A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

i

A Compendium of Chemical, Physical and 

Biological Methods for Assessing and 

Monitoring the Remediation of

Contaminated Sediment Sites

EPA Contract No. 68-W-99-033

Work Assignment 4-20

Submitted to 

U.S. Environmental Protection Agency

Prepared by

Battelle Memorial Institute

397 Washington Street

Duxbury MA 02332

February 17, 2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

ii

CONTACTS

James Lazorchak and Jon Josephs are the EPA contacts for the Compendium of Chemical, Physical, and

Biological Methods for Assessing and Monitoring the Remediation of Contam inated Sediment Sites. 

James Lazorchak, W ork Assignment Manager for the preparation of the Compendium , is an Aquatic

Ecotoxicologist assigned to the Ecological Exposure Research Division in Cincinnati, OH, which is under

the direction of the National Exposure Research Laboratory with headquarters in Research Triangle Park,

NC.

Jon Josephs, Deputy W ork Assignment Manager for preparation of the Compendium, is an Environmental

Engineer assigned to the Hazardous Substances Technical Liaison Program, which is under the direction

of the Office of Science Policy with headquarters in Washington, DC. He is stationed at the EPA Region 2

office in New York City, NY.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

iii

ABSTRACT

Considering the many organizations which have published methods for monitoring contaminated sediments

and the large number of documents on this subject, it can be a formidable task for a Superfund project

manager to find methods appropriate for his or her contaminated sediment site. This Compendium of

Chemical, Physical and Biological Methods for Assessing and Monitoring the Remediation of Contaminated

Sediment Sites has been prepared to inform Superfund project managers and others about appropriate

methods for monitoring and assessing the remediation of contaminated sediments. Although the document

can be printed as a text document, it is also intended to be viewed on a computer screen in order to take

advantage of its hypertext links to navigate the document and to access reference documents available on

the Internet. Search engines can also be utilized to locate information contained in the document.

The methods included in this document focus primarily on published or otherwise citeable chemical, physical,

and biological testing methodologies used by EPA at Superfund sites. The document summ arizes the

methods, including references to the methods and hypertext links to access those methods which are

available on the Internet. Without exception, it is intended that all of the methods presented will be suitable

for investigations at Superfund sites containing contaminated sediments. However, not all methods will be

suitable for all sites. The selection of methods for a particular site will depend on the site conditions,

remediation plans, budgetary constraints and other factors.

This report was submitted in fulfillment of Contract Number 68-W-99-033 by Battelle Memorial Institute under

the sponsorship of the United States Environmental Protection Agency. The report was prepared during a

period from July 6, 2001 to February 17, 2003 when the work was completed.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

iv

TABLE OF CONTENTS

Tables ............ ............ ............ ............ ............ ............ .... x

Table of Acronyms ............ ............ ............ ............ ............ ....... xi

1.0 Introduction ............ ............ ............ ............ ............ ........ 1

1.1 Background and Objectives ............ ............ ............ ............ ....... 1

1.2 Application and Uses of Field, Analytical, and Testing Data at Superfund Sites Containing Contaminated

Sediments ............ ............ ............ ............ ............ ........ 2

2.0 Monitoring Methods ............ ............ ............ ............ ............ . 8

2.1 Water ............ ............ ............ ............ ............ ............ 8

2.1.1 Field Sample Collection and Immediate Processing In Situ Data Acquisition ............ . 9

Fact Sheet No. 2.1.1-1 Method Title: In Situ sampling with the Hydrolab

Datasonde3® Unit ............ ............ .......... 11

Fact Sheet No. 2.1.1-2 Method Title: In Situ Dissolved Oxygen sampling with a

YSI Model 58 Dissolved Oxygen Meter and Probe ......... 12

Fact Sheet No. 2.1.1-3 Method Title: In Situ Sampling of Irradiance ............ . . 13

Fact Sheet No. 2.1.1-4 Method Title: In Situ Transparency Sampling ............ . 14

Fact Sheet No. 2.1.1-5 Method Title: Sample Collection Procedures for Marine

Water ............ ............ ............ ....... 15

Fact Sheet No. 2.1.1-6 Method No. LMMB 013, Method Title: In Situ Sample

Collection Using the Rosette Sampler ............ ...... 17

Fact Sheet No. 2.1.1-7 Method No. ERT SOP #2013, Method Title: W ater Sample

Collection with the Kemmerer Bottle and the Bacon Bomb Sampler

............ ............ ............ ........... 18

Fact Sheet No. 2.1.1-8 Method No. ERT SOP # 2013, Method Title: Dip

Sampler ............ ............ ............ ..... 19

Fact Sheet No. 2.1.1-9 Method Title: Sample and Preservation of W ater Specific

Parameters ............ ............ ............ . . 20

Fact Sheet No. 2.1.1-10 Method No. LMMB 014, Method Title: Sampling of

Particulate-Phase and Dissolved-Phase Organic Carbon

in Great Lakes Waters ............ ............ ...... 24

Fact Sheet No. 2.1.1-11 Method No. EPA Method 1669, Method Title: Sampling

Ambient Water for Trace Metals at EPA Water Quality

Criteria Levels ............ ............ ............ 25

Fact Sheet No. 2.1.1-12 Method No. LMMB 065, Method Title: ESS Method 340.2:

Total Suspended Solids, Mass Balance (Dried at

103-105°C) Volatile Suspended Solids (Ignited at 550°C) ... 26

Fact Sheet No. 2.1.1-13 Method Title: In situ peepers ............ ............ . 27

Fact Sheet No. 2.1.1-14 Method Title: Suction Samplers ............ ........... 28

Fact Sheet No. 2.1.1-15 Method Title: Physical Characterization of a Stream ....... 29

Fact Sheet No. 2.1.1-16 Method Title: Visual-Based Habitat Assessment .......... 30

Fact Sheet No. 2.1.1-17 Method No. LMMB 017, Method Title: USGS Field

Operation Plan: Tributary Monitoring ............ ....... 32
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

v

Fact Sheet No. 2.1.1-18 Method Title: Quality Assurance Plan for Discharge Measurements

Using Broadband Acoustic Doppler Current Profilers ....... 33

Fact Sheet No. 2.1.1-19 Method Title: Seepage Meters ............ ............ 34

Fact Sheet No. 2.1.1-20 Method Title: Caged Bivalve Deployment. ............ ... 35

2.1.2 Chemical and Physical Analysis ............ ............ ............ .......... 36

Fact Sheet No. 2.1.2-1 EPA Method No. 245.7, Method Title: Mercury in Water

by Cold Vapor Atomic Fluorescence Spectrom etry ........ 37

Fact Sheet No. 2.1.2-2 EPA Method No. 1631, Revision B, Method Title: Mercury

in Water by Oxidation, Purge and Trap, and Cold Vapor

Atomic Fluorescence Spectrom etry ............ ........ 38

Fact Sheet No. 2.1.2-3 EPA Method No. 1630, Method Title: Methyl Mercury in

Water by Distillation, Aqueous Ethylation, Purge and Trap,

and CVAFS ............ ............ ............ . 40

Fact Sheet No. 2.1.2-4 EPA Method No. 1639, Method Title: Determination of

Trace Elem ents in Ambient Waters by Stabilized

Tem perature Graphite Furnace Atomic Absorption ........ 41

Fact Sheet No. 2.1.2-5 EPA Method No. 1637, Method Title: Determination of

Trace Elem ents in Ambient Waters by Off-Line Chelation

Pre-concentration and Stabilized Tem perature Graphite

Furnace Atomic Absorption ............ ............ . . 42

Fact Sheet No. 2.1.2-6 EPA Method No. 1638, Method Title: Determination of

Trace Elem ents in Ambient Waters by Inductively Coupled Plasma

— Mass Spectrom etry ............ ............ ...... 43

Fact Sheet No. 2.1.2-7 EPA Method No. 1640, Method Title: Determination of

Trace Elem ents in Ambient Waters by On-Line Chelation

Pre-concentration and Inductively Coupled Plasma-Mass

Spectrom etry ............ ............ ............ . 45

Fact Sheet No. 2.1.2-8 EPA Method No. 1632, Method Title: Inorganic Arsenic in

Water by Hydride Generation Quartz Furnace Atomic

Absorption ............ ............ ............ ... 46

Fact Sheet No. 2.1.2-9 EPA Method No. 1632, Revision A, Method Title: Chemical

Speciation of Arsenic in Water and Tissue by Hydride

Generation Quartz Furnace Atomic Absorption

Spectrom etry ............ ............ ............ . 47

Fact Sheet No. 2.1.2-10 EPA Method No. 1636, Method Title: Determination of Hexavalent

Chromium by Ion Chromatography ............ ........ 48

Fact Sheet No. 2.1.2-11 EPA Method No. 1624b, Method Title: Volatile Organic Compounds

by Isotope Dilution GC/MS ............ ............ ... 49

Fact Sheet No. 2.1.2-12 Method No. OERR SOP #2109, Method Title: Photovac GC

Analysis for Soil, Water, and Air/Soil Gas ............ .... 50

Fact Sheet No. 2.1.2-13 EPA Method No. 1625, Method Title: Semi-volatile Organic

Compounds by Isotope Dilution GC/MS ............ ..... 51

Fact Sheet No. 2.1.2-14 Method Title: Quantitative Determination of Polynuclear

Arom atic Hydrocarbons by Gas Chromatography/Mass

Spectrom etry (GC/MS) - Selected Ion Monitoring

(SIM) Mode ............ ............ ............ . . 52

Fact Sheet No. 2.1.2-15 Method No. LMMB 041, Method Title: Analysis of Polychlorinated

Biphenyls and Chlorinated Pesticides by

Gas Chromatography with Electron Capture Detection ..... 53

Fact Sheet No. 2.1.2-16 Method No. LMMB, Method Title: PCBs and Pesticides in Surface

Water by XAD-2 Resin Extraction ............ ......... 54

Fact Sheet No. 2.1.2-17 EPA Method No. 1613, Method Title: Tetra- through

Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS ............ ............ ............ 55
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

vi

Fact Sheet No. 2.1.2-18 EPA Method No. 1668, Method Title: Toxic Polychlorinated

Biphenyls by Isotope Dilution High Resolution Gas

Chromatography/High Resolution Mass Spectrom etry ...... 57

Fact Sheet No. 2.1.2-19 EPA Method No. 1668, Revision A, Method Title:

Chlorinated Biphenyl Congeners in Water, Soil, Sediment,

and Tissue by HRGC/HRMS ............ ............ . 58

Fact Sheet No. 2.1.2-20 Method No. ESS Method 220.3, Method Title: Ammonia Nitrogen

and Nitrate+Nitrite Nitrogen, Automated Flow

Injection Analysis Method ............ ............ .... 59

Fact Sheet No. 2.1.2-21 Method No. ESS Method 230.1, Method Title: Total

Phosphorus and Total Kjeldahl Nitrogen, Semi-Automated

Method ............ ............ ............ ..... 60

Fact Sheet No. 2.1.2-22 Method No. ESS Method 310.2, LMMB 064, Method Title:

Phosphorus, Total, Low Level (Persulfate Digestion) ....... 61

Fact Sheet No. 2.1.2-23 Method No. ESS Method 310.1, LMMB 063, Method Title:

Ortho-Phosphorus, Dissolved Automated, Ascorbic Acid .... 62

Fact Sheet No. 2.1.2-24 Standard Method No. 5310, Method Title: Total Organic

Carbon ............ ............ ............ .... 63

Fact Sheet No. 2.1.2-25 Method No. LMMB 096, Method Title: Standard Operating

Procedure for the Analysis of Dissolved-Phase Organic

Carbon in Great Lakes Waters ............ ............ 65

Fact Sheet No. 2.1.2-26 Method No. LMMB 097, Method Title: Standard Operating

Procedure for the Analysis of Particulate-Phase Organic

Carbon in Great Lakes Waters ............ ............ 66

Fact Sheet No. 2.1.2-27 Method No. ESS Method 140.4, Method Title: Chloride - Automated

Flow Injection Analysis ............ ............ ...... 67

Fact Sheet No. 2.1.2-28 Method No. ESS Method 200.5, Method Title: Determination

of Inorganic Anions in Water by Ion Chromatography ...... 68

Fact Sheet No. 2.1.2-29 Method No. LMMB 092, Method Title: Standard Operating

Procedure for Electrometric pH ............ ........... 69

Fact Sheet No. 2.1.2-30 Method No. LMMB 091, Method Title: Standard Operating

Procedure for GLNPO Total Alkalinity Titration ............ 70

Fact Sheet No. 2.1.2-31 Method No. LMMB 094, Method Title: Standard Operating

Procedure for GLNPO Specific Conductance: Conductivity

Bridge ............ ............ ............ ...... 71

Fact Sheet No. 2.1.2-32 Method No. LMMB 090, Method Title: Standard Operating

Procedure for GLNPO Turbidity: Nephelometeric

Method ............ ............ ............ ...... 72

Fact Sheet No. 2.1.2-33 Method No. LMMB 065, Method Title: ESS Method 340.2:

Total Suspended Solids, Mass Balance (Dried at

103-105°C) Volatile Suspended Solids (Ignited at 550°C) ... 73

Fact Sheet No. 2.1.2-34 Method No. LMMB 095, Method Title: Total Hardness

Titration ............ ............ ............ ..... 74
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

vii

2.1.3 Biological Analysis Methods. .75

Fact Sheet No. 2.1.3-1 Method No. ERT SOP 2024, Method Title: Acute

Freshwater Crustacean Bioassay: 48 Hours ............ . 78

Fact Sheet No. 2.1.3-2 Method No. ERT SOP 2022, Method Title: Acute

Freshwater Fish Bioassay ............ ............ ... 80

Fact Sheet No. 2.1.3-3 Method No. ERT SOP 2027, Method Title: Chronic

Freshwater Algae Test ............ ............ ...... 81

Fact Sheet No. 2.1.3-4 Method No. ERT SOP 2025, Method Title: Chronic

Freshwater Crustacean Bioassay (7 Day) ............ ... 83

Fact Sheet No. 2.1.3-5 Method No. ERT SOP 2028, Method Title: Chronic

Freshwater Crustaceans Bioassay (10 days) ............ . 85

Fact Sheet No. 2.1.3-6 Method No. ERT SOP 2026, Method Title: Chronic

Freshwater Fish Bioassay ............ ............ ... 86

Fact Sheet No. 2.1.3-7 Method No. NHEERL-AED SOP 1.03.001, Method Title:

Chronic Marine Macroalgae, Champia parvula, Sexual

Reproduction Test ............ ............ ........ 87

Fact Sheet No. 2.1.3-8 Method No. NHEERL-AED SOP 1.03.003, Method Title:

Acute Marine Crustacean Bioassay ............ ........ 88

Fact Sheet No. 2.1.3-9 Method No. NHEERL-AED SOP 1.03.003, Method Title:

Acute Marine Fish Bioassay ............ ............ . . 89

Fact Sheet No. 2.1.3-10 Method No. NHEERL-AED SOP 1.03.005, Method Title:

Chronic Estuarine Survival, Growth and Fecundity Test ..... 90

Fact Sheet No. 2.1.3-11 Method No. NHEERL-AED SOP 1.03.006, Method Title:

Chronic Echinoderm Fertilization Test ............ ...... 91

Fact Sheet No. 2.1.3-12 Method No. NHEERL-AED SOP 1.03.004, Method Title:

Chronic Marine Fish Bioassay ............ ............ 93

Fact Sheet No. 2.1.3-13 Method Title: Toxicity Evaluations of Photoinduction of Polycyclic

Aromatic Hydrocarbons (PAH): In Situ

Analysis ............ ............ ............ ..... 94

Fact Sheet No. 2.1.3-14 Method Title: Toxicity Evaluations of Photoinduction of Polycyclic

Aromatic Hydrocarbons: Laboratory Analysis of Storm water 95

Fact Sheet No. 2.1.3-15 Method No. NHEERL-AED SOP 1.03.013, Method Title:

Growth and Scope for Growth Measurements with

Mytilus edulis ............ ............ ............ . 96

Fact Sheet No. 2.1.3-16 Method No. NHEERL-AED SOP 1.03.009, Method Title: Microtox®

tests ............ ............ ............ ....... 98

Fact Sheet No. 2.1.3-17 Com parative Toxicity of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin

to Seven Freshwater Fish Species During Early Life-Stage

Development ............ ............ ............ . 99

2.2 Sediments ............ ............ ............ ............ ............ ...... 101

2.2.1 Field Sample Collection and Processing, In Situ Data Acquisition ............ ........ 101

Fact Sheet No. 2.2.1-1 Method Title: Grab Sampling ............ ............ 102

Fact Sheet No. 2.2.1-2 Method Title: Core Samplers ............ ............ 106

Fact Sheet No. 2.2.1-3 Method Title: Hand Collection ............ ........... 110

Fact Sheet No. 2.2.1-4 Method Title: Hand Collection at Depth with SCUBA

Equipment ............ ............ ............ . . 111

Fact Sheet No. 2.2.1-5 Method Title: Sediment Traps ............ ........... 112

Fact Sheet No. 2.2.1-6 Method Title: Russian Peat Borer ............ ......... 113

Fact Sheet No. 2.2.1-7 Method Title: Split Core Sampler for Submerged

Sediments ............ ............ ............ . . 114

Fact Sheet No. 2.2.1-8 Method Title: Sediment Processing for Chem istry and

Toxicity Testing ............ ............ .......... 115
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

viii

Fact Sheet No. 2.2.1-9 Method Title: Sediment Processing for Elutriate Toxicity

Tests ............ ............ ............ ...... 116

Fact Sheet No. 2.2.1-10 Method No. ASTM E 1391-94, Method Title: Pore Water Extraction

through Centrifugation ............ ............ ..... 117

Fact Sheet No. 2.2.1-11 Method No. ASTM E 1391-94, Method Title: Pore Water Extraction

from Sediments through Squeezing ............ ....... 118

Fact Sheet No. 2.2.1-12 Method No. ASTM E 1391-94, Method Title: Pore water extraction

from sediment from Vacuum Filtration ............ ..... 119

Fact Sheet No. 2.2.1-13 Method No. DRP-2-03, Method Title: Acoustic Sub-bottom Profiling

Systems ............ ............ ............ .... 120

Fact Sheet No. 2.2.1-14 Method No. EEDP-01-10, Method Title: Side Scan

Sonar ............ ............ ............ ...... 121

Fact Sheet No. 2.2.1-15 Method No. DRP-2-3, Method Title: Settlement Phases .... 122

2.2.2 Chemical and Physical Analysis ............ ............ ............ ......... 123

Fact Sheet No. 2.2.2-1 Method No. Appendix to Method 1631, Method Title: Total Mercury

in Sludge, Sediment, Soil, and Tissue by Acid

Digestion and BrCl Oxidation ............ ............ 124

Fact Sheet No. 2.2.2-2 Method Title: Trace Element Quantification Techniques ... 125

Fact Sheet No. 2.2.2-3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively

Coupled Plasma Mass Spectrom etry ............ ...... 127

Fact Sheet No. 2.2.2-4 Method Title: Determination of Acid Volatile Sulfide and

Selected Sim ultaneously Extractable Metals in Sediment ... 129

Fact Sheet No. 2.2.2-5 Method No. OSW ER SOP # 2109 Method Title: Photovac

GC Analysis for Soil, Water, and Air/Soil Gas ........... 130

Fact Sheet No. 2.2.2-6 Method No. LMMB 040, Method Title: Extraction and

Clean-Up of Sediments for Semi-volatile Organics

Following the Internal Standard Method ............ .... 131

Fact Sheet No. 2.2.2-7 Method Title: Quantitative Determination of Polynuclear

Aromatic Hydrocarbons by Gas Chromatography/Mass

Spectrom etry (GC/MS) - Selected Ion Monitoring

(SIM) Mode ............ ............ ............ . 133

Fact Sheet No. 2.2.2-8 Method No. LMMB 041, Method Title: Analysis of Polychlorinated

Biphenyls and Chlorinated Pesticides by

Gas Chromatography with Electron Capture Detection .... 134

Fact Sheet No. 2.2.2-9 Method No. SW846 Method 4020, Method Title: Screening

for Polychlorinated Biphenyls by Immunoassay .......... 135

Fact Sheet No. 2.2.2-10 EPA Method No. 1613, Method Title: Tetra- through

Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HR MS ............ ............ ........... 136

Fact Sheet No. 2.2.2-11 EPA Method No. 1668, Method Title: Toxic Polychlorinated

Biphenyls by Isotope Dilution High Resolution Gas

Chromatography/High Resolution Mass Spectrom etry ..... 137
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

ix

Fact Sheet No. 2.2.2-12 EPA Method No. 1668 Revision A, Method Title:

Chlorinated Biphenyl Congeners in Water, Soil, Sediment,

and Tissue by HRGC/HRMS ............ ............ 138

Fact Sheet No. 2.2.2-13 Method Title: Butyltin in Sediments ............ ....... 139

Fact Sheet No. 2.2.2-14 Method Title: Procedures for Sediment Total Organic

Carbon (TOC) Determination ............ ............ 140

Fact Sheet No. 2.2.2-15 Method No. LMMB 084, Method Title: Determination of the Activity

of Lead-210 in Sediments and Soils ............ ....... 141

Fact Sheet No. 2.2.2-16 Method No. NHEERL-AED SOP 1.01.005, Method Title: Sediment

Grain Size Analysis ............ ............ ....... 142

Fact Sheet No. 2.2.2-17 Method Title: Procedures for Water Content

Determination ............ ............ ............ 143

Fact Sheet No. 2.2.2-18 Method No. ASTM D 2573, Method Title: Standard Test

Method for Field Vane Shear Test in Cohesive Soil ....... 144

Fact Sheet No. 2.2.2-19 Method No. ASTM D 854, Method Title: Standard Test

Method for Specific Gravity of Soil Solids by Water

Pycnometer ............ ............ ............ . 145

Fact Sheet No. 2.2.2-20 Method No. ASTM 2434, Method Title: Standard Test

Method for Permeability of Granular Soils (Constant Head) . 146

Fact Sheet No. 2.2.2-21 Method No. ASTM 2435, Method Title: Standard Test

Method for One-Dimensional Consolidation Properties of

Soil ............ ............ ............ ........ 147

Fact Sheet No. 2.2.2-22 Method No. ASTM 2487, Method Title: Standard Test

Method for Classification of Soils for Engineering Purposes (Unified

Soil Classification System) ............ ............ . . 148

Fact Sheet No. 2.2.2-23 Method No. ASTM 4318, Method Title: Standard Test

Method for Liquid Limit, Plastic Limit, and Plasticity Index

of Soils ............ ............ ............ ..... 149

Fact Sheet No. 2.2.2-24 Method No. 4020, Method Title: Field Portable X-Ray Fluorescence

Spectrom etry for the Determination of

Elemental Concentrations in Soil and Sediment .......... 150

Fact Sheet No. 2.2.2-25 Method Title: Sediment Age Dating Using Cesium-137 .... 151

Fact Sheet No. 2.2.2-26 Method Title: Beryllium-7 as a Tracer of Short Term Sediment

Deposition ............ ............ ............ . . 152

2.2.3 Biological Analysis Methods ............ ............ ............ ............ 153

Fact Sheet No. 2.2.3-1 Method Title: Acute Freshwater Crustacean Sediment

Bioassay: Flow-through ............ ............ .... 158

Fact Sheet No. 2.2.3-2 Method Title: Acute Freshwater Crustacean Sediment

Bioassay: In Situ Exposures ............ ............ . 159

Fact Sheet No. 2.2.3-3 Method Title: Acute Freshwater Crustacean Sediment

Bioassay: Static Laboratory Exposures ............ .... 160

Fact Sheet No. 2.2.3-4 EPA Method No. 100.1, Method Title: Acute/Chronic

Freshwater Amphipod and Freshwater Insect Larvae

Sediment Bioassay ............ ............ ....... 161

Fact Sheet No. 2.2.3-5 EPA Method No. 100.4, Method Title: Chronic Freshwater

Amphipod Sediment Bioassay ............ ........... 163

Fact Sheet No. 2.2.3-6 EPA Method No. 100.5, Method Title: Life-Cycle

Freshwater Midge Sediment Bioassay ............ ..... 164

Fact Sheet No. 2.2.3-7 Method Title: Acute Larval Bivalve Sediment

Bioassay ............ ............ ............ ... 165

Fact Sheet No. 2.2.3-8 Method Title: Acute Echinoderm Sediment

Bioassay ............ ............ ............ ... 166

Fact Sheet No. 2.2.3-9 Method Title: Acute Marine Crustacean Sediment

Bioassay ............ ............ ............ ... 167
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

x

Fact Sheet No. 2.2.3-10 EPA Method No. 100.4, Method Title: Acute Marine

Amphipod Crustacean Sediment Bioassay ............ . 168

Fact Sheet No. 2.2.3-11 Method No. ASTM E1611-00, Method Title: Acute Marine

Polychaete Sediment Bioassay ............ .......... 170

Fact Sheet No. 2.2.3-12 Method Title: Chronic Estuarine Amphipod Sediment

Bioassay ............ ............ ............ ... 171

Fact Sheet No. 2.2.3-13 Method No. ASTM E1611-00, Method Title: Chronic

Marine Polychaete Sediment Bioassay ............ .... 172

Fact Sheet No. 2.2.3-14 Method Title: Ames Mutagenicity Assay ............ .... 173

Fact Sheet No. 2.2.3-15 Method Title: Mutatox Genotoxicity Assay ............ . . 174

Fact Sheet No. 2.2.3-16 Method No. NHEERL-AED SOP 1.03.012, Method Title:

V79/ Sister Chromatid Exchange Assay ............ .... 175

Fact Sheet No. 2.2.3-17 EPA Method No. 100.3, Method Title: Bioaccumulation

Test for Marine, Estuarine and Freshwater Sediments ..... 177

2.3 Biota ............ ............ ............ ............ ............ ..... 179

2.3.1 Chemical and Physical Analyses ............ ............ ............ ........ 179

Fact Sheet No. 2.3.1-1 Method No. LMMB 023, Method Title: Phytoplankton

Sample Collection and Preservation in the

Great Lakes ............ ............ ............ . . 180

Fact Sheet No. 2.3.1-2 Method No. LMMB 015, Method Title: Chlorophyll-a

Sampling Method and Preservation: Field Procedure in the

Great Lakes ............ ............ ............ . . 181

Fact Sheet No. 2.3.1-3 Method Title: Chlorophyll a and Phaeophytin Field Filtering Protocols

............ ............ ............ ........... 182

Fact Sheet No. 2.3.1-4 Method No. LMMB 016, Method Title: Primary Productivity

Using 14C: Field Procedure in the Great Lakes ............ 183

Fact Sheet No. 2.3.1-5 Method No. LMMB 024, Method Title: Zooplankton Sample Collection

and Preservation in the Great Lakes ............ ....... 184

Fact Sheet No. 2.3.1-6 Method Title: Field-based Periphyton Survey in Wadeable Streams 185

Fact Sheet No. 2.3.1-7 Method Title: Laboratory-Based Periphyton Survey: Single

Habitat Sampling in Wadeable Streams ............ .... 186

Fact Sheet No. 2.3.1-8 Method Title: Laboratory-Based Rapid Periphyton Survey:

Multi habitat Sampling in Wadeable Streams ............ . 187

Fact Sheet No. 2.3.1-9 Method Title: Artificial Substrate Samplers of Macroinvertebrates in Wadeable Streams ............ ........ 188

Fact Sheet No. 2.3.1-10 Method Title: Algae and Macroinvertebrate Sampling with

Frames ............ ............ ............ ..... 190

Fact Sheet No. 2.3.1-11 Method No. NHEERL-AED SOP 1.02.001, Method Title:

Benthic Organism Collection from a Marine

Environment ............ ............ ............ . 191

Fact Sheet No. 2.3.1-12 Method Title: Benthic Macroinvertebrate Protocols in a

Wadeable Stream: Single Habitat Approach, 1-Meter

Kick Net ............ ............ ............ ..... 192
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

xi

Fact Sheet No. 2.3.1-13 Method Title: Benthic Macroinvertebrate Protocols in a

Wadeable Stream: Multi habitat Approach: D-Frame

Dip Net ............ ............ ............ ..... 193

Fact Sheet No. 2.3.1-14 Method Title: Photographic Habitat Documentation of the

Benthic Community ............ ............ ........ 194

Fact Sheet No. 2.3.1-15 Method Title: Sediment Profile Cam era ............ ..... 195

Fact Sheet No. 2.3.1-16 Method Title: Macroinvertebrate Drift Nets in Wadeable

Streams ............ ............ ............ ..... 196

Fact Sheet No. 2.3.1-17 Method Title: Stream-Net Samplers: Surber, Portable

Invertebrate Box Sampler, Hess Sampler, Hess Stream

Bottom Sampler, and Stream-Bed Fauna Sampler ........ 198

Fact Sheet No. 2.3.1-18 Method Title: Mussel Collection Using Brails ............ . 199

Fact Sheet No. 2.3.1-19 Method Title: Electrofishing ............ ............ . . 200

Fact Sheet No. 2.3.1-20 Method Title: Chemical Fishing ............ ........... 201

Fact Sheet No. 2.3.1-21 Method Title: Fish Collection Using Seine Nets ........... 202

Fact Sheet No. 2.3.1-22 Method Title: Entanglement Nets ............ .......... 203

Fact Sheet No. 2.3.1-23 Method Title: Entrapment Devices ............ ......... 204

Fact Sheet No. 2.3.1-24 Method Title: Pop Nets ............ ............ ..... 205

Fact Sheet No. 2.3.1-25 Method Title: Trawls ............ ............ ....... 206

Fact Sheet No. 2.3.1-26 Method No. LMMB 025, Method Title: Fish Processing

Method in the Great Lakes ............ ............ . . 207

Fact Sheet No. 2.3.1-27 Method Title: Fish Processing ............ ............ 208

Fact Sheet No. 2.3.1-28 Method Title: Swallows: Sampling Procedures ............ 210

Fact Sheet No. 2.3.1-29 Method Title: Sample Processing of Swallows ............ 211

2.3.2 Chemical and Physical Analysis ............ ............ ............ ......... 212

Fact Sheet No. 2.3.2-1 Method Title: Sample Preparation for Metal Contam inants

in Tissue ............ ............ ............ .... 213

Fact Sheet No. 2.3.2-2 Method No. Appendix to Method 1631, Method Title: Total Mercury

in Tissue, Sludge, Sediment, and Soil by Acid

Digestion and BrCl Oxidation ............ ............ . 214

Fact Sheet No. 2.3.2-3 Method No. LMMB 052, Method Title: Versatile

Combustion-Amalgamation Technique for the Photometric

Determination of Mercury in Fish and Environmental

Samples ............ ............ ............ .... 215

Fact Sheet No. 2.3.2-4 Method No. NS&T, Method Title: Trace Element

Quantification Techniques ............ ............ ... 216

Fact Sheet No. 2.3.2-5 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and

Inductively Coupled Plasma Mass Spectrom etry .......... 217

Fact Sheet No. 2.3.2-6 EPA Method No. 1632, Revision A, Method Title:

Chemical Speciation of Arsenic in Water and Tissue by

Hydride Generation Quartz Furnace Atomic Absorption

Spectrom etry ............ ............ ............ . 218

Fact Sheet No. 2.3.2-7 Method No. LMMB 043, Method Title: Extraction and Lipid Separation

of Fish Samples for Contaminant Analysis and

Lipid Determination ............ ............ ........ 219

Fact Sheet No. 2.3.2-8 Method No. NS&T, Method Title: Purification of Biological

Tissue Samples by Gel Permeation Chromatography of

Organic Analyses ............ ............ .......... 220
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

xii

Fact Sheet No. 2.3.2-9 Method Title: Quantitative Determination of Polynuclear

Aromatic Hydrocarbons by Gas Chromatography/Mass

Spectrom etry (GC/MS) - Selected Ion Monitoring

(SIM) Mode ............ ............ ............ . . 221

Fact Sheet No. 2.3.2-10 Method No. LMMB 041, Method Title: Analysis of

Polychlorinated Biphenyls and Chlorinated Pesticides by

Gas Chromatography with Electron Capture Detection ..... 222

Fact Sheet No. 2.3.2.11 EPA Method No. 1613, Method Title: Tetra- through Octa-Chlorinated

Dioxins and Furans by Isotope Dilution

HRGC/HRMS ............ ............ .... . . . . . . 223

Fact Sheet No. 2.3.2-12 EPA Method No. 1668, Method Title: Toxic Polychlorinated Biphenyls

by Isotope Dilution High Resolution Gas Chromatography/High

Resolution Mass Spectrom etry ............ ........... 225

Fact Sheet No. 2.3.2-13 EPA Method No. 1668 Revision A, Method Title:

Chlorinated Biphenyl Congeners in Water, Soil, Sediment,

and Tissue by HRGC/HRMS ............ ............ . 226

Fact Sheet No. 2.3.2-14 Method Title: Determination of Percent Dry Weight for

Tissues ............ ............ ............ ..... 227

Fact Sheet No. 2.3.2-15 Method Title: Determination of Percent Lipid in Tissue ..... 228

Fact Sheet No. 2.3.2-16 Method Title: Microwave Extraction of marine Tissue for Semivolatile

Organic Analyte ............ ............ ........... 229

2.3.3 Biological Analysis Methods ............ ............ ............ ........... 231

Fact Sheet No. 2.3.3-1 Method Title: Laboratory Identification, Enumeration and

Biomass Measurements of Periphyton in Wadeable

Streams ............ ............ ............ ..... 232

Fact Sheet No. 2.3.3-2 Method Title: Laboratory Periphyton Biomass

Determination ............ ............ ............ 234

Fact Sheet No. 2.3.3-3 Method Title: Laboratory Analysis of Benthic Macroinvertebrates in Wadeable Streams ............ ........ 235

Fact Sheet No. 2.3.3-4 Method Title: Laboratory Analysis of Water Column

Organisms ............ ............ ............ ... 237

Fact Sheet No. 2.3.3-5 Method No. LMMB 026 - Appendix 2 & LMMB 027 -

Appendix B, Method Title: SOP-2: Lab Analysis of Lake

Trout Stomachs and Data Entry; Appendix B. Standard

Operating Procedure for Lab Analysis of Coho Salmon

Stomachs and Data Entry ............ ............ ... 239

Fact Sheet No. 2.3.3-6 Method Title: Gonadal Analysis ............ ........... 240

Fact Sheet No. 2.3.3-7 Method Title: Histopathological Evaluations of Target and

Non Target Fish Species ............ ............ .... 241

Fact Sheet No. 2.3.3-8 Method No. NS&T, Method Title: Histopathology

Analysis ............ ............ ............ ..... 242

Fact Sheet No. 2.3.3-9 Method Title: Index of Biotic Integrity (IBI) ............ ... 243

Fact Sheet No. 2.3.3-10 Method Title: Fish Bioassessment I and II ............ ... 244

3.0 References ............ ............ ............ ............ ............ . 245

Index ............ ............ ............ ............ ............ .......... 252
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

xiii

TABLES

Table 2.1.1-1. A Summary of Sample Sizes, Containers, Preservation Techniques, and

Holding Times for Water ............ ............ ............ ....... 22

Table 2.1.3-1. A Summary of Test Types and Toxicological Endpoints for Liquid-Phase

Toxicity. ............ ............ ............ ............ ........ 76

Table 2.2.1-1. A Summary of Sediment Grab Devices ............ ............ ....... 104

Table 2.2.1-2. A Summary of Sediment Coring Devices ............ ............ ...... 108

Table 2.2.3-1. A Summary of Test Types and Toxicological Endpoints for Solid-Phase

Toxicity ............ ............ ............ ............ ....... 154

Table 2.3.1-1 A Summary of Stream Net Samplers Used to Collect Organisms from

Flowing Water. ............ ............ ............ ............ . 197
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

xiv

Table of Acronyms

ADCP Acoustic Doppler Current Profiler

AVS Acid Volatile Sulfide

AWQC Ambient Water Quality Criteria

BAFS Bioaccumulation Factors

BSAFS Biota-Sediment Accumulation Factors

CAD Confined Aquatic Disposal

CB Chlorinated Biphenyl

CDD Chlorinated Dibenzo-p-dioxins

CDF Chlorinated Dibenzofurans

CHV Chronic Value

COC Constituents of Concern

CVAA Cold Vapor Atomic Absorption

CVAFS Cold Vapor Atomic Fluorescence Spectrom etry

CTD Conductivity, Tem perature, Depth

CWT Coded Wire Tags

DBT Dibutyltin

DCM Dichloromethane

DMA Dimethylarsinic Acid

DO Dissolved Oxygen

DOC Dissolved-phase Organic Carbon

EGD Effluent Guidelines Division

EICP Extracted Ion Current Profile

EMAP Environmental Monitoring and Assessment Program

EMDL Estimated Method Detection Limit

ERL Effects Range Low

ET Ecotox Threshold

FAA Flame Atomic Absorption

4BT Tetra Butyltin

GC Gas Chromatography

GC/ECD Gas Chromatography using Electron Capture Detection
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

xv

GFAA Graphite Furnace Atomic Absorption

GPC Gel Permeation Chromatography

HIB Hilsenhoff’s Family Biotic Index

HOC Hydrophobic Organic Contam inants

HPLC High-performance Liquid Chromatography

IA Inorganic Arsenic

IBI Index of Biotic Integrity

IC Ion Chromatography

ICI Invertebrate Community Index

ICP-MS Inductively Coupled Plasma Mass Spectrom etry

INAA Instrumental Neutron Activation Analysis

IR Infrared

LC50 Lethal Concentration for 50%

LIR Load Increment Ratio

LOEC Lowest Observable Effects Concentration

MBT Monobutyltin

MDL Method Detection Limit

ML Minimum Level

MMA Monomethylarsonic Acid

MS Mass Spectrometer

NOEC No Observable Effects Concentration

NPDES National Pollutant Discharge Elimination System

NTU Nephelometeric Turbidity Units

PAH Polycyclic Aromatic Hydrocarbons

PAR Photosynthetically Active Radiation

PCB Polychlorinated Biphenyl

POC Particulate-phase Organic Carbon

PSDDA Puget Sound Dredged Disposal Analysis

RBP Rapid Bioassessment Protocols

SAI Sim ple Autecological Indices

SCE Sister Chromatid Exchange
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

xvi

SDS Soxhlet/Dean-Stark

SEC Sediment Exposure Chamber

SEM Sim ultaneously Extracted Metal

SFG Scope For Growth

SIM Selected Ion Monitoring

SOP Standard Operating Procedure

SPE Solid-phase Extraction

SPI Sediment Profiling Imaging

SPM Settling Particulate Matter

STPGFAA Stabilized Tem perature Platform Graphite Furnace Atomic Absorption

TBT Tributyltin

TEC Threshold Effects Concentrations

TIC Tentatively Identified Compounds

TIC Total Inorganic Carbon

TOC Total Organic Carbon

TCD Thermal Conductivity Detection

VSS Volatile Suspended Solids

XRF X-Ray Fluorescence
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

1

1.0 INTRODUCTION

1.1 Background and Objectives

The extent and severity of chemical contaminants in freshwater and marine sediments and their

impacts on ecological and human health have been studied by EPA and other federal, state,

tribal, and local organizations for over the past 30 years. Over this period of time, field and

laboratory tools and techniques have continuously improved and new methods have developed,

and a body of institutional knowledge has been accumulated and refined regarding suitable

methods for sample collection and field processing, laboratory processing and chemical analysis,

and toxicology testing and bioaccumulation or other effects studies. This body of knowledge

comprises methods manuals, guidance documents, standard methods, and published

governmental reports, as well as published manuscripts in the scientific literature. 

To disseminate information about current EPA methods and research on contaminated

sediments, EPA assembled this compendium of methods. Methods presented focus primarily on

published or otherwise citeable chemical, physical, and biological (toxicity and bioassessment)

testing methodologies used by EPA at Superfund sites to determine the effects of chemical

contaminants on aquatic life and human health. Although priority is given to those methods that

have demonstrated efficacy at Superfund sites, the document also includes methods employed by

other EPA and other federal and state programs at non-Superfund contaminated sediment sites. 

The following agencies and programs have contributed significantly to the study of contaminated

sediments and are the source of many of the methods or secondary references contained in this

compendium.

Federal

• Environmental Protection Agency

• Office of Research and Development

• Office of Water

• Office of Science and Technology

• Office of Wetlands Oceans and Watersheds

• Office of Wastewater Management

• Office of Solid Waste and Emergency Response

• Office of Pollution Protection and Toxics

• Office of Pesticide Programs

• Great Lakes National Program Office

• National Oceanic and Atmospheric Administration

• National Status and Trends Program

• Hazardous Material and Response Division

• Damage Assessment and Remediation Program

• U.S. Army Corps of Engineers

• Waterways Experiment Station

• New England Division

• Seattle District
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

2

• U.S. Navy

• U.S. Fish and Wildlife Service

• U.S. Geological Survey/National Biological Survey

Interagency

• National Dredging Team

• Puget Sound Dredged Disposal Analysis (PSDDA) Program

Considering the many organizations which have published methods for monitoring contaminated

sediments and the large number of documents on this subject, it can be a formidable task for a

Superfund project manager to find methods appropriate for his or her contaminated sediment site.

To the best of our knowledge, no compilation of such methods has been prepared to date. This

document summarizes many of the published methods from these agencies and programs related

to the characterization of contaminated sediments and contaminated sediment sites. Additionally,

related methods published as American Society for Testing and Materials Standards or Standard

Methods for the Examination of Water and Wastewater (APHA 1999) are also included, as

needed, to provide complete information in certain topic areas. Where a number of methods are

available for a given monitoring activity, not all methods will necessarily be included in this

document. If an EPA method is available, it will generally be given priority for inclusion. A method

published by another Federal agency will often be included if it is significantly different from the

EPA method or if an EPA method is not available. Methods published by other sources may be

included if Federal methods are not available or if the methods have special merit.

The compendium is divided in four sections. The first section addresses the application and uses

of monitoring data. While many of the monitoring methods have been developed for purposes

other than monitoring at Superfund sites, the first section addresses applications within the

Superfund decision making process, and not the broader area of marine or aquatic environmental

monitoring. The monitoring methods are presented in three sections by the matrix being

monitored — water, sediment, and biota. Each of these sections contains separate subsections

on sampling methods and immediate field processing, chemistry and physical analysis methods,

and biological analysis methods. In situ data collection methods are presented in the sampling

methods section. Some of the methods, particularly the chemistry methods, are applicable to

more than one matrix and, thus, have been presented more than once. Effort has been made in

these cases to reduce redundancy as much as possible.

1.2 Application and Uses of Field, Analytical, and Testing Data at Superfund Sites 

 Containing Contaminated Sediments

The collection of chemical, physical, and biological data at Superfund sites containing

contaminated sediments is used to support human health and ecological risk assessment. To

support human health risk assessment, the contaminated media to which humans may be

exposed must be characterized. Exposure may be through routes such as ingestion of (the edible

portions of) contaminated fish and shellfish, ingestion of contaminated drinking water, and dermal

contact during swimming and wading. (Monitoring to characterize air, terrestrial species and avian

species impacted by contaminated sediments is, with few exceptions, not addressed by this

document.) More monitoring methods may be needed to support ecological risk assessment than

to support human health risk assessment because of the variety of biota and the complexity of

interactions involving contaminants and aquatic ecosystems.

