METHOD

WASTE WATER

21st Edition

2005
PREPARED AND PUBLISHED JOINTLY BY
American Public Health Association
American Water Works Association
Water Environment Federation
JOINT EDITORIAL BOARD
. AndrewD. Eaton, AWWA, Chair
';....L?nore S. Clesceri, WEF
.,-j Eugene W. Rice, APHA
- Arnold E. Greenherg (deceased), APHA
MANAGING EDITOR
Mary Ann H. Franson
PUBLICATION OFFICE
American Public Health Association
8001 Street, NW
Washington, DC 20001-3 710

Centennial Edition

TABLE OF CONTENTS

PAGE

Part 2000 PHYSICAL AND AGGREGATE PROPERTIES

Part 1000 INTRODUCTION

1010 INTRODUCTION

1-1

A. Scope and Application of Methods. . . .

B. Statistics
C. Glossary
1020 QUALITY ASSURANCE

1-1
1-1
1-3
1-4

A. Introduction
B. Quality Control
C. Quality Assessment

1-4
1-5
1-12

1030 DATA QUALITY

1-13

A. Introduction
B. Measurement Uncertainty.
C. Method Detection Level
D. Data Quality Objectives
E. Checking Correctness of Analyses . . . .

1-13
1-13
1-17
1-18
1-21

1040 METHOD DEVELOPMENT AND EVALUATION . . .

1-22

A. Introduction
B. Method Validation
C. Collaborative Testing

1050 EXPRESSION OF-RESULTS

A. Units
B. Significant Figures . . . :

1-22
1-22
1-24
1-25

1-25
1-26

1060 COLLECTION AND PRESERVATION

OF SAMPLES

A. Introduction
B. Collection of Samples
C. Sample Storage and Preservation
1080 REAGENT WATER

A. Introduction
B. Methods for Preparation of Reagent
Water
C. Reagent Water Quality

1-27

1-27
1-29
1-34
1-36

1-36
1-36
1-37

1090 LABORATORY OCCUPATIONAL HEALTH

AND SAFETY

1-38

A. Introduction
1-38
B. Safe Laboratory Practices
1-39
C. Laboratory Facility/Fixed Equipment . . 1-44
D. Hazard Evaluation
1-45
E. Personal Protective Equipment
1-46
F. Worker Protection Medical Program. . . 1-49
G. Provisions for Work with Particularly
Hazardous Substances
1-50
H. Biological Safety
1-50
I. Radiological Safety
1-51
J. Chemical Hygiene Plan
1 -54
1100 WASTE MINIMIZATION AND DISPOSAL

A. Introduction
B. Waste Minimization
C. Waste Treatment and Disposal

1-55

1-55
1-55
1-55

2010 INTRODUCTION
2020 QUALITY ASSURANCE/QUALITY CONTROL

2-1
...

2110 APPEARANCE

2-1
2-1

2120 COLOR

2-1

A. Introduction
B. Visual Comparison Method
C. Spectrophotometric MethodSingleWavelength Method (PROPOSED) . .
D. Spectrophotometric MethodMultiWavelength Method
E. Tristimulus Spectrophotometric
Method
F. ADMI Weighted-Ordinate
Spectrophotometric Method
2130 TURBIDITY

2-1
2-2
2-3
2-4
2-7
2-7
2-8

A. Introduction
B. Nephelometric Method

2-8
2-9

2150 ODOR

2-11

A. Introduction
B. Threshold Odor Test

2-11
2-12

2160 TASTE

2-16

A. Introduction
2-16
B. Flavor Threshold Test (FTT)
2-16
C. Flavor Rating Assessment (FRA) . . . . 2-18
2170 FLAVOR PROFILE ANALYSIS

2-19

A. Introduction
B. Flavor Profile Analysis

2-19
2-19

2310 ACIDITY

A. Introduction
B. Titration Method

2-24

.__. . . 2-24
2-24

2320 ALKALINITY

A. Introduction
B. Titration Method
2330 CALCIUM CARBONATE SATURATION

A. Introduction
B. Indices Indicating Tendency of a Water
to Precipitate CaCO 3 or Dissolve
CaCO 3
C. Indices Predicting the Quantity of
CaCO 3 That Can Be Precipitated or
Dissolved
D. Diagrams and Computer Codes for
CaCO 3 Indices
2340 HARDNESS

A. Introduction
B. Hardness by Calculation
C. EDTA Titrimetric Method
2350 OXIDANT DEMAND/REQUIREMENT

2-27

2-27
2-27
2-30

2-30

2-31

2-34
2-35
2-37

2-37
2-37
2-37
2-40

TABLE OF CONTENTS

A. Introduction
B. Chlorine Demand/Requirement
C. Chlorine Dioxide
Demand/Requirement
D. Ozone Demand/RequirementBatch
Method
E. Ozone Demand/Requirement SemiBatch Method
2510 CONDUCTIVITY

A. Introduction
B. Laboratory Method

2-40
2-41
2-42

ANALYSIS

2-43
2-44

2810 DISSOLVED GAS SUPERSATURATION

2-44
2-47
2-48

2-48
2-49
2-50
2-50

Introduction
Electrical Conductivity Method
Density Method
Algorithm of Practical Salinity
'

A. Introduction
B. Paniculate Floatables
C. Trichlorotrifiuoroethane-Soluble
Floatable Oil and Grease
2540 SOLIDS

2-51

2-51
2-51
2-53
2-55

A.
B.
C.
D.

Introduction
2-55
Total Solids Dried at 103-105C
2-56
Total Dissolved Solids Dried at 180C . 2-57
Total Suspended Solids Dried
at 103-105C . . . ,
2-58
E. Fixed and Volatile Solids Ignited at
55OC
2-59
F. Settleable Solids
2-59
G. Total, Fixed, and Volatile Solids in
Solid and Semisolid Samples
2-60
2550 TEMPERATURE

A. Introduction
B. Laboratory and Field Methods
2560 PARTICLE COUNTING AND SIZE DISTRIBUTION .

A.
B.
C.
D.

Introduction
Electrical Sensing Zone Method
Light-Blockage Methods
Light-Scattering Method

