the presence of gram-negative, non-
spore-forming, rod-shaped bacteria in the agar culture may be considered a
satisfactorily completed test, demonstrating the positive presence of coliform