Monitoring and monitoring data are important during the decision making process, which includes
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

3

the following activities:

• Site Assessment (including Preliminary Assessment and Site Inspection)

• Planning and Implementing Removal Actions

• Remedial Investigation and Risk Assessment

• Feasibility Study/Remedy Selection (following NCP criteria as the basis for decision

making)

• Remedy Implementation and Monitoring Recovery

Remedial Investigation and Risk Assessment

During the initial site assessment, remedial investigation information on the extent and magnitude

of chemical contamination at the site is obtained. At contaminated sediment sites, water and

sediment samples are often collected and analyzed for Constituents of Concern (COCs).

Because the toxicity of many contaminants is dependent upon physical properties of media , as

well as the potential receptors, additional chemical, physical, and biological parameters are also

collected; for example:

• Water – hardness, alkalinity, pH, suspended organic matter, dissolved oxygen level,

dissolved organic matter, salinity, temperature, depth

• Sediment - pH, total organic carbon, clay content and type, grain size distribution,

redox potential, depth of sample

• Habitat structure -- bottom characteristics, grain size distribution, cobbles, boulders,

benthic community structure, fish community structure, vegetation, and debris, among

others

Data on the presence of endangered or threatened species, sensitive species, and species of

economic or recreational importance, and information on critical or sensitive habitats are also

collected.

These data will be used in the development of a screening-level problem formulation and

ecological effects evaluation and screening-level preliminary exposure estimates. Often,

appropriate Ecotox Thresholds (ETs) will be used during the preliminary stages. ETs are defined

as media-specific contaminant concentrations above which there is sufficient concern regarding

adverse ecological effects to warrant further site investigation (USEPA 1996a). At contaminated

sites, the benchmarks commonly used to assess preliminary risk (establish the ETs) include:

• Water – National Ambient Water Quality Criteria (AWQC). Water quality for both

water and pore water is evaluated by comparison to AWQC for protection of aquatic

life as specified by USEPA in numerous guidelines. These criteria, most recently

updated in 1999 (USEPA 1999a), are intended to accurately reflect the latest

scientific knowledge of the effects of these chemicals on aquatic life. The current

AWQC list recommended criteria for 157 pollutants.

• Sediment – (1) Effects Range Low values (ERL) (Long and Morgan 1990, USEPA

1992a, Long et al. 1995); (2) Threshold Effects Concentrations (TECs) (MacDonald

et al., 2000). ER-Ls have been developed from correlated biological and chemical
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

4

data from laboratory and field studies combined with modeling studies representing

marine and estuarine environments. TECs for 28 chemicals can be used to screen

freshwater sediment for risk to benthic organisms but are not necessarily protective

of higher trophic level organisms. 

EPA Superfund has developed an ET program that calculates site-specific ETs by adjusting for

pH and hardness in surface water and TOC in sediments (USEPA 1996a). The ET calculations

use as a baseline the risk-based benchmarks referenced above.

During the screening-level problem formulation/ecological effects evaluation and again during the

risk characterization, the ecological significance of potential ecological receptors, as described in

the Ecological Risk Assessment Guidance for Superfund (USEPA 1997a) is also estimated.

During the problem formulation phase, ecological attributes that will function as assessment

endpoints are identified and an assessment of the proposed endpoints is made that will help

determine risk.

During problem formulation, the significance of adverse toxicological, biological, and ecological

effects to receptors is considered as part of the selection process for assessment endpoints.

Examples of endpoints for contaminated sediment sites include:

• Individual Level -Endangered or threatened species known to be present

• Population Level - Sensitive fish population, sensitive macroinvertebrate population, or

sensitive bird population exposed to COCs

• Community Level - Distribution and abundance of fish and avian communities, benthic

macroinfauna communities, and aquatic plant communities

Distinguishing potential and current adverse effects due to releases of contaminants at population

and community levels from normal fluctuations requires knowledge of the natural variability

inherent in the ecosystem (population fluctuations, presence/absence, abundance, and diversity).

During the risk characterization phase, the likelihood, duration, and magnitude of risk to the

receptors represented by assessment endpoints, the spatial and temporal extent of the risk, and

the estimation of COCs below which contaminants would no longer be of concern are all

developed. As presented in USEPA 1994a, candidate assessment endpoints in field studies at

contaminated sediment sites can include

Type Measurement Endpoint

Populations Survival and reproduction of fish

Survival, growth, and reproduction of aquatic

insect eating or fish-eating birds and

mammals
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

5

Communities Biomass

Productivity and respiration

Species richness

Species density

Relative abundance

Dominance

Diversity

Evenness

Similarity/difference between Superfund site

and reference site

Similarity/difference in guild structure between

Superfund site and reference site Presence,

absence, or population density of indicator

species

The types of organisms/data collected and methods that may be used for collection include

Biota Method

Periphyton Scraping

Coring (or sampling with a grab)

Suction

Artificial substrate

Plankton Trapping

Pumping

Netting (towing)

Water sampling

Benthic Macroinvertebrates Dredging or digging

Stream netting, sweep netting

Coring (or sampling with a grab)

Artificial substrate

Fish Seining

Trawling

Passive netting (gill, trammel, or hoop nets)

Electrofishing

Chemical collection

Birds Auditory and visual studies

Nesting success

Trapping

Additionally, sediment toxicity tests provide another mechanism to determine if contaminated

sediments are causing adverse effects in organisms in a controlled laboratory setting. The tests

that are commonly used for testing contaminated sediments include, aquatic, sediment, and

microbial tests. Many are highly standardized and have the advantage of wide acceptance.

Standardization has also resulted in the advantage of multiple laboratories with qualifications to
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

6

perform the testing. Aquatic toxicity tests include both freshwater and marine acute and chronic

toxicity tests. Freshwater and marine sediment toxicity tests include acute and chronic tests of

whole sediment or sediment interstitial water. 

Several USEPA and other agency documents establishing testing methods for the suitability of

dredged material for disposal in the marine and freshwater environments have been prepared

(USEPA/USACE 1991, USEPA/USACE 1992, USEPA/USACE 1998, USEPA 1994a, USEPA

2000a, USACE/WDNR/WDEC 2000). While these documents do not specifically address the

Superfund program, they present the various approaches taken for the management of

contaminated sediments, including the use of both sediment chemistry and toxicity data in

contaminated sediment evaluation.

Remedy Selection/Feasibility Studies

While monitoring and field data acquisition can occur in the above phases of the Superfund risk

assessment process, the field data will also need to be acquired for the remedy selection. The

remedies to be considered at contaminated sediment sites include

• Monitored natural recovery

• Containment in-place (in situ capping)

• Treatment in-place

• Removal and disposal of contaminated sediments in confined aquatic disposal (CAD)

facility

• Removal and upland containment.

The extent to which one of these remedies is superior needs to be determined on a case by case

basis based on site data, appropriately acquired following sound project planning and the

application of sound monitoring methods and data assessment techniques. As presented in

Sediment Management Work Group (1999), the appropriate questions that need to be answered

to guide remedial at contaminated sediment sites include

• How long will it take natural recovery to return the site to acceptable conditions?

• Will dredging and removal accelerate this process?

• Will other remedial options (i.e., capping) accelerate the process?

• What are the risks (for example, from large storm events) of leaving the contaminated

sediment in place, or of removing the engineered clean sediment cap?

The answers to the above questions can be derived from an understanding of the site from site

characterization studies, the appropriate application of transport and fate models and ecological

and human health risk assessments, leading to the development of a site conceptual model of the

transport and fate of COCs at the site that will lead to a risk-based remedy decision. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

7

Specific site condition information needs to be collected and evaluated when considering an in situ

capping remediation alternative at contaminated sediment sites, including:

• Physical environment - For example, bathymetry will influence the ability to place an in

situ cap in many areas and will also influence dispersion at the site during dredging or

capping. Bottom slope needs to be considered. Moderate slopes can rule out the

ability to cap without the placement of a physical barrier to prevent downslope

movement. 

• Hydrodynamic conditions - Stability of the cap over time (typically 30 years) needs to

be determined. This determination will require data such as normal water column

currents or channel flow, tidal fluctuations, wave and storm induced bottom currents, or

flow during flood conditions need to be considered. Some of these data will feed into

modeling studies, which may be needed to understand conditions at open water sites.

• Hydrogeological conditions - Groundwater discharge to near shore areas of lakes,

rivers and estuaries is common in many areas of United States and groundwater flow

through a contaminated sediment site can transfer a fraction of the contaminants to

the overlying surface water. In this circumstance, a determination of the magnitude of

groundwater flow and thickness of the contaminated layer should be made.

Additionally, data on sediment physical characteristics, such as water content, grain

size and clay content and type, organic content, plasticity indices, and specific gravity,

are also required to evaluate site conditions and cap design. Engineering

measurements of sediment shear strength and compressibility are also required.

Monitoring activities associated with the evaluation of fate and transport processes for site

assessment, risk characterization, remedy selection, or post remedy monitoring can include those

with the purpose of:

• Characterizing stream flow and the potential for sediment deposition and/or scouring

• Assessing naturally occurring biodegradation of contaminants (which may include

methods to characterize sediment geochemistry, biodegradation products, microbial

populations, chiral analytes, etc.)

• Assessing diffusive transport of contaminants through sediments to the water column

• Assessing water flow across the sediment/water column interface

• Assessing the consolidation of sediments resulting from compressive forces and

biodegradation

• Determining changes in the location of the sediment/water interface

• Assessing the mobility of contaminants (e.g., speciation methods for metals, total

organic carbon, acid volatile sulfides, pH, redox potential, etc.)
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

8

• Characterizing the physical characteristics of sediments (e.g., sediment cohesion,

shear strength, particle size distribution, etc.)

Monitoring activities facilitating the implementation of remedial actions can include those with the

purpose of:

• Evaluating monitored natural recovery (e.g., by decrease in concentrations of COCs,

decrease in toxicity

• Facilitating installation of in situ caps

• Determining the long-term performance and condition of in situ caps (e.g., by

measuring cap thickness over time, measuring COCs in sediment cap porewater over

time)

• Evaluating aquatic system recovery following installation of in situ caps

• Facilitating remedial dredging

• Evaluating aquatic system recovery following completion of remedial dredging

• Ensuring that remedial construction activities do not have the potential to produce and

are not producing immediate adverse effects (e.g., by monitoring turbidity, dissolved

oxygen, acute toxicity testing)

The methods presented in the next several sections are frequently used for assessment of

environmental conditions at contaminated sediment sites and provide data to address many of the

data uses described above. Many of these methods were originally developed for the analysis of

water and wastewater or for the analysis of solid waste have been used without modification for

marine or aquatic investigations. Other methods originally developed for those purposes have

been modified for contaminated sediment investigations, often to lower detection limits needed for

risk assessments or to facilitate working with a high salinity water or sediment matrix. However,

many of the methods were specifically developed for working in the aquatic or marine

environment at contaminated sites, and elsewhere. Without exception, it is intended that all of the

methods presented will be suitable for investigations at Superfund sites containing contaminated

sediments. However, not all methods will be suitable for all sites. The selection of methods for a

particular site will depend on the site conditions, remediation plans, budgetary constraints and

other factors.

2.0 MONITORING METHODS

2.1 Water

Water sample collection methods, and chemical, physical, and biological analyses have

continuously been developed and implemented over the years to evaluate the health of our

nations streams, lakes, rivers, ponds, creeks, lagoons, estuaries, oceans and surface

impoundments. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

9

At Superfund sites, water samples are specifically collected with the following objectives in mind

(USEPA, 1994a):

• To determine if the contaminant is hazardous by identifying its composition and

characteristics;

• To determine if there is an imminent or substantial threat to public health or welfare or

to the environment;

• To determine the need for long-term action;

• To develop containment and control strategies;

• To evaluate appropriate disposal/treatment options; and,

• To verify treatment goals or clean up levels. 

In order to achieve the aforementioned objectives, field sampling and analytical strategies are

designed to provide site-specific information because characteristics and sampling strategies vary

widely. The following fact sheets relating to water are divided into sections pertaining to field

sample collection and processing, chemical and physical laboratory analyses and biological

laboratory analyses. These fact sheets intend to provide Superfund managers with a summary of

the existing methods that may be applicable to their site, their relative strengths, and their relative

weaknesses.

2.1.1 Field Sample Collection and Immediate Processing, In Situ Data Acquisition

Section 2.1.1 presents methods for field sample collection, field or immediate sample processing,

and in situ data acquisition. In situ data acquisition primarily collects those data easily and

economically collected with various sensors and includes parameters such as salinity,

conductivity, temperature, pH, dissolved oxygen, light transmission, and light attenuation. Data

collection in situ is often automated and allows for data acquisition over broad spatial and

temporal scales. Because many biological processes in the environment are affected, directly or

indirectly, by the physical characteristics of the environment, the data collected in situ are vital in

site assessments at aquatic sites. Water samples are also collected in numerous ways and

brought back to the laboratory to identify chemical contaminants that may disrupt existing physical

and biological processes and what effects those chemical contaminants may have on resident

organisms. These samples and analyses are crucial in determining potential exposure pathways

based on the concentration of the chemical in the environment and then the chemical’s behavior

in the environment based on biological, chemical, and physical parameters. Measurement and

collection methods vary based on the characteristics of the water body in question. Therefore,

numerous methods are provided. 

Many of the field sampling and data collection methods for water are routinely performed to

evaluate the health and biological integrity of our surface waters. Thus, they originate from

programs unrelated to contaminated sediments. Many of the methods have been taken from

USEPA Environmental Monitoring and Assessment Program documents. Other sources of

methods for water column field collection include:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

10

• The USEPA and the Puget Sound Water Quality Action Team

• The USEPA’s Lake Michigan Mass Balance Study Methods Compendium

• The USEPA’s Environmental Response Team

• The USEPA’s Environmental Research Laboratory-Narragansett

• Standard Methods for Examination of Water and Wastewater

• ASTM

• Journal publications

• The USEPA’s Rapid Bioassessment Protocols
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

11

Fact Sheet No. 2.1.1-1

Method Title In Situ Sampling with the Hydrolab Datasonde3® Unit

Purpose This procedure describes one of several CTD devices used to collect high-quality in

situ data for salinity, temperature, dissolved oxygen (DO) concentration, pH, and

water depth.

Method Summary The Hydrolab Datasonde3®unit is one of several comm ercial units that collect in

situ Conductivity, Temperature, and Depth (CTD) data, and also measures pH and

DO to obtain a vertical profile of water column conditions. 

After calibration, the Hydrolab is connected to a winch cable, and the protective

covers on the probes are removed and the stirrer is connected. The unit is lowered

over the side and allowed to equilibrate at the surface for at least two minutes.

While the unit is equilibrating, a QC check is performed with a YSI DO probe,

refractometer and a thermom eter to ensure that the readings from all equipment

agree.

The Hydrolab unit is lowered at intervals specific to the relative depth of the site,

allowing the unit to stabilize at each specified stop during descent. Data can be

acquired on the descent and ascent.

Data

Uses/Application

Salinity, conductivity, temperature and dissolved oxygen parameters are commonly

collected at all stations where samples for water quality are collected. 

Advantages The Hydrolab is a relatively sm all unit that may be hand deployed, if necessary.

Limitations The Hydrolab is much more expensive than alternate sampling equipment such as

the YSI data probe, refractometer and thermometer. Without proper calibration, the

Hydrolab or other CTD units may record erroneous information.

Reference Strobel, C.J. 2000. Coastal 2000 - Northeast Component: Field Operations Manual.

U.S. Environmental Protection Agency , National Health and Environmental Effects

Research Laboratory, Atlantic Ecology Division, Narragansett, R.I. EPA/620/R00/002.

Website http://www.epa.gov/emap/nca/html/docs/c2knefm .

html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

12

Fact Sheet No. 2.1.1-2

Method Title In Situ Dissolved Oxygen Sampling with a YSI Model 58 Dissolved Oxygen Meter

and Probe

Purpose This method describes the use of a YSI Dissolved Oxygen Meter and Probe to take

in situ oxygen measurem ents in the water column.

Method Summary After checking the batteries and replacing the probe mem brane, the YSI Model 58

Dissolved Oxygen(DO) Meter and Probe must be calibrated before sampling at

each station. The meter is then set to 0.1 mg/L mode. If measuring the DO in other

than surface water, collect water in a Go-Flo®bottle. Measure the salinity with a

refractometer. Insert the stirrer-probe unit directly into the Go-Flo bottle and turn on

the stirrer. When the meter reading has stabilized, record the oxygen value on the

data sheet. Turn the stirrer off, rinse the probe with freshwater, and store the unit

out of sunlight. 

Data

Uses/Application

DO is an important water quality parameter for surface water aquatic life. The YSI

Model 58 Dissolved Oxygen Meter and Probe can be used to take oxygen

measurem ents at the surface as a Quality Control check on DO measurem ents

determined by other methods. Probes are used mainly on in situ instruments for

providing continuous water-column profiles of dissolved oxygen.

Advantages The YSI Model 58 Dissolved Oxygen Meter and Probe is sm all and transportable. It

is a good, quick way to check on the accuracy of the Hydrolab. The measurement

of DO directly in sampling bottles reduces sampling bias associated with sample

transfer. 

Limitations The device must be calibrated at every station prior to use. The device could

produce erroneous data if the mem brane is at all damaged or dried out. The probe

method is not commonly used for oceanographic studies in which measurem ents

are made on discrete samples of seawater. 

Reference Strobel, C.J. 2000. Coastal 2000 - Northeast Component: Field Operations Manual.

U.S. Environmental Protection Agency , National Health and Environmental Effects

Research Laboratory, Atlantic Ecology Division, Narragansett, R.I. EPA/620/R00/002.

Website http://www.epa.gov/emap/nca/html/docs/c2knefm.html Last Accessed:

1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

13

Fact Sheet No. 2.1.1-3

Method Title In Situ Irradiance Measurements

Purpose This method describes the manner in which a vertical profile of light intensity can be

measured for the purpose of calculating a light attenuation coefficient at each

station.

Method Summary Photon flux, or Irradiance is the amount of light that diffuses the water referenced to

the input. Photosynthetically active radiation (PAR) is measured to determine the

absorption rate of that light by phytoplankton.

To obtain a PAR profile, a deck sensor and an underwater sensor are connected to

an independent data logger. The deck sensor is placed in a location where it is not

shaded. The underwater sensor is lowered on the sunny side of the boat to a depth

of about 10 cm (representing surface). Record the values from both sensors.

Lower the underwater sensor to 0.5 meters and record the values. Then lower the

sensor at given intervals depending on the relative depth of the sampling location

and record the values. Repeat the process on the upcast.

Data

Uses/Application

PAR is important in understanding the dynamics of the “photic zone” which helps to

understand lake health issues such as photosynthesis, and toxic algae blooms and

eutrophication. Irradiance and PAR are measured for the purpose of calculating a

light attenuation coefficient at each station which can be used in productivity

assessments or for site assessments. 

Advantages PAR sensors require no field calibration, but they should be returned to the

manufacturer prior to each field season for annual calibration. PAR sensors provide

more quantitative data to derive light attenuation data in a more exact manner.

Limitations PAR sensors are more expensive than other light attenuation methods. 

Reference Strobel, C.J. 2000. Coastal 2000 - Northeast Component: Field Operations Manual.

U.S. Environmental Protection Agency , National Health and Environmental Effects

Research Laboratory, Atlantic Ecology Division, Narragansett, R.I. EPA/620/R00/002.

Website http://www.epa.gov/emap/nca/html/docs/c2kn

efm.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

14

Fact Sheet No. 2.1.1-4

Method Title In Situ Transparency Measurements, Secchi disk profile

Purpose This method describes the manners in which a vertical profile of light can be

measured for the purposes of determining water column transparency. 

Method Summary To obtain a Secchi profile, a 20 cm black and white Secchi disk is lowered until it is

no longer visible. Note the depth using the markings on the line. Slowly raise the

disk until it just becomes visible and note the depth. Repeat this process 3 times and

record the average of the readings. 

Data

Uses/Application

The Secchi disk measures water column transparency and may be best used when

transparency is high. 

Advantages The Secchi disk can be used on any vessel, and computer data loggers are not

necessary. 

Limitations Secchi disks data may vary amongst investigators. More room for human error.

Reference PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water

Column, and Tissue in Puget Sound, Puget Sound Protocols and Guidelines. Puget

Sound Water Quality Action Team, Olympia, W A. 

Website http://wwwpsat.wa.gov/Publications/

protocols/protocol.htm l

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

15

Fact Sheet No. 2.1.1-5

Method Title Sample Collection Procedures for Marine Water

Purpose This method describes the methods and different type of water bottles used to

collect marine water column samples.

Method Summary Water samples should be collected prior to “bottom related” activities (sediment

grabs and trawling) to assure that bottom sediments are not re-suspended. These

samples should also be collected on a day when it is possible to ship samples on

the same or following day to meet toxicity testing holding times, if necessary.

The typical water bottle sampler consists of a cylindrical tube with stoppers at each

end, and a closing device that is activated from the surface by a messenger or an

electrical signal. Multiple water samplers can be attached sequentially to a vertical

hydrowire for sampling at multiple depths on a single cast, or they can be mounted

on a rosette frame (see fact sheet 2.1.1.-6; often in conjunction with an in situ sensor

array) which allows for collection of replicate samples at the same depth.

After the sampler is cocked, it is lowered to a designated depth. Avoid deploying

water bottles in surface slicks as these can contaminate samples with organic

compounds. If contamination by the microlayer is of concern, use samplers that are

designed to remain closed until they have descended below the microlayer (i.e., GoFloTM bottle from General Oceanics, Inc. Miami, Florida). TeflonTM-lined Go-FloTM

bottles are recommended when sampling marine water that will be analyzed for

ambient or trace levels of mercury or other metals upon proper cleaning Niskin

bottles are also often used to collect water samples, however these bottles remain

open as they descend through the water column, and the enclosure mechanism is

on the inside of the bottle, making them difficult to clean. 

Once the sampler reaches the desired depth, it should be allowed to equilibrate to

ambient conditions for approximately 1 minute before it is closed. If reversing

thermometers are involved, equilibration should be 5 minutes. After equilibrium, the

closing device is activated by a messenger or electric signal, and the sampler is

retrieved. To ensure that all samples are truly representative of the water column

within a specific water parcel, it is advisable that they be collected from a single cast.

As the water samplers are being brought on board, each bottle should be checked

immediately for leakage of sample water around the seals. If the sample has been

compromised, the cast should be repeated. 

Data

Uses/Application

Water bottle samplers are used to collect water column samples for laboratory

analyses of conventionals, metals, organics and microbiological analytical

procedures and toxicity tests
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

16

Fact Sheet No. 2.1.1-5 (contd.)

Advantages Both Niskin and Go-Flo bottles can be deployed individually or attached to a large

rosette sampler which collects in situ data while water samples are being collected. 

The Go-Flo design allows it to be deployed (and returned) in the closed position,

reducing the possibility of sample contamination from surface slicks and the

microlayer. These bottles also have external springs therefore there is no risk of

sample contamination. The Go-Flo bottles are commonly used to collect samples

later analyzed for metals or organics.

The EMAP Program provides similar guidance regarding marine water collection

(USEPA, 1990a).

Limitations Holding times for collecting samples must be observed. Niskin bottles are generally

not used to collect samples later analyzed for sensitive organics or metal analyses

since they are deployed in the open position and the spring used to close the cap is

located on the inside of the bottle.

Reference PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water

Column, and Tissue in Puget Sound, Puget Sound Protocols and Guidelines. Puget

Sound Water Quality Action Team, Olympia, W A. 

Website http://wwwpsat..wa.gov/Publications/p

rotocols/protocol.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

17

Fact Sheet No. 2.1.1-6

Method Title Field Sampling Using Rosette Sampler, LMMB 013

Purpose This procedure describes the collection of water samples using a Rosette Sampler.

Method Summary The Rosette sampler is the primary sampling instrument for the collection of all

nutrient parameters, phytoplankton, chlorophyll a, Phaeophytin a, and dissolved

oxygen, temperature, total suspended solids, turbidity, specific conductance, and

pH.

The system consists of 12 water sampling bottles (8 L Niskin or Go-Flo bottles), a

CTD (conductivity, temperature and depth sensor) attached at the bottom of the

Rosette, an A-frame, a multiconductor cable, a variable speed winch, and a deck

unit with attached computer. The bottles can be closed in any predetermined order,

remotely from the deck of the vessel while the array is submerged at the various

sampling depths. The CTD is built to provide real time information on a number of

water quality parameters as it moves through the water column. During sampling,

the Rosette/CTD system is lowered to the bottom to define the temperature profile of

the water column. The sampling depths are then selected. After collection of

samples with the Rosette sampler, the sampler is brought on board and the water

samples are transferred from the Niskin or Go-Flo bottles to various sample bottles,

depending on analysis, for storage until processing and analysis. During sampling,

each bottle is rinsed with sample water, emptied, and filled with sample water. The

cap is replaced after addition of the preservative, or immediately if no preservative is

added. Dissolved oxygen samples are processed immediately.

Data

Uses/Application

Rosette samplers are typically used in open water, both freshwater and marine

environments, where samples from multiple stations and depths will be required.

Advantages Real time in situ water column data can be collected to determine the best depths

for water sample collection. Each discrete water sample has a corresponding set of

water quality parameter data.

Limitations Dissolved nutrient samples must be filtered before analysis, and must be filtered

within one hour of collection.

Reference USEPA. 1997b. Lake Michigan Mass Balance Study Methods Compendium,

Volume 1: Sample Collection Techniques, EPA 905-R-97-012c. Great Lakes

National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmmb/meth

ods/mbross.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

18

Fact Sheet No. 2.1.1-7

Method Title Water Sample Collection with the Kemmerer Bottle and the Bacon Bomb Sampler,

ERT SOP #2013

Purpose Both the Kemmerer Bottle and the Bacon Bomb Sampler can be used to collect

surface water in situations where site access from a boat or structure is available

and where samples at depth are required. 

Method Summary A properly decontaminated, preset Kemmerer bottle or Bacon Bomb Sampler may

be lowered to a predetermined depth to collect an aqueous sample. 

The Kemmerer Bottle can be used for general purpose sampling at the surface or at

specified depths. The all-angle head locks the sampler in the open position and

unlocks when struck by the messenger, closing both end seals of the sampler. The

sampler is retrieved and samples are transferred to appropriate sampling container.

The Bacon Bomb Sampler opens when a protruded plunger strikes bottom. It

closes as the sampling device is hoisted back into the water column. By attaching a

cord to the upper end of the plunger, samples may also be taken from an

intermediate level as well. The sampler is then retrieved and the sample is

transferred to the appropriate sampling container.

Data

Uses/Application

These samplers can collect any sort of water sample that then may be used for

chemistry analysis, nutrients analysis, or toxicity studies.

Advantages These samplers can collect samples from a range of depths. The Kemmerer bottle

has few moving parts and needs little maintenance. 

Limitations The sampling stations need to be accessible by boat or from land. The samples

could be cross contaminated if sample equipment is not appropriately

decontaminated in between stations.

Reference USEPA. 1994b. Surface Water Sampling, SOP # 2013, in Compendium of ERT

Standard Operating Procedures. Environmental Response Team. Compendium of

ERT Standard Operating Protocols. Office of Solid Waste and Emergency

Response, U.S. Environmental Protection Agency, Edison NJ.

Website http://www.ert.org/products/2013.pdf Accessed: Last 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

19

Fact Sheet No. 2.1.1-8

Method Title Dip Sampler, ERT SOP # 2013

Purpose The Dip sampler is generally used to collect surface water samples in a situation

where the sample is to be recovered from an outfall pipe or along a lagoon bank

where direct access is limited.

Method Summary The dip sampler is extended into the sample location to collect sample. The

sampler is retrieved and the sample is transferred to the appropriate container.

Data

Uses/Application

These surface water samplers can collect any sort of liquid sample that then may be

used for water chemistry analysis, nutrients analysis, or toxicity studies.

Advantages The long handle on dip samplers allows them to collect samples from discrete

locations. Dip samplers are extremely durable. They can be useful in weedy

habitats where other samplers may not work.

Limitations The sampling location must be within reach of the investigator. The samples could

potentially be cross-contaminated if the sampler is not properly cleaned between

sample collections. 

Reference USEPA. 1994b. Surface Water Sampling, SOP # 2013, in Compendium of

Environmental Response Team Compendium of Standard Operating Procedures.

Office of Solid Waste and Emergency Response, U.S. Environmental Protection

Agency, Edison NJ.

Website http://www.ert.org/products/2013.pdf Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

20

Fact Sheet No. 2.1.1-9

Method Title Sample and Preservation of Water Specific Parameters

Purpose This method describes the sample collection procedures unique to conventionals,

metals, organics, and microbiological analytical parameters. See Table 2.1.1-1 for

details pertaining to recommended sample sizes, containers, preservation

techniques and holding times for water.

Method Summary For analysis of conventional parameters, those parameters subject to biological

alteration should be measured first and within 15 minutes of sampling. Dissolved

oxygen should be the first parameter collected, followed in order of priority by those

parameters which would be the most affected by subsampling delays.

For analysis of total metals or total mercury, the samples should be acidified to pH

<2 using ultrapure HNO3. Samples that will be analyzed for mercury speciation

should be preserved with HCl rather than HNO3

. Samples that will be analyzed for

both dissolved and particulate metals should be filtered as soon as possible, within

24 hours of collection. The filtrate, which contains the dissolved fraction, should be

preserved by acidifying to pH<2 using ultrapure HNO3. The particulate fraction,

which is retained on the filter, is frozen for preservation.

For organics analysis, the samples for analysis of volatile organic compounds are

collected first in 40 ml VOA vials leaving no head space. The samples should be

protected from possible contamination such as fuels, winch grease, exhaust, and

solvents that may be present on or around a research vessel. Preserve water

samples collected for organics analysis as soon as possible, according to the

guidelines summ arized in the attached table. 

For microbiological analyses, it is important to collect from the microlayer, or

surface-most layer of water. The microlayer is most easily included when using the

scoop method. The scoop method involves plunging an open bottle straight down to

a depth of 15 to 39 cm below the water surface, moving it horizontal to the surface

while tipping it slightly to let trapped air escape, and removing the bottle in a vertical

position. Approximately 2.5 cm of head space is required in the sample container.

Sample containers should be isolated from contact with wet ice as it could impart

contamination to the sample.

Data

Uses/Application

These samples are then shipped to analytical laboratories where the

aforementioned analyses are performed to determine water chemistry. 

Advantages Filtration is used for metals analysis since it is inexpensive and yields a sample that

is directly suitable for chemical analysis.

Limitations Holding times must be met in order to appropriately analyze the samples. Care

must be taken to avoid any sort of contamination of these samples since they will be

analyzed with sensitive procedures.

Sm all amounts of particulate metals are collected during filtration, which make it

difficult to use them for low-level metal analyses.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

21

Fact Sheet No. 2.1.1-9 (contd.)

Reference PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water

Column, and Tissue in Puget Sound, Puget Sound Protocols and Guidelines. Puget

Sound Water Quality Action Team, Olympia, W A. 

Website http://www.psat.wa.gov/Publications/

protocols/protocol.htm l

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

22

Table 2.1.1-1. A Summary of Sample Sizes, Containers, Preservation Techniques, and Holding Times for

Water (PSWQAT. 1997.)1

Parameter

Minimum

Sample

Size (ml) Container Preservation Technique Holding Time

Alkalinity 100 Glass or

Polyethylene

Refrigerate, 4

B C 14 Days

Total Hardness 100 Glass or

Polyethylene

Refrigerate, 4

B C

HNO3to pH<2

6 Months

Total Phosphorous 50 Glass or

Polyethylene

Refrigerate, 4

B C

H2SO4to pH<2

28 Days

Orthophosphate 50 Glass or

Polyethylene

Refrigerate, 4

B C

Filter on site

48 Hours

pH 25 Glass or

Polyethylene

None Analyze Immediately

Salinity 200 Glass or

Polyethylene

None 28 Days

Turbidity 100 Glass or

Polyethylene

Refrigerate, 4

B C 48 Hours

Total Suspended

Solids

1,000 to

4,000

Glass or

Polyethylene

Refrigerate, 4

B C 7 Days

Dissolved Oxygen

Winkler

125 Glass Bottle

with Glass Top

Fix with MnCl2 and Alk.

Iod. (2 ml ea.)

8 Hours (store in the dark)

Dissolved Oxygen

Probe

125 Glass Bottle

with Glass Top

None Analyze Immediately

Ammonia-N 100 Glass or

Polyethylene

Refrigerate, 4

B C

H2SO4

to pH<2

28 Days

Nitrite-N 100 Glass or

Polyethylene

Refrigerate, 4

B C 48 Hours

Nitrate-N 100 Glass or

Polyethylene

Refrigerate, 4

B C 48 Hours

Silica 200 Polyethylene Refrigerate, 4

B C 28 Days

Chlorophyll a 25 to 1,000 Glass or

Polyethylene

(Dark)

Store filters frozen (-20 B C)

in the dark

28 Days

Volatile Organics 80 Glass -2 40 ml

vials, No Head

space

Refrigerate, 4

B C

HCl to pH<2

14 Days
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

23

Table 2.1.1-1. (contd.)

Parameter

Minimum

Sample

Size (ml) Container Preservation Technique Holding Time

Semi-volatile

Organics

1,000 to

2,000

Glass Refrigerate, 4

B C 7 Days

Total Mercury and

Diss. Mercury

500 TeflonTM or

Glass with

TeflonTM Cap

Refrigerate, 4

B C

HNO3

to pH<2

28 Days

Total Metals and

Diss. Metals

1,000 Polyethylene

or TeflonTM

Refrigerate, 4

B C

HNO3

to pH<2

6 Months

Microbiology 500 HDPE

(Autoclaved)

Refrigerate, 4

B C 24 Hours

1 PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water Column, and Tissue in Puget

Sound, Puget Sound Protocols and Guidelines. Puget Sound Water Quality Action Team, Olympia, WA. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

24

Fact Sheet No. 2.1.1-10

Method Title Sampling of Particulate-Phase and Dissolved-Phase Organic Carbon in Great Lakes

Waters, LMMB 014

Purpose This method describes the sampling of water for particulate-phase organic carbon

(POC) and dissolved-phase organic carbon (DOC).

Method Summary Water samples are collected using either a submersible pump or Rosette sampler

(see Fact sheet 2.1.1-6). The volume of sample to be filtered is measured in a

graduated cylinder. Prior to filling, the cylinder is rinsed twice with sample water. 

The water sample for POC/DOC analysis is vacuum filtered through an ashed 47

mm diameter glass fiber filter (0.7 μm pore-size) in an all-glass filtration apparatus.

Samples are filtered simultaneously in duplicate. The samples are acidified with 0.2

N HCl during the filtration to remove inorganic carbonates. The POC is retained on

the filter, and the DOC is collected in the filtrate. The volume of sample required to

produce a reliable POC measurement varies with station location, depth, and time of

year. If the filter becomes visibly loaded with particles and the flow of water through

the filter slows considerably, sufficient particulate matter has been collected. The

POC filter is folded, placed in an aluminum foil pouch, and frozen at -10°C until

analysis. The filtrate is collected and promptly analyzed for DOC in a shipboard

laboratory. 

Data

Uses/Application

POC/DOC are parameters which are ancillary to the determination of hydrophobic

organic contaminants (HOCs). 

Advantages POC and DOC are important water quality measurem ents that add to the value of

water column assessments.

Limitations Shipboard or field processing is generally required to meet holding times for these

analyses.

Reference USEPA. 1997b. Lake Michigan Mass Balance Study Methods Compendium,

Volume 1: Sample Collection Techniques, EPA 905-R-97-012c. Great Lakes

National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

pocdoc2.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

25

Fact Sheet No. 2.1.1-11

Method Title Sampling Ambient Water for Trace Metals at EPA Water Quality Criteria Levels, EPA

Method 1669

Purpose This method is for the collection and filtration of ambient water samples for

subsequent determination of total and dissolved metals (antim ony, arsenic,

cadmium, chromium (III and VI), copper, lead, mercury, nickel, selenium, silver,

thallium, zinc).

Method Summary Method 1669 is "performance-based"; i.e., an alternate sampling procedure or

technique may be used, so long as neither samples nor blanks are contaminated

when following the alternate procedures. Before samples are collected, all sampling

equipment and sample containers are cleaned in a laboratory or cleaning facility

using detergent, mineral acids, and reagent water. After cleaning, sample containers

are filled with weak acid solution, individually double-bagged, and shipped to the

sampling site. If samples are to be collected for determination of trivalent chromium,

the sampling team processes additional QC aliquots are processed. Upon arrival at

the sampling site, one mem ber of the two-person sampling team is designated as

"dirty hands"; the second mem ber is designated as "clean hands." All operations

involving contact with the sample bottle and transfer of the sample from the sample

collection device to the sample bottle are handled by the individual designated as

"clean hands." "Dirty hands" is responsible for preparation of the sampler (except the

sample container itself), operation of any machinery, and for all other activities that do

not involve direct contact with the sample. All sampling equipment and sample

containers used for metals determinations must be nonmetallic and free from any

material that may contain metals. Sampling personnel are required to wear clean,

nontalc gloves at all times when handling sampling equipment and sample

containers. Samples for dissolved metals are filtered through a 0.45 :m capsule filter

at the field site. After filtering, the samples are double-bagged and iced immediately.

Acid preservation of samples is performed in the field or in the laboratory. Field

preservation is

necessary for determinations of trivalent chromium.

Data

Uses/Application

This method is applicable for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery Act,

the Com prehensive Environmental Response, Compensation and Liability Act, and

the Safe Drinking Water Act.

Advantages Clean sampling methods reduce/eliminate bias associated with sample collection

handling.

Limitations Samples may become contaminated by numerous routes. These methods are only

applicable for trace metal contaminants.

Reference USEPA. 1997b. Lake Michigan Mass Balance Study Methods Compendium, Volume

1: Sample Collection Techniques. EPA-821-C-01-001. Great Lakes National

Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/clariton/clhtml/pubtitleOW.html Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

26

Fact Sheet No. 2.1.1-12

Method Title ESS Method 340.2: Total Suspended Solids, Mass Balance (Dried at 103-105°C)

Volatile Suspended Solids (Ignited at 550°C), in LMMB 065

Purpose To measure the portion of total solid retained by a filter from drinking, surface, and

saline waters; domestic and industrial wastes.

Method Summary Water samples are collected by submersible pump or Rosette sampler (see Fact

sheet 2.1.1-6). A sample volume is selected that will yield 2 - 20,000 mg/L. total

suspended solids. For open-lake oligotrophic conditions, 2-4 liters will provide

enough particulate matter. For near-shore or eutrophic conditions, 200-500 ML may

be sufficient. A well-mixed sample is filtered through a preweighed standard glassfiber filter, and the residue retained on the filter is dried at 103 to 105 °C for at least

one hour. The increase in weight of the filter represents the total suspended solids.

If measuring TSS of estuarine water, the filter must be well rinsed with DI water to

remove salt residue. 

Following Method LMMB 098, water samples can be filtered in the field and then

frozen at -10°C until final weighing in the laboratory.

The Environmental Research Laboratory - Narragansett (NHEERL-AED) SOP

1.02.004 and Standard Method 2540D also describe similar methods for measuring

total suspended solids (USEPA and Naval Construction Battalion Center, 1992;

APHA, 1999).

Data

Uses/Application

TSS is an ancillary parameter to the determination of hydrophobic organic

contaminants (HOCs). TSS is also commonly measured to assess water clarity or

to assess sediment transport and to norm alize total (aqueous) contaminant data. 

Advantages The samples may be filtered in the field or in portable laboratory facilities. 

Limitations Excessive residue on the filter may form a water-entrapping crust. Sample size

should be limited to yield no more than 200 mg residue. 

Reference USEPA.1997c. Lake Michigan Mass Balance Study Methods Compendium,

Volume 3: Metals, Conventionals, Radio chemistry, and Biomonitoring Sample

Analysis Techniques, EPA 905-R-97-012c. Great Lakes National Program Office,

U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

methd340.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

27

Fact Sheet No. 2.1.1-13

Method Title In situ Peepers

Purpose This method describes an in situ method to collect pore water.

Method Summary A peeper, as described in this method, is a mesh-sided chamber that is inserted into

the sediment for the purpose of collecting pore water. Peepers are placed in situ

below the sediment surface with only the tubes emerging above the surface. The

chambers (constructed using 500-ml polyethylene bottles, mesh sides, tygon tubing

and a 50-ml syringe that slowly extracts the pore water) are buried in shallow waters

and sediment packed around the unit to ensure overlying waters are not in contact

with side mesh windows. Immediately following burial, water in the chamber is

evacuated to enhance entry and equilibration of pore water. After several minutes,

day-0 samples are collected. 

The syringe is capped and returned to the laboratory for analysis of the extracted

water. Pore water is collected on day-0 and over predetermined time periods of up

to a month.

Data

Uses/Application

In situ peepers are used to collect pore water for toxicological testing. The device

may also be modified to suitable collect samples for chemical contaminant analyses.

Advantages In situ sampling reduces sampling and laboratory-related errors that may affect

organism response (i.e., resuspension of sediments that organisms would otherwise

not be exposed to). 

Limitations There may be site-specific limitations that would prevent the deployment of a

peepers.

Reference Sarda, N. and G.A. Burton. 1995. Am monia Variation in Sediments: Spatial,

Temporal, and Method-Related Effects. Environmental Toxicology and Chemistry.

Vol. 14: 9. 

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

28

Fact Sheet No. 2.1.1-14

Method Title Suction Samplers

Purpose Suction samplers can be used to collect water that is then analyzed for various

chemicals of concern. This method is suitable for pore water samples analyzed for

amm onia. 

Method Summary The suction method uses an aquarium stone and a hand-operated vacuum pump to

extract pore water from surficial sediment in situ. The air stone is buried under the

surface of the sediment and the suction is applied. An imposed vacuum sucks pore

water into an in-ground porous cup. In simple systems, the water is stored in the

suction cell and is subsequently sucked or blown into a sample flask placed on the

ground surface. The vacuum is not maintained between samples.

Data

Uses/Application

Suction samplers are able to extract pore water in situ for chemical analysis. 

Advantages In situ sampling reduces sampling and laboratory-related errors that may affect

organism response (i.e., resuspension of sediments that organisms would otherwise

not be exposed to). 

Limitations There may be site-specific limitations that would prevent the deployment of a suction

sampler.

Reference Sarda, N. and G.A. Burton. 1995. Am monia Variation in Sediments: Spatial,

Temporal, and Method-Related Effects. Environmental Toxicology and Chemistry.

Vol. 14: 9. 

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

29

Fact Sheet No. 2.1.1-15

Method Title Physical Characterization of a Stream

Purpose These methods describe the data needed to characterize the physical attributes of a

stream needed for a habitat assessment.