2570 ASBESTOS

A. Introduction
B. Transmission Electron Microscopy
Method

2-61

2-61
2-61
2-62

2-62
2-66
2-68
2-69
2-70

2-70
2-71

2580 OXIDATION-REDUCTION POTENTIAL (ORP). . . 2-75

A. Introduction
B. Oxidation-Reduction Potential
Measurement in Clean Water
2710 TESTS ON SLUDGES

A.
B.
C.
D.
E.
F.

Introduction
Oxygen-Consumption Rate
Settled Sludge Volume
Sludge Volume Index
Zone Settling Rate
Specific Gravity

A. Introduction
B. Volumetric Method
C. Gas Chromatographic Method

A.
B.
C.
D.

2-75
2-76
2-79

2-79
2-79
2-80
2-81
2-82
2-83

2-83
2-85
2-86

2720 ANAEROBIC SLUDGE DIGESTER GAS

2-43

2520 SALINITY

2530 FLOATABLES

G. Capillary Suction Time

H. Time-to-Filter
I. Modified Settled Sludge Volume

A. Introduction
B. Direct-Sensing Membrane-Diffusion
Method
Part 3000

2-87

2-87
2-88
2-89
2-92

2-92
2-92

METALS

3010 INTRODUCTION

A. General Discussion
B. Sampling and Sample Preservation. . . .
C. General Precautions
3020 QUALITY ASSURANCE/QUALITY CONTROL . . .

A. Introduction
B. Quality Control Practices
3030 PRELIMINARY TREATMENT OF SAMPLES

A. Introduction
B. Filtration for Dissolved and Suspended
Metals
C. Treatment for Acid-Extractable Metals .
D. Digestion for Metals
E. Nitric Acid Digestion
F. Nitric Acid-Hydrochloric Acid
Digestion
G. Nitric Acid-Sulfuric Acid Digestion . . .
H. Nitric Acid-Perchloric Acid Digestion . .
I. Nitric Acid-Perchloric AcidHydrofluoric Acid Digestion
J. Dry Ashing
K. Microwave-Assisted Digestion .'~7 . . . .

3-1

3-1
3-2
3-3
3-4

3-4
3-4
3-5

3-5
3-6
3-6
3-7
3-8
3-9
3-10
3-10
3-10
3-11
3-11

3110 METALS BY ATOMIC ABSORPTION

SPECTROMETRY

3-13

3111 METALS BY FLAME ATOMIC ABSORPTION

SPECTROMETRY

3-13

A. Introduction
3-13
B. Direct Air-Acetylene Flame Method. . . 3-17
C. Extraction/Air-Acetylene Flame
Method
3-19
D. Direct Nitrous Oxide-Acetylene Flame
Method
3-20
E. Extraction/Nitrous Oxide-Acetylene
Flame Method
3-22
3112 METALS BY COLD-VAPOR ATOMIC ABSORPTION
SPECTROMETRY

A. Introduction
B. Cold-Vapor Atomic Absorption
Spectrometric Method

3-23

3-23
3-23

3113 METALS BY ELECTROTHERMAL ATOMIC

ABSORPTION SPECTROMETRY

3-25

TABLE OF CONTENTS

A. Introduction
B. Electrothermal Atomic Absorption
Spectrometric Method

3-25

3500-Ir
3500-Fe

3-27

SPECTROMETRY

35OO-Pb

3500-Li

SPECTROSCOPY

3500-Mg

MASS SPECTROMETRY

3-45

A. Introduction
3-45
B. Inductively Coupled Plasma/Mass
Spectrometry (ICPMS) Method . . . . 3-45
3130 METALS BY ANODIC STRIPPING

VOLTAMMETRY

3-53

A. Introduction
'
B. Determination of Lead, Cadmium,
and Zinc

3-54

35OO-A1

ALUMINUM

3-57

3500-Sb
3500-As

A. Introduction
B. Eriochrome Cyanine R Method
ANTIMONY
ARSENIC
A. Introduction
B. Silver Diethyldithiocarbamate Method. .

3-57
3-58
3-60
3-61
3-61
3-61

3500-Ba
35OO-Be
3500-Bi
3500-B

BARIUM
BERYLLIUM
BISMUTH
BORON

3-63
3-63
3-64
3-64

35OO-Cd
3500-Ca

CADMIUM
CALCIUM
.A. Introduction
B. EDTA Titrimetric Method

3-64
3-65
3-65
3-65

3500-Cs

CESIUM

3-66

3500-Cr

3500-Ga

CHROMIUM
A. Introduction
B. Colorimetric Method . . .'"
C. Ion Chromatographic Method
COBALT
COPPER
A. Introduction
B. Neocuproine Method
C. Bathocuproine Method
GALLIUM

3-67
3-67
3-67
3-69
3-71
3-71
3-71
3-72
3-73
3-74

3500-Ge

GERMANIUM

3-75

3500-Au

GOLD

3-75

3500-In

INDIUM

3-75

3500-Co
3500-Cu

'.

3500-Mn

MANGANESE

A. Introduction
B. Persulfate Method

3-39

3125 METALS BY INDUCTIVELY COUPLED PLASMA/

MAGNESIUM

A. Introduction
B. Calculation Method

3-38

3-38

LITHIUM

3-75
3-76

3-76
3-77
3-79

3-79
3-80
3-82

A. Introduction
3-82
B. Flame Emission Photometric Method . . 3-82

3120 METALS BY PLASMA EMISSION

A. Introduction
B. Inductively Coupled Plasma (ICP)
Method

LEAD

A. Introduction
B. Dithizone Method

3-32

A. Introduction
3-32
B. Manual Hydride Generation/Atomic
Absorption Spectrometric Method. . . 3-33
C. Continuous Hydride Generation/Atomic
Absorption Spectrometric Method. . . 3-37