Method Summary Physical characterization of stream habitat includes descriptions of the:

• General stream characteristics such as an analysis of the stream subsystem

(where relevant), stream type (i.e., cold-water vs. warm-water), and stream

origin (i.e., glacial, montane, swamp, bog);

• Watershed features such as the predominant land use type surrounding the

stream, local watershed nonpoint source pollution, and local watershed erosion;

• Riparian vegetation up to 18 meters from the stream bed;

• Instream features such as estimated reach length, estimated stream width,

sampling reach area, estimated stream depth, velocity, canopy cover, high water

mark, proportion of reach represented by stream morphological types,

channelization, and dam presence; 

• Surrounding woody debris in contact with the stream, noted by a wading visual

observer;

• The most dominant type of aquatic plants; 

• Water quality parameters such as temperature, conductivity, dissolved oxygen,

pH, turbidity, water odors, water surface oils, and turbidity; and,

• Sediment characteristics such as odors, oils, deposits, inorganic substrate

components, and organic substrate components. 

Data

Uses/Application

Evaluations of habitat quality via physical characteristics and water quality

parameters are pertinent to any assessments of ecological integrity. These types of

assessments are performed by many water resource agencies to determine if

degraded habitat is the result of toxicity and/or pollution. The full assessment

includes a general description of the site, the aforementioned physical

characterization and water quality assessment, and a visual assessment of instream

and riparian habitat quality (Fact Sheet 2.1.1-16). 

Advantages Most of the aforementioned data can be collected with field investigations in a quick

and relatively inexpensive manner. 

Limitations These types of assessments are not sufficiently comprehensive to adequately

identify all causes of impact. 

Reference Barbour, et al., 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second

Edition. EPA 841-B-99-002. Office of Water, U.S. Environmental Protection

Agency, Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/

ch05main.html#Section%205.2

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

30

Fact Sheet No. 2.1.1-16

Method Title Visual-Based Habitat Assessment

Purpose These methods describe the process for performing a visual-based habitat

assessment of a stream. Habitat parameters pertinent to the assessment of habitat

quality include those that characterize the stream “micro scale” habitat (e.g.,

estimation of embeddedness), the “macro scale” features (e.g., channel

morphology) and the riparian and bank structure features that are most often

influential in affecting the other parameters. 

Method Summary First, a 100 meter reach of a stream must be selected for the assessment. The

entire sampling reach is then evaluated for each of the following parameters listed

below:

• Epifaunal substrate/available cover;

• Embeddedness of rocks and snags;

• Pool substrate characterization;

• Velocity/depth combinations;

• Pool variability;

• Sediment deposition;

• Channel flow status;

• Channel alteration;

• Frequency of riffles (or bends);

• Channel sinuosity;

• Bank stability;

• Bank vegetative protection; and

• Riparian vegetative zone width.

An additional 7 general physical habitat attributes are also important in determining

stream ecology:

• Channel dimensions;

• Channel gradient;

• Channel substrate size and type;

• Habitat com plexity and cover;

• Riparian vegetation cover and structure;

• Anthropogenic alterations; and

• Channel-riparian interaction.

The habitat assessment process involves rating the parameters as optimal,

suboptim al, marginal, or poor based on criteria included in the data sheets. 

Data

Uses/Application

Habitat assessments based on visual observation can be separated into 2 basic

approaches- one designed for high-gradient streams and one designed for lowgradient streams. Some state programs have adapted this approach using

somewhat different or fewer parameters. 

Advantages Standardized parameters list and protocol allows for some intercomparison amongst

sites. 

Limitations Many of the data parameters are qualitative, thus assessments made at the same

location by different biologists may vary. The protocol suggests that a team of 2 or

more trained biologists should perform the assessment to enhance data quality. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

31

Fact Sheet No. 2.1.1-16 (contd.)

Reference Barbour, et al., 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second

Edition. EPA 841-B-99-002. Office of Water, U.S. Environmental Protection

Agency, Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/

ch05main.html#Section%205.2

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

32

Fact Sheet No. 2.1.1-17

Method Title USGS Field Operation Plan: Tributary Monitoring, in LMMB 017

Purpose This method describes the collection of water samples and field data from streams.

Method Summary At each proposed sampling station, a cross-section of the stream is measured, and

the data will be used to subdivide the cross-section into three equal flow areas. The

centroid of each of these areas is identified on a field map. At each centroid, water

samples and Hydrolab parameters (i.e., temperature, conductivity, dissolved

oxygen, pH) are collected at 0.2 and 0.8 times the depth. Samples are collected

during downstream flow, which is established for at least 1⁄2 hour prior to initiating

sample collection. Water samples from each of the 6 sampling locations are

composited. Water is collected for PCB, PAH, pesticide, and Atrazine analyses

using a submersible pump and passed through a 293 mm, stainless steel,

pentaplate filter holder. 2 - 5 glass fiber filters are used, depending on the

concentration of suspended material in the water column. Filters will be folded and

placed in aluminum foil pouches. The filtered samples will be stored in carboys until

analysis. Water for DOC, POC, and conventional constituents is also collected

using the pump. Secchi disk measurem ents are made at each centroid location for

each cross-section. Velocity and flow direction are recorded at each subsampling

location.

Several ASTM Methods deal with the measurement of open channel flow. These

methods include D1941, D3858, D4409, D5089, D5129, and D5130 (ASTM,

2001a).

Data

Uses/Application

Situations where samples are required for dissolved and organic contaminants.

Advantages This method provides a standardized approach for obtaining representative stream

samples. 

Limitations This collection method is limited to organic contaminants. Not applicable for trace

metal contaminants. Not applicable for chlorophyll, dissolved/particulate nutrients,

or TSS.

Reference USEPA. 1997b . Lake Michigan Mass Balance Study Methods Compendium,

Volume 1: Sample Collection Techniques, EPA 905-R-97-012c. Great Lakes

National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/metho

ds/field96.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

33

Fact Sheet No. 2.1.1-18

Method Title Quality Assurance Plan for Discharge Measurements Using Broadband Acoustic

Doppler Current Profilers

Purpose To measure velocities and discharge in the riverine and estuarine environm ent.

Method Summary The Acoustic Doppler Current Profiler (ADCP) is an electronic instrument that is

used to measure water velocities. The instrument functions by transmitting acoustic

signals into the water column. The velocity of particles in the water column, and

therefore the velocity of the water, is calculated by comparing the frequency of the

transmitted signals compared to the frequency of backscatter signals reflected off

the particles. The instrument can be mounted to the side of a boat and towed

through the water column. The ADCP measures the velocity of the water column

relative to the movement of the vessel to which it is attached. Multiple transects of

data are averaged to reduce variation due to turbulence and velocity surges. At

least 4 transects should be made at each site to ensure a valid determination of

discharge.

Additional information regarding the use of ADCPs is found in Dredging Research

Technical Note DRP-1-16 (U.S. Army Engineer Waterways Experiment Station

1994).

Data

Uses/Application

The ADCP can be used to measure water velocities and discharge in a variety of

aquatic environments.

Advantages Measurement time is reduced; data can be collected throughout the water column

and cross section; stationing devices are not necessary.

Limitations The ADCP and its associated software are complex systems that should be used

only be highly trained personnel. High initial cost is also a major disadvantage. This

instrumentation is also unable to function in shallow water. The ADCP can

accurately measure discharge for only a limited range of flow conditions.

Reference Lipscomb, SW. 1995. Quality Assurance Plan for Discharge Measurements Using

Broadband Acoustic Doppler Current Profilers. USGS Open-File Report 95-701.

Website http://il.water.usgs.gov/adcp/reports/OFR9

5-701.htm l

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

34

Fact Sheet No. 2.1.1-19

Method Title Seepage Meters

Purpose To determine the exchange of ground water to surface water and vice versa.

Method Summary The Krupaseep is a prototype seepage meter device constructed of translucent

polycarbonate plastic domes of varying size. Each dome has a vertical skirt to

anchor it into deep organic muds. A port in the top of each dome allows flowthrough of water. Flow meters are installed in this port, and water quality meters are

installed on the inside and outside of the dome. When in use, the domes are

pressed into the river bottom until the top of each dome is 14 inches above the

mudline. The monitoring equipment on each dome is tethered back to computers

onshore that record real-time water quality data, including photosynthesis-activated

radiation, on the inside and outside of the seepage meter and record the inflow or

outflow (flux) via heat pulse technology. Riverside solar panels charge the batteries

that power the flow meter computer and data logger. Water quality samples can

also be collected using hoses mounted on both the internal and external surfaces of

the dome and a onshore peristaltic pump.

Data

Uses/Application

A knowledge of the groundwater flux is needed to evaluate the significance of

contaminant flux into the water column from capped contaminated sediments.

Advantages This unit allows for the collection, both continuous and discrete, of water quality

parameters both inside and outside of the seepage meter. Measures of flux and

water quality can be made remotely.

Limitations The location of the meter is restricted to a radial operating distance of 160 feet from

the computers. Divers are required for installation and service of the units. Weekly

maintenance is required under harsh environmental conditions. 

Reference USGS. 2001. The Krupaseep. Next Generation Seepage Meter.

Website http://sofia.usgs.gov/sfrsf/entdisplays/krupa

seep/

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

35

Fact Sheet No. 2.1.1-20

Method Title Caged Bivalve Deployment, AED LOP 1.02.002, Revision 1

Purpose Deployment and retrieval of caged bivalves for environmental monitoring.

Method Summary Four mussel baskets are deployed along a trawl (similar to long-line). Each trawl

has a surface floatation (lobster buoy) and an anchor (cinder block) at either end.

The mussel baskets are attached by leaders to cinder blocks fixed along the trawl

and are suspended approximately one meter above the bottom with sub-surface

floatation. Four replicate mussel baskets placed 50' apart are adequate to

characterize an area for chemical bioaccumulation, though this number dependent

upon localized conditions. The desired length of the trawl line is 170' plus 2X the

high water depth at the deployment location.. 

Each basket is fabricated from a 18" x 12" rectangle of black polyethylene 1⁄2" mesh

netting rolled into a cylinder, and secured with cable ties. The ends of the cylinder

are also closed with cable ties. Each basket contains 25 blue mussels, 5-7 cm in

length. The baskets are prepare in the laboratory and the baskets and mussels are

transported to the field site in coolers.

In the field, the mussel baskets are attached to the trawl leaders with cable ties. The

trawl is deployed sequentially beginning with one surface buoy followed by each of

the cinder blocks and finally the second surface buoy. Retrieval is the reverse of

deployment, beginning with the “down wind” buoy. Mussel baskets should be

placed in coolers as they are removed from the line. 

Data

Uses/Application

Caged bivalve deployments are used to obtain tissue for chemical bioaccumulation

studies, frequently as part of environmental monitoring projects. 

Advantages Relatively low cost and simple method with potential for revealing different

biomarkers of toxicity, identifying the chemicals responsible for the effects

measured, delineating affected areas, and specifying risks to aquatic fauna of

environmental contamination.

Limitations Permits may be required from local harbor master and/or appropriate regulatory

agencies. Other users of waterway, such as boat captains ans comm ercial lobster

men, must be considered. Weather may inhibit or prohibit deployment and retrieval.

This procedure was written to meet the specific needs of the research program at

the U.S. EPA-Atlantic Ecology Division. It is not a U.S. EPA Standard Method and

must not be referred to as such. Mention of trade names or comm ercial products

does not constitute endorsement or recommendation for use.

Reference Unpublished laboratory SOP, EPA NHEERL-AED, Narragansett, RI

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

36

2.1.2 Chemical and Physical Analyses

Section 2.1.2 contains methods for sample preparation and chemical and physical analysis of

water. These methods characterize the chemical composition and physical properties of water

samples collected by methods described in Section 2.1.1. Samples are often analyzed for the

presence of various inorganic and organic contaminants that may pose a threat to human or

ecological health. Many of the methods described have been developed over time to optimize the

detection, identification, and quantification of potential chemicals of concern. Several are

performance-based and may be further modified to enhance the accuracy and precision of the

method.

A variety of methods may exist for the analysis of a particular chemical parameter, all with varying

levels of quantification or degrees of sensitivity. Less sensitive methods may be used as a

screening tool during the initial site assessment to identify potential chemicals of concern. Followup analysis may include the use of very precise methods that provide unequivocal identification

and trace level quantification of analytes. This variety also provides alternative methods useful in

the analysis of many types of water samples. Interferences from certain compounds in a water

sample may be avoided by the use of an alternative method.

Other than describing the water column itself, the physical properties of water often influence the

behavior of contaminants in the water column, and they may be helpful in further understanding

the fate of contaminants in the environment. Physical parameters may change the solubility and

chemical form of various chemical components in water. 

Many of the chemical and physical methods described in these fact sheets are routinely

performed and fairly standardized. As a result, more than one source of information is often cited

in each method description. Specifically, the following sources provided methods information for

section 2.1.2:

• The USEPA’s Office of Water

• The USEPA’s Lake Michigan Mass Balance Study Methods Compendium, 1997v The

USEPA’s Test Methods for Evaluating Solid Waste, Physical/Chemical Methods

(SW846 Methods)

• NOAA’s National Status and Trends Program, 1998

• Standard Methods for Examination of Water and Wastewater, 1999

• ASTM
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

37

Fact Sheet No. 2.1.2-1

Method Title Mercury in Water by Cold Vapor Atomic Fluorescence Spectrometry, EPA Method

245.7

Purpose To determine the concentration of mercury (Hg) in filtered and unfiltered

water by cold-vapor atomic fluorescence spectrometry (CVAFS).

Method Summary A 100- to 2000-mL sample is collected directly into a fluoropolymer

bottle using sample handling techniques specially designed for collection of mercury

at trace levels. For dissolved Hg, the sample is filtered through a 0.45-:m capsule

filter. The sample is preserved by adding 5 mL/L of pretested 12N HCl. Inorganic

Hg compounds and organic mercury species are oxidized by a potassium

bromate/potassium bromide reagent. After oxidation, the sample is sequentially

prereduced with NH2OHAHCl to destroy the excess bromine, then the ionic Hg is

reduced with SnCl2to convert Hg(II) to volatile Hg(0). The Hg(0) is separated from

solution by purging with high purity argon gas through a semipermeable dryer tube.

The Hg passes into an inert gas stream that carries the released Hg(0) into the cell

of a cold-vapor atomic fluorescence spectrometer (CVAFS) for detection. The

concentration of Hg is determined by atomic fluorescence spectrometry at 253.7 nm.

The method detection limit (MDL) and minimum level of quantization (ML) in this

method are 1.8 ng/L and 5.0 ng/L, respectively. This method may be used to

determine Hg up to 200 ng/L and may be extended by dilution of

the sample. The normal calibration range for ambient water monitoring is 5 ng/L to

100 ng/L.

A similar method for the detection of total mercury is ASTM Method D3223 (ASTM,

2001a).

Data

Uses/Application

This method is applicable to drinking water, surface and ground waters, marine

water, and industrial and municipal wastewater.

Advantages Wide analytical range makes method suitable for contaminated sites.

Limitations Ambient mercury levels frequently are below the detection limit provided by this

method.

Reference USEPA. 2001a. Method 245.7: Mercury in Water by Cold Vapor Atomic

Fluorescence Spectrometry, Draft, EPA 821-R-01-008. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

38

Fact Sheet No. 2.1.2-2

Method Title Mercury in Water by Oxidation, Purge and Trap, and Cold Vapor Atomic

Fluorescence Spectrometry, EPA Method 1631, Revision B

Purpose This method is for determination of mercury (Hg) in filtered and

unfiltered water by oxidation, purge and trap, desorption, and cold-vapor atomic

fluorescence spectrometry (CVAFS).

Method Summary A 100- to 2000-mL sample is collected directly into a fluoropolymer bottle. For

dissolved Hg, the sample is filtered through a 0.45-:m capsule filter. The sample is

preserved by adding either HCl or BrCl solution. If a sample will also be used for the

determination of methyl mercury, it should be preserved with HCl solution only. Prior

to analysis, a 100-mL sample aliquot is placed in a specially designed purge vessel,

and 0.2N BrCl solution is added to oxidize all Hg compounds to Hg(II). After

oxidation, the sample is sequentially prereduced with NH2OHAHCl to destroy the free

halogens, then reduced with SnCl2

to convert Hg(II) to volatile Hg(0). The Hg(0) is

separated from solution by purging with nitrogen onto a gold-coated sand trap. The

trapped Hg is thermally desorbed from the gold trap and carried into the cell of a

cold-vapor atomic fluorescence spectrometer (CVAFS) for detection.

This method is for determination of Hg in the range of 0.5–100 ng/L and may be

extended to higher levels by selection of a smaller sample size. The method

detection limit for Hg has been determined to be 0.2 ng/L when no interferences are

present. The minimum level of quantization (ML) has been established as 0.5 ng/L.

An MDL as low as 0.05 ng/L can be achieved for low Hg samples by using a larger

sample volume, a lower BrCl level (0.2%), and extra caution in sample handling.

For the analysis of water samples using Methods LMMB 048 and 049 (USEPA,

1997d), subsamples are oxidized with BrCl solution and heated for at least an hour

(preferably overnight) at 70°C before prereduction and analysis. Aliquots of 125-

500 mL are purged and trapped. For particulate samples, the filter is treated in a

similar fashion as the water samples, except that 2 mL of hydroxylamine HCl is used

to produce samples. Method LMMB 048 has a mean detection limit of

approximately 0.1 ng/L.

Data

Uses/Application

This method is for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery Act,

the Com prehensive Environmental Response, Compensation and Liability Act, and

the Safe Drinking Water Act.

Advantages The dual amalgam trap system and fluorescence detector provide greater sensitivity

and specificity in the presence of interferences, and this system must be used to

overcome interferences, if present, and to achieve the sensitivity required, if

necessary. The detection range of this method generally provides detection of

mercury in ambient surface waters.

Limitations This method does not distinguish between methyl mercury and inorganic mercury

species.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

39

Fact Sheet No. 2.1.2-2 (contd.)

Reference USEPA. 1999b. Method 1631, Revision B: Mercury in Water by Oxidation, Purge

and Trap, and Cold Vapor Atomic Fluorescence Spectrometry, EPA 821-R-99-005.

Office of Water, U.S. Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitleOW .

html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

40

Fact Sheet No. 2.1.2-3

Method Title Methyl Mercury in Water by Distillation, Aqueous Ethylation, Purge and Trap,

and CVAFS, EPA Method 1630

Purpose This method is designed for determination of CH3Hg in the range of 0.02-5 ng/L

and may be extended to higher levels by selection of a smaller sample size.

Method Summary A 100-2000 mL sample is collected directly into fluoropolymer or borosilicate

bottle(s). For dissolved CH3Hg, samples are filtered through a 0.45-:m capsule

filter. Fresh water samples are preserved by adding 11.6 M HCl, while saline

samples ([Cl - ] > 500 ppm) are preserved with 9 M H2SO4

. Prior to analysis, a

45-mL sample aliquot is placed in a specially designed fluoropolymer distillation

vessel, and 35 mL of the water is distilled into the receiving vessel at 125° C

under N2flow. After distillation, the sample is adjusted to pH 4.9 and ethylated

in a closed purge vessel. The ethyl analog of CH3Hg, methylethyl mercury, is

separated from solution by purging with N2 onto a graphitic carbon (Carbotrap ®

) trap. The trapped methylethyl mercury is thermally desorbed from the trap,

carried through a pyrolytic decomposition column, which converts organo

mercury forms to elemental mercury (Hg 0), and then into the cell of a coldvapor atomic fluorescence spectrometer (CVAFS) for detection.

The method detection limit for CH3Hg has been determined to be 0.02 ng/L

when no background elements or interferences are present. The minimum level

(ML) has been established as 0.06 ng/L. An MDL as low as 0.009 ng/L can be

achieved for low CH3Hg samples by using extra caution in sample handling and

reagent selection, particularly the use of “for ultra-low level

only” distillation equipment.

Data Uses/Application This method is for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery

Act, the Com prehensive Environmental Response, Compensation and Liability

Act, and the Safe Drinking Water Act.

Advantages Methyl mercury is frequently required for risk assessments. This method

provides the sensitivity to determine ambient levels in most aqueous samples.

Limitations Samples may become contaminated by numerous routes. Potential sources of

trace metals contamination include: metallic or metal-containing labware (e.g.,

talc gloves that contain high levels of zinc), containers, sampling equipment,

reagents, and reagent water; improperly cleaned or stored equipment, labware,

and reagents; and atm ospheric inputs such as dirt and dust. Even human

contact can be a source of trace metals contamination.

Reference USEPA. 2001b. Method 1630: Methyl Mercury in Water by Distillation,

Aqueous Ethylation, Purge and Trap, and CVAFS, EPA 821-R-01-020. Office

of Water, U.S. Environmental Protection Agency, Washington, DC.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

41

Fact Sheet No. 2.1.2-4

Method Title Determination of Trace Elem ents in Ambient Waters by Stabilized

Tem perature Graphite Furnace Atomic Absorption, EPA Method 1639

Purpose This procedure is for the determination of dissolved elements (antim ony, cadmium,

nickel, selenium, trivalent chromium, zinc) in ambient waters by Stabilized

Tem perature Graphite Furnace Atomic Absorption (GFAA). It may also be used for

determination of total recoverable element concentrations in these waters.

Method Summary For total recoverable analysis of an aqueous sample containing undissolved material,

analytes are first solubilized by gentle refluxing with nitric and hydrochloric acids.

After cooling, the sample is made up to volume, mixed, and centrifuged or allowed to

settle overnight prior to analysis. To determine dissolved analytes in a filtered

aqueous sample aliquot, the sample is prepared for analysis by the appropriate

addition of nitric acid, and then diluted to a predetermined volume and mixed before

analysis. The analytes listed in this method are determined by stabilized temperature

platform graphite furnace atomic absorption (STPGFAA).

Metal MDL (μg/L) ML (μg/L)

Antimony 1.9 5

Cadmium 0.023 0.05

Chromium (III) 0.1 0.2

Nickel 0.65 2

Selenium 0.83 2

Zinc 0.14 0.5

ASTM Method D1687 describes a similar (and alternative) method for the

measurement of hexavalent and total chromium in water (ASTM, 2001a). ASTM

Methods D3557 and D3859 describe the analysis of cadmium and selenium by

GFAA, respectively. ASTM Method D3919 and SW846 Method 7000A describe the

analysis of several elements by atomic absorption methods (ASTM, 2001a; USEPA

SW846.

Data

Uses/Application

This method is for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery Act,

the Com prehensive Environmental Response, Compensation and Liability Act, and

the Safe Drinking Water Act.

Advantages GFAA techniques in addition to ICP-MS methods are frequently required to obtain all

trace metal analytes of interest.

Limitations Samples may become contaminated by numerous routes. This method should be

used by analysts experienced in the use of graphite furnace atomic absorption

spectroscopy.

Reference USEPA. 1996b. Method 1639: Determination of Trace Elem ents in Ambient Waters

by Stabilized Tem perature Graphite Furnace Atomic Absorption, EPA 821-R-96-006.

Office of Water, U.S. Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitleO

W.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

42

Fact Sheet No. 2.1.2-5

Method Title Determination of Trace Elem ents in Ambient Waters by Off-Line Chelation Preconcentration and Stabilized Tem perature Graphite Furnace Atomic Absorption,

EPA Method 1637

Purpose This procedure is for the determination of dissolved elements in ambient

waters, namely cadmium and lead. It may also be used for determination of

total recoverable element concentrations in these waters.

Method Summary For total recoverable analysis of an aqueous sample containing undissolved

material, analytes are first solubilized with nitric acid. After cooling, the sample is

made up to volume, mixed, and centrifuged or allowed to settle overnight before

analysis. For the determination of dissolved analytes in a filtered aqueous

sample aliquot, the sample is made ready for analysis by the appropriate

addition of nitric acid, and then diluted to a predetermined volume and mixed

before analysis.

This method is used to preconcentrate trace elements using an iminodiacetate

functionalized chelating resin. After a sample is prepared, it is buffered using an

on line system before it enters the chelating column. Group I and II metals, as

well as most anions, are selectively separated from the analytes by elution with

amm onium acetate at pH 5.5. The analytes are subsequently eluded into a

simplified matrix consisting of 0.75 M nitric acid. The eluded sample is collected

and then analyzed by stabilized temperature platform graphite furnace atomic

absorption (STPGFAA).

The method detection limits for Cd and Pb have been determined to be 0.0075

μg/L and 0.036 μg/L, respectively. The minimum levels (ML) have been

established as 0.02 and 0.1 μg/L, respectively.

Similar methods include ASTM Methods D3557 for cadmium and D3559 for

lead (ASTM, 2001a).

Data Uses/Application This method is for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery

Act, the Com prehensive Environmental Response, Compensation and Liability

Act, and the Safe Drinking Water Act.

Advantages Off line Pre-concentration methods eliminate many sample handling

procedures, reducing sources of contamination. GFAA analysis affords

extremely low detection limits.

Limitations Due to its sensitivity, interferences can occur with GFAA analysis. This method

should be used by analysts experienced in the use of graphite furnace atomic

absorption spectroscopy. 

Reference USEPA. 1996c. Method 1637: Determination of Trace Elem ents in Ambient

Waters by Off-Line Chelation Pre-concentration and Stabilized Tem perature

Graphite Furnace Atomic Absorption, EPA 821-R-96-004. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubti

tleOW.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

43

Fact Sheet No. 2.1.2-6

Method Title Determination of Trace Elem ents in Ambient Waters by Inductively

Coupled Plasma — Mass Spectrometry, EPA Method 1638

Purpose This procedure is for the determination of dissolved elements (antim ony,

cadmium, copper, lead, nickel, selenium, silver, thallium, zinc) in ambient

waters. It may also be used for determination of total recoverable element

concentrations in these waters.

Method Summary For total recoverable analysis of an aqueous sample containing undissolved

material, analytes are first solubilized with nitric and hydrochloric acids. After

cooling, the sample is made to volume, mixed, and centrifuged or allowed to

settle overnight prior to analysis. For the determination of dissolved analytes in a

filtered aqueous sample aliquot, the sample is prepared for analysis by the

appropriate addition of nitric acid, and then diluted to a predetermined volume

and mixed before analysis. The digested sample is introduced into a radio

frequency plasma, where energy transfer processes cause desolvation,

atomization, and ionization. The ions are extracted from the plasma through a

differentially pumped vacuum interface and separated on the basis of their

mass-to- charge ratio (m/z) by a mass spectrometer. Ions transmitted through

the mass analyzer are detected by an electron multiplier or Faraday detector

and the resulting current is processed by a data handling system.

 1638 1638 LMMB 057

Metal MDL (μg/L) ML (μg/L) ML (ng/L)

Aluminum 25/15

Antimony 0.0097 0.02 

Arsenic 15/10

Cadmium 0.025 2.5/0.5

Chromium 20/8

Copper 0.087 0.2 8/4

Lead 0.015 0.05 3/0.5

Nickel 0.33 1

Selenium 0.45 1

Silver 0.029 0.1 1.5/0.3

Thallium 0.0079 0.02

Zinc 0.14 0.5 10/2.5

ICP-MS detection limits listed for LMMB 057 are for pneumatic and ultrasonic

nebulization, respectively (USEPA, 1997c).

Standard Method 3120B, ASTM Method D5673, and SW846 Method 6020

describe the analysis of metals by ICP-MS (APHA, 1999; ASTM, 2001a;

USEPA SW 846).

Data Uses/Application This method is for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery

Act, the Com prehensive Environmental Response, Compensation and Liability

Act, and the Safe Drinking Water Act.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

44

Fact Sheet No. 2.1.2-6 (contd.)

Advantages Analysis by ICP-MS provides a high level of sensitivity for some elements that

are difficult to determine by other methods. Up to 20 elements can be

determined from a single sample.

Limitations This method should be used by analysts experienced in the use of inductively

coupled plasma mass spectrometry (ICP-MS).

Reference USEPA. 1996d. Method 1638: Determination of Trace Elem ents in Ambient

Waters by Inductively Coupled Plasma-Mass Spectrometry, EPA

821-R-96-005. Office of Water, U.S. Environmental Protection Agency,

Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubti

tleOW.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

45

Fact Sheet No. 2.1.2-7

Method Title Determination of Trace Elem ents in Ambient Waters by On-Line Chelation

Pre-concentration and Inductively Coupled Plasma-Mass Spectrometry, EPA

Method 1640

Purpose This procedure is for the determination of dissolved elements (cadmium, copper,

lead, nickel) in ambient waters. It may also be used for determination of total

recoverable element concentrations in these waters.

Method Summary This method is used to preconcentrate trace elements using an iminodiacetate

functionalized chelating resin system that is connected directly to the ICP-MS.

Following acid solubilization, the sample is buffered prior to the chelating column

using an on line system. Group I and II metals, as well as most anions, are

selectively separated from the analytes by elution with amm onium acetate at pH 5.5.

The analytes are subsequently eluded into a simplified matrix consisting of dilute

nitric acid and are determined by ICP-MS using a directly coupled on line

configuration.

Metal MDL (μg/L) ML (μg/L)

Cadmium 0.0024 0.01

Copper 0.024 0.1

Lead 0.0081 0.02

Nickel 0.029 0.1

Data

Uses/Application

This method is for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery Act,

the Com prehensive Environmental Response, Compensation and Liability Act, and

the Safe Drinking Water Act.

Advantages The on line Pre-concentration system allows for reduced sampling handling,

minimizing the risk of sample contamination. Method 1640 is a convenient method

for the detection of a short list of toxic metals.

Limitations Neither mercury nor arsenic can be measured with this method.

Reference USEPA. 1996e. Method 1640: Determination of Trace Elem ents in Ambient

Waters by On-Line Chelation Pre-concentration and Inductively Coupled

Plasma-Mass Spectrometry, EPA 821-R-96-007. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitleOW

.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

46

Fact Sheet No. 2.1.2-8

Method Title Inorganic Arsenic in Water by Hydride Generation Quartz Furnace Atomic

Absorption, EPA Method 1632

Purpose This method is for determination of total inorganic arsenic (As) in filtered and

unfiltered water by hydride generation and quartz furnace atomic absorption

detection. This method is designed for measurement of dissolved and total arsenic

in the range of 10-200 ng/L.

Method Summary A 100-2000 mL sample is collected directly into a sample bottle. The sample is

either field or laboratory preserved by the addition of 10% HNO3

, depending on the

time between sample collection and arrival at the laboratory. An aliquot of sample is

placed in a specially designed reaction vessel and 6 M HCl is added. Before

analysis, 4% NaBH4solution is added to convert organic and inorganic arsenic to

volatile arsines. The arsines are purged from the sample onto a cooled glass trap

packed with 15% OV-3 on Chrom asorb ® WAW-DMCS0, or equivalent. The

trapped arsines are thermally desorbed, in order of increasing boiling points, and

carried into the quartz furnace of an atomic absorption spectrophotometer for

detection.

The first arsine to be desorbed will be AsH3

, which represents total inorganic

arsenic in the sample.

The method detection limit for total inorganic arsenic has been determined to be 3

ng/L when no background elements or interferences are present. The minimum

level (ML) has been established at 10 ng/L.

ASTM Method D2972B, Standard Method 3114B, and SW846 Method 7061A all

describe similar methods for the hydride generation atomic absorption detection of

arsenic in water (ASTM, 2001a; APHA, 1999; USEPA SW 846)

Data

Uses/Application

The method is for use in EPA's data gathering and monitoring programs associated

with the Clean Water Act.

Advantages Automation of this method reduces error and potential sources of contamination.

Limitations This method does not provide data on arsenic speciation, which is sometimes

required in risk assessments.

Reference USEPA. 1996f. Method 1632: Inorganic Arsenic in Water by Hydride Generation

Quartz Furnace Atomic Absorption, EPA 821-R-96-013. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitleO

W.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

47

Fact Sheet No. 2.1.2-9

Method Title Chemical Speciation of Arsenic in Water and Tissue by Hydride Generation Quartz

Furnace Atomic Absorption Spectrometry, EPA Method 1632, Revision A

Purpose This method is for determination of inorganic arsenic (IA), arsenite (As +3 ),

arsenate (As +5 ), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA)

in filtered and unfiltered water by hydride generation and quartz furnace atomic

absorption detection. This method is designed for measurement of As species in

water in the range 0.01-50 μg/L.

Method Summary Aqueous sample—A 500- to 1000-mL water sample is collected directly into a

cleaned sample bottle. Water samples are preserved in the field by the addition of

6M HCl. The recommended holding time is 28 days.

An aliquot of water sample is placed in a specially designed reaction vessel, and 6M

HCl is added. NaBH4solution is added to convert IA, MMA, and DMA to volatile

arsines. Arsines are purged from the sample onto a cooled glass trap packed with

15% OV-3 on Chrom osorb ® W AW-DMCS, or equivalent. The trapped arsines are

thermally desorbed, in order of increasing boiling points and carried into the quartz

furnace of an atomic absorption spectrophotometer for detection. To determine the

concentration of As +3, another aliquot of water sample or tissue digestate is placed

in the reaction vessel and Tris-buffer is added. The procedure is repeated to quantify

only the arsine produced from As +3. The concentration of As +5 is the

concentration of As +3 subtracted from the concentration of IA.

Analyte MDL ML 

IA (As +3 +As +5 ) 0.003 μg/L 0.01 μg/L 

Arsenite (As +3 ) 0.003 μg/L 0.01 μg/L 

MMA 0.004 μg/L 0.01 μg/L 

DMA 0.02 μg/L 0.05 μg/L 

Data

Uses/Application

The method is for use in EPA's data gathering and monitoring programs associated

with the Clean Water Act.

Advantages The relative amounts of carcinogenic arsenite (As +3) to total arsenic varies with

surface water body and varies with pH. This method directly quantifies arsenite.

Limitations Depending upon As levels at site, speciation may not be necessary.

Reference USEPA. 2001c. Method 1632, Revision A: Chemical Speciation of Arsenic in

Water and Tissue by Hydride Generation Quartz Furnace Atomic Absorption

Spectrometry, EPA 821-R-01-006. Office of Water, U.S. Environmental Protection

Agency, Washington, DC.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

48

Fact Sheet No. 2.1.2-10

Method Title Determination of Hexavalent Chromium by Ion Chromatography, EPA Method 1636

Purpose This method is for the determination of dissolved hexavalent chromium (as CrO4

2- )

in ambient waters at EPA water quality criteria (WQC) levels using ion

chromatography (IC).

Method Summary An aqueous sample is filtered through a 0.45 :m filter, and the filtrate is adjusted to

a pH of 9-9.5 with a concentrated buffer solution. A measured volume of the sample

(50-250 :L) is introduced into the ion chromatography. A guard column removes

organics from the sample before the Cr(VI), as CrO42-, is separated on a high

capacity anion exchange separator column. Post column derivatization of the Cr(VI)

with diphenylcarbazide is followed by detection of the colored complex at 530 nm.

The method detection limit (MDL), and the minimum level (ML) for

hexavalent chromium (Cr(VI)) are 0.23 μg/L and 0.5 μg/L, respectively.

ASTM Method D5257 and SW846 Method 7199 also describe the analysis of

hexavalent chromium by Ion Chromatography (ASTM, 2001a; USEPA SW 846).

Data

Uses/Application

The method is for use in EPA's data gathering and monitoring programs associated

with the Clean Water Act.

Advantages This method provides the rapid and reproducible isolation and analysis of Cr (VI)

without interference from other Cr species.

Limitations Samples containing high levels of anionic species, such as sulfate and chloride, may

cause column overload. Samples containing high levels of organics or sulfides

cause rapid reduction of soluble Cr(VI) to Cr(III). Samples must be stored at 4°C

and analyzed within 24 hours of collection unless preserved with sodium hydroxide.

Reference USEPA. 1996g. Method 1636: Determination of Hexavalent Chromium by Ion

Chromatography, EPA 821-R-95-029. Office of Water, U.S. Environmental

Protection Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtm l/pubtitle

OW.html

Last Accessed: 2/12/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

49

Fact Sheet No. 2.1.2-11

Method Title Volatile Organic Compounds by Isotope Dilution GC/MS, EPA Method 1624b

Purpose This method is designed to determine the volatile toxic organic pollutants.

Method Summary Stable isotopically labeled analogs of the compounds of interest are added to a 5 mL

water sample. The sample is purged at 20-25/C with an inert gas in a specially

designed chamber. The volatile organic compounds are transferred from the

aqueous phase into the gaseous phase where they are passed into a sorbent

column and trapped. After purging is completed, the trap is back flushed and heated

rapidly to desorb the compounds into a gas chromatography (GC). The compounds

are separated by the GC and detected by a mass spectrometer (MS). The labeled

compounds serve to correct the variability of the analytical technique. Identification

of a compound (qualitative analysis) is performed by comparing the GC retention

time and the background corrected characteristic spectral masses with those of

authentic standards. Quantitative analysis is performed by GC/MS using extracted

ion current profile (EICP) areas. Isotope dilution is used when labeled compounds

are available; otherwise, an internal standard method is used. The Minimum Level

for most VOC compounds is either 10 or 50 μg/L.

Similar methods for the determination of volatile organic compounds and volatile

aromatic organic compounds are presented in Standard Methods 6210 and 6220,

respectively (APHA, 1999). SW846 Methods 5030B, 5035, and 8260B and ASTM

Method D5790 describe the preparation and analysis of volatile organics by GC/MS

(USEPA SW 846); ASTM, 2001a).

Data

Uses/Application

The method is designed to meet the survey requirements of Effluent Guidelines

Division (EGD) and the National Pollutants Discharge Elimination System (NPDES)

under 40 CFR Parts 136.1 and 136.5. VOC in ambient waters is sometimes

monitored at sites of suspected contaminated groundwater inflow.

Advantages The combination of GC retention time and MS characterization provides

unequivocal identification of analytes.

Limitations The GC/MS portions of this method are for use only by analysts experienced with

GC/MS or under the close supervision of such qualified persons. Samples can be

contaminated by diffusion of volatile organic compounds (particularly methylene

chloride) through the bottle seal during shipment and storage. Contamination by

carry-over can occur.

Reference USEPA. 1989a. Method 1624, Revision B: Volatile Organic Compounds by Isotope

Dilution GC/MS, EPA440-1-89-023. Office of Water, U.S. Environmental Protection

Agency, Washington, DC.

Website Rev B not available online Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

50

Fact Sheet No. 2.1.2-12

Method Title Photovac GC Analysis for Soil, Water, and Air/Soil Gas, ERT SOP# 2109

Purpose This method is designed as a field screening procedure for the tentative

identification of various volatile organic compounds.

Method Summary Water samples are collected in triplicate in 40-mL VOA vials with TeflonTM-lined

silicone septum screw caps. The vials are filled completely, with no visible air

bubbles. A 20-mL aliquot of sample from one of the three sample triplicates is

pipetted into a second, clean VOA vial. The vial is capped, shaken vigorously

for one minute, and allowed to stand at room temperature for at least 30

minutes for vapor phase equilibration. An aliquot of the water head space is

then removed from the vial and injected into the GC using a gas-tight syringe.

The GC uses an ultraviolet light source and photoionization detector. The other

two vials are analyzed within seven days by another method to confirm the field

screening data.

Typical MDLs for this method range from 1 ppb to 5 ppb.

ERT SOPs # 2108 and #2107 describe the operation of specific models of

Photovac Gas Chromatographs.

Data Uses/Application Site assessment/characterization and health and safety surveys.

Advantages The data generated with this method allows for rapid evaluation of site

conditions.

Limitations Pollutant identification is only tentative.

Reference USEPA. 1994b. SOP # 2109: Photovac GC Analysis for Soil, Water, and

Air/Soil Gas. Environmental Response Team. Compendium of ERT Standard

Operating Protocols. Office of Solid Waste and Emergency Response, U.S.

Environmental Protection Agency, Edison NJ.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

51

Fact Sheet No. 2.1.2-13

Method Title Semi-volatile Organic Compounds by Isotope Dilution GC/MS, EPA Method 1625

Revision B

Purpose This method is used to determine the semi-volatile toxic organic pollutants (i.e.,

PAHs) in water.

Method Summary This method is performance-based. Stable isotopically labeled analogs of the

compounds of interest are added to a one liter water sample. The sample is

extracted at pH 12-13, then at pH <2 with methylene chloride using continuous

extraction techniques. The extract is dried over sodium sulfate and concentrated to

a volume of 1 mL. An internal standard is added to the extract, and the extract is

injected into the gas chromatography (GC). The compounds are separated by GC

and detected by a mass spectrometer (MS). The labeled compounds serve to

correct the variability of the analytical technique. Identification of a compound

(qualitative analysis) is performed by comparing the GC retention time and

background corrected characteristic spectral masses with those of authentic

standards. Quantitative analysis is performed by GC/MS using extracted ion current

profile (EICP) areas. Isotope dilution is used when labeled compounds are available;

otherwise, an internal standard method is used.

The method detection limit for most compounds of interest is 10 ug/L. The MDL for

some compounds is 20 or 50 ug/L.

Similar methods for the extraction and analysis of semi-volatile organic compounds

are Standard Method 6410B and SW846 Method 8270C (APHA, 1999).

Data

Uses/Application

The method is designed to meet the survey requirements of Effluent Guidelines

Division (EGD) and the National Pollutants Discharge Elimination System (NPDES)

under 40 CFR Part 136.1.

Advantages Mass spectral analysis, combined with gas chromatographic compound retention

time, provides unequivocal compound identification. Isotope dilution corrects

recovery and performance of each compound of interest.

Limitations The GC/MS portions of this method are for use only by analysts experienced with

GC/MS or under the close supervision of such qualified persons. The suites of

stable isotopes required for this analysis are often prohibitively expensive for use in

routine monitoring programs.

Reference USEPA. 1989b. Method 1625, Revision B: Semi-volatile Organic Compounds by

Isotope Dilution GC/MS, EPA440-1-89-023. Office of Water, U.S. Environmental

Protection Agency, Washington, DC.

Website Rev B not available online Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

52

Fact Sheet No. 2.1.2-14

Method Title Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS) - Selected Ion Monitoring (SIM)

Mode

Purpose To determine low concentrations of polycyclic aromatic hydrocarbons (PAHs) and

their alkylated homologues in extracts of water, sediments and biological tissues.

Method Summary Just prior to analysis, an aliquot of internal standard solution is added to the sample

extract producing a final internal standard concentration of approximately 40 ng/mL.

The analytical system includes a temperature programmable gas chromatography

with a DB-5MS fused silica capillary column. Helium is used as the carrier gas, and

the samples are handled by an auto sampler capable of making 1 - 4 μl injections.

A five point calibration curve is established to dem onstrate the linear range of the

detector. The effluent from the GC capillary column is routed directly into the ion

source of the mass spectrometer (MS). The MS is operated in the SIM mode using

appropriate windows to include the quantization and confirmation masses for target

PAHs. For all compounds detected at a concentration above the MDL, a

confirmation ion is checked to confirm its presence. The response factors of the

surrogate relative to each of the calibration standards are calculated, followed by the

calculation of the sample extract concentration. The sample concentration for each

compound is calculated by dividing the sample extract concentration by the sample

amount.

Data

Uses/Application

PAH concentrations, particularly pyrene, are one of the primary risk factors

associated with contaminated waters. PAH data obtained from this analysis are

used for site characterization and risk assessments.

Advantages GC/MS in the SIM mode provides unambiguous and sensitive detection for PAHs.

The PAH quantization method is very rigorous because PAHs have very strong

molecular ion peaks under the mass spectrometric conditions used. Also, the

availability of labeled surrogates internal standards of many of the analytes makes

very accurate determinations of analyte concentrations possible. Analysis of

alkylated PAH homologues can provide site-specific information that can be used in

source identification or product identification.

Limitations GC/MS in SIM mode cannot be used for simultaneous screening for other organic

contaminants of similar polarity or volatility; cannot be used to identify tentatively

identified compounds (TICs).

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and Trends

Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo NOS ORCA

130. National Oceanic and Atm ospheric Administration, Silver Spring, MD. 233 pp.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

53

Fact Sheet No. 2.1.2-15

Method Title Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

Purpose To quantify chlorinated hydrocarbons (i.e., chlorinated pesticides and PCBs) in

sample extracts.

Method Summary This method is based on high resolution, capillary gas chromatography using

electron capture detection (GC/ECD). Extracts normally have a holding time of 40

days. This method provides for initial, ongoing and final calibrations, which are done

as part of the analytical run. If the response for any peak exceeds the highest

calibration solution, the extract is diluted, a known amount of surrogate and TCMX

solution added, and the sample reanalyzed for those analytes that exceeded the

calibration range. Concentrations in the samples are calculated based on the

internal standard method. Data are reported as ng/g dry weight.

Other methods describing the analysis of PCBs and pesticides by GC/ECD are

NS&T methods, ASTM Methods D5317 and D3534, and SW846 Methods 8081A

and 8082 (NOAA, 1998; ASTM, 2001a).

Data

Uses/Application

PCBs and persistent pesticides (particularly DDT and metabolites) are two of the

primary risk factors of contaminated waters. Data are used in site characterization

and in risk analysis.

Advantages The ECD is very sensitive and allows for detection of the chlorinated hydrocarbons

at trace concentrations (ppb).

Limitations The detector does not have a linear response over a wide concentration

range and must be used by sufficiently trained personnel. Second column analysis

must be performed to provide unequivocal compound identification. These methods

do not measure the 12 World Health Organization congeners, which may be desired

data in some risk assessments.

Reference USEPA. 1997d. Lake Michigan Mass Balance Study Methods Compendium,

Volume 2: Organic and Mercury Sample Analysis Techniques. EPA 905-R-97-012c.

Great Lakes National Program Office, U.S. Environmental Protection Agency,

Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

sop-501.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

54

Fact Sheet No. 2.1.2-16

Method Title PCBs and Pesticides in Surface Water by XAD-2 Resin Extraction, LMMB 039

Purpose This method is used to determine congener specific PCB and pesticide

concentrations at trace levels in surface water.

Method Summary Water samples (80 - 160 L) are filtered for particulates and the dissolved PCBs are

collected using an XAD-2 resin column. Filters and resin columns are stored at 4°C

until analysis. The filters and resin are Soxhlet extracted using 50% acetone/50%

hexane for 16 hours. The water remaining in the samples is extracted with hexane.

The sample extracts are concentrated and dried with sodium sulfate. The samples

are run through a Florisil column and then through a silica column. Two fractions

are collected from the silica column. The first fraction is eluded with hexane and

contains the PCBs, HCB, and p,p’ DDE. The second fraction is eluded with 25%

ethyl ether in hexane and contains alpha-BHC, lindane, the chlordanes, nonachlors,

p,p’DDD, p,p’DDT, and toxaphene. These fractions are concentrated and further

cleaned with sulfuric acid. Sample extracts are analyzed by GC/ECD according to

Method LMMB 041. Confirmation of pesticides in the second fraction is performed

on a second column or by GC/MS. 

Preparation of XAD-2 Resin is presented in SW846 Method 0010: Appendix A

(USEPA SW 846).

Data

Uses/Application

Water quality assessments under Clean Water Act, RCRA, or CERCLA

Advantages Extraction of water through resin perm its the economical extraction of large volumes

(>20 L) with this method.

Limitations XAD-Z resin is difficult to prepare for trace PCB application.

Reference USEPA. 1997d. Method LMMB 039: PCBs and Pesticides in Surface Water by

XAD-2 Resin Extraction, Lake Michigan Mass Balance Study Methods

Compendium, Volume 2: Organic and Mercury Sample Analysis Techniques. EPA

905-R-97-012c. Great Lakes National Program Office, U.S. Environmental

Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

sec1293.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

55

Fact Sheet No. 2.1.2-17

Method Title Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA Method 1613

Purpose This method is for determination of tetra through octa-chlorinated dibenzo-p-dioxins

(CDDs) and dibenzo furans (CDFs) in water.

Method Summary This method is "performance-based.” Stable isotopically labeled analogs of 15 of

the 2,3,7,8-substituted CDDs/CDFs are spiked into a 1 L water sample, and the

sample is extracted by one of three procedures:

1. Samples containing no visible particles are extracted with methylene

chloride in a separatory funnel or by the solid-phase extraction technique. The

extract is concentrated for cleanup.

2. Samples containing visible particles are vacuum filtered through a glass-fiber

filter. The filter is extracted with toluene in a Soxhlet/Dean-Stark (SDS) extractor,

and the filtrate is extracted with methylene chloride in a separatory funnel. The

methylene chloride extract is concentrated and combined with the SDS extract prior

to cleanup.

3. The sample is vacuum filtered through a glass-fiber filter on top of a solid-phase

extraction (SPE) disk. The filter and disk are extracted in an SDS extractor, and the

extract is concentrated for cleanup. After extraction, 37Cl4-labeled 2,3,7,8-TCDD is

added to each extract to measure the efficiency of the cleanup process. Sample

cleanups may include back-extraction with acid and/or base, and gel permeation,

alumina, silica gel, Florisil and activated carbon chromatography. High-performance

liquid chromatography (HPLC) can be used for further isolation of the 2,3,7,8-

isomers or other specific isomers or congeners. After cleanup, the extract is

concentrated to near dryness. Immediately prior to injection, internal standards are

added to each extract, and an aliquot of the extract is injected into the gas

chromatography. The analytes are separated by the GC and detected by a highresolution ($10,000) mass spectrometer.

Minimum

CDD/CDF Level (pg/L) CDD/CDF ML(pg/L)

2,3,7,8-TCDF 10 1,2,3,4,7,8-HxCDD 50

2,3,7,8-TCDD 10 1,2,3,6,7,8-HxCDD 50

1,2,3,7,8-PeCDF 50 1,2,3,7,8,9-HxCDD 50

2,3,4,7,8-PeCDF 50 1,2,3,4,6,7,8-HpCDF 50

1,2,3,7,8-PeCDD 50 1,2,3,4,7,8,9-HpCDF 50

1,2,3,4,7,8-HxCDF 50 1,2,3,4,6,7,8-HpCDD 50

1,2,3,6,7,8-HxCDF 50 OCDF 100

1,2,3,7,8,9-HxCDF 50 OCDD 100

2,3,4,6,7,8-HxCDF 50

This method is also described in SW846 Method 8290 (USEPA SW846.

Data

Uses/Application

The method is for use in EPA's data gathering and monitoring programs associated

with the Clean Water Act, the Resource Conservation and Recovery Act, the

Com prehensive Environmental Response, Compensation and Liability Act, and the

Safe Drinking Water Act.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

56

Fact Sheet No. 2.1.2-17 (contd.)

Advantages Method 1613 is able to meet detection limits required for human health and

ecological risk assessments.

Limitations The GC/MS portions of this method are for use only by analysts experienced with

HRGC/HRMS or under the close supervision of such qualified persons.

Reference USEPA. 1994c. Method 1613: Tetra- through Octa-Chlorinated Dioxins and Furans

by Isotope Dilution HRGC/HRMS, EPA 821-B-94-005. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitleOW.h

tml

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

57

Fact Sheet No. 2.1.2-18

Method Title Toxic Polychlorinated Biphenyls by Isotope Dilution High Resolution

Gas Chromatography/High Resolution Mass Spectrometry, EPA Method 1668

Purpose This method is for determination of the toxic Polychlorinated Biphenyls (PCBs) in

water.

Method Summary This method is performance-based. Stable isotopically labeled analogs of the toxic

PCBs are spiked into a 1-L sample, and the sample is vacuum filtered through a

glass-fiber filter on top of a solid-phase extraction (SPE) disk. Sample com ponents

on the filter and disk are eluded with methylene chloride and the eluant is

concentrated for cleanup. After extraction, samples are cleaned up using backextraction with sulfuric acid and/or base, and gel permeation, silica gel, Florisil and

activated carbon chromatography. High-performance liquid chromatography (HPLC)

can be used for further isolation of specific isomers or congeners. After cleanup, the

extract is concentrated to near dryness. Immediately prior to injection, internal

standards are added to each extract, and an aliquot of the extract is injected into the

gas chromatography. The analytes are separated by the GC and detected by a highresolution ($10,000) mass spectrometer.

The Method Detection Limit (MDL) for PCB #126 has been determined as 40 pg/L in

water using this method.

 

IUPAC EMDL (pg/L) EML (pg/L)

77 5 20

123, 126 40 100

118/167/156/157/169/180/170/189 60 200

114 600 2000

105 400 1000

EMD: = Estimated Method Detection Limit; EML = Estimated Minimum Level

Data

Uses/Application

The method is for use in EPA's data gathering and monitoring programs associated

with the Clean Water Act, the Resource Conservation and Recovery Act, the

Com prehensive Environmental Response, Compensation and Liability Act, and the

Safe Drinking Water Act.

Advantages Method 1668 provides data for most, but not all, of the “dioxin-like” PCBs, including

those with the highest TEFs, as determined by the World Health Organization. This

method provides detection limits frequently required in risk assessments.

Limitations The GC/MS portions of this method are for use only by analysts experienced with

HRGC/HRMS or under the close supervision of such qualified persons. Method

1668 does not provide data for all of the “dioxin-like” PCBs, as does Method 1668A.

Reference USEPA. 1997e. Method 1668: Toxic Polychlorinated Biphenyls by Isotope Dilution

HRGC/HRMS, EPA-821-R-97-001. Office of Water, U.S. Environmental Protection

Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitleOW.h

tml

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

58

Fact Sheet No. 2.1.2-19

Method Title Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and Tissue by

HRGC/HRMS, EPA Method 1668 Revision A

Purpose This method is for congener-specific determination of more than 150 chlorinated

biphenyl (CB) congeners in water.

Method Summary This method is performance-based. Stable isotopically labeled analogs of the 12

PCBs designated as toxic by W HO and labeled congeners at each level of

chlorination are spiked into a 1-L sample. The sample is extracted using solid-phase

extraction, separatory funnel extraction, or continuous liquid/liquid extraction. After

extraction, a labeled cleanup standard is spiked into the extract which is then

cleaned up using back-extraction with sulfuric acid and/or base, and gel permeation,

silica gel, or Florisil chromatography. Activated carbon and high-performance liquid

chromatography (HPLC) can be used for further isolation of specific congener

groups. After cleanup, the extract is concentrated to 20 μL. Immediately prior to

injection, labeled injection internal standards are added to each extract and an

aliquot of the extract is injected into the gas chromatography (GC). The analytes are

separated by the GC and detected by a high-resolution ($10,000) mass

spectrometer.

The estimated method detection limit (EMDL) for congener 126 in water is 5 pg/L

with no interferences present. Without interferences, EMDLs and EMLs are,

respectively, 5 and 10 pg/L for aqueous samples, and EMLs for extracts are 0.5

pg/uL.

EMD: = Estimated Method Detection Limit; EML = Estimated Minimum Level

Data

Uses/Application

This Method is for use in data gathering and monitoring associated with the Clean

Water Act, the Resource Conservation and Recovery Act, the Com prehensive

Environmental Response, Compensation and Liability Act, and the Safe Drinking

Water Act.

Advantages Method 1668A provides congener data that can be used for source identification.

Listed PCBs include the 12 World Health Organization “dioxin-like” PCBs. The

HRMS method provides lower EMDLs compared to ECD or low resolution MS

analyses and provides unequivocal congener identification.

Limitations The GC/MS portions of this method are for use only by analysts experienced with

HRGC/HRMS or under the close supervision of such qualified persons. Solvents,

reagents, glassware, and other sample processing hardware may yield artifacts,

elevated baselines, and/or lock mass suppression causing misinterpretation of

chromatograms.

Reference USEPA 1999c. Method 1668, Revision A: Chlorinated Biphenyl Cogeners in Water,

Soil, Sediment, and Tissue by HRGC/HRMS, EPA-821-R-00-002. Office of Water,

U.S. Environmental Protection Agency, Washington, DC.

Website Rev A not available online Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

59

Fact Sheet No. 2.1.2-20

Method Title ESS Method 220.3: Am monia Nitrogen and Nitrate+Nitrite Nitrogen, Automated

Flow Injection Analysis Method, LMMB 061

Purpose This method is for the simultaneous determination of ammonia and nitrate/nitrite in

surface, drinking, and ground waters, and domestic and industrial wastes.

Method Summary Water samples are collected and preserved with sulfuric acid. Samples are

analyzed directly using an automated flow injection analyzer. For amm onia, alkaline

phenol and sodium hypochlorite react with ammonia to form a blue compound that is

proportional to the ammonia concentration. Am monia is measured colorimetrically at

630 nm. For nitrate+nitrite-N, nitrate is quantitatively reduced to nitrite using a

copperized cadmium column. The sample solution (total nitrite) then reacts with

sulfanilamide and N-(1-naphthyl)ethylenediamine dihydrochloride to form a magenta

solution. Total nitrite is measured colorimetrically at 520 nm. Nitrite alone can be

determined by removing the cadmium column. Nitrate is quantified by subtracting

the measured nitrite concentration from the measured total nitrite concentration. 

Samples with a concentration of 0.02-10.0 mg NH3-N/L and 0.02-35.0 mg NO3+NO2-

N/L can be analyzed with this method. These ranges can be extended through the

use of a digital diluter.

ASTM Method D1426 and Standard Method 4500-NH3C also describe the

colorimetric determination of ammonia (ASTM, 2001a; APHA, 1999). ASTM

Method D 3867 and Standard Methods 4500-NO2

-

.B and 4500-NO3

-

.F describe the

analysis of nitrite and nitrate.

Data

Uses/Application

The spatial and temporal variations in nutrients concentrations are often critical

parameters for understanding aquatic productivity and conditions of estuarine

habitat.

Advantages EDTA used in this procedures inhibits precipitation of residual calcium and

magnesium ions. The detection range of this method includes most concentrations

found in the environm ent.

Limitations Since a straight line calibration curve is not obtained, a greater number of standards

is needed.

Reference USEPA. 1997c. Method LMMB 061: ESS Method 220.3: Am monia Nitrogen and

Nitrate+Nitrite Nitrogen, Automated Flow Injection Analysis Method, Lake Michigan

Mass Balance Study Methods Compendium, Volume 3: Metals, Conventionals,

Radio chemistry, and Biomonitoring Sample Analysis Techniques, EPA 905-R-97-

012c. Great Lakes National Program Office, U.S. Environmental Protection Agency,

Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

methd220.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods for Assessing and Monitoring the 

Remediation of Contaminated Sediment Sites February 17, 2003 

Fact Sheet No. 2.1.2-21 

Method Title ESS Method 230.1: Total Phosphorus and Total Kjeldahl Nitrogen, Semi-Automated 

Method, LMMB 062 

Purpose This method is for the determination of total Kjeldahl nitrogen and total phosphorus 

in drinking, surface and was te waters. 

Method Sum m ary 

W ater samples are collected and preserved in the field using sulfuric acid. In this method, 

organic nitrogen and phosphorus compounds are digested using a sulfuric acid solution 

containing potassium sulfate. Mercuric sulfate is used as a catalyst in the digestion. 

H2SO4 + organic nitrogen Hg (NH4)2SO4 K2SO4 

H2SO4 + organic phosphorus Hg K3PO4 K2SO4 

Tubes of sam ple aliquots and acid are placed in a block digestor, where they are heated 

at 200°C for about 1 hour and then at 380°C for 75 minutes. The digestate is analyzed 

spectrophotom etrically as am m onia and phosphate using an Auto Analyzer. Total 

phosphorus and total Kjeldahl nitrogen concentrations are obtained directly from the 

plotter. 

Method LMMB 058 describes the general operating and maintenance procedures for using 

the Auto Analyzer. The Auto Analyzer is comprised of a sam pler, proportioning pump, 

manifold, colorimeter, and printer/plotter. The flow of reag ents and samples are 

proportioned by the pump, and air bubbles introduced into the tubing help to sep arate 

samples, mix reagents, and cleanse tubing. 

The operating range for this method is 0.1 to 10.0 mg N/L and 0.02 to 2.00 mg P/L. 

Standard Method 4500-N orgB and ASTM Method D3590 describe the determination of 

Kjeldahl nitrogen (APHA, 1999; ASTM, 2001a). 

Data This method details the conversion of nitrogen compounds such as amino acids, 

Uses/Application proteins and peptides to am m onia and can be used to evaluate drinking, surface 

and was te waters. 

Advantages Many water quality assessm ents require the measurem ent of total kjeldahl nitrogen. 

Limitations The digestion process may not convert all compounds (amines, nitro compounds, 

hydrazones, oximes, semicarbazones, and some tertiary amines) to amm onia. 

Reference USEPA. 1997c. Method LMMB 062: ESS Method 230.1: Total Phosphorus and 

Total Kjeldahl Nitrogen, Semi-Automated Method, Lake Michigan Mass Balance 

Study Methods Compendium, Volume 3: Metals, Conventionals, Radio chem istry, 

and Biomonitoring Sam ple Analysis Techniques. EPA 905-R-97-012c. Great Lakes 

National Program Office, U.S. Environmental Protection Agency, Chicago, IL. 

W ebs ite http://www.epa.gov/grtlakes/lmm b/methods/ Last Accessed: 1/31/2003 

methd230.pdf 

60 
A Compendium of Chemical, Physical and Biological Methods for Assessing and Monitoring the 

Remediation of Contaminated Sediment Sites February 17, 2003 

Fact Sheet No. 2.1.2-22 

Method Title ESS Method 310.2: Phosphorus, Total, Low Level (Persulfate Digestion), LMMB 

064 

Purpose This method is for the determination of total phosphorus in surface waters. 

Method Summary W ater samples are collected and preserved in the field with sulfuric acid. They are stored 

at 4°C until analysis. To determine dissolved phosphorus, samples are filtered through a 

0.45μm filter before digestion. The samples are digested with ammonium persulfate and 

sulfuric acid in an autoclave for 30 minutes at 121°C. All phosphorus is converted to 

orthophosphate. After the digestion, any particulate matter is allowed to settle overnight. 

Orthophosphate is then analyzed spectrophotometrically using an Auto Analyzer. The 

phosphorus concentration is obtained directly from the plotter. 

Method LMMB 058 describes the general operating and maintenance procedures for using 

the Auto Analyzer. The Auto Analyzer is comprised of a sampler, proportioning pump, 

manifold, colorimeter, and printer/plotter. The flow of reagents and samples are 

proportioned by the pump, and air bubbles introduced into the tubing help to separate 

samples, mix reagents, and cleanse tubing. 

The operating range for this method is 0.002-0.200mg P/L. 

The persulfate digestion procedure is also described in Standard Method 4500-P.B.5 

(APHA, 1999). 

Data These measurem ents are often required for water quality studies. 

Uses/Application 

Advantages The automated analysis allows economical analyses of multiple samples. 

Limitations This method describes only the phosphorus method. 

Reference USEPA. 1997c. Method LMMB 064: ESS Method 310.2: Phosphorus, Total, Low Level 

(Persulfate Digestion), Lake Michigan Mass Balance Study Methods Compendium, 

Volume 3: Metals, Conventionals, Radio chemistry, and Biomonitoring Sample Analysis 

Techniques. EPA 905-R-97-012c. Great Lakes National Program Office, U.S. 

Environmental Protection Agency, Chicago, IL. 

W ebsite http://www.epa.gov/grtlakes/lmm b/methods/ Last Accessed: 1/31/2003 

methd310.2.pdf 

61 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

62

Fact Sheet No. 2.1.2-23

Method Title ESS Method 310.1: Ortho-Phosphorus, Dissolved Automated, Ascorbic Acid, LMMB

063

Purpose This method is for the determination of orthophosphate in most waters and

wastewater.

Method Summary Water samples are collected, filtered through a 0.45 μm filter, cooled to 4°C, and

analyzed as soon as possible. The samples are analyzed spectrophotometrically

using an auto analyzer. In the instrument, amm onium molybdate and antimony

potassium tartrate react in an acid medium with dilute solutions of orthophosphatephosphorus to form an antimony-phospho-molybdate complex. This complex is

reduced to an intensely blue-colored complex by ascorbic acid. The color is

measured at 880 nm and is proportional to the phosphorus concentration. The

phosphorus concentration is obtained directly from the plotter.

 

Method LMMB 058 describes the general operating and maintenance procedures

for using the auto analyzer. The auto analyzer is comprised of a sampler,

proportioning pump, manifold, colorimeter, and printer/plotter. The flow of reagents

and samples are proportioned by the pump, and air bubbles introduced into the

tubing help to separate samples, mix reagents, and cleanse tubing.

The operating range for this method is 0.002-0.200mg P/L. This range may be

extended to 0.2-2.00 mg P/L by utilizing a dilution loop.

The automated ascorbic acid reduction method is also described in Standard

Method 4500-P F (APHA, 1999).

Data

Uses/Application

These measurem ents are useful for productivity assessments and site

characterizations.

Advantages The auto analyzer method provides fast, reproducible nutrient results.

Limitations Barium, lead, and silver may interfere with the analysis by forming a precipitate.

Reference USEPA. 1997c. Method LMMB 063: ESS Method 310.1: Ortho-Phosphorus,

Dissolved Automated, Ascorbic Acid, Lake Michigan Mass Balance Study Methods

Compendium, Volume 3: Metals, Conventionals, Radio chemistry, and

Biomonitoring Sample Analysis Techniques. EPA 905-R-97-012c. Great Lakes

National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

methd310.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

63

Fact Sheet No. 2.1.2-24

Method Title Total Organic Carbon, Standard Method 5310

Purpose This method is for the determination of total organic carbon in a wide variety of

water samples.

Method Summary TOC methods utilize heat and oxygen, ultraviolet irradiation, chemical

oxidants, or combinations of these oxidants to convert organic carbon to

carbon dioxide (CO2). The CO2 may be measured directly by a nondispersive

infrared analyzer, reduced to methane and measured with a flame ionization

detector, or CO2 may be titrated chemically.

In the Combustion-Infrared Method (5310B), the sample is homogenized and

diluted as necessary, and a microportion is injected into the heated reaction

chamber of a carbon analyzer, which is packed with an oxidative catalyst. The

water is vaporized, and the carbon is oxidized to CO2 and H2O. The CO2

from

oxidation of organic and inorganic carbon is measured by means of a

nondispersive infrared analyzer. This gives the measure of total carbon. TOC

is obtained by the difference of total carbon and inorganic carbon (IC). IC is

measured by injecting the sample into a separate reaction chamber packed

with phosphoric acid-coated quartz beads. Under acidic conditions, all IC is

converted to CO2

, which is measured. Under these conditions, organic carbon

is not oxidized and only IC is measured. Alternatively, TOC can be measured

by first acidifying the sample and purging the inorganic carbon from the sample

and then measuring the remaining carbon.

Other methods for measuring TOC exist, such as the Persulfate-Ultraviolet

Oxidation Method (5310C) and the Wet-Oxidation Method (5310D). In both of

these methods, organic carbon is oxidized to CO2 using persulfate. ASTM

Methods D6317, D2579, D4129, D4839, and D5790 and SW846 Method 9060

also describe various methods for the analysis of total organic carbon (ASTM,

2001a; USEPA SW 846).

Dissolved organic carbon (DOC) can be measured by first filtering the sample

through a 0.45-μm-pore-diam filter. Particulate organic carbon (POC) is the

fraction of TOC retained by this filter and is analyzed using a CHN elemental

analyzer.

The minimum detectable concentrations are 1 mg carbon/L, 0.05 mg organic

carbon/L, and 0.10 mg organic carbon/L can be measured with methods

5310B, 5310C, and 5310D, respectively. Method LMMB 067 (USEPA 1997c)

has an MDL of 5μg for POC.

Data Uses/Application Site characterization. TOC is also used in the assessment of trace metal and

organic contaminant data.

Advantages One of several standard methods for TOC analysis.

Limitations Acidification, purging, and sample blending may result in the loss of volatile

organic substances. Large organic particles may fail to enter the needle used

for sample injection or may oxidize slowly.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

64

Fact Sheet No. 2.1.2-24 (contd.)

Reference APHA. 1999. Standard Methods for the Examination of Water and

Wastewater, 20th Edition.

Website N/A Last Accessed: 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

65

Fact Sheet No. 2.1.2-25

Method Title Standard Operating Procedure for the Analysis of Dissolved-Phase Organic Carbon

in Great Lakes Waters, LMMB 096

Purpose To measure dissolved organic carbon (DOC) in the filtrates of water samples.

Method Summary Samples are filtered immediately after collection, and stored at 4°C until analysis on

the ship. Samples that are sent to the lab for analysis are frozen after arrival.

Inorganic carbon is removed from the filtrate by the addition of sulfuric acid and

purging of the sample with organic-free air. The samples are analyzed by

conversion of organic carbon to CO2 by an ultraviolet (UV) digester. The resulting

CO2is detected by a non-dispersive infrared (IR) analyzer. The concentration of

dissolved organic carbon is calculated using the peak height method.

Alternatively, Method LMMB 066 measures DOC by high temperature (680°C)

catalytic oxidation (USEPA 1997c). Inorganic carbon is first removed by

acidification and purging. The CO2

resulting from organic carbon is detected by a

nondispersive infrared (IR) analyzer. This method is applicable to organic carbon

concentrations from 0.2 to 50 mg/L and inorganic carbon concentrations less than

1000 mg/L.

Standard Method 5310 also describes the analysis of dissolved organic carbon

(APHA, 1999).

Data

Uses/Application

Dissolved and particulate metals and organic contaminant data are frequently

compared to DOC.

Advantages Both methods provide a rapid, reproducible analytical method for DOC analysis.

Limitations Results for volatile organic compounds using this method may be low.

Reference USEPA. 1997b. Method LMMB 096: Standard Operating Procedure for the

Analysis of Dissolved-Phase Organic Carbon in Great Lakes Waters, Lake Michigan

Mass Balance Study Methods Compendium, Volume 1: Sample Collection

Techniques. EPA 905-R-97-012c. Great Lakes National Program Office, U.S.

Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

docanal2.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

66

Fact Sheet No. 2.1.2-26

Method Title Standard Operating Procedure for the Analysis of Particulate-Phase Organic

Carbon in Great Lakes Waters, LMMB 097

Purpose To measure particulate organic carbon (POC) in the filtrates of water samples.

Method Summary Samples are filtered immediately after collection through glass fiber filters. Four

12mm discs are cut from each filter and allowed to dry. The disks are folded and

placed into individual tin sample containers. The disks are analyzed by catalytic

combustion using an elemental analyzer (CHNS analyzer). The sample container

with the disk is placed into a 1000°C furnace with a catalytic reactor tube. The

sample is oxidized, and the sample gases pass through a packed chromatographic

column for separation. The sample com ponents are separated as CO2, H2, N2, and

H2S. The com ponents are detected by thermal conductivity detection (TCD). To

calculate POC concentration in mg/L, the resulting mass of carbon from the four

discs per sample are summ ed, multiplied by an area correction factor, and divided

by the volume of water filtered.

Alternatively, Method LMMB 067 first treats the sample filters with sulfurous acid,

dries the filters at 60°C for 20-30 minutes, acidifies the filters again and dries them

again for 1 hour prior to analysis (EPA 905-R-97-012c). The entire filter is analyzed

using a CHN elemental analyzer. The method detection limit for this procedure is 5

ug of organic carbon remaining on a GF/F filter. The maximum amount of carbon

measurable is approximately 5 mg of carbon.

Standard Method 5310 also describes the analysis of particulate organic carbon

(APHA, 1999).

Data

Uses/Application

Normalization of trace metal and organic contaminant data; flux measurements.

Advantages This method can be highly automated.

Limitations Results for volatile organic compounds using this method may be low.

Reference USEPA. 1997b. Method LMMB 097: Standard Operating Procedure for the

Analysis of Particulate-Phase Organic Carbon in Great Lakes Waters, Lake

Michigan Mass Balance Study Methods Compendium, Volume 1: Sample Collection

Techniques. EPA 905-R-97-012c. Great Lakes National Program Office, U.S.

Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

pocanal2.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

67

Fact Sheet No. 2.1.2-27

Method Title ESS Method 140.4: Chloride - Automated Flow Injection Analysis, LMMB 060

Purpose This method is for the determination of chloride in drinking water, surface water,

saline water, domestic and industrial wastes.

Method Summary Water samples are collected and analyzed within 28 days. Samples are analyzed

directly using an automated flow injection analyzer. This method is based on the

interaction of chlorine ions and mercuric thiocyanate. As a result, a highly colored

solution is formed, which is measured colorimetrically. 

Samples with a concentration of 1.0-100 mg Cl/L can be analyzed directly. This

range can be extended through the use of a digital diluter.

Standard Method 4500-Cl- E also describes the automated analysis of chloride

(APHA, 1999). Alternative methods for measuring chlorine include titration and use

of ion-selective electrode. These are described in ASTM Method D512 and

Standard Methods 4500-Cl- B, C, and D (ASTM, 2001a; APHA, 1999). The ionselective electrode method can measure chloride concentrations up to 1000 mg

Cl/L.

Data

Uses/Application

The EPA recognizes chloride in drinking water as a secondary standard. 

Advantages This method is capable of analyzing up to 100 samples per hour.

Limitations Since a straight line calibration curve is not obtained, a greater number of standards

is needed.

Reference USEPA. 1997c. Method LMMB 060: ESS Method 140.4: Chloride - Automated

Flow Injection Analysis, Lake Michigan Mass Balance Study Methods Compendium,

Volume 3: Metals, Conventionals, Radio chemistry, and Biomonitoring Sample

Analysis Techniques. EPA 905-R-97-012c. Great Lakes National Program Office,

U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

methd140.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

68

Fact Sheet No. 2.1.2-28

Method Title ESS Method 200.5: Determination of Inorganic Anions in Water by Ion

Chromatography, LMMB 059

Purpose This method is for the determination of chloride, nitrate-N, and sulfate in drinking

water, surface water, and mixed domestic and industrial wastewater.

Method Summary Water samples are collected and preserved as follows:

Chloride: No preservation required. Analyze within 28 days.

Nitrate-N: Cool to 4°C. Analyze within 48 hours.

Sulfate: Cool to 4°C. Analyze within 28 days.

Samples are filtered through a 0.45 μm filter to remove particulate matter. A portion

of the sample (usually 5 mL) is injected into the ion chromatography, comprised of a

guard column, separator column, suppressor column, and conductivity detector .

The anions are separated based on their affinity for the exchange sites of the resin

in the analytical and guard column. Anions are identified base on their retention

times compared to known standards. Results are reported as mg/L.

Alternative methods for the determination of anions (including additional anions,

such as phosphate and nitrite) by Ion Chromatography are Standard Method 4110B,

ASTM D4327, and SW846 9056 (APHA, 1999; ASTM, 2001a;USEPA SW 846).

Data

Uses/Application

Site characterization/site assessment.

Advantages The suppressor column reduces background conductivity.

Limitations Phosphate is not analyzed with this method. Nitrite can interfere with the detection

of chloride.

Reference USEPA. 1997c. Method LMMB 059: ESS Method 200.5: Determination of

Inorganic Anions in Water by Ion Chromatography, Lake Michigan Mass Balance

Study Methods Compendium, Volume 3: Metals, Conventionals, Radio chemistry,

and Biomonitoring Sample Analysis Techniques. EPA 905-R-97-012c. Great Lakes

National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

methd200.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

69

Fact Sheet No. 2.1.2-29

Method Title Standard Operating Procedure for Electrometric pH, LMMB 092

Purpose To measure pH in drinking, surface, and saline waters; domestic and industrial

wastes.

Method Summary The working range of this method is 6.0 to 10.0 pH units. Samples are collected in

clean glass or plastic containers and stored at 4°C until analysis. The pH meter is

calibrated with 7.0 and 10.0 buffers. The sample is brought to 25°C before analysis.

An aliquot of the sample is placed into a suitable container, which is then placed on

a stirrer. The pH meter electrode is submerged into the sample and the pH reading

is taken once the meter stabilizes.

ASTM Method D1293, Standard Method SM 4500-H+

.B, and SW846 Method 9040B

also describe the measurement of pH of water (ASTM, 2001a; APHA, 1999). 

Data

Uses/Application

The pH of water is an important parameter in the solubility of trace minerals in water

and the suitability of the water to sustain life.

Advantages Little skill and training is needed to perform this analysis.

Limitations Tem perature affects the electrometric response and must be compensated for.

Extremely acidic waters require a different calibration range (i.e., 4.0 and 7.0

buffers). 

Reference USEPA. 1997c. Methods LMMB 092: Standard Operating Procedure for

Electrometric pH, Lake Michigan Mass Balance Study Methods Compendium,

Volume 1: Sample Collection Techniques. EPA 905-R-97-012c. Great Lakes

National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

phydrion.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

70

Fact Sheet No. 2.1.2-30

Method Title Standard Operating Procedure for GLNPO Total Alkalinity Titration, LMMB 091

Purpose To measure alkalinity in drinking, surface, and saline waters; domestic and industrial

wastes.

Method Summary This method is designed for waters in the range of 10 - 250 mL/L total alkalinity as

CaCO3. The pH meter is calibrated with 4.0 and 7.0 buffers. 100mL of sample are

titrated with 0.0200 N sulfuric acid to pH 4.5. Total alkalinity is calculated as CaCO3

in mg/L by multiplying the volume of titrant (in mL) by 10.

Similar methods for the measurement of alkalinity are described in ASTM Methods

D1067 and D3875 and Standard Method 2320B (ASTM, 2001a; APHA, 1999).

Data

Uses/Application

Alkalinity is a measure of water’s natural buffering capacity, thus an important

parameter measured to assess overall water quality.

Advantages This method can be automated or semi-automated for multiple analyses.

Limitations Oil, grease, and high mineral content may interfere with the alkalinity determination.

Reference USEPA. 1997b. Methods LMMB 091: Standard Operating Procedure for GLNPO

Total Alkalinity Titration, Lake Michigan Mass Balance Study Methods

Compendium, Volume 1: Sample Collection Techniques. EPA 905-R-97-012c.

Great Lakes National Program Office, U.S. Environmental Protection Agency,

Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

alkali.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

71

Fact Sheet No. 2.1.2-31

Method Title Standard Operating Procedure for GLNPO Specific Conductance: Conductivity

Bridge, LMMB 094

Purpose To measure conductance of drinking and surface waters.

Method Summary Samples are collected and stored at 4°C until analysis. The specific conductance of

the samples is measured using a self-contained conductivity meter. Care should be

taken to assure that not air bubbles are present in the conductivity cell. The

temperature is adjusted to 25°C and the conductivity read. 

The approximate working range of this method of 10-500 mhos/cm.

ASTM Method D1225, Standard Method 2510B, and SW846 Method 9050A

describe similar methods for the measurement of conductivity of water (ASTM,

2001a; APHA, 1999). The test range of Method D1225A is 10-200000 μS/cm.

Data

Uses/Application

This method is applicable to the quantitative measurement of ionic constituents

dissolved in water.

Advantages The procedure and equipment are simple and easy to operate.

Limitations Sample temperatures other than 25°C will cause incorrect results. Oil, grease,

algae, or dirt may interfere with the readings.

Reference USEPA. 1997b. Method LMMB 094: Standard Operating Procedure for GLNPO

Specific Conductance: Conductivity Bridge, Lake Michigan Mass Balance Study

Methods Compendium, Volume 1: Sample Collection Techniques. EPA 905-R-97-

012c. Great Lakes National Program Office, U.S. Environmental Protection

Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmm b/methods/

conducti.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

72

Fact Sheet No. 2.1.2-32

Method Title Standard Operating Procedure for GLNPO Turbidity: Nephelometeric Method,

LMMB 090

Purpose To measure turbidity in drinking, surface, and saline waters.

Method Summary Turbidity samples are analyzed immediately or stored at 4°C. The working range of

the turbidimeter is 0-20 nephelometeric turbidity units (NTU). Dilutions can be

performed to measure turbidities greater than 20 NTU. The instrument is calibrated

with a geometric series of calibration standards. An aliquot of the sample is warmed

to 25°C and placed into the turbidimeter for measurem ent. The instrument

measures turbidity by comparing the intensity of light scattered by the sample with

the intensity of light scattered by a standard reference suspension.

ASTM Method D1889 and Standard Method 2130B describe a similar method of

measuring the turbidity of water (ASTM, 2001a; APHA, 1999). These methods have

a working range of 1.0-40 NTU. 

Data

Uses/Application

Turbidity is measured as part of overall site characterization, as an indirect measure

of light penetration. Turbidity can also be monitored during dredging or other

sediment excavations to measure amounts of suspended material entering the

environment for site activities.

Advantages Turbidity measured with a nephelometer provides a much more rapid and

reproducible measurement compared to the filtration/color chart method.

Limitations Floating debris and air bubbles in the sample may give high readings.

Condensation or scratches on the sample vial and sample color may give erroneous

readings.

Reference USEPA. 1997b. Methods LMMB 090: Standard Operating Procedure for GLNPO

Turbidity: Nephelometeric Method, Lake Michigan Mass Balance Study Methods

Compendium, Volume 1: Sample Collection Techniques. EPA 905-R-97-012c.

Great Lakes National Program Office, U.S. Environmental Protection Agency,

Chicago, IL.

Website http://www.epa.gov/grtlakes/lmmb/meth

ods/turbid.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

73

Fact Sheet No. 2.1.2-33

Method Title ESS Method 340.2: Total Suspended Solids, Mass Balance (Dried at 103-105°C)

Volatile Suspended Solids (Ignited at 550°C), LMMB 065

Purpose To measure the portion of total solid retained by a filter from drinking, surface, and

saline waters; domestic and industrial wastes.

Method Summary Water samples are collected by submersible pump or Rosette sampler. A sample

volume is selected that will yield 2 - 20,000 mg/L. total suspended solids. For

open-lake oligotrophic conditions, 2-4 liters will provide enough particulate matter.

For near-shore or eutrophic conditions, 200-500 mL may be sufficient. A well-mixed

sample is filtered through a preweighed standard glass-fiber filter, and the residue

retained on the filter is dried at 103 to 105 °C for at least one hour. The increase in

weight of the filter represents the total suspended solids. 

After determining TSS, the filters may be placed in a muffle furnace and ignited at

550°C for 30 minutes to determine volatile suspended solids (VSS).

Following Method LMMB 098, water samples can be filtered in the field and then

frozen at -10°C until final weighing in the laboratory.

Standard Method 2540D, ASTM Method D5907, and Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies describe similar methods for measuring total suspended solids

(APHA, 1999; ASTM, 2001a; USEPA 1992b, respectively).

Data

Uses/Application

TSS is an ancillary parameter to the determination of hydrophobic organic

contaminants (HOCs). 

Advantages Glass fiber filters can be ignited without damage, allowing TSS and VSS to be

performed on the same set of filters.

Limitations Excessive residue on the filter may form a water-entrapping crust. Sample size

should be limited to yield no more than 200 mg residue. Glass fiber filters are not

appropriate for measurement of TSS in estuarine waters. 

Reference USEPA. 1997c. Method LMMB 065: ESS Method 340.2: Total Suspended Solids,

Mass Balance (Dried at 103-105°C) Volatile Suspended Solids (Ignited at 550°C).

Lake Michigan Mass Balance Study Methods Compendium, Volume 3: Metals,

Conventionals, Radio chemistry, and Biomonitoring Sample Analysis Techniques.

EPA 905-R-97-012c. Great Lakes National Program Office, U.S. Environmental

Protection Agency, Chicago, IL.

Website http://www.epa.gov/grtlakes/lmmb/meth

ods/methd340.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

74

Fact Sheet No. 2.1.2-34

Method Title Total Hardness Titration, LMMB 095

Purpose To measure the total concentration of the calcium and magnesium ions expressed

as calcium carbonate.

Method Summary A water sample is collected at mid depth during unstratified conditions, or on the

mid-epilimnion and mid hypo-limnion when stratification is present. A 100 mL water

sample is buffered to pH 10.1, and an indicator (such as Chrome Black T3) is then

added to the buffered sample. The indicator turns red in the presence of Ca and Mg

ions. The sample is titrated with 0.01M EDTA, which complexes with Mg and Ca

cations, removing them from association with the indicator. When all the Mg and Ca

are complexed with EDTA, the indicator will turn blue. The volume of titrant is

recorded. Total Hardness is calculated as 10 X mL of titrant and reported as mg/L

as CaCO3. Standard Method 2340C and ASTM Method D1126 also describe the

titration analysis of hardness (APHA, 1999; ASTM, 2001a). 

An alternative method for the analysis of hardness is to determine the amount of

calcium and magnesium ions separately and then sum them to calculate total

hardness. Standard Method 2340B and ASTM Method D1126 both describe this

alternative method (APHA, 1999; ASTM, 2001a). When determined as separate

ions, hardness is calculated as: 2.497[Ca, mg/L] + 4.118 [Mg, mg/L]

Data

Uses/Application

Hardness is measured in aquatic systems because it is known to mitigate metals

toxicity in fish.

Advantages The reagents and chemicals required for this analysis can be obtained as

prepackage test kits. The titration method affords a means of rapid analysis, where

as the calculation method is more accurate.

Limitations This test method is not suitable for highly colored waters, which obscure the color

change of the indicator. A limit of 5 minutes is set for the duration of the titration to

minimize the tendency toward CaCO3 precipitation.

Reference USEPA. 1997c. Method LMMB 095: Total Hardness Titration. Lake Michigan Mass

Balance Study Methods Compendium, Volume 3: Metals, Conventionals, Radio

chemistry, and Biomonitoring Sample Analysis Techniques. EPA 905-R-97-012c.

Great Lakes National Program Office, U.S. Environmental Protection Agency,

Chicago, IL.

Website http://www.epa.gov/grtlakes/lmmb/meth

ods/titratio.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

75

2.1.3 Biological Analysis Methods

Section 2.1.3 provides a compendium of water-related biological analyses. As mentioned

previously, once contaminants enter into an aqueous system, the contaminant’s chemical nature

and the biological, chemical, or physical characteristics of the receiving water body will determine

whether it remains in the water column, becomes buried in sediment, or is ingested by organisms.

The negative impacts of contaminant exposure can be examined in laboratory toxicity tests, which

use site-specific effluents, leachates or elutriates prepared from sediments collected from the site.

All liquid-phase toxicity tests are performed to ultimately determine the lowest observable effect

concentration, the no observable effect concentration and other related parameters. These

results are then compared with chemistry data to identify and compare toxicity effects data with

contaminant exposure data to determine risk to ecological or human receptors. While elutriate

toxicity testing is performed to evaluated sediment toxicity, they are included in this section

because the same aqueous phase methods are also used to evaluate water and wastewater.