IRON

A. Introduction
B. Phenanthroline Method

3114 ARSENIC AND SELENIUM BY'HYDRIDE

GENERATION/ATOMIC ABSORPTION

IRIDIUM

3-83

3-83
3-84
3-84

3-84
3-84

3500-Hg

MERCURY

3-86

3500-Mo

MOLYBDENUM

3-86

3500-Ni
3500-Os
3500-Pd

NICKEL

3-87

OSMIUM

3-87

PALLADIUM

3-87

3500-Pt

PLATINUM

3-87

3500-K

POTASSIUM

A. Introduction
B. Flame Photometric Method
C. Potassium-Selective Electrode
Method

3-53

3500-Re
3500-Rh
3500-Ru
35OO-Se

RHENIUM

3500-Sr

3-89
3-90

RHODIUM

3-90

RUTHENIUM

3-90

SELENIUM

3-91

A.
B.
C.
D.
E.
3500-Ag
35OO-Na

3-88

3-88
3-88

Introduction
3-91
Sample Preparation
3-92
Colorimetric Method
3-94
Determination of Volatile Selenium . . . 3-96
Determination of Nonvolatile Organic '
Selenium Compounds
3-96

SILVER
SODIUM

3-98
3-98

A. Introduction
B. Flame Emission Photometric
Method

3-98
3-99

STRONTIUM

3-101

A. Introduction
3-101
B. Flame Emission Photometric Method . . 3-101
3500-Te
3500-T1
3500-Th
3500-Sn
3500-Ti
3500-U
. 3500-V

TELLURIUM

3-103

THALLIUM

3-103

THORIUM
TIN
TITANIUM

3-104

URANIUM

3-104

VANADIUM

A. Introduction
B. Gallic Acid Method
3500-Zn

3-103

3-103

ZINC

3-104

3-104
3-105
3-106

TABLE OF CONTENTS

A. Introduction
B. Zincon Method

3-106
3-106

F. Cyanide-Selective Electrode Method. . . 4-43

G. Cyanides Amenable to Chlorination
after Distillation
. . 4-44
H. Cyanides Amenable to Chlorination
without Distillation
(Short-Cut Method)
4-46
I. Weak Acid Dissociable Cyanide
4-47
J. Cyanogen Chloride
4-48
K. Spot Test for Sample Screening
4-49
L. Cyanates
4-50
M. Thiocyanate
4-51
N. Total Cyanide after Distillation, by
Flow Injection Analysis
4-53
0 . Total Cyanide and Weak Acid
Dissociable Cyanide by Flow
Injection Analysis
4-55

Part 4000 INORGANIC NONMETALLIC

CONSTITUENTS
4010 INTRODUCTION

4-1

4020 QUALITY ASSURANCE/QUALITY CONTROL . . .

4-1

A. Introduction
B. Quality Control Practices

4-1
4-1

4110 DETERMINATION OF ANIONS BY ION

CHROMATOGRAPHY

4-3

A. Introduction
B. Ion Chromatography with Chemical
Suppression of Eluent Conductivity . .
C. Single-Column Ion Chromatography
with Direct Conductivity Detection . .
D. Ion Chromatographic Determination of
Oxyhalides and Bromide

4-3

4-7

4120 SEGMENTED CONTINUOUS FLOW ANALYSIS . . .

4-11

4-3
4-6

4500-C1

CHLORINE (RESIDUAL)

A.
B.
C.
D.
E.

A. Introduction
4-11
B. Segmented Flow Analysis Method. . . . 4-11
4130 INORGANIC NONMETALS BY FLOW INJECTION
ANALYSIS

'

A. Introduction
B. Quality Control

F.
G.
H.
1.

4-13

4-13
4-13

4140 INORGANIC ANIONS BY CAPILLARY ION

ELECTROPHORESIS

A. Introduction
B. Capillary Ion Electrophoresis with
Indirect UV Detection
4500-B

4500-Br"

4-14

4-14

BORON

4-23

A. Introduction
B. Curcumin Method
C. Carmine Method

4-23
4-23
4-25

BROMIDE

4-25

A.
B.
C.
D.

Introduction
4-25
Phenol Red Colorimetric Method . . . . 4-26
(Reserved)
4-27
Flow Injection Analysis
4-27

4500-CO 2 CARBON DIOXIDE

A. Introduction
B. Nomographic Determination of Free
Carbon Dioxide and the Three Forms
of Alkalinity
C. Titrimetric Method for Free Carbon
Dioxide
D. Carbon Dioxide and Forms of Alkalinity
by Calculation
4500-CN" CYANIDE

A.
B.
C.
D.
E.

4500-C1"

4-14

A.
B.
C.
D.
E.

4-28

4500-F~

4-28
4-33
4-34
4-34

Introduction
4-34
Preliminary Treatment of Samples . . . . 4-37
Total Cyanide after Distillation
4-39
Titrimetric Method
4-41
Colorimetric Method
4-41

CHLORIDE

4500-H +

4500-1

4-70

A. Introduction
4-70
B. Argentometric Method
. 4-70
C. Mercuric Nitrate Method
4-71
D. Potentiometric Method
4-73
E. Automated Ferricyanide Method
4-74
F. (Reserved)
4-75
G. Mercuric Thiocyanate Flow Injection
Analysis
"7 . . . . 4-75

4500-ClO 2 CHLORINE DIOXIDE

4-28

4-56

Introduction
4-56
Iodometric Method I
4-58
Iodometric Method II
4-60
Amperometric Titration Method
4-62
Low-Level Amperometric Titration
Method
4-64
DPD Ferrous Titrimetric Method
4-64
DPD Colorimetric Method
4-67
Syringaldazine (FACTS) Method . . . . 4-68
Iodometric Electrode Technique
4-69

Introduction
Iodometric Method
Amperometric Method I
(Reserved)
'
Amperometric Method II

4-77

4-77
4-77
4-78
4-79
4-79

FLUORIDE

4-82

A.
B.
C.
D.
E.
F.
G.

4-82
4-83
4-84
4-85
4-87
4-88

Introduction
Preliminary Distillation Step
Ion-Selective Electrode Method
SPADNS Method
Complexone Method
(Reserved)
Ion-Selective Electrode Flow Injection
Analysis

4-88

P H VALUE

4-90

A. Introduction
B. Electrometric Method

4-90
4-90

IODINE

4-95

TABLE OF CONTENTS

XIX

A. Introduction
B. Leuco Crystal Violet Method
C. Amperometric Titration Method
4500-r

IODIDE

A.
B.
C.
D.

NITROGEN

4-103

A. Introduction
B. In-Line UV/Persulfate Digestion and
Oxidation with Flow Injection
Analysis
C. Persulfate Method
D. Conductimetric Determination of
Inorganic Nitrogen