The advantages and limitations associated with each test are provided in their respective fact

sheets. However, there are general limitations in interferences associated with all liquid-phase

toxicity tests. These potential interferences are identified by some of the source documents

(USEPA, 1994d; Weber, 1991). They are listed below:

• Toxicity tests do not reflect temporal fluctuations in effluent toxicity;

• Non-target chemicals can cause adverse effects to the organisms giving false

results;

• dissolved oxygen depletion due to biological and chemical oxygen demand and

metabolic wastes can be a problem;

• The toxicant may be lost through volatilization and adsorption to the exposure

chamber; and,

• The effect of the toxicant is organism dependant.

Liquid-phase toxicity tests vary considerably in test length, endpoints, and test species. Table

2.1.3-1 summarizes those toxicity tests described in the fact sheets, however it highlights the

differences between specific organisms to aid the Superfund manager with selecting the most

appropriate test for his/her site. 

The toxicity test fact sheets provide methods described in USEPA guidance documents, Dredging

Manuals and ASTM reports. Specifically, the following sources provided methods information for

section 2.1.3:

• The USEPA’s Environmental Response Team SOP’s (USEPA , 1994b); ERT SOPs

are available online at: http://www.ert.org 

www.epa.gov/waterscience/WET/disk1/

• The USEPA’s Methods document for Effluents and Receiving waters (Weber, 1991);

• The USEPA’s guidance document for contaminated sediment assessment in the Great

Lakes (USEPA, 1993a);

• The Inland Lakes Testing Manual (USEPA and USACE, 1998);

• The USEPA’s Environmental Research Laboratory-Narragansett (USEPA and the

Naval Construction Battalion Center, 1992)
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

76

Table 2.1.3-1. A Sum mary of Test Types and Toxicological Endpoints for Liquid-Phase Tox icity 1, 2

Test Type/Fact

Sheet Num ber Test Organism Scientific N am e Endpoints Test Specifics Comments

Acute Freshwater

2.1.3-1

Crustacean Da phnia magna Survival Static, staticrenewal, 24, 48 or 96

hours

Co mmonly used for bioassays;

optimum pH 6.8-8.5

Acute Freshwater

2.1.3-1

Crustacean Da phnia Pulex Survival Static, staticrenewal, 24, 48 or 96

hours

Co mmonly used for bioassays;

optimum pH 6.8-8.5

Acute Freshwater

2.1.3-2

Fish Pimephales

Promelas

Survival Static, static-renewal,

24, 48 or 96 hours

Other commonly used freshwater

fish in bioassays include the

Oncorhynchus mykiss

Ch ronic

Freshwater

2.1.3-3

Algae Selanastrum

capricornutum

Growth,

biostimu latory

effects (cell density

Static, 96 hours Originally designed as a

eutrophication test, now used in the

Superfund program for effluents.

Ch ronic

Freshwater

2.1.3-4

Crustacean Ce riodaphnia

dubia

Survival,

reproduction

Static renewal, 7

days

Co mmonly used for bioassays

Ch ronic

Freshwater

2.1.3-5

Crustacean Da phnia magna Reproduction,

growth

Static-renewal, 10

days

Co mmonly used for bioassays;

optimum pH 6.8-8.5

Ch ronic

Freshwater

2.1.3-5

Crustacean Da phnia Pulex Reproduction,

growth

Static-renewal, 10

days

Co mmonly used for bioassays;

optimum pH 6.8-8.5

Ch ronic

Freshwater

2.1.3-6

Fish Pimephales

promelas

Larval Survival and

growth

Static-renewal, 7

days

Other commonly used freshwater

fish in bioassays include the

Oncorhynchus mykiss

Ch ronic

Freshwater

2.1.3-17

Fish Pimephales

promelas,

Ictalurus

punctatus,

Coregonus artedii,

Oryzias latipes,

Catastomus

comm ersoni, Esox

lucius, Da nio

danio

Survival, growth Single exposure, 32

days (100 days for

Coregonus artedii)

One of the few, published early life

stage methods

Acute Marine

2.1.3-7

Macroalgae Ch ampia parvula Sexual

reproduction

Static, 48 hours Used for whole effluent toxicity

testing in the NPDES Program

Acute Marine

2.1.3-8

Crustacean Mysid oposis

bahia

1-25 ppt Static-renewal, 96

hours

Other commonly used

estuarine/marine mysid shrimp in

bioassays include the

Ho lmesimysis costata and the

Ne omysis americana

Acute Marine

2.1.3-9

Fish Cyprinodon

variegatus

Survival Static-renewal, 24, 48

or 96 hours

Co mmonly used in bioassays

Acute Marine

2.1.3-9

Fish Menidia beryllina Survival Static-renewal, 24, 48

or 96 hours

Co mmonly used in bioassays
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

77

Table 2.1.3-1. (contd.)

Test Type Test Organism Scientific N am e Endpoints Test Specifics Comments

Ch ronic Marine

2.1.3-10

Crustacean Mysid opsis bahia Survival, growth,

fecundity

Static-renewal, 7

days

Other commonly used

estuarine/marine mysid shrimp in

bioassays include the

Ho lmesimysis costata and

Ne omysis americana

Ch ronic Marine

2.1.3-11

Echinoderm Arb acia

punctulata

Toxicity to eggs and

sperm/ %

fertilization

One hour Ch ronic test but test time is quite

short. 

Ch ronic Marine

2.1.3-12

Fish Cyprinodon

variegatus

Larval survival and

growth

static-renewal, 7 days Co mmonly used in bioassays

Ch ronic Marine

2.1.3-12

Fish Menidia beryllina Larval survival and

growth

static-renewal, 7 days Co mmonly used for bioassays

1

Three species are generally recommended for water column bioassays. They should represent different phyla whe re possible.

2 Specific characteristics to consider when selecting water-column test species include availability year-round, tolerance to handling and laboratory conditions,

consistent and reproducible responses to toxicants, phylogenic similarities to species characteristic of those inhabiting water column at impacted area,

availability of standardized test protocols, ability to test as juveniles or larvae to increase sensitivity, appropriate sensitivity.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

78

Fact Sheet No. 2.1.3-1

Method Title Acute Freshwater Crustacean Bioassay: 48 Hours, ERT SOP 2024

Purpose This static toxicity test measures the survival of the freshwater crustaceans, Daphnia

Magna and Daphnia Pulex, after exposure to leachates, effluents, or liquid phases of

sediments (i.e., elutriates or pore water) for 48 hours in the laboratory. 

Method Summary Larval daphnids, Daphnia Magna and Daphnia Pulex, are exposed to various

concentrations of liquid-phase test media for 48 hours in 100-mL containers (or 250

mL, USEPA 1993a). The media concentrations levels intend to span a range of

those causing zero mortality to those causing com plete mortality. If the test medium

is a liquid, dilution may be made directly for the required test concentrations. If the

test medium is a sediment, preliminary filtration and dilutions are required to produce

a liquid phase that will then be diluted to attain the above test concentrations.

Once all the exposure chambers are set up with their designated test

concentrations, the test organisms are added after being acclimated to the dilution

water in separate chambers. The experiment officially begins when half of the

organisms are in the exposure chambers. Test temperature is 20.0 +/- 2

NC for the

daphnids. The test is static; water will not be renewed throughout the duration of the

test. The endpoints are survival at 1-hour, 24-hours, and 48-hours. Organisms are

not fed during the test.

At the termination of the test, mortality and water quality parameters are recorded.

Range-finding tests may be performed prior to these analyses to determine the

appropriate test media dilutions. Reference toxicant tests will also be performed

simultaneous to these tests. 

The acute toxicity tests described for use in the NPDES Permits Program indicate

that this test may be static or static-renewal and it is run for 24, 48 or 96 hours

(Weber, 1991). 

A similar method for acute toxicity using Daphnia spp. is described in the

Environment Canada Method EPS 1/RM/11.

Data

Uses/Application

The results are used to determine the lethal concentration for 50% of the test

species (LC50). The Lowest Observable Effects Concentration (LOEC) and the No

Observable Effects Concentration (NOEC) are also recorded.

Advantages Both of these species of daphnids are considered sensitive benchmark species

(USEPA and USACE 1998). Benchmark species comprise a substantial data base,

represent the sensitive range of a variety of ecosystems and provide comparative

data on the relative sensitivity of local test species. 

Liquid-phase toxicity testing is often simpler to run than bulk sediment testing;

excellent correspondence between bulk sediment contaminant concentrations and

pore water toxicity has been observed. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

79

Fact Sheet No. 2.1.3-1 (contd.)

Limitations Daphnia are freshwater crustaceans, therefore they cannot be used in estuarine and

marine settings. 

The optimum pH range for Daphnids is 6.8 to 8.5; therefore, the pH of the dilution

water or the concentrations may have to be adjusted prior to the start of the test. 

Reference USEPA. 1994b. 48-Hour Acute Toxicity Test Using Daphnia Magna and

Daphnia Pulex, SOP #2024. Compendium of ERT Standard Operating Protocols.

Office of Solid Waste and Emergency Response, U.S. Environmental Protection

Agency, Edison NJ.

Website http://www.ert.org/products/2024.

pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

80

Fact Sheet No. 2.1.3-2

Method Title Acute Freshwater Fish Bioassay, ERT SOP 2022

Purpose These methods describe a static-renewal toxicity test using the freshwater fish,

Pimephales promelas. These tests are effective when testing the acute toxicity of

effluents, leachates and liquid phases of sediments for 96 hours in the laboratory.

Method Summary The larval Pimephales promelas (fathead minnow) is exposed to various

concentrations of test media over a 96-hour period in 1-L test containers. The

medium concentration level is planned to span a range of those causing zero

mortality to those causing com plete mortality. If the test medium is a liquid, dilution

may be made directly for the required test concentrations. If the test medium is a

sediment, preliminary filtration and dilutions are required to produce a liquid phase

that will then be diluted to attain the above test concentrations. 

The test temperature is 25 +/- 2

NC. Fish will be fed during the acclimation period

and during the toxicity test. Test solutions will be replaced daily in exposure

chambers. Record survival at one hour and then daily thereafter. After the 96 hours

has past, final mortality and water quality measurem ents are recorded.

Range-finding tests may be performed prior to these analyses to determine the

appropriate test media dilutions. Reference toxicant tests will also be performed

simultaneous to these tests. 

The acute freshwater fish bioassay used in the NPDES perm its program indicates

that this test may be static or static-renewal and run for 24, 48 or 96 hours (Weber.

1991).

The rainbow trout, Oncorhynchus mykiss, is another benchmark freshwater fish

specie used for water column toxicity tests (USEPA and USACE. 1998). 

Data

Uses/Application

This test may be conducted on effluents, leachates, or liquid phase of sediments.

The results will be used to determine the lethal concentration of test media that

causes 50% mortality (LC50). The Lowest Observable Effect Concentration (LOEC)

and the No Observable Effects Concentration (NOEC) is also recorded. 

Advantages Pimephales promelas are easily reared in the laboratory and they are important

forage fish in the food chain. Pimephales promelas are considered benchmark

species indicating that they comprise a substantial database, represent the sensitive

range of a variety of ecosystems, and provide comparative data on the relative

sensitivity of local test species (USEPA and USACE, 1998).

Limitations Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 1994b. 96-Hour Acute Toxicity Test Using Larval Fathead Minnows

(Pimephales proomelas) SOP #2022. Environmental Response Team.

Compendium of ERT Standard Operating Protocols. Office of Solid Waste and

Emergency Response, U.S. Environmental Protection Agency, Edison NJ.

Website http://www.ert.org/products/2022

.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

81

Fact Sheet No. 2.1.3-3

Method Title Chronic Freshwater Algae Test, ERT SOP 2027

Purpose This test measures the biostimulatory capabilities of leachates, effluents, or

liquid phases of sediments (i.e., elutriates or pore water) on Selenastrum

capricornutum for 96-hour exposure of in the laboratory.

Method Summary The freshwater algae, Selenastrum capricornutum is exposed to various

concentrations of test media over a 96-hour period in 100-mL test containers.

The media concentration levels intend to span a range of those causing zero

mortality to those causing com plete mortality. If the test medium is a liquid,

dilution may be made directly for the required test concentrations. If the test

medium is a sediment, preliminary filtration and dilutions are required to

produce a liquid phase that will then be diluted to attain the above test

concentrations. 

Test temperature is 25 +/- 2

N C. The endpoint is growth. There is no water

renewal throughout the test. 

Growth is measured at the end of the test by cell counts, chlorophyll content or

turbidity (light absorbance), or biomass. Cell counts are determined using an

autom atic particle counter or manually under a microscope. Chlorophyll content

may be measured using in-vivo or in-vitro fluorescence or in-vitro

spectrophotometry. Turbidity is measured by spectrophotometry at 750 nm.

Biomass is measured by multiplying the cell count by the mean cell volume or

by direct gravimetric dry weight analysis. 

This test can also be conducted for 24-hours using 25-mL glass borosilicate

tubes with the following dilutions: 0%, 10%, 25%, 50% and 94% elutriate. 

This test protocol is also described in EPA guidance documents for effluents

and receiving waters and assessing contaminated sediments in the Great

Lakes (Weber, 1993; USEPA, 1993a).

Environment Canada methods EPS 1/RM/25 and EPS 1/RM/37 describe

similar tests using Selenastrum capricornutum and Lemna minor.

Data Uses/Application This method was originally designed to test for eutrophication, however it has

been recommend for use in testing the toxicity of complex effluents and has

been widely used to test single chemicals.

The results of this test will be used to determine the No Observable Effect

Concentration (NOEC), Lowest Observable Effect Concentration (LOEC) and

the Chronic Value (CHV). These results will determine the long term effects of

those sediment samples on the surrounding biotic community. 

Advantages There is currently no method for exposing algae directly to whole sediments,

thus an elutriate most closely simulates the most likely exposure conditions for

natural algal populations. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

82

Fact Sheet No. 2.1.3-3 (contd.)

Limitations The concentration of natural nutrients in the test media may affect the results.

This is not a benchmark test species used for toxicity testing (USEPA and

USACE, 1998).

Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 1994b. 96-Hour Static Toxicity Test Using Selenastrum

capricornutum, SOP #2027, Environmental Response Team. Compendium of

ERT Standard Operating Protocols. Office of Solid Waste and Emergency

Response, U.S. Environmental Protection Agency, Edison NJ. 

Website http://www.ert.org/products/2027.pdf Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

83

Fact Sheet No. 2.1.3-4

Method Title Chronic Freshwater Crustacean Bioassay (7 Day), ERT SOP 2025

Purpose These methods describe a 7-day, static-renewal toxicity test used to measure

chronic effects from liquid-phase test media on the freshwater crustacean,

Ceriodaphnia dubia. 

Method Summary The freshwater water flea, Ceriodaphnia dubia, are exposed to various

concentrations of test media over a 7-day period in 30mL test chambers. The

media concentration levels intend to span a range of those causing zero

mortality to those causing com plete mortality. If the test medium is a liquid,

dilution may be made directly for the required test concentrations. If the test

medium is a sediment, preliminary filtration and dilutions are required to

produce a liquid phase that will then be diluted to attain the desired test

concentrations. 

Test temperature is 25 +/- 2

N C. New test media concentrations are prepared

daily. The organisms are physically transferred to newly prepared exposure

chambers. Organisms are fed daily. 

Survival is recorded over a 7-day period as well as the number of broods and

the brood size. The number of males surviving are counted at test termination. 

Range-finding tests may be performed prior to these analyses to determine the

appropriate test media dilutions. Reference toxicant tests will also be

performed simultaneous to these tests. 

Similar freshwater crustacean species such as the Daphnia magna and the

Daphnia pulex may also be used for similar assays.

The EPA guidance documents for toxicity testing of effluents and receiving

waters also describes this method (Weber. 1991), as does Environment

Canada’s method EPS 1/RM/21.

Data Uses/Application The data from these tests will be used to determine the Lowest Observable

Effect Concentration (LOEC), the No Observable Effect Concentration (NOEC),

the EC50 and the chronic value of the test media.

Advantages Ceriodaphnia dubia are commonly used test species for freshwater toxicity

testing, therefore resulting data will be comparable to a large number of

previous studies. 

These organisms are benchmark species indicating that they comprise a

substantial database, represent the sensitive range of a variety of ecosystems,

and provide comparative data on the relative sensitivity of local test

species(USEPA and USACE, 1998). 

Limitations Ceriodaphnia dubia are freshwater species; they can only be used when salinity

is <1 ppt.

Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

84

Fact Sheet No. 2.1.3-4 (contd.)

Reference USEPA. 2002. 7-Day Static Renewal Toxicity Test Using Ceriodaphnia dubia.

Rev 1. SOP #2025, Environmental Response Team. Compendium of ERT

Standard Operating Protocols. Office of Solid Waste and Emergency

Response, U.S. Environmental Protection Agency, Edison NJ.

Website http://www.ert.org/products/2

025.pdf

Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

85

Fact Sheet No. 2.1.3-5

Method Title Chronic Freshwater Crustaceans Bioassay (10 days), ERT SOP 2028

Purpose These methods describe a 10-day, static renewal toxicity test used to measure

chronic effects from contaminated test media on the freshwater daphnids, Daphnia

magna or Daphnia pulex

Method Summary Larval Dapnia magna or Daphnia pulex are placed in individual 100-mL containers

and exposed to different concentrations of liquid-phase test media over a 10-day

period. If the test medium is a liquid, dilution may be made directly for the required

test concentrations. If the test medium is a sediment, preliminary filtration and

dilutions are required to produce a liquid phase that will then be diluted to attain the

desired test concentrations. 

Test temperature is 25 degrees +/- 2

N C. Test media concentrations are renewed

every other day for the duration of the test; organisms are physically transferred into

new exposure chambers. Organisms are fed every other day. The endpoints of the

test are mortality, reproduction and growth. 

Ceriodaphnia dubia is also recommended as a freshwater daphnid that may be

used for similar assays (USEPA and USACE, 1998).

Data

Uses/Application

This test is applicable to leachates, effluents, and liquid phases of sediments. The

data from these tests will be used to determine the Lowest Observable Effect

Concentration (LOEC), the No Observable Effect Concentration (NOEC), the EC50

and the chronic value of the test media. 

Advantages All three of these species of daphnids are considered sensitive benchmark species

(USEPA and USACE 1998). Benchmark species comprise a substantial data base,

represent the sensitive range of a variety of ecosystems and provide comparative

data on the relative sensitivity of local test species. 

Limitations Daphnia are freshwater crustaceans, therefore they cannot be used in estuarine and

marine settings. 

The optimum pH range for daphnids is 6.8 to 8.5; therefore, the pH of the dilution

water or the concentrations may have to be adjusted prior to the start of the test. 

Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA.1994b. 10-day Chronic Toxicity Test using Daphnia Magna or Daphnia

Pulex. SOP #2028. Environmental Response Team. Compendium of ERT

Standard Operating Protocols. Office of Solid Waste and Emergency Response,

U.S. Environmental Protection Agency, Edison NJ. 

Website http://www.ert.org/products/2028.pdf Last Accessed: 1/23/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

86

Fact Sheet No. 2.1.3-6

Method Title Chronic Freshwater Fish Bioassay, ERT SOP 2026

Purpose These methods describes a 7-day, static-renewal toxicity test using the larval

freshwater fish, Pimephales promelas.

Method Summary The Pimephales promelas are exposed to various concentrations of liquid-phase

test media over a 7-day period in 500 mL - 1L test containers. The media

concentration levels intend to span a range of those causing zero mortality to those

causing com plete mortality. If the test medium is a liquid, dilution may be made

directly for the required test concentrations. If the test medium is a sediment,

preliminary filtration and dilutions are required to produce a liquid phase that will then

be diluted to attain the desired test concentrations. 

Test temperature is 25 +/- 2

NC. The fish are fed daily at 4 hour intervals. New

dilutions of test media are prepared daily. The old solution is drawn out with a

siphon and the newly prepared solutions are added to each chamber. The

endpoints are survival and growth.

Range-finding tests may be performed prior to these analyses to determine the

appropriate test media dilutions. Reference toxicant tests will also be performed

simultaneous to these tests. 

The rainbow trout, Oncorhynchus mykiss, is another benchmark freshwater fish

specie used for water column toxicity tests. A method for testing acute toxicity using

rainbow trout can be found in Environment Canada’s method EPS 1/RM/9.

EPA guidance for toxicity testing of effluents and receiving waters also describes

this method (Weber, 1991, USEPA, 1990b), as does Environment Canada’s method

EPS 1/RM/22.

Data

Uses/Application

The results of this test will be used to determine the No Observable Adverse Effect

Concentration (NOAEC), Lowest Observable Adverse Effect Concentration

(LOAEC) and the Chronic Value (CHV). These results will determine the long term

effects of those sediment samples on the surrounding biotic community. 

Advantages Pimephales promelas are easily reared in the laboratory and they are important

forage fish in the food chain. Pimephales promelas are considered benchmark

species indicating that they comprise a substantial database, represent the sensitive

range of a variety of ecosystems, and provide comparative data on the relative

sensitivity of local test species (USEPA and USACE, 1998).

Reference USEPA. 2002. 7-Day Static Toxicity Test Using Larval Pimephales promelas. Rev

1. SOP #2026. Environmental Response Team. Compendium of ERT Standard

Operating Protocols. Office of Solid Waste and Emergency Response, U.S.

Environmental Protection Agency, Edison NJ. 

Website http://www.ert.org/products/2026.pdf Last Accessed: 2/12/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

87

Fact Sheet No. 2.1.3-7

Method Title Chronic Marine Macroalgae, Champia parvula, Sexual Reproduction Test,

NHEERL-AED SOP 1.03.001

Purpose This toxicity test measures the effects of toxic substances in effluents and receiving

waters on the sexual reproduction of the marine macroalga, Champia parvula,

during a forty-eight hour exposure.

Method Summary Macroalga are exposed to different concentrations of test concentrations of effluent

test medium over a 48-hour period. The selection of the effluent test concentrations

should be based on the objectivity of the test, however the maximum effluent

concentration which can be tested is 50% and the dilution factors may range from

0.5 to 0.3.

Three test chambers are devoted to effluent treatment and three chambers are

controls. Each test chamber will be filled with 100 mL of control or treatment water

and five female branches and one male branch. The water is not renewed

throughout the test, however each chamber is hand-swirled twice a day to mix the

water column. After 48 hours, the organisms are removed and placed in recovery

bottles. Investigators then count the cystocarps under a stereomicroscope.

NPDES Guidance suggest a 5-7 day test duration (Weber, 1991).

Data

Uses/Application

This is a laboratory test applicable to testing toxicity of effluents and receiving

waters. The results are used to determine the sexual reproduction ability of the

macroalgae exposed to different dilutions of the test medium.

Advantages This is one of few toxicity methods developed using marine plants.

Limitations The salinity of the test water must be 30 ppt +/- 2 ppt.

Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296, Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.03.001.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

88

Fact Sheet No. 2.1.3-8

Method Title Acute Marine Crustacean Bioassay, NHEERL-AED SOP 1.03.003

Purpose This static toxicity test measures the survival of the marine crustacean, Mysidopsis

bahia, after exposure to effluents and receiving waters for 96 hours in the laboratory.

Method Summary Mysids are exposed to various concentrations (minimum of 5) of effluent test

medium over a 96-hour period in 250 mL containers. The effluent concentrations

are commonly selected to approximate a geometric series (i.e., a dilution factor of

0.5). At least 20 organisms of a given species are exposed to each effluent

concentration. 

Test temperature is 20 +/- 2

N C. Organisms will be fed during the acclimation

period and during the toxicity test. Record survival at one hour and then daily

thereafter. After the 96 hours has past, final mortality and water quality

measurem ents are recorded.

Range-finding tests may be performed prior to these analyses to determine the

appropriate test media dilutions. Reference toxicant tests will also be performed

simultaneous to these tests. 

Other mysid shrimp species used for water column bioassays include Neomysis

americana and the Holmesimysis costata. 

Data

Uses/Application

This test may be conducted on effluents, leachates, or liquid phase of sediments.

The results will be used to determine the lethal concentration of test media that

causes 50% mortality (LC50).

Advantages Mysid shrimp are considered sensitive benchmark species (USEPA and USACE,

1998). Benchmark species comprise a substantial data base, represent the

sensitive range of a variety of ecosystems and provide comparative data on the

relative sensitivity of local test species. 

Limitations Mysid shrimp are near coastal species; they are used for testing in marine/estuarine

systems with salinities between 15 and 30 ppt (ASTM Standard Method E1191;

ASTM, 2001b.) 

Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296, Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.03.003.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

89

Fact Sheet No. 2.1.3-9

Method Title Acute Marine Fish Bioassay, NHEERL-AED SOP 1.03.003

Purpose These methods describe a 96-hour, static renewal acute effluent toxicity test using

the marine fish species Menidia beryllina and Cyprinodon variegatus

Method Summary Marine fish species, Menidia beryllina and Cyprinodon variegatus are exposed to

various concentrations (minimum of 5) of liquid-phase test media over a 96-hour

period in 250 mL containers. The test media concentrations are commonly selected

to approximate a geometric series (i.e., a dilution factor of 0.5). At least 20

organisms of a given species are exposed to each effluent concentration. 

Both species are generally used in salinities greater than 25 ppt.

Test temperature is 20 +/- 2

"C. Organisms will be fed during the acclimation period

and during the toxicity test. Test solutions must be replaced daily in the exposure

chambers. Survival is recorded at one hour and then daily thereafter. After the 96

hours have passed, final mortality and water quality measurem ents are recorded.

Range-finding tests may be performed prior to these analyses to determine the

appropriate test media dilutions. Reference toxicant tests will also be performed

simultaneous to these tests. 

A similar effluent toxicity test is also described in EPA guidance pertaining to

effluents and receiving waters (Weber, 1991).

Data

Uses/Application

This test may be conducted on effluents, leachates, or liquid phase of sediments.

The results will be used to determine the lethal concentration of test media that

causes 50% mortality (LC50).

Advantages Both fish species are considered benchmark species indicating that they comprise a

substantial database, represent the sensitive range of a variety of ecosystems, and

provide comparative data on the relative sensitivity of local test species (USEPA and

USACE, 1998).

Limitations Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296, Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.03.003.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

90

Fact Sheet No. 2.1.3-10

Method Title Chronic Estuarine Survival, Growth and Fecundity Test, NHEERL-AED SOP

1.03.005

Purpose These methods describe a toxicity test used to measure chronic effects from

effluents and receiving waters on the estuarine mysid, Mysidopsis bahia during a

seven-day, static-renewal exposure.

Method Summary The estuarine mysid shrimp, Mysidopsis bahia, are exposed to various

concentrations of test media over a 7-day period in 200 mL glass beakers. The

media concentration levels intend to span a range of those causing zero mortality to

those causing com plete mortality. 150 mL of the appropriate effluent dilution is

added to each beaker. The test can be run with smaller volumes of water as well.

(Ho, 2000).

Test temperature range is 26 - 27N C. New test media concentrations are prepared

daily. The test organisms are fed daily. 

Mortality is recorded over a 7-day period. Following the test, the live animals are

examined for eggs and the sexes are determined within 12 hours of the test

termination.

Other mysid shrimp species used in similar analyses include Neomysis americana

and Holmesimysis costata. 

A similar effluent toxicity test is also described in EPA guidance pertaining to

effluents and receiving waters (Weber, 1991)

Data

Uses/Application

The data from these tests will be used to determine the Lowest Observable Effect

Concentration (LOEC), the No Observable Effect Concentration (NOEC), the EC50

and the chronic value of the test medium.

Advantages Mysidopsis bahia are benchmark species indicating that they comprise a substantial

database, represent the sensitive range of a variety of ecosystems, and provide

comparative data on the relative sensitivity of local test species (USEPA and

USACE, 1998).

Limitations Mysidopsis bahia are near coastal species, They are used for salinities between 15

and 30 ppt (ASTM Method E1191; ASTM, 2001b).

Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296, Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.03.005.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

91

Fact Sheet No. 2.1.3-11

Method Title Chronic Echinoderm Fertilization Test, NHEERL-AED SOP 1.03.006

Purpose These methods describe a toxicity test used to measure chronic effects from

effluent and receiving waters to the gametes of the sea urchin, Arbacia

punctulata, during a 48 hour exposure.

Method Summary Arbacia punctulata are exposed to various effluent test concentrations that

should be based on the objectives of the study. A dilution factor of 0.5 is used

with this procedure, starting with a high concentration of 70% effluent . If the

effluent is known or suspected to be highly toxic, a lower range of effluent

concentrations should be used. 

Four females and four males are placed in shallow bowls, barely covering the

animals with seawater. Both females and males will be stimulated to release

their respective eggs or sperm. The egg stock and sperm are collected.

Sperm are diluted and mixed with seawater. This sperm suspension is then

distributed to vials and the number of sperm/mL are determined. 

The eggs are washed, diluted and counted. The test begins when diluted

sperm are added to each test vial containing eggs and various dilutions of the

effluent. All test vials are incubated for one hour at 20 NC. The suspension is

then mixed and incubated again for 20 minutes. Fertilization is then

determined using a Sedgwick-Rafter counting chamber. Fertilization is

indicated by the presence of a fertilization mem brane surrounding the egg.

Larval development may also be measured in this test. The egg suspension

is mixed and incubated for a longer period of time: 48 hours at 20 NC. At the

termination of the test, the total number of larvae and the appropriately

developed larvae are counted to determine survival and development per

treatm ent. 

A similar toxicity test is described in the EPA guidance pertaining to effluents

and receiving waters (Weber, 1991) and in Environment Canada’s method

EPS 1/RM/27.

Data Uses/Application This sperm cell toxicity test determines the concentration of a test substance

that reduces fertilization of exposed gametes relative to that of the control. 

This test may also be modified and used to assess pore water toxicity once

the pore water is extracted from whole sediments. 

Advantages Sea urchin toxicity tests have been proven to be extremely sensitive

indications of toxicity effects. The pore water toxicity tests with gametes and

embryos of sea urchins are approximately an order -of-magnitude more

sensitive than the 10-day solid-phase test with amphipods (Carr 2001). 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

92

Fact Sheet No. 2.1.3-11 (contd.)

Limitations Sea urchin toxicity tests are not considered standard toxicity tests, therefore

fewer laboratories currently perform the test. Sea urchin tests have a limited

salinity regime; therefore, they will be useful for samples from a marine

environm ents and select estuarine environments. 

Please note the list of general limitations for all liquid-phase toxicity tests in

the introduction to this section. 

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard

Operating Procedures and Field Methods Used for Conducting Ecological

Risk Assessment Case Studies. Technical Document 2296. Naval

Command, Control and Ocean Surveillance Center, RDT&E Division,

San Diego, CA. 

Website http://www.duxbury.battelle.org/co

mpendium/methods/NHEERLAED-SOP-1.03.006.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

93

Fact Sheet No. 2.1.3-12

Method Title Chronic Marine Fish Bioassay, NHEERL-AED SOP 1.03.004

Purpose This method describes a 7-day, static renewal chronic aqueous toxicity testing using

the marine fish species, Menidia beryllina and Cyprinodon variegatus.

Method Summary The fish species, Menidia beryllina and Cyprinodon variegatus, are exposed to

various concentrations of effluent over a 7-day period in 1-L test containers. The

media concentration levels span a range of those causing zero mortality to those

causing com plete mortality. To determine effluent concentrations, one of two

dilution factors is commonly used: approximately 0.3 or 0.5. 

Menidia beryllina and Cyprinodon variegatus are near coastal fish and are generally

used in salinities greater than 25 ppt.

Test temperature is 25o

+ 2

oC. The fish are fed daily. New dilutions of test media

are prepared daily. The old solution is drawn out with a siphon and the newly

prepared solutions are added to each chamber. The endpoints are survival and

growth.

Range-finding tests may be performed prior to these analyses to determine the

appropriate test media dilutions. Reference toxicant tests will also be performed

simultaneous to these tests.

NPDES Guidance also outlines a 9-day static-renewal test with Cyprinodon

variegatus to determine effluent effects on embryo larval survival and teratogenicity

(Weber, 1991). 

Data

Uses/Application

The results of this test will be used to determine the No Observable Effect

Concentration (NOEC), Lowest Observable Effect Concentration (LOEC) and the

Chronic Value (CHV). These results will determine the long term effects of those

sediment samples on the surrounding biotic community. 

Advantages Both fish species are considered benchmark species indicating that they comprise a

substantial database, represent the sensitive range of a variety of ecosystems, and

provide comparative data on the relative sensitivity of local test species (USEPA and

USACE, 1998).

Limitations Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296, Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.03.004.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

94

Fact Sheet No. 2.1.3-13

Method Title Toxicity Evaluations of Photoinduction of Polycyclic Aromatic Hydrocarbons (PAH):

In Situ Analysis

Purpose This method was designed to evaluate the degree of photoinduced toxicity during

wet-weather events via in situ experiments. 

Method Summary In situ chambers are constructed with two long rectangular windows to allow water

flow (holding 200 mL of water) and UV exposure (about 70% UV penetration) to

organisms inside the chambers. Cham bers containing Ceriodaphnia dubia are

placed on the sediment surface in the shade or in the sunlight (four replicates each)

at both a reference site and a test site. Dark mesh screens are placed over

chambers on the shaded devices to further block out UV wavelengths. 

The chambers are retrieved 48-hours after being placed in the river. At test

termination, the chambers are placed in coolers with site water and transported to

the laboratory. Percent survival is determined within 6 hours of chamber collection. 

Water quality analysis is conducted at both the reference site and the test sites.

Ultraviolet measurem ents are made at the surface and the bottom of the river.

Water is collected also at the surface and the bottom in 1-L amber polyethylene

sample bottles for PAH analysis. 

Similar freshwater crustacean species such as Daphnia magna and Daphnia pulex

may also be used for similar assays.

Data

Uses/Application

The results of this in situ evaluation are analyzed to determine the acute toxicity of

photoinduced PAHs. In particular, these tests are helpful in determine whether PAH

effects are more prevalent after major storm events in both agricultural and urban

environments. 

Advantages In situ toxicity testing reduces sampling and laboratory-related errors from the

assessment process. Fluctuating field conditions that may affect organism

response cannot be mimicked in the laboratory. 

These organisms are benchmark species indicating that they comprise a substantial

database, represent the sensitive range of a variety of ecosystems, and provide

comparative data on the relative sensitivity of local test species (USEPA and

USACE, 1998). 

Limitations Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference Ireland, D.S., Burton, G.A. and G.G. Hess. 1996. In Situ Toxicity Evaluations of

Turbidity and Photoinduction of Polycylic Aromatic Hydrocarbons. Environmental

Toxicology and Chemistry. Vol. 15: 4. 

p 574-581.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

95

Fact Sheet No. 2.1.3-14

Method Title Toxicity Evaluations of Photoinduction of Polycyclic Aromatic Hydrocarbons:

Laboratory Analysis of Storm water

Purpose This method was designed to evaluate the degree of photoinduced toxicity during wetweather events with in situ toxicity testing.

Method Summary Storm water runoff samples are collected in the field, returned to the laboratory and

fractionated to measure toxicity. After an increase in toxicity is observed in the

presence of UV radiation, a modified Toxicity Identification Evaluation procedure is

performed via the following protocol (no standard protocol exists for a TIE with Storm

water). 

Suspended solids are removed using glass fiber filters and the runoff is also filtered

through B and J Solid Phase ExtractionTM to remove all organics. With some

modifications, the 7-day Ceriodaphnia dubia chronic toxicity test follows the U.S.

Environmental Protection Agency’s standard methods. 

In each 30 mL beaker, 25 mL of sample is placed in various dilutions intended to span

a range of those causing zero mortality to those causing com plete mortality. One

organism is placed in each chamber. Water is renewed on days 3, 5, and 7.

Organisms are fed daily.

These tests are conducted in the presence of UV radiation. Lamps are constructed to

emit amounts of visible and UV radiation, by using two cool-white fluorescent lamps: a

350-nm and a 300-nm photreactor lamp. Survival, reproduction and PAH

concentrations are determined at the end of the 7-day test period. 

Data

Uses/Application

The results of this evaluation are analyzed to determine the acute toxicity of

photoinduced PAHS. In particular, these tests are helpful in determine whether PAH

effects are more prevalent after major storm events in both agricultural and urban

environments. 

Advantages Laboratory tests, in comparison with in situ tests, can control other parameters such

as temperature, pH etc in order to isolate effects as a result of contaminant levels.

These organisms are benchmark species indicating that they comprise a substantial

database, represent the sensitive range of a variety of ecosystems, and provide

comparative data on the relative sensitivity of local test species (USEPA and USACE,

1998). 

Limitations Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference Ireland, D.S., Burton, G.A. and G.G. Hess. 1996. In Situ Toxicity Evaluations of

Turbidity and Photoinduction of Polycylic Aromatic Hydrocarbons. Environmental

Toxicology and Chemistry. Vol. 15: 4. 

p 574-581. 

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

96

Fact Sheet No. 2.1.3-15

Method Title Growth and Scope for Growth Measurements with Mytilus edulis, NHEERL-AED

SOP 1.03.013

Purpose This method describes a test used to determine growth and the scope for growth

(SFG) index using the blue mussel, Mytilus edulis. To derive the SFG index, the

test measures the mussel’s clearance rate, respiration rate and assimilation

efficiency throughout the procedure.

Method Summary Mussels collected in different field conditions are sorted by size and placed in

individual clearance rate chambers. In these chambers, they are allowed to feed

overnight on algae pumping through the system at a set concentration. A Coulter

Counter is used to measure particles in order to determine the clearance rate.

The mussels are moved from the clearance rate chambers to chambers where

respiration measurement tools are set up. The respiration rate is measured in these

chambers with a radiometer and 450 mL glass respirometer vessels, which are

placed in the chambers with the individual mussels.

Assimilation efficiency is measured in the clearance rate chambers after the

chambers have been cleaned of fecal and algal matter and the mussels have feed

overnight. Fecal pellets are collected to determine the dry weight and ash weight of

feces. A similar procedure is completed with the cultured algae to obtain the dry

weight and ash weight of the food.

Mussel growth from pre exposure to post exposure is also determined using a

vernier caliper. Lastly, the mussel tissues are excised, dried and weighed.

Data

Uses/Application

The above method measures the following three physiological parameters in

mussels collected from different field conditions: clearance rate, respiration rate and

food assimilation efficiency. Clearance rate and assimilation efficiency

measurem ents are used to determine total amount of energy available, while

respiration rate is used to estimate metabolic energy costs.

These data are used to calculate the SFG index, which is a measure of the energy

available to an organism for somatic and reproductive growth after accounting for

routine metabolic costs. Mussels of similar physiological condition should

dem onstrate similar SFG responses under standardized conditions; therefore,

differences in SFG are attributed to persistent physiological effects of field exposure.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

97

Fact Sheet No. 2.1.3-15 (contd.)

Advantages Investigators have found reduced growth and ultim ately reduced fecundity and

fitness in Mytilus edulis after sustained reduction in SFG. The SFG index, therefore,

provides an additional way in which to quantify potential chronic effects from

changes in field conditions.

Mytilus edulis can withstand salinities ranging from 1 ppt to > 25 ppt. Mytilis is also

considered a benchmark species indicating that they comprise a substantial

database, represent the sensitive range of a variety of ecosystems, and provide

comparative data on the relative sensitivity of local test species (USEPA and

USACE, 1998).

Limitations Please note the list of general limitations for all liquid-phase toxicity tests in the

introduction to this section. 

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296, Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.03.013.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

98

Fact Sheet No. 2.1.3-16

Method Title Microtox® tests, NHEERL-AED SOP 1.03.009

Purpose This method determines acute toxicity in effluents, receiving waters and elutriates

through use of a bioluminescent bacteria. These tests are also applicable to wholephase sediment samples. 

Method Summary Microtox® tests measure acute toxic effects in luminescent bacteria

(Photobacterium phosporeum) after exposure to effluents, receiving waters or

elutriate samples (aqueous phase of a 4:1 water to sediment mixture). Metabolic

inhibition in the luminescent organisms occurs if a sample is toxic, and the

subsequent reductions in light output are used to derive a dose-response curve from

which the effective concentration of the sample is determined. 

Water samples are evaluated at 45, 22.5, 11.3, and 5.6 % dilutions of the fullstrength samples. The Microtox® reagent (Photobacterium phosphreum) are

placed in the Microtox® turrets to measure initial light levels. The reagent is then

added to the respective dilutions. Generally, the reagent reacts quickly to organic

compounds and toxicity is elicited within 5 minutes. Metals take longer to elicit

toxicity; up to 15 minutes should be allowed. After a set exposure time, the reagent

and dilution are placed in a turret of the Microtox machine and the light levels are

recorded. The Microtox® test is also conducted with undiluted samples (e.g. 100%

test media).

Data

Uses/Application

Microtox® tests can be used to determine if the toxicity of liquid or solid phase

samples extracted from a contaminated environm ent. They can either be used

alone or in concert with other analyses screening samples as a preliminary step to

identifying potential contamination.

Advantages Microtox® tests are much quicker than traditional toxicity tests and they may be

performed in a field laboratory. They can be performed with liquid phase samples

from both freshwater and marine environments.

The apparent toxicity of elutriates can be a function of extraction solvent and overall

procedure. 

Limitations Bacteria response to potential toxicity in water or sediment may not be

representative of the response of a larger organism encountering the same medium

in the wild.

Reference USEPA. 1993a. Biological and Chemical Assessment of Contaminated Great

Lakes Sediment, EPA 905-R93-006. Great Lakes National Program Office, U.S.

Environmental Protection Agency, Chicago, IL. 

Website http://www.duxbury.battelle.org/compendium/met

hods/NHEERL-AED-SOP-1.03.009.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

99

Fact Sheet No. 2.1.3-17

Method Title Com parative Toxicity of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin to Seven

Freshwater Fish Species During Early Life-Stage Development

Purpose To determine and compare the toxicity of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin

(TCDD) to early life stages of freshwater fish on the basis of waterborne

exposure of fertilized eggs.

Method Summary Fish species suitable for this method include northern pike, white sucker, lake

herring, fathead minnow, channel catfish, medeka, and zebrafish. Eggs are

obtained by stripping the adult fish and artificially fertilizing the eggs in clean

water. The exposure system consists of three tanks, one egg control tank, one

solvent control tank and one test egg tank. Solvent and TCDD are added to the

tanks (minus the control tank) 30 minutes prior to the start of each egg

exposure. The recirculating flow rate in the tanks is adjusted to approximately

80 mL/m in for all tests. Eggs are checked for fungus daily during the incubation

period and dead eggs are removed and recorded. Following the hatch, all

organisms are released into clean-water tanks and observed daily for signs of

TCDD toxicity, which include edema, hemorrhaging, head and spinal

deformities, lethargy, loss of equilibrium, skin discoloration, and mortality.

The concentrations of TCDD in the test tanks are also measured and recorded.

GC/MS analysis is used to determine the spercific activity and radiopurity of

TCDD concentrations at the beginning and end of each exposure.

A similar method for early life-stage toxicity testing is described in Environment

Canada Method EPS 1/RM/28, 1

st and 2

nd editions. Available from

Environmental Protection Publications, Environmental Protection Service,

Environment Canada, Ottawa, Ontario, K1A OH3, Canada.

Data Uses/Application TCDD is the most toxic of the hydrophobic, halogenated aromatic comounds

that include polychlorinated dibenzodioxins (PCDDs), dibenzofurans (PCDFs),

and polychlorinated biphenyls (PCBs). In addition, the toxic effects of TCDD

are bioaccumulatable in aquatic systems. Because of its association with

aquatic sediments, TCDD poses a potential risk to aquatic organisms. This

method describes the determination of toxicity in the early-life stages of several

fish species.