4-103

4500-NH3 NITROGEN (AMMONIA)

NITROGEN (NITRATE)

4-118
4-118
4-120

4500KMnO 4

. . :

Introduction
Macro-Kjeldahl Method
Semi-Micro-Kjeldahl Method
Block Digestion and Flow Injection
Analysis

4-120
4500-SiO 2

4-127
4-130

4-130
4-131
4-132
4-133

4-144

4-144
4-144

PHOSPHORUS

4-146

A. Introduction . . .
B. Sample Preparation
C. Vanadomolybdophosphoric Acid
Colorimetric Method
D. Stannous Chloride Method
E. Ascorbic Acid Method
F. Automated Ascorbic Acid Reduction
Method
G. Flow Injection Analysis for
Orthophosphate
H. Manual Digestion and Flow Injection
Analysis for Total Phosphorus . . .
I. In-line UV/Persulfate Digestion
and Flow Injection Analysis for
Total Phosphorus
J. Persulfate Method for Simultaneous
Determination of Total Nitrogen and
Total Phosphorus
POTASSIUM PERMANGANATE

4500-S 2 "

4-151
4-152
4-153
4-155
4-156
. 4-158

4-159

4-160
4-162

SILICA

SULFIDE

4-146
4-149

4-162
4-163
- r . . 4-164

Introduction
(Reserved)
Molybdosilicate Method . . . .'
Heteropoly Blue Method
Automated Method for MolybdateReactive Silica
F. Flow Injection Analysis for MolybdateReactive Silicate

4-121
4-122
4-123

4-126

OZONE (RESIDUAL)

A.
B.
C.
D.
E.

4-120

4-125
4-126

4-141
4-141

A. Introduction
B. Spectrophotometric Method

4-120

A. Introduction
B. Ultraviolet Spectrophotometric
Screening Method
C. Second-Derivative Ultraviolet
Spectrophotometric Method
(PROPOSED)
D. Nitrate Electrode Method
E. Cadmium Reduction Method.
F. Automated Cadmium Reduction
Method
G. (Reserved)
H. Automated Hydrazine Reduction
Method
I. Cadmium Reduction Flow Injection
Method
A.
B.
C.
D.

. .

4-118

A. Introduction
B. Colorimetric Method
C. (Reserved)

4500-Norg NITROGEN (ORGANIC)

4500-P

4-108

NITROGEN (NITRITE)

4-136
4-136
4-138
4-140
4-141

A. Introduction
B. Indigo Colorimetric Method

.4-107

Introduction
4-108
Preliminary Distillation Step
4-109
Titrimetric Method
4-110
Ammonia-Selective Electrode Method. . 4-111
Ammonia-Selective Electrode Method
Using Known Addition
4-112
F. Phenate Method .'
;
4-114
G. Automated Phenate Method
4-115
H. Flow Injection Analysis
4-116

4500-NO3~

4500-O 3

. 4-104
4-105

A.
B.
C.
D.
E.

4-136

Introduction
Iodometric Methods
Azide Modification
Permanganate Modification
Alum Flocculation Modification
Copper Sulfate-Sulfamic Acid
Flocculation Modification
G. Membrane Electrode Method

4-97
4-98
4-99
4-100
4-102
4-102

OXYGEN (DISSOLVED)

A.
B.
C.
D.
E.
F.

4-102

A. Introduction . . . . : :
~B. Polarographic Method

4500-NO2"

4500-O

4-97

Introduction
Leuco Crystal Violet Method
Catalytic Reduction Method
Voltammetric Method

4500-KV IODATE

4500-N

4-95
4-95
4-97

A. Introduction
B. Separation of Soluble and Insoluble
Sulfides
C. Sample Pretreatment to Remove
Interfering Substances' or to
Concentrate the Sulfide
D. Methylene Blue Method
E. Gas Dialysis, Automated Methylene
Blue Method
F. Iodometric Method

4-164
4-165
4-165
4-167
4-168
4-169
4-170

4-170
4-172

4-173
4-174
4-175
4-176

TABLE OF CONTENTS

XX

G. Ion-Selective Electrode Method

H. Calculation of Un-ionized Hydrogen
Sulfide.. I. Distillation, Methylene Blue Flow
Injection Analysis Method
J. Acid-Volatile Sulfide
4500-SO 3 2 "

4-179
4-181
4-182

SULFITE

4-183

A. Introduction
B. Iodometric Method
C. Phenanthroline Method
4500-SO 4 2 "

4-177

4-183
4-183
4-184

SULFATE

A. Introduction
B. (Reserved)
C. Gravimetric Method with
Residue .
D. Gravimetric Method with
Residue
E. Turbidimetric Method
F. Automated Methylthymol
Method
G. Methylthymol Blue Flow
Analysis

4-186

4-186
4-186
Ignition of
Drying of
4-187
4-188
Blue

5560 ORGANIC AND VOLATILE ACIDS

5-1

5020 QUALITY ASSURANCE/QUALITY CONTROL . . .

5-1

5210 BIOCHEMICAL OXYGEN DEMAND ( B O D ) . . . .

Introduction
5-Day BOD Test
Ultimate BOD Test
Respirometric Method

5220 CHEMICAL OXYGEN DEMAND (COD)

Introduction
Open Reflux Method
Closed Reflux, Titrimetric Method. . . .
Closed Reflux, Colorimetric Method. . .

A. Introduction
B. Adsorption-Pyrolysis-Titrimetric
Method
5510 AQUATIC HUMIC SUBSTANCES

5-2

5-2
5-2
5-7
5-10
5-14

5-14
5-15
5-16
5-18

A. Introduction
B. Chromatographic Separation Method for
Organic Acids
C. Distillation Method
D. Gas Chromatographic Method

5-55

5-55
5-55
5-56

5-56
5-57
5-58
5-59

5710 FORMATION OF TRIHALOMETHANES AND OTHER

DISINFECTION BY-PRODUCTS

A. Introduction
B. Trihalomethane Formation Potential
(THMFP)
C. Simulated Distribution System
Trihalomethanes (SDS-THM)
D. Formation of Other Disinfection ByProducts (DBPs)
r
5910 UV-ABSORBING ORGANIC CONSTITUENTS . . .