Advantages For many fish species, the toxicity of TCDD is increased when the eggs are

exposed prior to the hatch. The mechanism involved in TCDD uptake is known

to be extremely functional in the very early life stages of fish and results of

toxicity (other than mortality) are measurable in the post-hatch population.

Limitations Comparisons of TCDD toxicity using this method to previously tested fish

species are difficult because test conditions (exposure regimes and life stages)

may vary so much. Also, within a fish species, sensitivity to TCDD is dependant

on the age and size of the organism, and the exposure time and stage of

development.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

100

Fact Sheet No. 2.1.3-17 (cont’d)

Reference Elonen, Gregory E., RL Spehar, GW Holcombe, RD Johnson, JD Fernandez,

RJ Erickson, JE Tietge, and PM Cook. 1998. Com parative Toxicity of 2,3,7,8-

Tetrachlorodibenzo-p-Dioxin to Seven Freshwater Fish Species During Early

Life-Stage Development. Environmental Toxicolgy and Chemistry, Vol. 17, No.

3, pp 472-483.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

101

2.2 Sediments

Sediment samples are collected at Superfund sites with the same objectives in mind as when

collecting water samples. These objectives, as stated in Section 2.1, are the following:

• To determine if the contaminant is hazardous by identifying its composition and

characteristics;

• To determine if there is an imminent or substantial threat to public health or welfare or

to the environment;

• To determine the need for long-term action;

• To develop containment and control strategies;

• To evaluate appropriate disposal/treatment options; and,

• To verify treatment goals or clean up levels (USEPA, 1994a).

To adequately characterize a site, the plans for sediment sampling and related analyses must be

developed in consideration of the site characteristics. Therefore, the following fact sheets relating

to sediment are divided into sections pertaining to field sample collection and processing,

chemical and physical analyses, and biological analyses. These fact sheets intend to provide

Superfund managers with a summary of the existing methods that may be applicable to their site,

the method’s relative strengths, and the method’s relative weaknesses.

2.2.1 Field Sample Collection and Processing, In Situ Data Acquisition

Section 2.2.1 provide field sample collection and processing methods for sediments. Sediments

are collected at Superfund sites for sediment chemistry, toxicity and benthic community analyses

to determine the extent of chemical contamination and impact of contamination on the site . The

two primary methods for sediment collection include sediment grab samplers and sediment core

samplers. However, there are quite a few different types of grabs and cores. These different

samplers are summarized following their respective fact sheets in Tables 2.2.1-1 and 2.2.1-2. 

Other sediment and processing collection methods are also provided for situations where grab

and core deployment is un necessary or impossible due to physical interferences. These sample

collection methods were gathered from the following information sources:

• The USEPA’s Office of Water

• The USEPA’s Office of Research and Development

• ASTM

• Standard Methods for Examination of Water and Wastewater, 1999

• Puget Sound Water Quality Action Team

• The USEPA’s Coastal EMAP Program

• The USEPA’s Great Lakes Program Office

Field observations and preliminary identification of sediment type are pertinent to all

sediment collections. Guidelines for making visual observations of sediment type can be

found in the US Army Corps of Engineers manual on Soil Sampling, EM 1110-1-1906.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

102

Fact Sheet No. 2.2.1-1

Method Title Grab Sampling

Purpose To collect samples of the benthos for quantitative or qualitative sampling

procedures, intended to determine sediment chemistry, toxicity and/or benthic

com munity composition and abundance.

Method Summary Grabs are to be lowered slowly from a boat or by hand into the water column.

When most grabs reach the bottom their weight will cause them to penetrate the

substrate (areas of 0.02 to 0.5 m

2

and depths ranging from 5 to 15 cm). The

slack on the cable allows the locking lever to release, therefore permitting the

movement that allows the horizontal locking bar to drop out of the locking notch

and allow the jaws to close as the device is raised. Other grabs are closed by

spring action or some other mechanical device after penetrating the substrate. 

After the grabs are brought to the surface, they are examined for acceptability.

Collection of undisturbed sediment requires that the sampler :

-create a minimal pressure wave when descending

-form a leakproof seal when the sediment sample is taken

-prevent winnowing and excessive sample disturbance when ascending

-allow easy access to the sample surface in order that undisturbed subsamples

may be taken (USEPA, 1992c).

The required amount of sediment is removed for sub-sampling and placed in the

appropriately cleaned sample containers. (See Fact Sheet 2.2.1-8.) 

Other USEPA, ASTM and APHA documents provide grab sample collection

information and details pertaining to different grab samplers (USEPA, 1992c;

ASTM Method E1391 (ASTM 2001b); Standard Method 10500 (APHA 1999).

There are many types of grabs that vary in penetration depth, surface area

sample and sampling various substrate types. The various grabs and their

respective advantages and disadvantages are detailed on the following matrix

(Table 2.2.1-1).

Data

Uses/Application

Grab sampling devices collect sediments that may be used to analyze for

sediment chemistry, sediment toxicity, and/or the samples may be sieved to

determine benthic com munity composition and abundance. They are commonly

used in estuarine and marine monitoring programs due to their ability to provide

reliable quantitative data at a relatively low cost. 

Advantages Since there are many types of grabs, it is easy to find one that will be effective in

various site conditions (Table 2.2.1-1). Grabs are able to sample a larger surface

area than most coring devices.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

103

Fact Sheet No. 2.2.1-1 (contd.)

Limitations Grabs have a relatively shallow and variable depth of penetration depending on

the sediment properties. As the grab sampler bites into the sediment, the

sediment is inevitably folded resulting in the loss of information concerning the

vertical structure of sediments. 

The shock wave that results from the grab’s deployment also results in a loss of

the fine surface sediments and water-soluble compounds and volatile organic

compounds present on the surface of the sediment. 

Reference USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating

the Biological Integrity of Surface Waters. EPA/600/4-90/030. Office of Research

and Development, U.S. Environmental Protection Agency, Washington, D.C.

Website http://www.epa.gov/bioiweb1/pdf/benthos_

methods_ch5.pdf

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

104

Table 2.2.1-1. A Summary of Sediment Grab Devices (USEPA, 1992c; APHA, 1999; USEPA, 1990b; Murdoch and Azcue,

1995)

Sediment Grab Habitat

Substrate Type, Surface

Area and Penetration

Depth Advantages Disadvantages

Van Veen Open sea and

large lakes

Sand, silt, clay or similar

substrates

Surface Area: 0.25 m

2

Penetration Depth: 5-7 cm

This grab can sample

most sediment types.

Large enough to permit

sub-sampling.

Shock wave from descent

may disturb “fines.” Possible

incomplete closure of jaws

results in sample loss.

Possible contamination from

metal frame construction.

Sample must be further

prepared for analysis. 

Young Grab

(fluorocarbon

plastic or kynar

lined modified

0.1 m2van

veen)

Lakes and

marine areas

Sand, silt, clay or similar

substrates

Surface Area: .1 m

2

Lined grabs eliminate

metal contamination.

Small size reduces

pressure wave.

Expensive, requires winch.

Orange Peel Marine

environments

and deep lakes

Sandy substrates

Round grab; collects up to

1600 cm3of sediment

Comes in a range of

sizes, works in deep

water, closes relatively

well to prevent sample

loss, good for

reconnaissance.

Very heavy, requires power

winch. Does not sample

constant area and depth.

Smith McIntyre Marine,

estuarine,

adaptable to

large rivers,

lakes and

reservoirs

Used on most substrates;

designed specifically to

sample hard substrates

Surface Area: 0.2 or 0.1m2

This grab is stable and

easy to control in rough

water.

Loss of fines. Possible

contamination from metal

frame construction. Very

heavy, requires a power

winch.

Shipek Used primarily

in marine

waters and

large inland

lakes and

reservoirs

Sand, gravel, mud and clay

Surface Area: 0.4m2

Penetration Depth: 10 cm

This grab is good for

collecting a small sample

in deep water. It has a

sample bucket from which

a sub-sample may be

obtained. It retains finegrained sediments

effectively.

Possible contamination from

metal frame construction.

Very heavy, requires a power

winch. 

Petersen Freshwater

lakes,

reservoirs,

rivers and

estuaries

Useful on most substrates;

especially hard substrates

with swift currents and

deep water.

This grab can obtain a

large sample and it can

penetrate most substrate. 

Heavy, may require winch.

No cover lid to permit

subsampling. All other

disadvantages of Ekman and

Ponar.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

105

Table 2.2.1-1. (contd.)

Petite Ponar Deep rivers,

lakes and

reservoirs

Moderately hard

sediments, such as sand,

silt and mud. 

Surface Area: .02 m

2

This grab has a good

penetration for a small

grab, side plates and

screens to prevent

washout, and it can be

operated by hand without

a boat or a winch.

It is not effective for sampling

deep burrowing organisms or

for sampling clay substrate. 

Ponar Deep rivers,

lakes and

reservoirs

Fine to coarse textured

sediments such as clay,

hard pan, sand, gravel and

muck. Less effective in

softer sediments.

Surface Area: .05 m

2

This grab is very efficient

for hard sediments;

considered universal

sampler due to ability to

collect adequate samples

from most substrate

types.

Shock wave from descent

may disturb “fines.” Possible

incomplete closure of jaws

results in sample loss.

Possible contamination from

metal frame construction.

Sample must be further

prepared for analysis. 

Ekman or Box

Dredge

Can be used

from boat,

bridge, or pier

in waters of

various depths

Consolidated, fine textured

sediments. Efficient in soft

sediments, such as silt,

muck and sludge in water

with little current.

Surface Area: .05 m

2

This grab is light weight; it

can be operated by hand.

It is commonly used for

benthic evaluations.

Obtains a larger sample

than coring tubes. Can be

subsampled through box

lid.

Shock wave from descent

may disturb “fines.” Possible

incomplete closure of jaws

results in sample loss.

Possible contamination from

metal frame construction.

Sample must be further

prepared for analysis. Difficult

to use in rocky or sandy

bottoms
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

106

Fact Sheet No. 2.2.1-2

Method Title Core Samplers

Purpose To collect an undisturbed sediment sample from varying depths in any

substrate. 

Method Summary Prior to deployment, the sampling device is inspected to see that the sediment

retainer behind the cutting edge will provide a good seal. For a box corer, the

cable must feed through the pulley system properly and the spade must rotate

freely. All portions of the sampling device that will be in contact with the sample

(i.e., the core tube and the core liner, where applicable) should be constructed

of noncontaminating material. 

Cores are deployed from a suitable vessel. Cores use either inertia (i.e.,

gravity cores, piston cores) or mechanical motion (hammering or vibration) as

the primary driving force to achieve the desired penetration depth depending

on the specified depth and the sediment properties.

The amount of pull that is required to extract a core tube from the substrate

depends on the specific gravity of the device and its contents, plus the amount

of frictional force against the surface of the core tube walls that must be

overcome. During the extraction, the wire strain should be steady and

continuous; the vessel should be held stationary directly above the coring

device. Once clear of the bottom, winch take-up speed may increase. 

Once the sampling device is onboard the vessel, one or both ends of the core

tube is capped if possible. Overlying water is siphoned off at the top of the

core tube (after allowing for settling time). The length of the sediment core

should be determined by comparing measurem ents of the length of the core

material against the overall penetration depth. The ratio of penetration depth

to core material length is calculated to determine the compaction of the

sediment during coring.

If the core is acceptable (for example, acceptable depth of penetration, surface

layer intact), the core tube (or liner) should be labeled with the core

identification number, collection date, core orientation, and length of core

material collected. Until the core sample can be extruded or split into sections,

the core tube should be secured in an upright position, taking care not to avert

the core. Cores should be split within 24 hours of collection.

Other USEPA, ASTM and APHA method documents provide specifics

regarding core sampling and determining what type of core device is

applicable (USEPA, 1990b; ASTM Method E1391(ASTM 2001b; Standard

Method 10500 (APHA, 1999).

There are also several types of cores; these cores and their respective

advantages and disadvantages are detailed on the following table (Table 2.2.1-

2).
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

107

Fact Sheet No. 2.2.1-2 (contd.)

Data Uses/Application Core samples can be analyzed for sediment chemistry, sediment toxicity

and/or benthic com munity analyses. 

Advantages Core sampling removes sediments with less disruption than grab and dredge

sampling. Gravity corers or hand-driven corers can collect sediments up to 1

to 2 m in depth. When vibratory corers or others using hydralics for sediment

penetration, corers can collect sediments up to 10 m in depth. Corers are

more efficient than grabs. Corers are the most accurate samplers of benthic

macroinvertebrate populations. 

Limitations Core sampling provides an imprecise estimate of the standing crop of

macrobenthos, because of the sm all area sampled. Gravity operated samples

have limited surface area, and they require a boat and powered winch. Cores

generally sample an area 13 to 26 cm2

.

Cores are more difficult to handle than grabs in rough water. They do not work

well in sandy sediments. 

Reference PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment,

Water Column, and Tissue in Puget Sound, Puget Sound Protocols and

Guidelines. Puget Sound Water Quality Action Team, Olympia, W A. 

Website http://www.psat.wa.gov/Publications/

protocols/protocol_pdfs/field.pdf

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

108

Table 2.2.1-2. A Summary of Sediment Coring Devices (USEPA, 1992c; APHA, 1999; USEPA, 1990b; Murdoch and

Azcue, 1995)

Sediment Core

Core liner

diameter;

Penetration

Depth

Substrate Type;

Habitat Advantages Disadvantages

Fluorocarbon

plastic or Glass

tube

Core diameter:

Varies

Penetration

Depth: Up to 50

cm

Shallow

wadeable waters

or deep waters if

SCUBA

available

Preserves layering and permits

historical study of sediment

deposition. Minimal risk of

contamination.

Small sample size requires

repetitive sampling. 

Hand Corer Core diameter:

3.5 to 7.5 cm ID

Penetration

Depth: 50 to

120 cm

Soft or semicompacted

sediment in

shallow

wadeable

waters; deep

waters if SCUBA

available

Handles provide for greater ease

of substrate penetration.

Preserves layering and permits

historical study of sediment

deposition. Minimal risk of

contamination.

Small sample size requires

repetitive sampling. Careful

handling necessary to prevent

spillage. Requires removal of

liners before repetitive sampling.

Slight risk of metal

contamination from barrel and

core cutter.

Push Core Varies Soft or semicompacted

sediment in

shallow water

Push cores with shallow

penetration and in relatively

shallow water do not require a

winch. 

They are difficult to deploy in

areas with strong currents and

deep water. They may be

difficult to retrieve if the

penetration depth exceeds 50

cm.

Phleger Core

(Gravity Core)

Core diameter:

3.5 cm ID

Penetration

Depth: Up to 50

cm

Soft substrates,

semi-compacted

substrates, peat

and vegetated

roots in shallow

lakes and

marshes

There is low risk of undisturbed

sample contamination. This

sampler maintains sample

integrity relatively well. It does

not require a winch.

Careful handling is necessary to

avoid sediment spillage. Small

sample, requires repetitive

operation and removal of liners.

Time consuming. 

KB Core

(Gravity Core)

Core diameter:

3.5 cm ID and 5

cm

Penetration

Depth: Up to 70

cm

Soft, fine-grained

substrates

Useful for obtaining estimates of

the standing crop of

macrobenthos in soft substrates.

Winch is not required.

The messenger system used to

close the core is sometimes

ineffective when the core does

not penetrate the sediment

vertically.

Box Core

(Gravity Core)

Surface area:

typically 0.04

and 0.1 m

2

Penetration

Depth: Up to

1 m

Soft sediments This core samples the same

surface area as a grab, but it

disturbs the surface less. Allows

for subsampling.

This core does not penetrate

sediments deeply. Hard to

handle, very heavy.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

109

Table 2.2.1-2. (contd.)

Benthos Gravity

Core

Core diameter:

6.6 cm and 7.1

cm

Penetration

Depth: Up to

3 m

Soft, fine-grained

substrates

It has stabilizing fins that

promote vertical penetration. It

also has a valve system that

prevents sample loss. It can

sample substrates from great

depths.

More difficult to deploy and

retrieve, large device, very

heavy. 

Alpine Gravity

Core

Core diameter:

3.5 cm

Penetration

depth: 0.6, 1.2

and 1.8 m

Compacted

substrates from

depth

It can sample substrates from

great depths.

Lack of stabilizing fins makes

vertical penetration difficult. This

core also disturbs surface

sediments significantly. Very

heavy.

Multi-Gravity

Core

Core diameter

and penetration

depth vary.

Varies Multiple samples can be taken

from one site for comparative

studies, evaluation of sediment

samples and determination of

sediment heterogeneity over a

small area.

Large and difficult to deploy and

retrieve.

Piston Core Core diameter:

typically 3, 5, or

6"

Penetration

depth: 3-20

meters

Soft substrates This core can be used for

samples requiring significant

penetration depths. Relatively

undisturbed samples

A heavy crane is needed with

lifting capacity over 2000kg to

deploy and retrieve piston cores.

This device is not suitable for

sediment profiles since it

disturbs the top layer.

VibratoryHammer Core

Core diameter:

typically 3, 5, or

6"

All types This core can be used in all

types of sediments. Method of

choice for many environmental

dredging studies.

Large and difficult to deploy and

retrieve.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

110

Fact Sheet No. 2.2.1-3

Method Title Hand Collection

Purpose This method describes how sediment samples can be collected by hand in the

intertidal zone with a favorable tide.

Method Summary Sediment samples may be collected by hand with a variety of sampling implements

such as spoons or trowels for surface sediments, or with hand augers or corers for

collecting sediments at discrete depths. Any sampling implement that comes in

contact with the sample should be constructed of stainless steel or TeflonTM. If

individual sample collection kits are not available for each sampling location,

sampling equipment should be thoroughly decontaminated between stations by

scrubbing with a phosphate-free detergent solution, followed with a thorough rinse

with analyte-free water. If heavy contamination by metals or organic contaminants is

expected at the site, sampling equipment may be rinsed with methanol, acetone or a

50:50 acetone/hexane mix for organics or 10% HNO3

for metals. 

Once the samples have been collected with one of the aforementioned tools, the

samples should be homogenized in a stainless steel bowl with a stainless steel or

TeflonTM spoon or spatula. Sample aliquots are transferred to appropriate laboratory

supplied containers and preserved as required.

Data

Uses/Application

These methods are used in shallow waters where a core is not needed. Sediment

samples can be analyzed for sediment chemistry, sediment toxicity and/or benthic

com munity analyses. 

Advantages Hand collection is less expensive and labor intensive than other sediment collection

techniques.

Limitations Sediments can only be collected by hand in shallow waters or locations where the

tide has exposed the desired sampling area. This collection procedure also

increases human exposure to potential contaminants present in the samples. 

Reference PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water

Column, and Tissue in Puget Sound, Puget Sound Protocols and Guidelines. Puget

Sound Water Quality Action Team, Olympia, W A. 

Website http://www.psat.wa.gov/Publications/pro

tocols/protocol_pdfs/field.pdf

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

111

Fact Sheet No. 2.2.1-4

Method Title Hand Collection at Depth with SCUBA Equipment

Purpose To sample benthos in locations where conventional sampling devices are not

practical and the water depth is too deep for hand-collection.

Method Summary The collection of sediment samples by a diver should be considered when

undisturbed samples are required, particularly for studies of the sediment-water

interface. SCUBA certified professionals can conduct the following types of benthos

sampling: placement and retrieval of artificial substrate; use of suction samplers;

sampling with a quadrate frame; and , perhaps most importantly, identifying and

delineating substrate types for purposes of determining sampling effort (stratified

sampling) and choice of samplers. 

Data

Uses/Application

SCUBA divers can use traditional sediment collection devices that are then analyzed

for sediment chemistry, sediment toxicity, and/or the samples may be sieved to

determine benthic com munity composition and abundance.

Advantages Allows for benthic sampling in locations that are inaccessible by conventional grab

sampling. Allows for more precise sampling of the sediment-water interface.

Limitations Requires certified SCUBA divers who will comply with rigid safety standards.

Somewhat expensive. The diver’s visibility can be obscured if fine-grained

sediments are disturbed or if the water is turbid. 

Reference USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating

the Biological Integrity of Surface Waters. EPA/600/4-90/030. Office of Research

and Development, U.S. Environmental Protection Agency, Washington, D.C.

Website http://www.epa.gov/bioiweb1/pdf/bentho

s_methods_ch5.pdf

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

112

Fact Sheet No. 2.2.1-5

Method Title Sediment Traps

Purpose Sediment traps collect settling particulate matter (SPM) which provides useful data

for studies of sedimentation rates and resuspension of bottom sediments.

Method Summary If collecting SPM for chemical analysis, the traps should be cleaned with a

phosphate-free detergent solution, then sequentially rinsed with hot water, 10

percent HNO3, analyte-free, pesticide grade acetone, and finally, wrapped in

aluminum foil until deployment in the field. If the sediment trap is constructed of

plexiglass, the acetone rinse should be avoided as acetone will damage the

plexiglass. Prior to deployment, the traps should be filled with two liters of highsalinity, analyte-free water containing a preservative to reduce microbial degradation

of the samples during deployment period. 

SPM samples are collected by retrieving the traps and removing the overlying water

in the collection cylinders using a peristalic pump. The water immediately overlying

the trapped sediment is analyzed to determine the salinity and the presence of

preservative to determine if the trap was disturbed during the deployment. The SPM

is then transferred to sample containers and taken to an analytical laboratory for

processing. The particulate fraction of the SPM is removed by centrifuge and split

into sample aliquots for chemical analysis.

Sediment trap designs vary. The trap must be made suitable for the study area.

Considerations on biofouling must be addressed and controlled appropriately with

regard to the type of samples being collected.

Data

Uses/Application

SPM data are used for studies of sedimentation rates and for sediment transport

studies.

Advantages Sediment traps can collect relatively large volumes of suspended matter for

transport studies (compared to filtration).

Limitations Construction, deployment, and retrieval often require resources beyond typical field

studies. Biological invasions of traps typically occur. Controlling agents are not

suitable for all applications.

Reference PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water

Column, and Tissue in Puget Sound, Puget Sound Protocols and Guidelines.

Puget Sound Water Quality Action Team, Olympia, W A. 

Website http://www.psat.wa.gov/Publications/pro

tocols/protocol_pdfs/field.pdf

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

113

Fact Sheet No. 2.2.1-6

Method Title Russian Peat Borer

Purpose To collect discrete, relatively uncompressed sediment samples. 

Method Summary The Russian Peat Borer is a manually driven, chambered-type, side-filling core

sampler, and its com ponents include a stainless-steel core tube, aluminum

extension rods, a stainless-steel turning handle, and a Delrin ® core head and

bottom point that support a stainless-steel cover plate. 

To collect a sediment sample, the bottom point of the Russian Peat Borer is

manually inserted into sediment, with the blunt edge of the core tube turned against

the cover plate to prevent sediment from entering the tube during penetration. A

slide-hammer mechanism can be used to drive the sampler through highly

consolidated sediment or peat that is hard to penetrate. Once the sampler is driven

into the sediment to the desired depth, the core tube is turned 180 degrees

clockwise. This allows the core tube to rotate and its sharp edge to longitudinally cut

through the sediment, collecting a semicylindrical sediment core. While the core

tube is manually turned, the stainless-steel cover plate provides support so that the

collected material is retained in the core tube. The sampler is then rotated and

placed on the sampling platform in such a way that the core tube is above the cover

plate. The core tube is then manually turned counterclockwise, rotating the tube and

exposing the semicylindrical core sample on the cover plate.

Data

Uses/Application

The sampler can collect discrete, relatively uncompresssed core samples from

shallow to deep depth intervals with out disturbing the sediment stratification. 

Advantages The sampler is lightweight and easy to operate, requiring minimal training and skill.

It does not require disassembly to extrude the sample. It requires no support

equipment other than two sawhorses for support during sample extrusion.

Limitations For deployment in deep water applications, the sampler requires extension rods.

This sampler is not equipped with disposable core liners. During deployment, the

cover plate is exposed to different layers of sediment. Also, partially decomposed

plant matter or sm all stones may cause the core tube to remain in the open position

during sampling retrieval. To use this sampler, sediment must offer enough support

to keep the cover plate stationary and allow rotation of the tube core.

Reference USEPA. 1999d. Innovative Technology Verification Report: Aquatic Research

Instruments Russian Peat Borer, EPA/600/R-01/010. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/ordntrnt/ORD/SITE/r

eports/600r01010.htm

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

114

Fact Sheet No. 2.2.1-7

Method Title Split Core Sampler for Submerged Sediments

Purpose To collect undisturbed core samples of sediment up to a maximum depth of four feet

below sediment surface.

Method Summary The fully assembled sampler is manually lowered into the water in such a way that

the coring tip is placed on the sediment surface. The sampler can then be either

manually pushed with the cross handle or driven with the slide-hammer or an

electric hamm er to the desired sediment depth. The sampler is removed from the

sediment either manually by reverse hamm ering or with the tripod-mounted winch.

Once the sampler has been retrieved, either the interlocking split core tubes are

disassembled or the coring tip or top cap is removed to allow removal of the core

tube liner.

Data

Uses/Application

This sampler is designed to collect undisturbed, cylindrical core samples of various

types of sediment, including saturated sands and silts, to a maximum depth of 48

inches below the sediment surface.

Advantages The sampler is lightweight and easy to operate, requiring minimal skills and training.

An (SOP) accompanies the sampler when it is purchased. A combination of

stainless-steel split core tubes can be used to collect 6- to 48-inch-long sediment

cores. Plastic core tube liners can be used with the sampler. The sampler design

uses a ball check valve-vented top cap. This feature: (1) allows air and water to exit

the sampler during deployment, (2) prevents water from entering the sampler during

retrieval, and (3) creates a vacuum to help retain a sediment core during sampler

retrieval.

Limitations The core tube liner is exposed to different layers of sediment contamination during

sample collection. The ball check valve-vented top cap may become clogged if the

sampler is deployed in such a way that the top cap is below the sediment surface.

The sampler cannot collect discrete samples from various sediment depths and is

subject to core shortening. Because an external power source is required to operate

the electric hammer, the sampling platform must be able to accommodate the

weight and size of a portable generator. Use of the tripod-mounted winch or similar

device limits the sampling platform locations from which the sampler can be

deployed.

Reference USEPA. 1999e. Sediment Sampling Technology Art’s Manufacturing and Supply,

Inc., Split Core Sampler for Submerged Sediments, Superfund Innovative

Technology, Evaluation Program, EPA/600/R-01/009. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/ORD/SITE/reports/6

00r01009.htm

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

115

Fact Sheet No. 2.2.1-8

Method Title Sediment Processing for Chem istry and Toxicity Testing

Purpose These methods describe protocols for removing sediment samples from sampling

collection devices and processing them for laboratory analyses. 

Method Summary A clean stainless steel or TeflonTM spoon is used to remove sediments from grab

samples for these analyses. Surficial sediments are removed (usually 0-2 cm) and

placed in a stainless pot. The pot is then placed in a cooler on ice (not dry ice) and

stored at 4

oC. This process is repeated until sufficient quantity of sediment has

been collected and composited with the other sediments (approximately 4 L).

Sediments from sediment cores are extruded and subsampled also for the following

analyses.

For organic analysis, 250 cc of sediment is placed into a 500 mL pre-cleaned, glass

bottle for chemical analysis. The sample number is recorded and the jar is wrapped

in bubble wrap (to prevent breakage) and packed in ice. 

For metals, approximately 100 cc of sediment is placed into pre-cleaned, plastic

(HDPE) sampling jars. The sample number is recorded and the sample is kept on

ice at 4

B C. 

For Total Organic Carbon, approximately 100 cc of sediment is placed into precleaned, glass sampling jars. The sample number is recorded and the sample is

kept on ice at 4

B C. 

For sediment grain size, approximately 100 cc of sediment is placed into a clean

plastic (HDPE) sampling jar or whirl-pak bag. The sample number is recorded and

the sample is kept on ice at 4

B C.

For sediment toxicity, the volume of sediments collected will vary depending on the

objectives and methods of the specific toxicity tests.

Data

Uses/Application

These methods describe the steps necessary to prepare samples for toxicological or

chemical analyses. The applicability of these tests and analyses are described in

their respective fact sheets (Sections 2.2.2 and 2.2.3).

Advantages Compositing the surface 2 cm of sediment from multiple grabs or cores allows a

representative samples to be collected. This process provides sufficient sediment

volume for toxicity testing and supporting chemistry and physical measurements. 

Limitations Extreme care must be taken to ensure that the samples are not contaminated during

sampling or processing procedures.

Reference USEPA. 2000b. Coastal 2000 Northeast Component: Field Operations Manual,

Environmental Monitoring and Assessment Program (EMAP). EPA/620/R-00/002.

Office of Research and Development, U.S. Environmental Protection Agency,

Washington, DC.

Website http://www.epa.gov/emap/nca/html/docs/c2kn

efm.pdf

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

116

Fact Sheet No. 2.2.1-9

Method Title Sediment Processing for Elutriate Toxicity Tests

Purpose This method describes how elutriate samples are prepared for toxicity tests from the

original whole-phase sediment samples. 

Method Summary Elutriates are prepared with one part sediments (from sampling location) and four

parts reconstituted dilution water. A 200 g sub-sample of homogenized sediment is

removed from the containers and placed in a centrifuge bottle with 800 g of dilution

water (usually site water). The contents are weighed, mixed and centrifuged. The

overlying water is removed and the elutriate sample is sub-sampled and stored in 1-

L amber bottles equipped with TeflonTM-lined lids until testing. Test organisms are

exposed to varying concentrations of the elutriate material (0%, 12.5 %, 25%, 50%

and 100%) for a designated period of time

Data

Uses/Application

Elutriate toxicity tests provide information that can be used to support inferences

about the potential toxicity of the contaminated sediments from which the elutriates

are prepared and to identify the biologically active constituents of the contaminated

sediment. 

Elutriate tests are commonly used to evaluate proposals to discharge dredged

material into ocean waters, and to evaluate the potential of the dredged materials to

impact ocean ecology. 

Advantages Elutriate tests allow the investigator to assess the potential hazard of contaminated

sediments to aquatic organisms, to compare the relative toxicity of contaminated

sediments from different locations, and to study the biological availability of the

contaminants associated with sediments. 

Limitations Elutriate toxicity tests do not necessarily reflect the toxicity of in-place sediment. 

Reference USEPA. 1993b. Assessment and Remediation of Contaminated Sediments

(ARCS) Program: Biological and Chemical Assessment of Contaminated Great

Lakes Sediment, EPA 905-R93-006. Great Lakes National Program Office, U.S.

Environmental Protection Agency,

Chicago, IL.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

117

Fact Sheet No. 2.2.1-10

Method Title Sediment Processing for Pore Water Extraction through Centrifugation, E1391-02

Purpose This method describes a process for separation and collection of interstitial pore

water from sediment samples to provide either a matrix for toxicity testing or an

indication of the concentration and partitioning of contaminants within the sediment

matrix. 

Method Summary Centrifugation may be used to isolate interstitial water for chemical or toxicological

analyses. The centrifugation conditions (i.e., speed and temperature) will vary

considerably depending on the contaminants potentially present in the pore water.

Sim ilarly, the filtration scheme will depend on the sediment composition and the

analytes of interest. For dissolved metals and dissolved organic carbons, sediments

should be centrifuged at high speed and filtered with a 0.2-um mem brane filter.

For other dissolved organic contaminants, collodial matter, and aquatic bacteria,

sediments should be centrifuged at a lower speed and filtered through a 0.45-um

mem brane filter. 

Generally, 30 minutes of centrifugation at 10,000 x g is recommended for routine

toxicity testing of interstitial waters. The temperature should be set to reflect ambient

temperature at time of collection. 

Data

Uses/Application

Interstitial water is analyzed to either provide a matrix for toxicity testing or an

indication of the concentration and partitioning of contaminants within the sediment

matrix. There is some indication that interstitial water may be as useful as whole

sediment for evaluating the toxicity of some sediment-associated compounds.

Advantages Centrifugation will extract a relatively large volume of interstitial water as compared

to other separation techniques.

Limitations Centrifugation procedures vary depending on the various compositions of the

sediments, therefore it is difficult to have an established protocol. Manipulation and

centrifugation changes the redox potential of the pore water from the in situ

conditions. Filtration may also remove toxicity. Sometimes a double centrifugation is

required to remove fine particles.

Reference ASTM. 2001b. ASTM Book of Standards. Section 11.05. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

118

Fact Sheet No. 2.2.1-11

Method Title Pore Water Extraction from Sediments through Squeezing, ASTM E1391-02

Purpose This method describes a process for separation and collection of interstitial pore

water from sediment samples to provide either a matrix for toxicity testing or an

indication of the concentration and partitioning of contaminants within the sediment

matrix. 

Method Summary The apparatus used for isolation of pore water by squeezing includes a filter. The

characteristics of filters should be considered carefully based on the types of

contaminants expected. 

An example method for squeezing can be found in Manheim (1966). Briefly, the

apparatus consists of a standard laboratory press and a filter unit containing a

stainless steel screen, perforated steel plate, steel filter holder and filters. W et

sediment is transferred into a cylinder at the top of the apparatus. The whole unit is

placed in a press and pore water is removed through a bottom filter. The extraction

time will vary depending on the amount of sediment placed in the unit. 

Data

Uses/Application

Squeezing is used to extract pore water from loose seabed sediments. Interstitial

water is analyzed to either provide a matrix for toxicity testing or an indication of the

concentration and partitioning of contaminants within the sediment matrix. There is

some indication that interstitial water may be as useful as whole sediment for

evaluating the toxicity of some sediment-associated compounds. 

Advantages This is a rapid, reproducible method for pore water collection. 

Limitations Squeezing can produce artifacts due to shifts in equilibrium from pressure,

temperature, and gradient changes. It can also affect the electrolyte concentration

and redox potential compared to in situ conditions.

Filtrations may remove toxicity from water samples. It is difficult to choose a filtering

scheme without knowing the cause of toxicity or the mixture of the toxicant.

Reference ASTM. 2001b. ASTM Book of Standards. Section 11.05. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

119

Fact Sheet No. 2.2.1-12

Method Title Pore water extraction from sediment from Vacuum Filtration, ASTM E1391-02

Purpose This method describes a process for separation and collection of interstitial pore

water from sediment samples to provide either a matrix for toxicity testing or an

indication of the concentration and partitioning of contaminants within the sediment

matrix. 

Method Summary Vacuum filtration is one of several methods (including gas pressurization and

displacement) that can be used to remove pore water from sediments for chemical

analysis when only a sm all volume is required. A vacuum extractor consists of a

fused-glass air stone connected to a syringe with tubing. The syringe is then used to

extract the water until sufficient volume has been collected.

Data

Uses/Application

Interstitial water is analyzed to either provide a matrix for toxicity testing or an

indication of the concentration and partitioning of contaminants within the sediment

matrix. There is some indication that interstitial water may be as useful as whole

sediment for evaluating the toxicity of some sediment-associated compounds. 

Advantages These methods are useful for collecting sm all amounts of water for chemical

analysis.

Limitations Problems comm on to this type of method are a loss of equilibration between the

interstitial water and the solids, filter clogging, and oxidation. Further research is

needed to dem onstrate the utility of this method to in situ collection of sediment pore

water. 

Reference ASTM. 2001b. ASTM Book of Standards. Section 11.05. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

120

Fact Sheet No. 2.2.1-13

Method Title Acoustic Sub-Bottom Profiling Systems, DRP-2-03

Purpose To collect sub-bottom data for sea-floor mapping.

Method Summary High-resolution acoustic profiling systems use high power signals to penetrate the

sediments of the sea floor. The signal is reflected from interfaces between

sediment strata of different acoustic impedance. These data are printed on a

graphic recorder as a continuous two-dimensional profile. The amount of

penetration will depend on the combination of the frequency and power of the

profiler being used. A 3.5 - 14 kHz frequency pulse is typically used. Penetration

also depends on the material type which composes the bottom and sub-bottom. For

instance, differences in soil types, density, water content, and degree of

solidification greatly influence the reflecting properties of the sub-bottom strata.

Most Sub-bottom surveys are conducted in conjunction with bathym etric and/or side

scan sonar surveys. The survey uses a predetermined grid pattern with lines

spaced at variable distances, depending on objectives of the survey. Sm all survey

boats (30 to 65 ft long) are adequate for performing a com plete multi system survey.

Over-the-side, surface-towed, and hull-mounted source/receiver arrays can be

used. Data acquisition must be interfaced with the navigation system so that

accurate information between position and data is recorded at all times.

Environmental Effects of Dredging Technical Notes EEDP-01-5 and EEDP-01-10

also discuss Sub-bottom profilers (US Army Engineer Waterways Experiment

Station, 1989; US Army Engineer Waterways Experiment Station, 1988).

Sub-bottom profiling is also discussed in the Guidelines for the conduct of benthic

studies at aggregate dredging sites by the U.K. Centre of Environm ent, Fisheries

and Aquaculture, available online (www.planning.odpm.gov.UK/benthic Last

accessed 2/10/03).

Data

Uses/Application

Typical applications include the monitoring of sediment disposal sites to detect

stratification within and just below deposits.

Advantages Primary advantages associated with acoustic sub-bottom profiling are continuous

documentation of reflecting strata, rapid coverage, and relatively low cost.

Limitations The quality of records obtained in seismic reflection studies depends greatly on the

presence of subsurface horizons which will reflect acoustic energy. Records in

deep water will tend to show average conditions over an area rather than a specific

profile directly below the ship. Effective use of this instrumentation requires a

trained operator.

Reference US Army Engineer Waterways Experiment Station. 1991. Hydrologic Surveys

Applicable to Dredging Operations. DRP-2-03.

Waterways Experiment Station, U.S. Army Corps of Engineers, Vicksburg MS.

Website http:/el.erdc.usarce.army.mil/elpubs/pdf/drp2-

03.pdf

Last Accessed: 1/28/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

121

Fact Sheet No. 2.2.1-14

Method Title Side Scan Sonar, EEDP-01-10

Purpose To qualitatively map the surface characteristics of the sea floor.

Method Summary During side scan sonar analysis, an acoustic towfish projects acoustic energy at a

frequency of 100-500 kHz in a lateral direction using a pair of transducers. The

acoustic signal bounces off of the sea floor back to the transducers on the towfish.

The received signal is transmitted through the tow cable to the shipboard receiver,

which processes the signal and produces a sonograph.

A frequency of 100 or 500 kHz is generally used for monitoring disposal sites. The

lower frequency gives a greater range (i.e., 200-400 m of bottom compared to 100

m at 500 kHz) but provides less detail. At 500 kHz, the sonar is able to distinguish

differences in bottom texture that can be used to map grain size. For example, lowbackscatter indicates a silty bottom. Sand ripples in the image can be used to

interpret grain-size variation and the movements of sediment.

Environmental Effects of Dredging Technical Notes EEDP-01-5 briefly discusses

side scan sonar (US Army Engineer Waterways Experiment Station, 1989).

Side scan sonar is also discussed in the Guidelines for the conduct of benthic

studies at aggregate dredging sites by the U.K. Centre of Environm ent, Fisheries

and Aquaculture, available online (www.planning.odpm.gov.UK/benthic Last

accessed 2/10/03).

Data

Uses/Application

Typical applications include the monitoring of sediment disposal sites before and

after disposal. 300 kHz or higher frequencies can also be used for habitat mapping

(i.e., sea grass beds). The information gathered with side-scan sonar may be used

to direct subsequent monitoring studies.

Advantages Overlapping coverage allows precise and continuous mapping of the sea floor. Side

scan sonar delineates the edge of disposal deposits more accurately than

bathym etric data.

Limitations The interpretation of side scan images requires some training and experience. 

Reference US Army Engineer Waterways Experiment Station. 1988. Acoustic Tools and

Techniques for Physical Monitoring of Aquatic Dredged Material Disposal Sites.

EEDP-01-10. Waterways Experiment Station, U.S. Army Corps of Engineers,

Vicksburg MS.

Website http://www.wes.army.mil/el/dots/pdfs/eedp01-

10.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

122

Fact Sheet No. 2.2.1-15

Method Title Settlement Plates, DRP-2-03

Purpose To monitor changes in thickness of various layers of dredged and capping material

in confined upland or aquatic disposal sites.

Method Summary

ISC

Telescoping settlement plates are used to measure changes in height of individual

material layers at disposal site. The lower tier plate is placed on the foundation

sediment before the dredged material is deposited. After dredged material disposal,

a second tier settlement plate is slipped over the riser pipe of the lower tier and

comes to rest on the surface of the dredged material. After placement of the cap, a

third tier settlement plate is placed over the riser pipe of the second tier, and the

plate rests on the surface of the cap. Readings are made to determine changes in

individual layer thickness.

Riser pipes and settling plate can be constructed from a variety of materials. For

aquatic disposal sites, settlement plates can be designed and constructed to have a

unit weight approximating that of water so that the plates do not sink through the soft

dredged material or cause consolidation of the underlying material. 

Data

Uses/Application

Monitoring sediment consolidation at dredged material disposal sites, consolidation

of cap at ISC sites, and consolidation of sediment and/or cap at CAD sites.

Advantages Exact changes in thickness (i.e., settlem ent) of various layers of deposited material

can be directly measured. Settlement of the foundation sediments can be

determined using a stationary benchmark relative to the lower tier riser pipe. 

Limitations Divers must be used to place the plates and make the settlement readings. The

riser pipes/settlement plates are vulnerable to accidental disturbance, removal, or

damage by boating, fishing, or dredge disposal activities.

Reference US Army Engineer Waterways Experiment Station. 1989. Monitoring Dredged

Material Consolidation and Settlement at Aquatic Disposal Sites, Environmental

Effects of Dredging Technical Notes EEDP-01-5. Waterways Experiment Station,

U.S. Army Corps of Engineers, Vicksburg MS.

Website http://www.wes.army.mil/el/dots/pdfs/eedp0

1-5.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

123

2.2.2 Chemical and Physical Analysis

Section 2.2.2 contains methods for sample preparation and the chemical and physical analysis of

sediment and soil. These methods characterize the chemical composition and physical properties

of sediment/soil samples collected by methods described in Section 2.2.1. Samples are often

analyzed for the presence of various inorganic and organic contaminants that may pose a threat

to human or ecological health. Many of the methods described have been developed over time to

optimize the detection, identification, and quantification of potential chemicals of concern. Several

are performance-based and may be further modified to enhance the accuracy and precision of the

method.

A variety of methods may exist for the analysis of a particular chemical parameter, all with varying

levels of quantification or degrees of sensitivity. Less sensitive methods may be used as a

screening tool during the initial site assessment to identify potential chemicals of concern. Followup analysis may include the use of very precise methods that provide unequivocal identification

and trace level quantification of analytes. This variety also provides alternative methods useful in

the analysis of many types of sediment or soil samples. Interferences from certain compounds in

a sample may be avoided by the use of an alternative preparation or analytical method.

The physical properties of soil/sediment often influence the behavior of soil/sediment in the

environment, and they may be helpful in further understanding the fate of contaminants

associated with soil/sediment.

Many of the chemical and physical methods described in these fact sheets are routinely

performed and fairly standardized. As a result, more than one source of information is often cited

in each method description. Specifically, the following sources provided methods information for

section 2.1.2:

• The USEPA’s Office of Water

• The USEPA’s Lake Michigan Mass Balance Study Methods Compendium, 1997v

• The USEPA’s Test Methods for Evaluating Solid Waste, Physical/Chemical Methods

(SW846 Methods)

• NOAA’s National Status and Trends Program, 1998

• Standard Methods for Examination of Water and Wastewater, 1999

• ASTM
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

124

Fact Sheet No. 2.2.2-1

Method Title Total Mercury in Sludge, Sediment, Soil, and Tissue by Acid Digestion and BrCl

Oxidation, Appendix to Method 1631

Purpose These procedures may be used in conjunction with EPA Method 1631B for

determination of mercury in sludge, sediment, soil, tissue, industrial samples, and

certified reference materials.

Method Summary Digestion I—This procedure is preferred for matrices containing organic materials,

such as sludge and plant and animal tissues, because the organic matter is

completely destroyed. In this procedure, a 0.2 - 1.5 g sample is digested with

HNO3/H2SO4. The digestate is diluted with BrCl solution to destroy the remaining

organic material.

Digestion II—This procedure is preferred for geological materials because of rapid

and com plete dissolution of cinnabar (HgS), which is otherwise more slowly

attacked by the BrCl in Digestion I. In this procedure, a 0.5 - 1.5 g sample is

digested with aqua regia (HCl/HNO3

) to solubilize inorganic materials.

The Hg concentration in the digestate is determined using EPA Method 1631B.

These procedures, in conjunction with Method 1631B, allow determination of Hg at

concentrations ranging from 1.0 to 5000 ng/g in solid and semi-solid matrices.

The method detection limit for Hg has been determined to be in the range of 0.24 to

0.48 ng/g when no interferences are present. The minimum level of quantization

(ML) has been established as 1.0 ng/g. These levels assume a sample size of 0.5 g.

Method LMMB 050 (USEPA 1997b, EPA 905-R-97-012c) describes the automated

digestion and analysis of total mercury in sediment samples using the Cold Vapor

technique.

Data

Uses/Application

Mercury is one of the primary risk factors in many contaminated sediments and is

one of the primary contaminants measured as part of a chemical assessment.

Advantages The dual amalgam trap system and fluorescence detector provide greater sensitivity

and specificity in the presence of interferences, and this system must be used to

overcome interferences, if present, and to achieve the sensitivity required, if

necessary. For some site monitoring programs, total mercury is measured because

it is a more rapid method compared to methyl mercury.

Limitations In cases where total mercury exceeds threshold values, samples may need to be

analyzed for toxic methyl mercury to determine risk.

Reference USEPA. 2001d. Appendix to Method 1631: Total Mercury in Sludge, Sediment, Soil,

and Tissue by Acid Digestion and BrCl Oxidation, EPA-821-R-01-013. Office of

Water U.S. Environmental Protection Agency, Washington, DC.

Website http://www.brooksrand.com/FileLib/1

631.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

125

Fact Sheet No. 2.2.2-2

Method Title Trace Element Quantification Techniques

Purpose To determine major and trace elements in sediment and biological tissue

samples utilizing atomic absorption and neutron activation techniques.

Method Summary Sediment samples are homogenized and freeze dried, and the dry aliquot is

homogenized. Approximately 0.2 g of dried sediment is weighed and

transferred to a TeflonTM bomb. Samples are digested by adding 3 mL HNO3

and placing the bombs in a 130°C oven for approximately 12 hours. The

bombs are removed, 2 mL of concentrated HF are added, and the bombs are

returned to the oven for 12 hr. After cooling, 18 mL of 4% boric acid are added

and the bombs are returned to the oven for another 12 hr. Solution volume is

determined, and a 20-fold dilution is made for FAAS analysis of Al, Fe, Mn, Si,

and Zn. For analysis of Hg, sediment samples are digested using a modified

version of EPA method 245.5. Samples were analyzed using the following

instrumentation:

Analyte Method

Hg Cold vapor atomic absorption (CVAA)

Al, Fe, Mn, Zn Flame atomic absorption (FAA)

Ag, As, Cd, Cr, Cu, Ni, Pb, Graphite furnace atomic absorption

Se, Sn (GFAA)

Al, Cr, Fe, Mn Instrumental neutron activation 

 analysis (INAA)

ASTM Methods D1971, D3974, and D4698 and SW846 Method 3050B

describe various digestion methods for determination of metals in sediments

(ASTM, 2001a).

Standard Method 3030K, ASTM Method D5258, and SW846 Method 3051

describe the microwave digestion method (APHA, 1999; ASTM, 2001a).

Several methods describe the analysis of metals using various atomic

absorption methods, such as Standard Method 3112 B for CVAA, 3111 for

FAA, 3113B for GFAA and SW846 7000 series of methods. 

Data Uses/Application Chemical screening of trace metals in sediments against contaminant

guidelines provides an indication that adverse effects may or may not be

occurring.

Advantages Tissue sample digestion in a TeflonTM bomb is a standard method for “clean”

digestion for metals analysis. The instrumental suite employed in this method

takes advantage of the know strengths of each instrument for trace analysis.

For example, GFAA is much more sensitive than FAA, requiring only a sm all

volume of sample for trace analysis, and CVAA is very sensitive for mercury.

Limitations Processing of samples for trace levels required for risk assessments requires

class-100 clean room, or other suitable environm ent.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

126

Fact Sheet No. 2.2.2-2 (contd.)

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and

Trends Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo

NOS ORCA 130. National Oceanic and Atm ospheric Administration, Silver

Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications/

tm130.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

127

Fact Sheet No. 2.2.2-3

Method Title Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence, Atomic

Absorption and Inductively Coupled Plasma Mass Spectrom etry

Purpose To determine the concentration of 17 metals in sediment and biological tissue

samples utilizing atomic absorption, inductively coupled plasma mass spectrometry

(ICP-MS), and energy dispersive X-ray fluorescence (XRF).

Method Summary Sediment samples are weighed and freeze-dried. The dried sample is ground in a

ball mill. 0.5 g aliquots are used directly for XRF analysis or are further digested for

AA or ICP-MS analysis. 

200 mg of dried sediment is placed in a TeflonTM bomb, to which 1 mL of 4:1

HNO3/HClO4is added. The bombs are heated in a 130°C oven for 4 hours. After

cooling, 3 mL of concentrated HF are added to the bomb, and the bombs are

heated again in a 130°C oven for 8 hours. After cooling, the digestates are diluted to

approximately 20 mL with deionized water. Solution volumes are calculated, and the

digestates are analyzed directly by GFAA or CVAA. For ICP-MS analysis, a 10-mL

aliquot of the digestate is dried in a perchloric acid hood. The dried digestate is

dissolved in 1 mL of 10% HNO3 and dried again. The dried digestate is dissolved

again in 1 mL of 10% HNO3 and 9 mL of deionized water.

AnalyteMethod

Hg Cold vapor/gold foil amalgam

Cd, Se, Ag Graphite furnace atomic 

absorption(GFAA)

Ag, Al, Cr, Cd, Ni, Pb, Sb, Sn ICP-MS

Al, As, Cr, Cu, Fe, Mn, Ni, Pb, Si, Zn XRF

Standard Method 3030K, ASTM Method D5258, and SW846 Method 3051 describe

the microwave digestion method (APHA, 1999; ASTM, 2001c). Selected methods

for the analysis of metals include: Standard Methods 3112B for CVAA, 3113B for

GFAA, and 3120B for ICP-MS; SW846 Method 6020 for ICP-MS and SW846 7000

series for atomic absorption (APHA, 1999). 

Data

Uses/Application

Chemical screening of trace metals in sediments against contaminant guidelines

provides an indication that adverse effects may or may not be occurring.

Advantages XRF analysis does not require digestion of the sample. Crustal elements such as

Al, Cr, Fe, Ni, and Si, that can be difficult to dissolve from sediment, can be

quantified by XRF. ICP-MS has the advantage of simultaneous analysis of many

elements with detection limits much lower than the XRF and similar to those of

GFAA. ICP-MS is particularly sensitive for Al, Cr, Ni, Ag, Cd, Sn, Sb, and Pb.

CVAA is very sensitive and reliable for Hg analysis.

Limitations Leakage at high pressure can cause loss of Hg from the sample during digestion.

Analysis of GFAA requires the use of matrix modifiers and standardization of the

instrument by method of addition to the sample matrix to provide accurate results.

XRF and total HF digestion measure total metals, which may not reflect levels that

are bioavailable.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

128

Fact Sheet No. 2.2.2-3 (contd.)

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and Trends

Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo NOS ORCA

130. National Oceanic and Atm ospheric Administration, Silver Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications

/tm130.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

129

Fact Sheet No. 2.2.2-4

Method Title Determination of Acid Volatile Sulfide and Selected Sim ultaneously Extractable

Metals in Sediment

Purpose For the determination of acid volatile sulfide (AVS) and for selected metals that are

solubilized during the acidification step (simultaneously extracted metal, SEM).

Method Summary Sediment samples must be protected from exposure to oxygen during collection and

storage. If possible, the head space in the sample container should be filled with

oxygen-free nitrogen or argon. Appropriate storage conditions: frozen (preferred if

sediments to be used for chemical analyses only) or refrigerated to 4 degrees

Celsius (if sediments to be used for biological tests as well); glass containers if

refrigerated, but plastic is acceptable if frozen. About 10 gm of sediment is acidified

with hydrochloric acid at room temperature to convert the AVS to hydrogen sulfide.

The H2S is then purged from the sample and trapped in an aqueous solution, which

varies depending upon the analytical method being used. Using the colorimetric

method, the H2S is trapped in sodium hydroxide. The sulfide reacts with N-Ndimethyl-p-phenylenediamine to form methylene blue, which is then measured

colorimetrically at 670 nm. Using the gravimetric method, the sulfide is trapped in

silver nitrate, forming a silver sulfide precipitate. The silver sulfide is isolated onto a

1.2 micron filter by filtration. The filter is dried and the amount of silver sulfate is

weighed. Using the third method, the sulfide is trapped in an antioxidant buffer, and

the sample is analyzed using an ion-selective sulfide electrode.

After the determination of AVS, the acidified sediment sample is filtered through a

0.2μ mem brane filter. The filtrate is analyzed for SEM (commonly, cadmium,

copper, lead, silver, nickel, and zinc) by atomic absorption or inductive coupled

plasma spectrometric methods.

Both AVS and SEM are expressed on a μm ole per gram dry sediment basis. The

ratio of SEM to AVS is the sum of the concentrations of SEM metals divided by the

acid volatile sulfide concentration.

Data

Uses/Application

Sulfide is important in controlling the bioavailability of metals in anoxic sediments.

The amounts of SEM and AVS are important in predicting the bioavailability of

metals.

Advantages The gravimetric procedure can be used with samples that have a moderate or high

AVS concentration. The colorimetric method is capable of determining AVS

concentrations over a range of 0.01-1000 μmoles/gram dry weight. The sample

purging and trapping apparatus may consist of either Erlenmeyer flasks (less costly)

or ground glass stoppered flasks (better sealing).

Limitations Sulfide ion is unstable in the presence of oxygen. Sulfide can be formed or lost due

to biological activity during storage. Leakage of the Erlenmeyer flasks may cause

low recovery of AVS.

Reference USEPA. 1991. Draft Analytical Method for Determination of Acid Volatile Sulfide in

Sediment, EPA-821-R-91-100. Office of Water, U.S. Environmental Protection

Agency, Washington, DC. Proper storage of samples will mitigate these limitations.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

130

Fact Sheet No. 2.2.2-5

Method Title Photovac GC Analysis for Soil, Water, and Air/Soil Gas, OSW ER SOP# 2109

Purpose This method is designed as a field screening procedure for the tentative

identification of various volatile organic compounds.

Method Summary Soil samples are collected in 40-mL VOA vials with TeflonTM-lined silicone septum

screw caps. A 5 gm aliquot of sample is weighed into a second, clean VOA vial.

Reagent water is added to the sample to bring the volume in the vial to 20 mL. The

vial is capped, shaken vigorously for one minute, and allowed to stand at room

temperature for at least one hour for vapor phase equilibration. An aliquot of the soil

head space is then removed from the vial and injected into the GC using a gas-tight

syringe. The GC uses an ultraviolet light source and photoionization detector.

Concentrations are reported as μg/kg.

Typical MDLs for this method range from 1 ppb to 5 ppb.

SOPs # 2108 and #2107 describe the operation of specific models of Photovac Gas

Chromatographs.

Data

Uses/Application

Site assessment/characterization and health and safety surveys.

Advantages The data generated with this method allows for rapid evaluation of site conditions.

Limitations Pollutant identification is only tentative.

Reference USEPA. 1994b. SOP # 2109: Photovac GC Analysis for Soil, Water, and Air/Soil

Gas. Compendium of Emergency Response Team Standard Operating Protocols.

Office of Solid Waste and Emergency Response, U.S. Environmental Protection

Agency, Edison NJ.

Website http://www.ert.org/products/2109.PDF LastAccessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

131

Fact Sheet No. 2.2.2-6

Method Title Extraction and Clean-Up of Sediments for Semi-volatile Organics Following the

Internal Standard Method, LMMB 040

Purpose To prepare the sediment samples for the measurement of organic

contaminants, such as polychlorinated biphenyls, polynuclear aromatic

hydrocarbons, and chlorinated pesticides.

Method Summary Following Method LMMB 040 (USEPA 1997d: EPA 905-R-97-012c), 15 - 30 g

of sediment are chemically dried with sodium sulfate, spiked with surrogate

standards, and extracted with DCM using a 30°C sonication bath (sonicate for

60 minutes, let stand in bath overnight [24 hours] and sonicate again for 60

minutes). During a silica/alumina column clean-up, two separate fractions are

collected. The first fraction, which is eluded with hexane, contains PCBs,

HCB, 4, 4'-DDE, aldrin, and heptachlor. The second fraction, which is eluded

with 10% diethyl ether in hexane, contains alpha- and gamma-BHC,

chlordanes, 4,4'-DDT, 4,4'-DDD, and all PAHs. The two fractions are

concentrated. The first fraction is treated with activated copper and frozen at -

15°C until analysis. The second fraction is solvent exchanged into hexane and

refrigerated until analysis.

Following NS&T procedures, samples are stored frozen at approximately -

15°C until extraction. A 10 - 30 gram aliquot of the homogenized sediment

sample is chemically dried with sodium sulfate, spiked with surrogate

standards, and extracted with dichloromethane (DCM) using a Soxhlet

apparatus for 8 hours. The extract is concentrated, filtered if necessary, and

solvent changed to hexane. The sample is cleaned-up using purified silica

gel/alumina column chromatography before instrumental analysis. Activated

copper in the column removes elemental sulfur that may be present. The

sample is concentrated to 1 mL in hexane for analysis. Chemical surrogates

are used to monitor extraction and cleanup efficiency.

Several methods for sample extraction and clean-up are described in SW846

Methods 3500B and 3600C. ASTM Method D3976 describes the preparation

of sediment samples for volatile, semi-volatile, and nonvolatile analyses

(ASTM, 2001c).

Data Uses/Application Polychlorinated biphenyls and high molecular weight PAHs are primary risk

factors in many contaminated sediments and are measured as part of the

chemical assessment of a site characterization and to assess remediation

effectiveness.

Advantages Both of the above listed methods, or variations based on performance, are

sufficient for the analysis of PCB by GC-ECD or GC/MS (low resolution) or the

analysis of PAH by GC/MS (low resolution).

Limitations Method interferences may be caused by contaminants in solvents, reagents,

glassware, and other sample processing hardware. Matrix interferences result

from co-extraction of compounds other than the analytes of interest, such as

elemental sulfur and lipids.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

132

Fact Sheet No. 2.2.2-6 (contd.)

Reference USEPA. 1997d. Method LMMB 040: Extraction and Clean-Up of Sediments for

Semi-volatile Organics Following the Internal Standard Method in Lake Michigan

Mass Balance Study Methods Compendium, Volume 2: Organic and Mercury

Sample Analysis Techniques, EPA 905-R-97-012c. Great Lakes National Program

Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/glnpo/lmmb/methods/sop401.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

133

Fact Sheet No. 2.2.2-7

Method Title Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS) - Selected Ion Monitoring (SIM)

Mode

Purpose To determine low concentrations of Polycyclic aromatic hydrocarbons (PAHs) and

their alkylated homologues in extracts of water, sediments and biological tissues.

Method Summary Just prior to analysis, an aliquot of internal standard solution is added to the sample

extract producing a final internal standard concentration of approximately 40 ng/mL.

The analytical system includes a temperature programmable gas chromatography

with a fused silica capillary column. Helium is used as the carrier gas, and the

samples are handled by an auto sampler capable of making 1 - 4 μl injections. A

five point calibration curve is established to dem onstrate the linear range of the

detector. The effluent from the GC capillary column is routed directly into the ion

source of the mass spectrometer (MS). The MS is operated in the SIM mode using

appropriate windows to include the quantization and confirmation masses for target

PAHs. For all compounds detected at a concentration above the MDL, a

confirmation ion is checked to confirm its presence. The response factors of the

surrogate relative to each of the calibration standards are calculated, followed by the

calculation of the sample extract concentration. The sample concentration for each

compound is calculated by dividing the sample extract concentration by the sample

amount.

Data

Uses/Application

PAH concentrations are primary risk factors associated with contaminated

sediments. PAH data obtained from this analysis are used for site characterization

and risk assessments.

Advantages GC/MS in the SIM mode provides unambiguous and sensitive detection for PAHs.

The PAH quantization method is very rigorous because PAHs have very strong

molecular ion peaks under the mass spectrometric conditions used. Alkylated PAH

homolog data can be used for source identification. Also, the availability of labeled

surrogates internal standards of many of the analytes makes very accurate

determinations of analyte concentrations possible.

Limitations GC/MS in SIM mode cannot be used for simultaneous screening for other organic

contaminants of similar polarity or volatility; cannot be used to identify tentatively

identified compounds (TICs).

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and Trends

Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo NOS ORCA

130. National Oceanic and Atm ospheric Administration, Silver Spring, MD. 233 pp.

Website http://ccma.nos.noaa.govfpublications

/tm30.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

134

Fact Sheet No. 2.2.2-8

Method Title Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

Purpose To quantify chlorinated hydrocarbons (i.e., chlorinated pesticides and PCBs) in

sample extracts.

Method Summary This method is based on high resolution, capillary gas chromatography using

electron capture detection (GC/ECD). Extracts normally have a holding time of 40

days. The instrument’s detector is calibrated before the sample in injected.

Pesticide/PCB calibration is done also as part of the analytical run. If the response

for any peak exceeds the highest calibration solution, the extract is diluted, a known

amount of surrogate and TCMX solution added, and the sample reanalyzed for

those analytes that exceeded the calibration range. Concentrations in the samples

are calculated based on the internal standard method. Data is reported as ng/g dry

weight.

Other methods describing the analysis of PCBs and pesticides by GC/ECD are

NS&T methods, ASTM Method E697, and SW846 Methods 8081A and 8082

(NOAA, 1998; ASTM, 2001d).

Data

Uses/Application

PCBs and persistent pesticides (particularly DDT and metabolites) are two of the

primary risk factors of contaminated sediments. Data are used in site

characterization and in risk analysis.

Advantages The ECD is very sensitive and allows for detection of the chlorinated hydrocarbons

at trace concentrations (ppb).

Limitations The detector does not have a linear response over a wide concentration

range and must be used by sufficiently trained personnel. Second column analysis

must be performed to provide unequivocal compound identification. These methods

do not measure the 12 World Health Organization PCB congeners, which may be

desired data in some risk assessments. A separate analysis using a different GC

column is required for peak confirmation.

Reference USEPA. 1997d. Method LMMB 041: Analysis of Polychlorinated Biphenyls and

Chlorinated Pesticides by Gas Chromatography with Electron Capture Detection, in

Lake Michigan Mass Balance Study Methods Compendium, Volume 2: Organic and

Mercury Sample Analysis Techniques, EPA 905-R-97-012c. Great Lakes National

Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/glnpo/lmmb/methods/

sop-501.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

135

Fact Sheet No. 2.2.2-9

Method Title Screening for Polychlorinated Biphenyls by Immunoassay, SW846 Method 4020

Purpose This method is used to screen soils and non-aqueous waste liquids for the presence

of polychlorinated biphenyls (PCBs).

Method Summary This method is used to determine when PCBs are present at concentrations above

5, 10 or 50 mg/kg. The method is most often performed using a sample extract.

Determining the presence of PCBs above concentrations other than 5, 10 or 50

mg/kg is possible by dilution of the sample extract. The sample extract and an

enzyme conugate reagent are added to immobilized antibody. The enzyme

conjugate “competes” with PCB present in the sample for binding to immobile antiPCB antibody. Test kits are commercially available for this method. Each

commercially-available test kit will supply or specify the apparatus and materials

necessary for successful completion of the test. The manufacturer’s directions

should be followed. Method 4020 provides an estimate for the concentration of

PCBs by comparison with a standard.

Data

Uses/Application

Site characterization, screening.

Advantages Test kits are commercially available for this method.

Limitations This method does not provide information regarding congener or Aroclor

concentrations. High levels of chemically-similar compounds may register a false

positive. Method is intended for screening, not for quantitative analysis. In cases

where the exact concentrations of PCBs are required, quantitative techniques

should be used. If the proportions of PCB congeners in the calibration standard are

not similar to the proportions present at the site, accuracy can be compromised.

Reference USEPA. 1996h. Screening of Polychlorinated Biphenyls by Immunoassay, SW846

Method 4020. Office of Solid Waste, U.S. Environmental Protection Agency,

Washington, DC.

Website http://www.epa.gov/epaoswer/hazwaste/

test/pdfs/4020.pdf

Last Accessed: 2/13/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

136

Fact Sheet No. 2.2.2-10

Method Title Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA Method 1613

Purpose This method is for determination of tetra- through octa-chlorinated dibenzo-p-dioxins

(CDDs) and dibenzo furans (CDFs) in solids (not tissue).

Method Summary This method is "performance-based". The labeled compounds are spiked into a

sample containing 10 g (dry weight) of solids. Samples containing multiple phases

are pressure filtered and any aqueous liquid is discarded. Coarse solids are ground

or homogenized. Any non aqueous liquid from multi phase samples is combined

with the solids and extracted in an extractor. The extract is concentrated for cleanup.

After extraction, 37Cl4-labeled 2,3,7,8-TCDD is added to each extract to measure the

efficiency of the cleanup process. Sample cleanups may include back-extraction

with acid and/or base, and gel permeation, alumina, silica gel, Florisil and activated

carbon chromatography. High-performance liquid chromatography (HPLC) can be

used for further isolation of the 2,3,7,8-isomers or other specific isomers or

congeners. After cleanup, the extract is concentrated to near dryness. Immediately

prior to injection, internal standards are added to each extract, and an aliquot of the

extract is injected into the gas chromatography. The analytes are separated by the

GC and detected by a high-resolution ($10,000) mass spectrometer.

CDD/CDF Minimum Level (ng/kg) CDD/CDF ML(ng/kg)

2,3,7,8-TCDF 1 1,2,3,4,7,8-HxCDD 5

2,3,7,8-TCDD 1 1,2,3,6,7,8-HxCDD 5

1,2,3,7,8-PeCDF 5 1,2,3,7,8,9-HxCDD 5

2,3,4,7,8-PeCDF 5 1,2,3,4,6,7,8-HpCDF 5

1,2,3,7,8-PeCDD 5 1,2,3,4,7,8,9-HpCDF 5

1,2,3,4,7,8-HxCDF 5 1,2,3,4,6,7,8-HpCDD 5

1,2,3,6,7,8-HxCDF 5 OCDF 10

1,2,3,7,8,9-HxCDF 5 OCDD 10

2,3,4,6,7,8-HxCDF 5

This method is also described in SW846 Method 8290 and NS&T methods (NOAA

1998).

Data

Uses/Application

The method is for use in EPA's data gathering and monitoring programs associated

with the Clean Water Act, the Resource Conservation and Recovery Act, the

Com prehensive Environmental Response, Compensation and Liability Act, and the

Safe Drinking Water Act.

Advantages Method 1613 is able to meet detection limits required for human health and

ecological risk assessments.

Limitations The GC/MS portions of this method are for use only by analysts experienced with

HRGC/HRMS or under the close supervision of such qualified persons.

Reference USEPA. 1994c. Method 1613: Tetra- through Octa-Chlorinated Dioxins and Furans

by Isotope Dilution HRGC/HRMS, EPA 821-B-94-005. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/waterscience/methods/1613.pdf Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

137

Fact Sheet No. 2.2.2-11

Method Title Toxic Polychlorinated Biphenyls by Isotope Dilution High Resolution

Gas Chromatography/High Resolution Mass Spectrometry, EPA Method 1668

Purpose This method is for determination of the toxic polychlorinated biphenyls (PCBs) in

solids (not tissue).

Method Summary This method is performance-based. The labeled compounds are spiked into a

sample containing 10 g (dry weight) of solids. Samples containing multiple phases

are pressure filtered and any aqueous liquid is discarded. Coarse solids are ground

or homogenized. Any non-aqueous liquid from multi-phase samples is combined

with the solids and extracted in an SDS extractor. The extract is concentrated for

cleanup. After extraction, samples are cleaned up using back-extraction with

sulfuric acid and/or base, and gel permeation, silica gel, Florisil and activated carbon

chromatography. High-performance liquid chromatography (HPLC) can be used for

further isolation of specific isomers or congeners. After cleanup, the extract is

concentrated to near dryness. Immediately prior to injection, internal standards are

added to each extract, and an aliquot of the extract is injected into the gas

chromatography. The analytes are separated by the GC and detected by a highresolution ($10,000) mass spectrometer.

Extract

IUPAC EMDL (ng/kg) EML (ng/kg) EML (pg/μL)

77 0.5 2 1

123 4 10 5

126 10 4 5

118/167/156/157/169/180/170/189 6 20 10

114 60 200 100

105 40 100 50

EMD: = Estimated Method Detection Limit; EML = Estimated Minimum Level

Data

Uses/Application

The method is for use in EPA's data gathering and monitoring programs associated

with the Clean Water Act, the Resource Conservation and Recovery Act, the

Com prehensive Environmental Response, Compensation and Liability Act, and the

Safe Drinking Water Act.

Advantages Method 1668 provides data for most, but not all, of the “dioxin-like” PCBs, including

those with the highest TEFs, as determined by the World Health Organization. This

method provides detection limits frequently required in risk assessments.

Limitations The GC/MS portions of this method are for use only by analysts experienced with

HRGC/HRMS or under the close supervision of such qualified persons. Method

1668 does not provide data for all of the “dioxin-like” PCBs, as does Method 1668A.

Reference USEPA. 1997e. Method 1668: Toxic Polychlorinated Biphenyls by Isotope Dilution

HRGC/HRMS, EPA-821-R-97-001. Office of Water, U.S. Environmental Protection

Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitl

eOW.html

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

138

Fact Sheet No. 2.2.2-12

Method Title Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and Tissue by

HRGC/HRMS, EPA Method 1668 Revision A

Purpose This method is for congener-specific determination of more than 150 chlorinated

biphenyl (CB) congeners in solids (not tissue).

Method Summary This method is performance-based. The labeled compounds are spiked into a

sample containing 10 g (dry weight) of solids. Samples containing multiple phases

are pressure filtered and any aqueous liquid is discarded. Coarse solids are ground

or homogenized. Any non-aqueous liquid from multi phase samples is combined

with the solids and extracted in a Soxhlet/Dean-Stark extractor. The extract is

concentrated for cleanup. After extraction, a labeled cleanup standard is spiked into

the extract which is then cleaned up using back-extraction with sulfuric acid and/or

base, and gel permeation, silica gel, or Florisil chromatography. Activated carbon

and high-performance liquid chromatography (HPLC) can be used for further

isolation of specific congener groups. After cleanup, the extract is concentrated to

20 μL. Immediately prior to injection, labeled injection internal standards are added

to each extract and an aliquot of the extract is injected into the gas chromatography

(GC). The analytes are separated by the GC and detected by a high-resolution

($10,000) mass spectrometer.

Without interferences, EMDLs and EMLs will be, respectively, 0.5 and 1.0 ng/kg for

soil, tissue, and mixed-phase samples, and EMLs for

extracts will be 0.5 pg/uL.

EMD: = Estimated Method Detection Limit; EML = Estimated Minimum Level

Data

Uses/Application

This Method is for use in data gathering and monitoring associated with the Clean

Water Act, the Resource Conservation and Recovery Act, the Com prehensive

Environmental Response, Compensation and Liability Act, and the Safe Drinking

Water Act.

Advantages Method 1668A provides congener data that can be used for source identification.

Listed PCBs include the 12 World Health Organization “dioxin-like” PCBs. The

HRMS method provides lower EMDLs compared to ECD or low resolution MS

analyses and provides unequivocal congener identification.

Limitations The GC/MS portions of this method are for use only by analysts experienced with

HRGC/HRMS or under the close supervision of such qualified persons. Solvents,

reagents, glassware, and other sample processing hardware may yield artifacts,

elevated baselines, and/or lock mass suppression causing misinterpretation of

chromatograms.

Reference USEPA. 1999c. Method 1668, Revision A: Chlorinated Biphenyl Cogeners in Water,

Soil, Sediment, and Tissue by HRGC/HRMS, EPA-821-R-00-002. Office of Water,

U.S. Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/region08/water/wast

ewater/biohome/biosolidsdown/methods

/1668a5.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

139

Fact Sheet No. 2.2.2-13

Method Title Butyltin in Sediments

Purpose The extraction and analysis of organotin compounds in soil and sediment.

Method Summary A 15 g aliquot of freeze-dried sediment sample is spiked with surrogate standards

and shaker extracted four times with 0.2% tropolone in dichloromethane. The

extract is concentrated by Kuderna-Danish technique and solvent exchanged to

hexane. Organotin compounds are hexylated with hexylmagnesium bromide

(Grignard reagent) by adding the reagent to the sample and heating the sample at

70°C for 30 minutes. The excessive reagent is removed with HCl and the organic

phase of the sample removed. The remaining aqueous phase is extracted twice

with pentane: CH2Cl2(3/1, v/v). The combined hexylated extract is dried with

sodium sulfate and concentrated. The sample is loaded onto a silica gel/alumina

column and eluded with pentane. The cleaned sample is concentrated and

analyzed by high resolution, capillary gas chromatography using flame photometric

detection (GC/FPD). This method quantitatively determines Tetra butyltin (4BT),

tributyltin (TBT), dibutyltin (DBT), and monobutyltin (MBT). Results are reported as

ng Sn/g. 

Data

Uses/Application

Butylin is a principal risk factor in many freshwater and marine harbor sediments.

Advantages FPD is a sensitive detector that is specific to tin. Hexylation of the organotin anions

provides compounds amenable to the GC/FPD technique and provides reliable

quantization of organotins at low concentrations (ng Sn/g).

Limitations Organotins are ubiquitous laboratory contaminants. Clean methods must be

observed.

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and Trends

Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo NOS ORCA

130. National Oceanic and Atm ospheric Administration, Silver Spring, MD. 233 pp.

Website http://ccma.nos.noaa.govfpublications

/tm130.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

140

Fact Sheet No. 2.2.2-14

Method Title Procedures for Sediment Total Organic Carbon (TOC) Determination

Purpose Determination of the TOC content in a sample by combustion.

Method Summary This method describes TOC determination by combusting preacidified samples at

high temperature and measuring the volume of carbon dioxide gas produced.

Samples are homogenized and 5 g wet weight subsamples dried for approximately

48 hours in a covered evaporation dish. The dried sediment sample is then ground

with a porcelain pestle. Approximately 20 to 30 mg of the dried and ground

sediment sample are placed in small beakers and acidified to remove sources of

inorganic carbon. Samples are then dried again and then exposed to a precombusted stream of oxygen. The CO2 evolved is measured by an infrared gas

analyzer and the resulting gas peak is integrated. Integrator units are compared to a

standard curve to convert to organic carbon. 

Alternatively, using the NS&T method, 0.1 to 0.5 ± 0.001 g of oven-dried, finely

ground homogenized sediment is weighed into a combustion crucible (NOAA,

1998). Approximately 1.4 g each of copper and iron chip accelerators are added to

the crucible. The crucible is placed and sealed within the oven combustion tube.

Total carbon compounds in samples are decomposed by pyrolysis in the presence

of oxygen, and the CO2

that is formed is quantified by infrared detection. Total

organic carbon (TOC) is determined by acidifying the sample with 10% HCl and

drying the sample overnight at 50°C. Acidification converts carbonate carbon to

carbon dioxide, which is purged from the acidified sample prior to analysis.

Carbonate carbon, or total inorganic carbon (TIC), is determined by the difference

between total carbon and total organic carbon.

Data

Uses/Application

TOC is used to norm alize the concentration of nonionic organic contaminants in the

development of equilibrium partitioning sediment guidelines (ESGs), and this is an

important chemical parameter for sediment quality.

Advantages The EMAP method is one of several combustion methods for TOC determination.

Limitations The TOC determination is not a substitute for the determination of biological oxygen

demand or chemical oxygen demand, should those parameters be needed. 

Reference USEPA. 1995. Environmental Monitoring and Assessment Program Laboratory

Methods Manual, Estuaries, Volume 1-Biological and Chemical Analyses,

EPA/620/R-95/008. Environmental Monitoring and Assessment Program , U.S.

Environmental Protection Agency, Washington DC.

Website http://www.epa.gov/emap/html/pubs/docs/grou

pdocs/estuary/field/lab_man.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

141

Fact Sheet No. 2.2.2-15

Method Title Determination of the Activity of Lead-210 in Sediments and Soils, LMMB 084

Purpose This procedure measures the activity of the Pb-210 granddaughter, Po-210.

Method Summary Sediment is collected with a gravity or box corer. The samples are extruded at

known intervals and placed into preweighed bottles. The samples are dried in a

60°C oven, and the dry weight calculated. The samples are then ground to a fine

powder and stored until used. 0.50 g of dried sediment is digested with a

combination of HCl and hydrogen peroxide. During the digestion, the sample is

heated on a hot plate to 90-95°C for a total of four hours. After sitting overnight, the

sample is filtered through a Whatman No. 42 filter paper into a flask. The sample is

heated until its volume is reduced to 5 mL. The pH of the sample is adjusted to 0.5

to 1.0. Ascorbic acid is added to prevent interference from ferric iron. A copper disk

is added to the sample, which is heated overnight in a 95°C oven. The Po-210

concentration on the disk is determined by alpha spectrometry using silicon surface

barrier detectors . A yield monitor, Po-208, is added to each sample to determine

the exact activity of Po-210.

Data

Uses/Application

The activity of Pb-210 can be used to estimate dates of sediment deposition. Such

dating can be used in establishing chronologies of sediment contamination, rates of

sediment accumulation and rates of contaminant attenuation.

Advantages Straight forward and accurate analytical method.

Limitations Pb-210 has a half life of 22 years, limiting age dating using Pb-210 to approximately

the last 100 years. If sediments have been disturbed (e.g., by past dredging,

scouring, bioturbation), accurate dating may not be possible. It should be noted that

other analyses may be more accurate for dating sediment deposition which occurred

more recently than the last 100 years. For example, analysis for Cesium-137 may

be suitable for dating sediments deposited in the previous 50 years, while Beryllium7 can date sediments deposited in the previous two years. Using more than one

dating method can provide added assurance in date estimates.

Reference USEPA. 1997c. Method LMMB 084: Determination of the Activity of Lead-210 in

Sediments and Soils. Lake Michigan Mass Balance Study Methods Compendium,

Volume 3: Metals, Conventionals, Radio chemistry, and Biomonitoring Sample

Analysis Techniques. EPA 905-R-97-012c. U.S. Environmental Protection Agency,

Washington, DC.

Website http://www.epa.gov/glnpo/lmmb/methods/lead210.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

142

Fact Sheet No. 2.2.2-16

Method Title Sediment Grain Size Analysis, NHEERL-AED SOP 1.01.005

Purpose To determine the percentages by weight of sand, silt, and clay in sediment samples

Method Summary 15 - 20 gm of sediment is treated with 50 - 100 mL of 30% hydrogen peroxide for 12

hours to oxidize any organic matter present. The sample is then washed with

distilled water to remove salts. 400 mL of sodium hexametaphosphate solution is

used to disperse the particles in the sediment. The sample is shaken for 24 hours.

The gravel/sand fraction is wet-sieved through a .063 mm sieve into an underlying

evaporation dish to separate the sands (>.063 mm) from the silt and the clay

(<.063mm). The material remaining on the sieve(gravel/sand) is washed into a preweighed beaker. This fraction is dried at 100 - 130°C for 24 hours and then weighed

to the nearest 0.1 g.

The silt and clay in the evaporation dish is then transferred to a graduated cylinder

and 10 mL of dispersant from stock solution of sodium hexametaphosphate is

added. The solution is stirred and stored for 12 hours. If flocculation occurs, the

sample is treated with more dispersant and mixed. The silt and clay fractions are

measured using the pipette method (Folk, 1974). The solution is stirred and aliquots

are pipetted out at specified times and depths. Silt and clay fraction: After 20

seconds of stirring, the pipette is inserted to 20 cm and at the end of 20 seconds, 20

mL is removed. This fraction is placed into a beaker. Clay fraction: After 2 hours

and 3 minutes, the pipette is inserted to a depth of 5 cm and 20 mL is withdrawn.

This fraction is placed into a beaker.

The removed fractions are dried in an oven at 100 - 130°C for at least 24 hours.

Total dry weight = wt. sand + wt. silt + wt. clay. The weight of silt+clay is multiplied

by 50 and the weight of one dispersant is subtracted to obtain the total weight of

silt+clay. This process is repeated for the clay fraction. Total dry weight = wt. sand

+ wt. silt + wt. clay. The respective percentages of sand, silt and clay are derived by

dividing the individual weights by the total weight and multiplying by 100.

Grain size analysis is also described in ASTM Method D422, NOAA NS&T, and

EMAP Lab methods (ASTM, 2001e; NOAA, 1998).

Data

Uses/Application

Grain size is an important characteristic of sediments that may be correlated with

contaminant concentrations. Data may also be used in sediment transport methods.

Advantages This is one of several methods derived from Folk (1974) that has gained wide

acceptance for grain size determination. 

Limitations This method does not provide M classification data; these data are important in

some transport models.

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296. Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.01.005.pdf

Last Accessed:

Fact Sheet No. 2.2.2-17
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

143

Method Title Procedures for Water Content Determination

Purpose Determine the percent water content of sediment samples.

Method Summary Sediments are placed in a 250 mL glass beaker and homogenized. Approximately

5-10 grams wet weight of sediment will be placed in a clean, tared 50 mL glass

beaker and the weight are recorded. The sample is dried for 24 hours and then

reweighed. Percent water content is then calculated. 

Alternatively, using the NS&T method, sediments are homogenized using a solvent

rinsed spatula (NOAA, 1998). Approximately five grams of sample is placed in a

pre-weighed scintillation vial (combusted for 4 hr at 400°C) and the weight recorded.

The samples are dried for 24 hours in a drying oven set at 63-65°C. Samples are

placed in a desiccator and allowed to cool to room temperature for at least 30

minutes. The samples are weighed. The samples are put back in the oven for at

least 2 hr after which they are removed from the oven and allowed to cool for at

least 30 min in a desiccator. The sample is reweighed. If the difference between

the first and second weighting wis less than ± 0.02 g, the dry weight percent is

calculated based on the last weighing.

 [Vial wt. + Dry sample wt.] - [Vial wt.]

Percent dry weight = [Vial wt. + W et sample wt.] - [Vial wt.] X 100

The percent water content is calculated as 100 - percent dry weight.

Data

Uses/Application

Dry weight measurem ents are used to calculate sediment analyte concentrations on

a dry weight basis. In addition, water content is one of a suite of parameters

typically used in settlement calculations for contaminated sediment capping studies.

The EPA Region I functional guidelines use percent solids data as follows. If the

percent solids content of a soil/sediment sample falls below 30%, all positive and

non-detect results in that sample are qualified (J for positive results, R for nondetects). When the percent solids content of a soil/sediment sample falls below

10%, all results in that sample are rejected (R).

Advantages Dry weight measurem ents determined in this manner can be compared directly to

NOAA NS&T database.

Limitations This method has not been widely accepted as the method for drying to constant

weight at 105° C.

Reference USEPA. 1995. Environmental Monitoring and Assessment Program Laboratory

Methods Manual, Estuaries, Volume 1-Biological and Chemical Analyses,

EPA/620/R-95/008. Environmental Monitoring and Assessment Program , U.S.

Environmental Protection Agency, Washington DC.

Website http://www.epa.gov/emap/html/pubs/docs/grou

pdocs/estuary/field/lab_man.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

144

Fact Sheet No. 2.2.2-18

Method Title Standard Test Method for Field Vane Shear Test in Cohesive Soil, ASTM Method

D2573

Purpose To determine the shear strength of cohesive soils.

Method Summary A four-bladed vane is placed in the undisturbed soil and is rotated from the surface

to determine the torsional force required to cause a cylindrical surface to be sheared

by the vane. Friction of the vane rod and instrument must be accounted for, so as

not to record friction as soil strength. Shear is calculated as torque multiplied by the

inverse of a constant, depending on the dimensions and shape of the vane.

Data

Uses/Application

Field vane shear testing measures the in situ undrained shear strength of sediments

underlying the surficial low- bearing capacity sediments. In-situ contaminated

sediments to be capped are predominately fine-grained, and may have high water

contents and low shear strengths. The shear strength of sediments will influence

their resistance to localized bearing capacity or sliding failures, which may cause

localized mixing of capping and contaminated materials.

Advantages There are cost and schedule advantages to performing this procedure in the field.

Limitations This test should not be performed in any soil such as sand or silt that will permit

drainage during the test period or in soils where stones or shells are encountered by

the vane.

Reference ASTM. 2001e. ASTM Book of Standards. Volume 04.08. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

145

Fact Sheet No. 2.2.2-19

Method Title Standard Test Method for Specific Gravity of Soil Solids by Water Pycnometer,

ASTM Method D854

Purpose To determine the specific gravity of soil solids that pass the 4.75-mm sieve, by

means of a water pycnometer..

Method Summary Two alternative methods exist for measuring specific gravity: procedure for moist

specimens and procedure for oven-dry specimens. Guidelines exist that

recommend the dry soil mass versus soil type and pycnometer size. After these

guidelines have been consulted and the sample size selected, the pycnometer

volume is calibrated. The pycnometer is then weighed. Following the procedure for

moist samples, the water content of the sample is determined, and the sample is

mixed with 100 mL of water to form a slurry. The slurry is transferred into the

pycnometer. Following the procedure for oven-dry specimens, the sample is dried

to a constant mass in a 100°C oven and then transferred to the pycnometer. 