A. Introduction
B. Ultraviolet Absorption Method

5-62

5-62
5-64
5-68
5-69
5-71

5-71
5-72

5-19

5-19
Part 6000 INDIVIDUAL ORGANIC COMPOUNDS
5-21

6010 INTRODUCTION

5-23
5-25

A. General Discussion
B. Sample Collection and Preservation . . .
C. Analytical Methods

5-26

6020 QUALITY ASSURANCE/QUALITY CONTROL . . .

5-26
5-27
5-32

A. Introduction
5-32
B. Diethylaminoethyl (DEAE) Method . . . 5-33
C. XAD Method
5-34
5520 OIL AND GREASE

5-47

Introduction
5-47
Surfactant Separation by Sublation. . . . 5-48
Anionic Surfactants as MBAS
5-50
Nonionic Surfactants as CTAS
5-53

4-190

5-1
5-1

5320 DISSOLVED ORGANIC HALOGEN

A.
B.
C.
D.

A. Introduction
B. Colorimetric Method

A. General Discussion .,
B. Sample Collection and Preservation . . .

A. Introduction
B. High-Temperature Combustion
Method
C. Persulfate-Ultraviolet or HeatedPersulfate Oxidation Method
D. Wet-Oxidation Method

5-43
5-44
5-45
5-47

Introduction
Cleanup Procedure
Chloroform Extraction Method
Direct Photometric Method

5550 TANNIN AND LIGNIN

Injection

5310 TOTAL ORGANIC CARBON (TOC)

5-43

A.
B.
C.
D.

4-189

5010 INTRODUCTION

A.
B.
C.
D.

5530 PHENOLS

5540 SURFACTANTS

4-186

Part 5000 AGGREGATE ORGANIC CONSTITUENTS

A.
B.
C.
D.

A. Introduction
5-35
B. Liquid-Liquid, Partition-Gravimetric
Method
5-37
C. Partition-Infrared Method
5-39
D. Soxhlet Extraction Method
5-40
E. Extraction Method for Sludge Samples . . . 5-41
F. Hydrocarbons
5-41
G. Sold-Phase, Partition-Gravimetric Method. . 5-42

5-35

A. Introduction
B. Quality Control Practices

6-1

6-1
6-3
6-4
6-6

6-6
6-6

6040 CONSTITUENT CONCENTRATION BY GAS

EXTRACTION

A. Introduction
B. Closed-Loop Stripping, Gas
Chromatographic/Mass Spectrometric
Analysis

6-8

6-8

6-8

TABLE OF CONTENTS

B. Liquid-Liquid Extraction Gas

Chromatographic Method
C. Liquid-Liquid Extraction Gas
Chromatographic/Mass Spectrometric
Method

C. Purge and Trap Technique

6-19
D. Solid-Phase Microextraction (SPME) . . 6-19
6200 VOLATILE ORGANIC COMPOUNDS

6-23

A. Introduction
B. Purge and Trap Capillary-Column Gas
Chromatographic/Mass Spectrometric
Method
C. Purge and Trap Capillary-Column Gas
Chromatographic Method

6-23

A. Introduction
B. Liquid-Liquid Extraction
Chromatographic Method
C. Liquid-Liquid Extraction Gas
Chromatographic/Mass Spectrometric
Method

6-25
6-31
6-35

A. Introduction
6-35
B. Combustible-Gas Indicator Method . . . 6-35
C. Volumetric Method
6-37
6231 1,2-DlBROMOETHANE (EDB) AND 1,26-37

6-40
6-40

'

6-40

6-40
6-41
6-46

6-92
6-100

6-100
6-100

6-107
6-108

6-115

6640 ACIDIC HERBICIDE COMPOUNDS

. .

6-46

6-46

6651

6-115
6-116

GLYPHOSATE HERBICIDE

6-126

A. Introduction
B. Liquid Chromatographic Post-Column
Fluorescence Method

6-47

6252 DISINFECTION BY-PRODUCTS: ALDEHYDES

(PROPOSED)
A. Introduction
.B. PFBHA Liquid-Liquid Extraction Gas
Chromatographic Method

6-115

A. Introduction
B. Micro Liquid-Liquid Extraction Gas
Chromatographic Method

6-46

A. Introduction
B. Micro Liquid-Liquid Extraction Gas
Chromatographic Method

6-126
6-127

6-57
6-57
Part 7000
6-58
. .

A. Introduction . . ._
B. Liquid-Liquid Extraction Gas
Chromatographic/Mass Spectrometric
Method

6-65

6-65

6420 PHENOLS

RADIOACTIVITY

7010 INTRODUCTION

6-66
6-79
6-79

6-85
6-85

7-1
7-2

. . .

7-3

A. Basic Quality Control Program

B. Quality Control for Wastewater
Samples
C. Statistics
D. Calculation and Expression of Results. .

7-3

7030 COUNTING INSTRUMENTS

A. Introduction
B. Description and Operation
of Instruments
7040 FACILITIES

6-85

7-1

A. General Discussion
B. Sample Collection and Preservation . . .
7020 QUALITY ASSURANCE/QUALITY CONTROL

6-79

A. Introduction
B. Liquid-Liquid Extraction Gas
Chromatographic Method
C. Liquid-Liquid Extraction Gas
Chromatographic/Mass Spectrometric
Method
A. Introduction

6-91

6-91

A. Introduction
B. Liquid-Liquid Extraction Gas
Chromatographic Method 1
AppendixStandardization of
Magnesia-Silica Gel Column by
Weight Adjustment Based on
Adsorption of Laurie Acid
C. Liquid-Liquid Extraction Gas
Chromatographic Method II
D. Liquid-Liquid Extraction Gas
Chromatographic/Mass Spectrometric
Method

6-37

6251 DISINFECTION BY-PRODUCTS: HALOACETIC

6431 POLYCHLORINATED BlPHENYLS (PCBS)

6-91

6630 ORGANOCHLORINE PESTICIDES

A. Introduction
B. Liquid-Liquid Extraction Gas
Chromatograpriic Method
C. Purge and Trap Gas Chromatographic/
Mass Spectrometric Method
D. Purge and Trap Gas Chromatographic
Method

6410 EXTRACTABLE BASE/NEUTRALS AND ACIDS

6-86

A. Introduction
B. High-Performance Liquid
Chromatographic Method

6232 TRIHALOMETHANES AND CHLORINATED

ACIDS AND TRICHLOROPHENOL

6-86

6610 CARB AM ATE PESTICIDES

DIBROMO-3-CHLOROPROPANE (DBCP) . . . 6-37

ORGANIC SOLVENTS

6-85

6 4 4 0 POLYNUCLEAR AROMATIC HYDROCARBONS . . . 6 - 8 6

6211 METHANE

A. Introduction
"
B. Liquid-Liquid Extraction Gas
Chromatographic Method
C. Purge and Trap Gas Chromatographic/
Mass Spectrometric Method
D. Purge and Trap Gas- Chromatographic
Method

6-85

A. Counting Room

7-6
7-6
7-7
7-8

7-8
7-8
7-13

7-13

TABLE OF CONTENTS

XXII

B.
C.
D.
E.

Radiochemistry Laboratory
Laboratory Safety
Pollution Prevention
Waste Management

7-14
7-14
7-14
7-15

7110 GROSS ALPHA AND GROSS BETA

RADIOACTIVITY
(TOTAL, SUSPENDED, AND DISSOLVED) . . . .

8020 QUALITY ASSURANCE AND QUALITY CONTROL

7-15

A. Introduction
7-15
B. Evaporation Method for Gross AlphaBeta
7-15
C. Coprecipitation Method for Gross Alpha
Radioactivity in Drinking Water. . . . 7-19
7120 GAMMA-EMITTING RADIONUCLIDES

A. Introduction
B. Gamma Spectroscopic Method
7500-Cs

RADIOACTIVE CESIUM

A. Introduction
B. Precipitation Method

/
7500-1

RADIOACTIVE IODINE

A.
B.
C.
D.
7500-Ra

7500-Rn

7500-Sr

7500-3H

7500-U

Introduction
Precipitation Method
Ion-Exchange Method
Distillation Method

7-20

7-20
7-20
7-24

7-24
7-24
7-25

7-25
7-26
7-27
7-28

RADIUM

7-29

A.
B.
C.
D.
E.

7-29
7-29
7-32
7-38
7-40

Introduction
7
Precipitation Method
Emanation Method
Sequential Precipitation Method
Gamma Spectroscopy Method

RADON

7-43

A. Introduction
B. Liquid Scintillation Method

7-43
7-43

TOTAL RADIOACTIVE STRONTIUM

AND STRONTIUM-90

7-46

A. Introduction
B. Precipitation Method

7-46
7-46

TRITIUM

7-49

A. Introduction
B. Liquid Scintillation Spectrometric
Method

7-49
7-50

URANIUM

7-51

A. Introduction
B. Radiochemical Method
C. Isotopic Method

7-51
7-51
7-53

Part 8000 TOXICITY

8010 INTRODUCTION

A.
B.
C.
D.
E.

General Discussion
Terminology
Basic Requirements
Conducting Toxicity Tests
Preparing Organisms for Toxicity
Tests
F. Toxicity Test Systems, Materials,
and Procedures

G. Calculating, Analyzing, and Reporting

Results of Toxicity Tests
8-20
H. Interpreting and Applying Results of
Toxicity Tests
8-24
I. Selected Toxicological Literature . . . . 8-25

8-1

8-1
8-2
8-3
8-4
8-7

IN LABORATORY TOXICITY TESTS

A. General Discussion
B. Elements of QA/QC
8030 MUTAGENESIS

A. Introduction
B. Salmonella Microsomal Mutagenicity
Test
8050 BACTERIAL BIOLUMINESCENCE

A. Introduction
B. Bacterial Bioluminescence Test

8-30

8-30
8-31
8-37

8-37
8-37

8070 P450 REPORTER GENE RESPONSE TO DIOXINLIKE ORGANIC COMPOUNDS

A. Introduction
B. The P450 RGS Test

8-40

8-40
8-40

8071 COMET/SINGLE-CELL GEL ELECTROPHORESIS

ASSAY FOR DETECTION OF DNA
DAMAGE

A. Introduction
B. Comet/Single-Cell Gel Electrophoresis
Assay
8080 SEDIMENT POREWATER TESTING

A.
B.
C.
D.

8-42

8-42
8-43
8-46

Introduction
8-46
Sediment Collection and Storage
8-47
Extraction of Sediment Pore Water . . . 8-47
Toxicity Testing Procedures
8-49

8110 ALGAE

8-50

8111 BIOSTIMULATION (ALGAL PRODUCTIVITY) . . .

8-51

A. General Principles
8-51
B. Planning and Evaluating Algal Assays . 8-51
C. Apparatus
V
8-52
D. Sample Handling
8-53
E. Synthetic Algal Culture Medium
8-53
F. Inoculum
8-53
G. Test Conditions and Procedures
8-54
H. Effect of Additions
8-55
I. Data Analysis and Interpretation
8-56
8112 PHYTOPLANKTON

A. Introduction
B. Inoculum
C. Test Conditions and Procedures
8113 MARINE MACROALGAE

A. Introduction
B. Selecting and Preparing Macrocystis
pyrifera Sporophylls
C. Toxicity Test Procedures
D. Data Evaluation
8200 AQUATIC FLOWERING PLANTS
8211 DUCKWEED. . .'

8-15

8-26

8-26
8-27

A. Introduction

8-57

8-57
8-57
8-57
8-58

8-58
8-59
8-60
8-62
8-63
8-63

8-63

TABLE OF CONTENTS

B. Selecting and Preparing Test

Organisms
C. Toxicity Test Procedure
8220 AQUATIC EMERGENT PLANTS

A. Introduction
B. Selecting and Preparing Test
Organisms
C. Toxicity Test Procedure
8310 CILIATED PROTOZOA

A. Introduction
- B. Growth Inhibition Test with Freshwater
Ciliate Colpidium campylum
C. Chemotactic Test with Freshwater
Ciliate Tetrahymena thermophila . . .
D. Growth Inhibition Test with the Soil
Ciliate Colpoda inflata
8420 ROTIFERS

8714 MYSIDS

8-66

8-66

A. Introduction
.
B. Selecting and Preparing Test
Organisms
C. Toxicity Test Procedures
D. Sediment Test Procedures .Using the
Marine Polychaete Neanthes
arenaceodentata
E. Sediment Test Procedures Using the
Freshwater and Marine Oligochaetes
Pristina leidyi, Tubifex tubifex, and
Lumbriculus variegatus . . . . . . . .
F. Data Evaluation
8610 MOLLUSKS

A. Introduction
B. Selecting and Preparing Test
Organisms
C. Short-Term Test Procedures Using
Marine Mollusk Larvae
D. Sediment Test Procedures Using
Marine Bivalves
E. Field Test Procedures Using Freshwater
and Marine Bivalves
8710 ARTHROPODS
8711 DAPHNIA

A. Introduction
B. Selecting and Preparing Test
Organisms
C. Procedures
8712 CERIODAPHNIA

A. Introduction
B. Selecting and Preparing Test
Organisms
C. Procedures

8-111
8-112
8-117

8740 DECAPODS

8-70

8-70
8-71

8-121
8-121
8-128
8-132

8750 AQUATIC INSECTS

8-133

A. Introduction
B. Selecting and Preparing Test
Organisms
C. Toxicity Test Procedures
D. Data Evaluation

8-73
8-75
8-76

8-133
8-133
8-135
8-137

8810 ECHINODERM FERTILIZATION AND

DEVELOPMENT

8-137

A. Introduction
B. Selecting and Preparing Test
Organisms
C. Echinoderm Fertilization Test
D. Echinoderm Embryo Development
Test

8-80

8-80
8-81
8-86

8-121

A. Introduction
B. Selecting and Preparing Test
Organisms
C. Toxicity Test Procedures
D. Data Evaluation

8-67
8-69

A. Introduction
8-76
B. Selecting and Preparing Testing
Organisms
8-77
C. Aquatic Toxicity Test,Procedures . . . . 8-78
8510 ANNELIDS. . .

8-111

A. Introduction
B. Selecting and Preparing Test
Organisms
C. Toxicity Test Procedures

8-64
8-65

8910 FISH

8-138
8-140
8-143
8-146

A. Introduction
8-146
B. Fish Selection and Culture Procedures. . 8-146
C. Test Procedures
8-151

8-89

8921 FATHEAD MINNOW

8-157

A. Introduction .
B. Culture and Maintenance of Test
Organisms
C. Procedures

8-91
8-91
8-92

8-137

8930 AMPHIBIANS (PROPOSED)

8-157
8-158
8-159
8-166

A. Introduction
. . - - . . . 8-166
B. Culture and Maintenance of Test
Organisms
8-167
C. Procedures
8-169

8-92
8-93
8-94
8-96

Part 9000 MICROBIOLOGICAL EXAMINATION

9010 INTRODUCTION

8-97

A. Introduction
B. Intralaboratory Quality Control
Guidelines
C. Interlaboratory Quality Control

8-101
8-101

8-101
8-103
8-104

9-1

9020 QUALITY ASSURANCE/QUALITY CONTROL . . .

9030

9-2

9-2
9-3
9-14

9-15

LABORATORY APPARATUS

8-106

A. Introduction
B. Equipment Specifications

9-15
9-15

8-106

9040 WASHING AND STERILIZATION

9-17

9050 PREPARATION OF CULTURE MEDIA

8-107
8-109

A. General Procedures
B. Water

9-18

9-18
9-19

TABLE OF CONTENTS

C. Media Specifications

9-19

9060 SAMPLES

9-20

A. Collection
B. Preservation and Storage

9-20
9-22

9211 RAPID DETECTION METHODS

A. Introduction
B. Seven-Hour Fecal Coliform Test
C. Special Techniques

9-23

9-23
. . . . 9-23
9-23

9212 STRESSED ORGANISMS

A. Introduction
B. Recovery Enhancement
9213 RECREATIONAL WATERS

A.
B.
C.
D.
E.

Introduction
'
Swimming Pools
Whirlpools
Natural Bathing Beaches
Membrane Filter Technique for
Pseudomonas aeruginosa
F. Multiple-Tube Technique for
Pseudomonas aeruginosa

9215 HETEROTROPHIC PLATE-COUNT .

A.
B.
C.
D.

Introduction
Pour Plate Method
Spread Plate Method . . .
Membrane Filter Method

9216 DIRECT TOTAL MICROBIAL COUNT

9-25

9-25
9-26
9-28

9-28
9-29
9-31
9-31

A. Introduction
B. Pseudomonas fluorescens Strain P-17,
Spirillum Strain NOX Method
9218 AEROBIC ENDOSPORES

A. Introduction
B. Membrane Filter Method
MEMBERS OF THE COLIFORM GROUP

9-33
9-34

9-34
9-37
9-38
9-40
9-41

9-42

9-42
9-44
9-47

9-47
9-48
9-48

9-48
9-49
9-53
9-55
9-56
9-57

9222 MEMBRANE FILTER TECHNIQUE FOR MEMBERS

OF THE COLIFORM GROUP

A. Introduction
B. Standard Total Coliform Membrane
Filter Procedure
C. Delayed-Incubation Total Coliform
Procedure
D. Fecal Coliform Membrane Filter
Procedure

9-72

A. Introduction
B. Enzyme Substrate Test

9-72
9-72

9224 DETECTION OF COLIPHAGES

9-74

A. Introduction
9-74
B. Somatic Coliphages Assay
9-75
C. Male-Specific Coliphage Assay Using
Escherichia coli Famp
9-77
D. Male-Specific Coliphage Assay Using
S a l m o n e l l a t y p h i m u r i u m W G 4 9 . . . . 9-78
E. Single-Agar-Layer Method
9-80
F. Membrane Filter Method
9-81
9225 DIFFERENTIATION OF THE COLIFORM

9221 MULTIPLE-TUBE FERMENTATION TECHNIQUE FOR

A. Introduction
B. Standard Total Coliform Fermentation
Technique
C. Estimation of Bacterial Density
D. Presence-Absence (P-A) Coliform
Test
E. Fecal Coliform Procedure
F. Escherichia coli Procedure
(PROPOSED)

9223 ENZYME SUBSTRATE COLIFORM TEST

9-33

A. Introduction
;
9-41
B. Epifluorescence Microscopic Method . . 9-41
9217 ASSIMILABLE ORGANIC CARBON

E. Delayed-Incubation Fecal Coliform

Procedure
9-68
F. Klebsiella Membrane Filter Procedure. . 9-69
G. MF Partition Procedures
9-70

9-59

9-59
9-60
9-65
9-66

BACTERIA

A.
B.
C.
D.

Introduction
Culture Purification
Identification
Media, Reagents, and Procedures . . . .

9-82

9-82
9-83
9-83
9-84

9230 FECAL STREPTOCOCCUS AND ENTEROCOCCUS

GROUPS

A. Introduction
B. Multiple-Tube Technique
C. Membrane Filter Techniques
9240 IRON AND SULFUR BACTERIA

A.
B.
C.
D.
E.

9-86

9-86
9-88
9-88
9-91

Introduction
9-91
Iron Bacteria
9-91
Sulfur Bacteria
9-95
Bacteria Living in Acidic Environments. 9-97
Enumeration, Enrichment, and Isolation
of Iron and Sulfur Bacteria
9-101

9245 NITRIFYING BACTERIA

A. Introduction
B. Multiple-Tube Method . . :

9-106

9-106
9-107

9250 DETECTION OF ACTINOMYCETES

9-109

A. Introduction
B. Actinomycete Plate Count

9-109
9-110

9260 DETECTION OF PATHOGENIC BACTERIA

9-111

A. Introduction
9-111
B. General Qualitative Isolation and
Identification Procedures for
Salmonella
9-113
C. Immunofluorescence Identification
Procedure for Salmonella
9-116
D. Quantitative Salmonella Procedures . . . 9-118
E. Shigella
9-118
F. Pathogenic Escherichia coli
9-120
G. Campylobacter jejuni
9-121
H. Vibrio cholerae
9-123
I. Leptospira
9-125
J. Legionella
9-127
K. Yersinia enterocolitica
9-130

TABLE OF CONTENTS

L. Aeromonas
[Link]
9510 DETECTION OF ENTERIC VIRUSES

xxv
9-131
9-134

A. Introduction
B. Pour Plate Technique
C. Spread Plate Technique.
D. Membrane Filter Technique
E. Technique for Yeasts
F. Zoosporic Fungi
G. Aquatic Hyphomycetes
H. Fungi Pathogenic to Humans
9711 PATHOGENIC PROTOZOA

A.
B.
C.
D.
E.

9-136

A. Introduction
9-136
B. Virus Concentration from Small Sample
Volumes by Adsorption to and
Elution from Microporous Filters . . . 9-138
C. Virus Concentration from Large Sample
Volumes by Adsorption to and
Elution from Microporous Filters . . . 9-141
'- D. Virus Concentration by Aluminum
Hydroxide Adsorption-Precipitation . . 9-145
E. Hydroextraction-Dialysis with
Polyethylene Glycol
9-147
F. Recovery of Viruses from Suspended
Solids in Water and Wastewater. . . . 9-148
G. Assay and Identification of Viruses in
Sample Concentrates
9-149
9610 DETECTION OF FUNGI

10400 MACROPHYTES

9-153

9-153
9-156
9-157
9-158
9-158
9-159
9-160
9-160

A.
B.
C.
D.

A.
B.
C.
D.
E.

10-1

A. Introduction
10-2
B. Sample Collection
10-3
C. Concentration Techniques
10-10
D. Preparing Slide Mounts
10-11
E. Microscopes and Calibrations
10-12
F. Phytoplankton Counting Techniques. . . 10-13
G. Zooplankton Counting Techniques. . . . 10-17
H. Chlorophyll
10-18
I. Determination of Biomass
(Standing Crop)
10-26
J. Metabolic Rate Measurements
10-28
10300 PERIPHYTON

A. Introduction
B. Sample Collection
C. Sample Analysis
D. Primary Productivity
E. Interpreting and Reporting R e s u l t s . . . .

10-76

Introduction
Data Acquisition
Sample Preservation
Analysis of Collections
Investigation of Fish Kills

10-78

10-78
10-79
10-88
10-89
10-93
10-94

10750 NEMATOLOGICAL EXAMINATION

10-95

A. Introduction
B. Collection and Processing Techniques
forNematodes
C. Illustrated Key to Freshwater
Nematodes

10-95
10-97
10-98

10900 IDENTIFICATION OF AQUATIC

ORGANISMS

10-113

A. Procedure in Identification
10-114
B. Key to Major Groups of Aquatic
Organisms (Plates 1-35)
10-114
Acknowledgments
10-118
C. Key for Identification of Freshwater
Algae Common in Water Supplies
and Polluted Waters
(Plates 1A, IB, 28-35)
-'. . 10-152
D. Index to Illustrations
10-157
E. Selected Taxonomic References . . . . 10-160

SUPPLEMENT TO THE 21ST EDITION

6610 CARBAMATE PESTICIDES

A. Introduction
B. High-Performance Liquid
Chromatographic Method
6710 TRIBUTYLTIN

A. Introduction
B. Gas Chromatographic/Mass
Spectrometric Method
C. Gas Chromatographic/Flame
Photometric Method

10-32

10-32
10-32
10-34
10-37
10-45

10-63

10-63
10-65
10-74

10700 BENTHIC MEIOFAUNA

Part 10000 BIOLOGICAL EXAMINATION

10-2

Introduction
Sample Collection
Sample Processing and Analysis
Data Evaluation, Presentation,
and Conclusions

10600 FISHES

A. Introduction
9-161
B. Giardia and Cryptosporidium Methods . 9-162
C. Acanthamoeba
9-168

10200 PLANKTON

10-47

10-47
10-48
10-49
10-51
10-54

10500 BENTHIC MACROINVERTEBRATES

9-161

10010 INTRODUCTION

Introduction
Preliminary Survey
Vegetation Mapping Methods
Population Estimates
Productivity

INDEX

S-l

S-l
S-l
S-10

S-10
S-10
S-15

1-1