Water is then added to the pycnometer to form a slurry, and the slurry is deaired

using either heat, vacuum, or a combination of both. The pycnometer is filled with

deaired water and placed into an insulated container, where it is left overnight to

thermally equilibrate. The following day, the mass of the pycnometer, soil, and water

are determined, and the temperature of the slurry is measured. The soil slurry is

transferred to a tared pan and dried to a constant mass at 110°C. The specific

gravity of soil solids at test temperature is calculated using the following equation:

 Gt = Ms

 

 (Mpw,t - (Mpws,t - Ms

)), where

Ms = the mass of the oven dry soil solids (g)

Mpw,t = the mass of pycnometer and water at the test temperature (g)

Mpws,t = the mass of pycnometer, water, and soil solids at the test temperature (g)

The specific gravity of soil solids at 20°C is calculated by multiplying the specific

gravity at the test temperature by a temperature coefficient.

Data

Uses/Application

Specific gravity is used to calculate the phase relationship of soils, such as void ratio

and degree of saturation, and soil solid density. In addition, specific gravity is one of

a suite of parameters typically used in settlement calculations for contaminated

sediment capping studies.

Advantages This is the standard, accepted method for measuring specific gravity in soil.

Limitations This analysis cannot be performed on soil solids that may be altered by this method

or on highly organic soil solids. 

Reference ASTM. 2001e. ASTM Book of Standards. Volume 04.08. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

146

Fact Sheet No. 2.2.2-20

Method Title Standard Test Method for Permeability of Granular Soils (Constant Head), ASTM

Method D2434

Purpose To determine the coefficient of perm eability by a constant-head method.

Method Summary A representative sample of soil is analyzed for particle size before the perm eability

test. Any particles larger than 19 mm are separated out by sieving and the

percentage of oversize material is recorded. Of the sieved sample, a subsample of

approximately twice the amount required to fill the parameter chamber is selected by

the method of quartering. First, the cross-sectional area of the sample is calculated.

A subsample of the soil is analyzed for water content. The air-dried soil sample is

spread and compacted in successive layers into the parameter until the device is

filled to the proper level. The unit weights, void ratio, and relative density of the test

specimen are calculated. Air adhering to soil particles and present in the voids is

removed with a vacuum pump or aspirator. The sample is then saturated with

water, preferably native water. The inlet valve from the filter tank is opened and

time, head, quantity of flow and water temperature are recorded when a stable head

condition is attained. Test runs are repeated at heads increasing by 0.5 cm in order

to accurately establish the region of laminar flow with velocity directly proportional to

hydraulic gradient. The coefficient of perm eability is calculated as follows:

 k = QL/Ath, where

k = coefficient of permeability,

Q = quantity of water discharged,

L = distance between manom eters,

A = cross-sectional area of specimen,

t = total time of discharge, and

h = difference in head on manom eters.

The perm eability is corrected to that for 20°C.

Data

Uses/Application

Permeability measurem ents are needed for engineering sediment capping

alternatives.

Advantages This is the standard and accepted method for measuring perm eability in soil.

Limitations This procedure is limited to disturbed granular soils containing not more than 10%

soil passing the 75-μm sieve. 

Reference ASTM. 2001e. ASTM Book of Standards. Volume 04.08. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

147

Fact Sheet No. 2.2.2-21

Method Title Standard Test Method for One-Dimensional Consolidation Properties of Soil, ASTM

Method D2435

Purpose To determine the magnitude and rate of consolidation of soil.

Method Summary This test procedure is usually performed on undisturbed samples of fine grained

soils naturally sedimented in water. The sample is trimm ed and placed in the tared

consolidation ring. The sample is then trimm ed flush with the plane ends of the ring.

The initial wet mass, height, volume, and water content of the sample are

determined. The sample is loaded into the consolidometer. The standard loading

schedule consists of a load increment ration (LIR) of one which is obtained by

doubling the pressure on the soil to obtain values of approximately 12, 25, 50, 100,

200, etc. kPa. The standard unloading schedule is selected by halving the pressure

on the soil. Before each pressure increment is applied, the height or change in

height of the sample is recorded. Two alternative procedures exist for the analysis

of soil consolidation. Test Method A is performed with constant load increment

duration of 24 hours, or multiples thereof. Time-deformation readings are required

on a minimum of two load increments. Test Method B requires time-deformation

readings on all load increments. Successive load increments are applied after

100% primary consolidation is reached, or at constant time increments as described

in Test Method A. The deformation results are plotted (void ratio or strain), and the

plot is used to determine the value of the preconsolidation pressure.

Data

Uses/Application

The data from the consolidation test are used to estimate the magnitude and rate of

both differential and total settlement of a structure or earthfill.

Advantages This is the standard and accepted method for measuring consolidation in soil.

Limitations The test results can be greatly affected by sample disturbance and greatly

dependent on the competence of the personnel performing the test. 

Reference ASTM. 2001e. ASTM Book of Standards. Volume 04.08. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

148

Fact Sheet No. 2.2.2-22

Method Title Standard Test Method for Classification of Soils for Engineering Purposes (Unified

Soil Classification System), ASTM Method D2487

Purpose To classify mineral and organo-mineral soils based on laboratory determination of a

variety of parameters.

Method Summary This classification system identifies three major soil divisions: coarse-grained soils,

fine-grained soils, and highly organic soils. These three divisions are further

subdivided into a total of 15 basic soil groups. 

The minimum amount of test sample required for this test method depends on which

of the laboratory tests need to be performed. The laboratory tests include particlesize determination, liquid limit, and plasticity index. The percentage of fines found in

the sample dictates which tests are required. The results of these tests are used to

classify soils by group.

Data

Uses/Application

Soil classification is used to describe a soil and to aid in the evaluation of its

significant properties for engineering use.

Advantages Provides visual classification of soils for engineering purposes.

Limitations This classification system is dependent on the competence of trained personnel. 

Reference ASTM. 2001e. ASTM Book of Standards. Volume 04.08. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

149

Fact Sheet No. 2.2.2-23

Method Title Standard Test Method for Liquid Limit, Plastic Limit, and Plasticity Index of Soils,

ASTM Method D4318

Purpose To determine the liquid limit, plastic limit, and plasticity index of soils.

Method Summary The liquid and plastic limits of soils (collectively referred to as Atterberg limits)

distinguish the boundaries of the several consistency states of plastic soils. These

analyses are performed only on the portion of soil that passes through the 425-μm

(No. 40) sieve. Two alternative methods exist for determining the liquid limit: the

multipoint test (the recommended method) and the one-point test. The liquid limit is

determined by conducting trials in which a portion of the sample is spread in a brass

cup, divided in two by a grooving tool, and allowed to flow together which the cup is

repeatedly dropped in a standard mechanical device. In the multipoint test, three or

more trials are conducted over a range of water contents. In the one-point test, two

trials at one water content is conducted. 

The plastic limit is determined by alternately pressing and rolling a portion of plastic

soil into a 3.2 mm diameter thread until its water content is reduced to a point at

which the thread crumbles and can no longer be pressed together and recoiled.

The water content of the soil at this point is reported as the plastic limit. The

plasticity index is calculated as the difference between the liquid limit and the plastic

limit.

Data

Uses/Application

Results from these analyses are used in the soil classification process to

characterize the fine-grained fractions of soils.

Advantages This is the standard, accepted method for measuring the plasticity of soils.

Limitations The one-point test is not recommended for inexperienced analysts. The one-point

test may not be valid for certain types of soil, such as organic or marine soils. 

Reference ASTM. 2001e. ASTM Book of Standards. Volume 04.08. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

150

Fact Sheet No. 2.2.2-24

Method Title Field Portable X-Ray Fluorescence Spectrom etry for the Determination of Elemental

Concentrations in Soil and Sediment

Purpose Site characterization, screening

Method Summary Under this method, inorganic analytes of interest are identified and quantified

directly by a field portable X-Ray fluorescence spectrometer. Radiation from one or

more radioisotope sources or an electrically excited X-Ray tube is used to generate

the characteristic X-Ray emissions in a sample. Each source emits a specific set of

primary x-rays that excite a corresponding range of elements in a samples. When

more than one source can excite the element of interest, the source is selected

according to its excitation efficiency for the element of interest.

For measurem ent, the sample is positioned in front of the probe window. Samples

may be analyzed in one of two manners: in situ or intrusive. If in situ mode, the

probe window is placed in direct contact with the soil or sediment. In the intrusive

mode, the sample is collected, prepared, and placed in a sample cup, which is then

placed on top of the probe window inside a protective cover for analysis.

Most FPXRF instruments are menu-driven from software developed by the

manufacturer. The measurement time for each source is user-selectable. Shorter

source measurement times (30 seconds) are used for initial screening or hot spot

delineation. Longer times (up to 300 seconds) are used to meet higher precision

and accuracy requirements.

 

Data

Uses/Application

Twenty-six elements can be measured by this method. Field-based detection limits

established for Sb, As, Ba, Cr, Co, Cu, Pb, Mn, Mo, Ni, Rb, Sr, Sn, Zn, and Zr. Field

screening of Cu, Pb, and Zn at metal contaminated sites has been demonstrated by

the Navy (NFESC, 2000). Method 6200 is intended for dry samples. The Navy has

demonstrated good results on unprepared, wet sediment (NFESC, 2000).

Advantages Rapid on-site screening for selected elements allows rapid characterization of many

metal-contaminated sites.

Limitations High levels of V and Fe will interfere with the quantitation of Cr and Co, respectively.

A high ratio of Pb to As may result in no As being reported regardless of actual

concentration. Trained operators must understand the limitations of the method.

Confirmatory laboratory analysis required.

Reference USEPA. 2000c. Method 6200, Field Portable X-Ray Fluorescence Spectrom etry for

the Determination of Elemental Concentrations in Soil and Sediment. Office of Solid

Waste, U.S. Environmental Protection Agency, Washington DC.

Website http://www.epa.gov/epaoswer/hazwaste/

test/pdfs/6200.pdf

Last Accessed: 1/30/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

151

Fact Sheet No. 2.2.2-25

Method Title Sediment Age Dating Using Cesium-137

Purpose To determine chronological timescales in sediments using the radioisotope Cesium137

Method Summary Sediment is collected with a coring device, as warranted by the sampling location.

The samples are extruded from the corer in 5-cm segments and collected in

sampling jars. A 100g aliquot is used for 137Ce analysis. The sediment is

homogenized, weighed and an aliquot is freeze-dried prior to analysis to determine

percent solids. Sediment for analysis can be either wet or dry and does not require

any special storage conditions. Sediment for 137Ce analysis is gamma counted on a

Ge-diode detector. The sample is placed in front of or on top of the detector,

depending on the configuration of the system. The sample is placed in exactly the

same position as the standard and this position remains constant for all samples in

the batch. The sample is generally counted for 24 hours, depending on the size and

activity of the sample. 

Data

Uses/Application

The activity of Cesium 137 can be used to estimate dates of sediment deposition.

Such dating can be used to establish chronologies of sediment contamination, rates

of sediment accumulation and rates of contaminant attenuation.

Advantages Cesium 137 has a relatively short residence time in natural waters.

Limitations Cesium 137 is a thermonuclear byproduct. Its presence in natural systems is

directly related to thermonuclear activity and therefore its useful in detecting effects

since the 1950's only (onset of aboveground nuclear weapons testing). Particle size

distribution (PSD, or grain size analysis) should be conducted concurrently to

support the assumption that uniform sediment processes were occurring during the

time of interest. 

Reference Battelle. 2001. Natural Recovery of Persistent Organics in Contaminated Sediments

at the Sangamo-Weston/Twelvemile Creek/Lake Hartwell Superfund Site. USEPA

National Risk Management Research Laboratory Cincinnati, OH.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

152

Fact Sheet No. 2.2.2-26

Method Title Beryllium-7 as a Tracer of Short Term Sediment Deposition

Purpose To aid in the estimation of particle resuspension and deposition rates in sediments.

Method Summary Sediments are collected in the field with the appropriate sampler (e.g. core, grab, or

sediment trap). Sediments are returned to the lab for analysis within 48 hours of

collection. In the lab, the samples are dried to constant weight, ground, and

homogenized using a mortar and pestle. The dried sediments are then compressed

into pellets, weighed, and measured. The activity of 7Be is determined on either a

lithium-drifted or intrinsic germanium detector coupled to a multichannel

autoanalyzer. The sample prep and analysis should take place in a shielded clean

room. Prior to sample analysis the detectors are calibrated and a calibration curve

is generated based on the sample size (in height). Measured counts per minute

(cpm) are divided by the detector efficiency yielding disintegrations per minute

(dpm). The activity is converted to picocuries per gram of dry weight and corrected

for the detector efficiency. 

Data

Uses/Application

7Be is a naturally occurring, atomospherically derived radioisotope that can help

provide actual rates of short-term deposition and resuspension of sediments. 

7Be

can be used to elucidate types and rates of processes that directly affect the cycling

of certain contaminants in the sediments, such as PCBs.

Advantages Because of it’s relatively short half-life, 7Be allows definition of short-term deposition

and resuspension rates.

Limitations 7Be tracer analysis is useful in describing processes (short-term deposition and

resuspension rates), and not particle type and composition.

Reference Fitzgerald, S.A., J Val Klump, PW Swarzenski, RA Mackenzie, and KD Richards.

2001. Beryllium-7 as a Tracer of Short-Term Sediment Deposition and

Resuspension in the Fox River, Wisconsin. Environmental Science and Technology

35/ 300-305.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

153

2.2.3 Biological Analysis Methods

Section 2.2.3 provides a compendium of sediment-related biological analyses. Table 2.2-1 lists

numerous acute freshwater, chronic freshwater, acute marine, chronic marine and

bioaccumulation tests that can be used to assess the effects that sediments may have on

endemic organisms. Attention was paid to include ***reference all standard test species, different

periods, endpoints. After performing physical analyses, the Superfund managers may select the

test that best suits the physical, chemical and biological parameters at their site. These tests are

performed to determine the lethal (acute) and sub-lethal (chronic) effects of the sediment on

resident organisms. These results are then compared with chemistry data to identify and

compare effects data with contaminant exposure data to determine the sediment’s risk to

ecological or human health.

The advantages and limitations associated with each test are provided in their respective fact

sheet. However, there are general limitations and interferences associated with all

active/chronic/freshwater/marine solid-phase toxicity tests. These potential interferences are

identified by some of the source documents (ASTM Method E1706 (ASTM, 2001b), USEPA,

2000d). They are listed below:

• Sediment collection handling and storage may alter bioavailability;

• Natural geochemical characteristics of sediment may affect the response of test

organism;

• Indigenous animals may be present in field-collected sediments;

• Route of exposure may be uncertain and data generated in sediment toxicity tests

may be difficulty to interpret if factors controlling the bioavailability of contaminants

in sediments are unknown;

• Tests applied to field samples may not discriminate effects of individual chemicals;

• Few comparisons have been made of methods or species;

• Only a few chronic methods for measuring sublethal effects have been developed

or extensively evaluated; and

• Laboratory tests have inherent limitations in predicting ecological effects.

The toxicity test fact sheets provide methods described in USEPA guidance documents, Dredging

Manuals, and ASTM reports from the following agencies and offices: 

• The USEPA’s Office of Water

• The USEPA’s Office of Research and Development

• The USEPA and USACOE Dredging Teams

• The Puget Sound Water Quality Action Team

• The Puget Sound Dredged Disposal Analysis Program

• The USEPA’s EMAP Program

• ASTM

• The USEPA’s Environmental Research Laboratory-Narragansett,

• The USEPA’s Great Lakes Program Office
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

154

Table 2.2.3-1. A Summary of Test Types and Toxicological Endpoints for Solid-Phase Toxicity

Test Type Test Organism Scientific Name Endpoints Test

Specifics

Comments

Acute Freshwater

2.2.3-1

2.2.3-2

2.2.3-3

Amphipod

Crustacean

Ceriodaphnia dubia Survival Static, Flow

through and in

situ, 48 hours

Commonly used for bioassays

Acute Freshwater

2.2.3-4

Crustacean

other than

amphipods

Hyallela azteca Survival and

growth

10 days Short generation time, contact

with sediment

Acute Freshwater

2.2.3-4

Insect Larvae Chironomus

tentans

Survival and

growth

10 days Short generation time, contact

with sediment

Chronic

Freshwater

2.2.3-5

Amphipod

Crustacean

Hyallela azteca Survival growth

and reproduction

growth

42 days Short generation time, contact

with sediment

Chronic

Freshwater

2.2.3-5

Insect Larvae Chironomus

tentans

Survival, growth,

reproduction,

emergence of

adults, egg

number, and

hatching success

Up to 60 days Short generation time, contact

with sediment

Acute Marine

2.2.3-7

Bivalve Mytilus edulis Larval Survival 48 hours May need to substitute if

sediments contain greater than

60% fines. Recommended at

dredged material sites 

Acute Marine

2.2.3-7

Bivalve Crassostrea gigas Larval survival,

abnormal shell

development

48 hours May need to substitute if

sediments contain greater than

60% fines. Recommended at

dredged material sites 

Acute Marine

2.2.3-8

Echinoderm Strongylocentrotus

purpuratus

Embryonic

survival

48-96 hours Recommended at dredged

material sites 

Acute Marine

2.2.3-8

Echinoderm Strongylocentrotus

droebachiensis

Acute/Embryonic

survival

48-96 hours Recommended at dredged

material sites

Acute Marine

2.2.3-8

Echinoderm Dendraster

excentricus

Embryonic

survival

48-96 hours Recommended at dredged

material sites

Acute

Estuarine/Marine

2.2.3-9

Crustaceans

other than

amphipods

Mysidopsis bahia Survival 96 hours Wide tolerance to grain size,

salinity and temperature.

Epibenthos. Filter-feeder,

deposit-feeder

Acute Estuarine/

Marine

2.2.3-9

Crustaceans

other than

amphipods

Penaeus sp. Survival 96 Hours Deposit-feeder, burrower
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

155

Table 2.2.3-1. (contd.)

Test Type Test Organism Scientific Name Endpoints Test

Specifics

Comments

Acute Marine

2.2.3-10

Amphipod

Crustacean

Ampelisca abdita Survival 10 days Wide tolerance to grain size,

salinity and temperature. May

be used if test sediment

contains greater than 60%

fines. Tube dweller, depositfeeder, burrower.

Recommended at dredged

material sites

 

Acute

Marine/Estuarine

2.2.3-10

Amphipod

Crustacean

Eohaustorius

estuarius

Survival 10 days Wide tolerance to grain size,

salinity and temperature. May

be considered for use over

grain size distributions ranging

from 100% sand to 0.6% sand,

as long as the clay fraction

<30%; Free burrowing, depositfeeder. Recommended at

dredged material sites

 

Acute Marine

2.2.3-10

Amphipod

crustacean

Rhepoxynius

abronius

Survival 10 days Preferred species for coarsergrained sediments (i.e., fines

<60%) Free burrowing, depositfeeder. Recommended at

dredged material sites

 

Acute Marine

2.2.3-11

Polychaete Neanthes

arenaceodentata

Survival 10 days Size class must be uniform for

biomass estimates. Tube

dweller. Deposit-feeder,

burrower. Recommended at

dredged material sites

Identified for bioaccumulation

studies based on feeding type,

biomass, salinity tolerance,

pollution tolerance, culture

potential, bioaccumulation

toxicity information, commercial

availability, and historic use in

other programs.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

156

Table 2.2.3-1. (contd.)

Test Type Test Organism Scientific Name Endpoints Test

Specifics

Comments

Acute Marine

2.2.3-11

Polychaete Neanthes virens Survival 10 Days Must be held under flowthrough conditions. Depositfeeder, burrower.

Identified for bioaccumulation

studies based on feeding type,

biomass, salinity tolerance,

pollution tolerance, culture

potential, bioaccumulation

toxicity information, commercial

availability, and historic use in

other programs.

Chronic Marine

2.2.3-12

Amphipod

crustacean

Leptocheirus

plumulosus

Survival, Growth,

and Reproduction

28 days Deposit-feeder, burrower

Chronic Marine

2.2.3-13

Polychaete Neanthes

arenaceodentata

Survival 28 days same as above

Bioaccumulation

Freshwater

2.2.3-17

Oligochaete Lumbrculus

variegatus

Bioaccumulation 28 days Easy to culture and handle,

tolerant of a wide range of

sediment characteristics, and it

is adaptable to long-term test

exposures.

Bioaccumulation1

Marine

2.2.3-17

Bivalve Macoma balthica Bioaccumulation 28 days Identified based on feeding

type, biomass, salinity

tolerance, pollution tolerance,

culture potential,

bioaccumulation toxicity

information, commercial

availability, and historic use in

other programs.

Bioaccumulation 1

Marine

2.2.3-17

Bivalve Macoma nasuta Bioaccumulation 28 days Identified based on feeding

type, biomass, salinity

tolerance, pollution tolerance,

culture potential,

bioaccumulation toxicity

information, commercial

availability, and historic use in

other programs.

Bioaccumulation 1

Marine

2.2.3-17

Bivalve Yoldia limatula Bioaccumulation 28 days Identified based on feeding

type, biomass, salinity

tolerance, pollution tolerance,

culture potential,

bioaccumulation toxicity

information, commercial

availability, and historic use in

other programs.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

157

Table 2.2.3-1. (contd.)

Test Type Test Organism Scientific Name Endpoints Test

Specifics

Comments

Bioaccumulation1

Marine

2.2.3-17

Polychaete Neanthes virens Bioaccumulation 28 days Must be held under flowthrough conditions. Depositfeeder, burrower.

Identified for bioaccumulation

studies based on feeding type,

biomass, salinity tolerance,

pollution tolerance, culture

potential, bioaccumulation

toxicity information, commercial

availability, and historic use in

other programs.

Bioaccumulation1

Marine

2.2.3-17

Polychaete Nereis diversicolor Bioaccumulation 28 days Identified for bioaccumulation

studies based on feeding type,

biomass, salinity tolerance,

pollution tolerance, culture

potential, bioaccumulation

toxicity information, commercial

availability, and historic use in

other programs.

1

For bioaccumulation tests, it is recommended that a deposit-feeding bivalve mollusk and a burrowing polychaete are used. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

158

Fact Sheet No. 2.2.3-1

Method Title Acute Freshwater Crustacean Sediment Bioassay: Flow-through

Purpose This test is used to determine the effects of a 48-hour exposure of the freshwater

crustacean, Ceriodaphnia dubia, to sediments in a laboratory flow-through system. 

Method Summary 200-mL aliquots of sediment (by weight) are placed in 1-L glass beakers, and 800

mL of site water is added slowly. Ceriodaphnia neonates are then placed in

sediment exposure chamber (SEC) units and the units are placed in one of the test

beakers containing sediment. The inlet port of the SEC unit connects to a reservoir

(20 L) of site water. The flow-through rate is controlled by a metering pump

calibrated to match the retention time of water flowing through the chambers in the

field. Survival of Ceriodaphnia neonates is determined at the end of the 48-hour test.

Data

Uses/Application

The flow-through test allows investigators to better assess sediment contamination

by simulating field conditions with water flow and controlling parameters such as pH

and dissolved oxygen to examine for direct effects from contaminants.

Advantages Ceriodaphnia dubia have been used widely as a test species. It has shown to be a

sensitive and useful test species for assessments of sediment toxicity.

Limitations Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference Sasson-Brickson, G. and G.A. Burton, Jr. 1991. In Situ and Laboratory Sediment

Toxicity Testing with Ceriodaphnia Dubia. Environmental Toxicology and

Chemistry. Vol 10. P 201-207. 

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

159

Fact Sheet No. 2.2.3-2

Method Title Acute Freshwater Crustacean Sediment Bioassay: In Situ Exposures

Purpose This test is used to determine the effects of a 48-hour exposure of the freshwater

crustacean, Ceriodaphnia dubia, to sediments in situ. 

Method Summary Ceriodaphnia dubia are transported to the test location and placed in one of five

sediment exposure chambers (SEC). These acrylic chambers are tied together and

placed onto the sediment surface. The inlet port of the SEC is directed upstream,

so that it receives water that flows into the chamber, circulates and then exits via the

outlet port, which is directed downstream. 

The SEC units are collected after 48-hours, placed in a polyethylene bucket

containing site water, covered, and transported back to the laboratory, where

surviving organisms were enumerated within 1.5 hours.

Test-site water toxicity is differentiated from sediment toxicity by placing SEC units in

situ with a plastic barrier between the unit base and the sediment surface. Water

samples are also collected simultaneously in high-density polyethylene bottles and

returned to the laboratory for toxicity testing. 

Data

Uses/Application

In situ sediment exposures prove to be sensitive indicators of both degraded and

nondegraded stream conditions. These types of experiments provide a way to

compare and validate laboratory results to determine the accuracy of these

laboratory experiments.

Advantages Ceriodaphnia dubia have been used widely as a test species. It has shown to be a

sensitive and useful test species for assessments of sediment toxicity.

Limitations It is more difficult in the field to test effects exclusively from contaminants, since the

investigators are unable to control all other parameters such as temperature, pH,

and dissolved oxygen levels. Although the test is more representative of actual

conditions, it is less definitive in correlating contaminant presence and ecological

effects. 

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference Sasson-Brickson, G. and G.A. Burton, Jr. 1991. In Situ and Laboratory Sediment

Toxicity Testing with Ceriodaphnia Dubia. Environmental Toxicology and

Chemistry. Vol 10. P 201-207. 

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

160

Fact Sheet No. 2.2.3-3

Method Title Acute Freshwater Crustacean Sediment Bioassay: Static Laboratory Exposures

Purpose This test is used to determine the effects of a 48-hour exposure of the freshwater

crustacean, Ceriodaphnia dubia, to sediments in a static laboratory system. 

Method Summary To assess the partitioning of sediment-bound toxicants, the sediment is tested as

solid phase, interstitial phase and elutriate phase. For solid-phase tests, sediments

are homogenized in the laboratory with a hand paddle for approximately 5 minutes.

The solid-phase test is prepared by placing 30 mL of wet sediment (by weight) into

test beakers, then slowly adding 120 mL of reconstituted hard water with a syringe.

Test beaker contents are allowed to settle for one hour prior to adding the test

organisms. 

For interstitial phase tests, 400 to 450 mL of wet sediment is placed into a 500 mL

polycarbonate bottle. The samples are centrifuged at 9000 rpm for 15 minutes. The

resultant interstitial water is immediately siphoned off and placed in the test beakers.

For elutriate phase tests, sediment and water are mixed in a 1:4 ratio by volume.

Sediment is placed in a 500-mL polycarbonate bottle; 300 mL of reconstituted water

is added to the bottle. The bottles are shaken on a Eberback shaker table for 30

minutes. After the mixture has settled, the liquid portion is siphoned off and

centrifuged at 9000 rpm for 15 minutes. The supernatant water is siphoned off the

top and added to the test beakers. 

Ceriodaphnia neonates are then exposed to test media (i.e., whole sediment,

interstitial water or elutriates) in 250-mL glass beakers containing 150-mL of the test

solution. Beakers are maintained at 25 +/- 1

N C. Survival numbers are recorded at

24 and 48 hours. Tests were considered valid when control mortality was < 10%. 

Data

Uses/Application

This test method may be useful in assessing sediment contamination, registration

of pesticides, assessments of new and existing chemicals, Superfund site

assessment, and assessment and cleanup of hazardous waste treatm ent, storage,

and disposal facilities. 

Advantages Ceriodaphnia dubia have been used widely as a test species. It has shown to be a

sensitive and useful test species for assessments of sediment toxicity.

Limitations Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference Sasson-Brickson, G. and G.A. Burton, Jr. 1991. In Situ and Laboratory Sediment

Toxicity Testing with Ceriodaphnia Dubia. Environmental Toxicology and

Chemistry. Vol 10. P 201-207. 

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

161

Fact Sheet No. 2.2.3-4

Method Title Acute Freshwater Amphipod and Freshwater Insect Larvae Sediment Bioassay, EPA

Method 100.1

Purpose This test measures the survival and growth of the estuarine amphipod crustacean,

Hyallela azteca, and the freshwater midge, Chironomus tentans after exposure to

sediments for 10 days in the laboratory. 

Method Summary Hyalella azteca and/or Chironomous tentans are exposed to sediments for 10-days

in a 300 mL test chamber containing 100 mL of sediment and 175 mL of overlying

water that is renewed. Test temperature is 23 °+/- 1 °C. There are 10 organisms per

chamber. The endpoints are survival and growth. Minimum mean control survival

must be 70% and measurable growth of test organisms. Test organisms are fed

daily.

ASTM E1706-00 describes similar methods for conducting whole sediment toxicity

tests with Cladocerans, mayflies, Chironomus riporius, Diporeia spp and Tubifex

(ASTM Method E1706, ASTM 2001b).

Environment Canada also describes a similar method using the test species Hyalla

azteca in EPS/ 1/RM/33 and Chironomus riparies in EPS/1/RM/32.

Data

Uses/Application

Sediments tests can be used to determine the relationship between toxic effects and

bioavailability, investigate interactions among chemicals, compare the sensitivities of

different organisms, determine spatial and temporal distribution of contamination,

evaluate hazards of dredged material, measure toxicity as part of product licensing or

safety testing, rank areas for clean up and estimate the effectiveness of remediation

or managem ent practices. 

Advantages Amphipods and midges are commonly used as appropriate test species to determine

acute toxicity because they are relatively sensitive to contaminants associated with

sediments, they have a short generation time, they have lots of contact with the

sediment (i.e., they are both deposit-feeders and burrowers), they are relatively easy

to culture in the laboratory and they tolerate varying physico-chemical characteristics

of sediment. 

Both the amphipod and midges are considered benchmark organisms by the USEPA

and USACE Inland Testing Manual Standards (1998) and testing data comprise a

substantial database. The organisms represent the sensitive range in a variety of

ecosystems. These organisms provide comparative data on the relative sensitivity of

local test species. 

The amphipod, Hyallela azteca, can also be used for estuarine toxicity studies; they

are tolerant to salinities up to 25ppt. 

Limitations Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

162

Fact Sheet No. 2.2.3-4 (contd.)

Reference USEPA. 2000d. Methods for Measuring the Toxicity and Bioaccumulation of

Sediment-Associated Contam inants with Freshwater Invertebrates. EPA/600/R99/064. Office of Science and Technology, U.S. Environmental Protection Agency,

Washington, D.C.

Website http://www.epa.gov/waterscience/cs/freshmanua

l.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

163

Fact Sheet No. 2.2.3-5

Method Title Chronic Freshwater Amphipod Sediment Bioassay, EPA Method 100.4

Purpose This test measures the survival, growth and reproduction of the freshwater

crustacean, Hyalella azteca, after exposure to sediments for 42 days in the

laboratory. 

Method Summary The freshwater invertebrate, Hyalella azteca, is exposed to sediments for 42-days in

a 300 mL container containing 100 mL of sediment and 175 mL of overlying water.

Test temperature is 23 +/- 1

NC. 100 mL of overlying water will be renewed every 12

hours. The organisms are fed daily. The endpoints are 28-day survival and growth,

35-day survival and reproduction; and 42-day survival, growth and reproduction.

Survival is measured by counting the number of alive vs. dead amphipods at 28, 35

and 42 days. Reproduction is measured by exposing amphipods up until a few days

before the release of the first brood. The amphipods are then sieved from the

sediment and held in water to determine the number of young produced. Length

and weight are measured to provide data for the growth endpoint. 

Data

Uses/Application

This is a laboratory method for determining the chronic toxicity of contaminants

associated with sediments collected from freshwater environments. 

Advantages The amphipod, Hyallela azteca, is considered benchmark organism by the USEPA

and USACE Inland Testing Manual Standards (1998) and testing data comprise a

substantial database. The amphipod sensitivity corresponds to the sensitive range

in a variety of ecosystems. This organism provide comparative data on the relative

sensitivity of local test species. 

Limitations The methodology recommended for measuring reproduction may not be accurate;

the amphipods may recover from effects of sediment exposure during the holding

period in clean water.

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 2000d. Methods for Measuring the Toxicity and Bioaccumulation of

Sediment-Associated Contam inants with Freshwater Invertebrates, EPA/600/R99/064. Office of Water, U.S. Environmental Protection Agency, Washington, D.C.

Website http://www.epa.gov/waterscience/cs/freshman

ual.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

164

Fact Sheet No. 2.2.3-6

Method Title Life-Cycle Freshwater Midge Sediment Bioassay, EPA Method 100.5

Purpose This test measures the survival, growth and reproduction characteristics of the

freshwater midge, Chironomus tentans, after exposure to sediments for 40 or 50

days in the laboratory. 

Method Summary Chironomus tentans, a freshwater invertebrate, is exposed to sediments for up to 60

days in a 300 mL beaker containing 100 mL of sediment and 175 mL of overlying

water. Test temperature is 23 +/- 1 °C. The endpoints are 20-day and end of test

(50-60 days) survival, 20-day growth, and reproduction is monitored daily after day

23, examining endpoints such as emergence of adults, egg number and hatchling

success.

The end of the life-cycle test depends upon the sediments being evaluated. In clean

sediments, the test typically requires 40 to 50 d from initial setup to completion.

However, test duration will increase in the presence of environmental stressors

which act to reduce growth or delay emergence. Where a strong gradient of

sediment contamination exists, emergence patterns between treatm ents will likely

become asynchronous, in which case each treatment needs to be ended separately.

 

Data

Uses/Application

This is a laboratory method for determining the chronic toxicity of contaminants

associated with sediments collected from freshwater environments.

Advantages C. tentans is a good candidate for long-term toxicity testing because it has a short

life cycle and a variety of developmental (growth, survivorship) and reproductivity

(fecundity) endpoints can be monitored. 

The midge, C. tentans, is considered a benchmark species by the USEPA and

USACE Inland Testing Manual Standards (1998) and midge testing data form a

substantial database. Midge sensitivity is in the range of a many ecosystems, and

midges provide comparative data on the relative sensitivity of local test species.

They are burrowers and deposit-feeders.

Limitations Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 2000d. Methods for Measuring the Toxicity and Bioaccumulation of

Sediment-associated Contam inants with Freshwater Invertebrates, EPA/600/R99/064. Office of Water, U.S. Environmental Protection Agency, Washington, D.C.

Website http://www.epa.gov/waterscience/cs/freshmanu

al.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

165

Fact Sheet No. 2.2.3-7

Method Title Acute Larval Bivalve Sediment Bioassay

Purpose This test measures the survival of the marine bivalve larvae, Crassostrea gigas, and

Mytilus edulis, after exposure to sediments for 48 hours in the laboratory.

Method Summary Adult bivalves, conditioned as necessary in the laboratory, are induced to spawn

with selected thermal and biological (i.e., sperm) stimulation. Selected densities of

the resulting embryos are exposed to the test or reference area sediments for 48

hours, during which the embryos normally will develop into prodissoconch I larvae.

Exposure time should not exceed 60 hours for an acceptable test. Toxicity test

endpoints are based on abnormal shell development and larval death. 

Data

Uses/Application

This toxicity test can be used to assess the toxicity of marine sediments, especially

dredged material. It may be used alone as a screening tool or in combination with

sediment chemistry and in situ biological indices, and in laboratory experiments

addressing a variety of sediment and water quality manipulations. These sediment

bivalve bioassays are performed as a part of the Dredged Material Evaluation and

Disposal Procedures in the Puget Sound (Puget Sound Dredged Disposal Analysis

(PSDDA) Program, 2000).

Advantages Refer to table 2.2.3-1 to compare alternative methods for acute sediment bioassays.

Limitations Data from tests with longer exposures (> 48 hours) may not be comparable to those

tests conducted using the standard 48-hour exposure. 

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water

Column, and Tissue in Puget Sound, Puget Sound Protocols and Guidelines. Puget

Sound Water Quality Action Team, Olympia, W A.

Website http://www.psat.wa.gov/Publicat

ions/protocols/protocol_pdfs/field.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

166

Fact Sheet No. 2.2.3-8

Method Title Acute Echinoderm Sediment Bioassay

Purpose This test measures the survival of the marine echinoderms, Dendraster excentricus,

Strongylocentrotus purpuratus and S. Droebachiensis, after exposure to sediments

for 48-96 hours in the laboratory. 

Method Summary Adult Echinoderms, Dendraster excentricus, Strongylocentrotus purpuratus and S.

droebachiensis, are induced to spawn with chemical stimulation. The resulting

embryos are exposed to test sediment for 48 to 96 hours during which the embryos

will develop into the four-armed pluteus stage. The toxicity test endpoint is based on

failure to develop normal pluteus larvae. These sediment echinoderm bioassays are

performed as a part of the Dredged Material Evaluation and Disposal Procedures in

the Puget Sound. (USACE/WDNR/WDEC, 2000)

Data

Uses/Application

This toxicity test can be used to assess the toxicity of marine sediments, particularly

dredged material. It may be used alone as a screening tool or in combination with

sediment chemistry and in situ biological indices, and in laboratory experiments

addressing a variety of sediment and water quality manipulations.

Advantages Refer to table 2.2.3-1 to compare alternative methods for acute sediment bioassays.

Limitations The three species may show different levels of sensitivity; therefore, the results for

corresponding endpoints may not be comparable between the three species. 

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water

Column, and Tissue in Puget Sound, Puget Sound Protocols and Guidelines. Puget

Sound Water Quality Action Team, Olympia, W A.

Website http://www.psat.wa.gov/Publications/pro

tocols/protocol_pdfs/field.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

167

Fact Sheet No. 2.2.3-9

Method Title Acute Marine Crustacean Sediment Bioassay

Purpose This test measures survival of the marine mysid shrimp, Mysidopsis bahia and the

marine penaeid shrimp, Penaeus sp., after a 96-hour exposure to sediments in the

laboratory. 

Method Summary The mysid shrimp, Mysidopsis bahia, and Penaeid shrimp, Penaeus sp., are

placed in 1-L glass chambers containing 175 mL sediment and about 800 mL of

overlying water for 96-hours. Test temperature 20°C and the recommended

overlying water salinity is 20 ppt. The test chambers will be lightly aerated, but

water will not be renewed. Test species will be fed once daily. The endpoints in the

toxicity test are survival. Performance criteria established for this test include the

average survival of organisms in negative control treatment must be $ 90%.

A miniaturized method of this test also exists (Ho, 2000) and will be incorporated

into EPA’s TIE guidance document.

Data

Uses/Application

This test method may be useful in assessing sediment contamination, registration of

pesticides, assessments of new and existing chemicals, Superfund site

assessment, and assessment and cleanup of hazardous waste treatm ent, storage,

and disposal facilities. 

Advantages Mysid shrimp are filter- and deposit-feeders commonly found in marine sediments;

therefore, exposure to contaminated sediments is likely. Penaeid shrimp are

deposit-feeders and burrowers, so they are also likely exposed to contaminants

through there feeding regime. Mysid shrimp tolerate a wide range of salinities.

Limitations Amphipod crustaceans are more commonly used for short term sediment

bioassays. They are considered benchmark species by the USEPA and USACE

Inland Testing Manual (1998) in that they comprise a substantial database,

represent the sensitive range of a variety of ecosystems, and provide comparative

data on the relative sensitivity of local test species. 

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 1995. Environmental Monitoring and Assessment Program Laboratory

Methods Manual, Estuaries, Volume 1-Biological and Chemical Analyses,

EPA/620/R-95/008. Environmental Monitoring and Assessment Program , U.S.

Environmental Protection Agency, Washington DC.

Website http://www.epa.gov/emap/html/pubs/doc

s/groupdocs/estuary/field/lab_man.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

168

Fact Sheet No. 2.2.3-10

Method Title Acute Marine Amphipod Crustacean Sediment Bioassay, EPA Method 100.4

Purpose This test measures the survival of the marine amphipod crustaceans, Ampelisca

abdita, Eohaustorius estuarius, Leptocheirus plumulosus or Rhepoxynius abronius,

after exposure to sediments for 10 days in the laboratory. 

Method Summary Infaunal amphipods, Ampelisca abdita, Eohaustorius estuarius, Leptocheirus

plumulosus and Rhepoxynius abronius are used in toxicity studies assessing

sediments from marine and estuarine environments. The toxicity test is conducted for

10 d in 1-L glass chambers containing 175 mL sediment and about 800 mL of

overlying water. Test temperature is 15°C for E. estuarius, 20°C for A. Abdita and

25°C for L. plumulosus, and the recommended overlying water salinity is 20 ppt for

E. estuarius and L. plumulosus and 28 ppt for A. abdita and R. abronius. There will

be no feeding during the test and no renewal of overlying water. The endpoints in the

toxicity test are survival of amphipods. Performance criteria established for this test

include the average survival of amphipods in negative control treatment must be $

90%.

The Rhepoxynius abronius is the preferred species for coarser-grained sediments

(i.e., fines <60%) with a salinities >25 ppt. The Ampelisca abdita may be used when

test sediment contains > 60% fines and in a wide range of salinities. The

Eohaustorius estuarius may be used when grain size ranges from 0.6 % sand to

100% sand and salinities range from 1 ppt to 25 ppt. 

Puget Sound Dredged Disposal Analysis Program (USACE/WDNR/WDEC, 2000)

also recommends a similar 10-day acute toxicity test with the marine amphipods,

Ampelisca abdita, Eohaustorius estuarius, and Rhepoxynius abronius. ASTM E1367

and NHEERL-AED SOP 1.03.002 also describe methods for sediment bioassays

with amphipods (ASTM, 2001b and USEPA and Naval Construction Battalion

Center, 1992, respectively).

A miniaturized method of this test also exists (Ho, 2000) and will be incorporated into

EPA’s TIE guidance document.

Data

Uses/Application

This is a sediment toxicity method used to evaluate the effects (reduction in survival)

of marine and estuarine sediments on the marine amphipods. The test method may

be useful in assessing sediment contamination, registration of pesticides,

assessments of new and existing chemicals, Superfund site assessment, and

assessment and cleanup of hazardous waste treatm ent, storage, and disposal

facilities. 

The choice of these amphipod species as test organisms is based on sensitivity to

sediment-associated contaminants, availability and ease of collection, tolerance of

environmental conditions (e.g., temperature, salinity, grain-size), ecological

importance, and ease of handling in the laboratory. Either alone or in combination

they may be used to measure toxicity of any commonly encountered marine

sediment. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

169

Fact Sheet No. 2.2.3-10 (contd.)

Advantages Amphipods are among the first taxa to disappear from benthic comm unities impacted

by pollution, and have been shown to be more sensitive to contaminated sediments

than several other major taxa. All of these organisms are considered benchmark

species by the USEPA and USACE’s Inland Testing Manual (1998) indicating that

they comprise a substantial database, represent the sensitive range of a variety of

ecosystems, and provide comparative data on the relative sensitivity of local test

species.

Limitations Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 1994d. Methods for Assessing the Toxicity of Sediment-Associated

Contam inants with Estuarine and Marine Amphipods, EPA/600/R-94/025. Office of

Research and Development, U.S. Environmental Protection Agency, Washington,

D.C.

Website http://www.epa.gov/waterscience/cs/freshmanua

l.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

170

Fact Sheet No. 2.2.3-11

Method Title Acute Marine Polychaete Sediment Bioassay, ASTM Method E1611-00

Purpose This test measures the survival of the marine polychaetes, Neanthes

arenaceodentata and Neanthes virens, after exposure to sediments for 10 days in

the laboratory. 

Method Summary Infaunal polychaetes Neanthes arenaceodentata and Neanthes virens are used in

toxicity studies assessing sediments from marine and estuarine environments. The

toxicity test is conducted for 10 d in 1-L glass chambers with a sediment depth of 2

to 3 cm and aerated overlying water. Water is not renewed during the 10-day

exposure and there is no feeding. The endpoints in the toxicity test is survival of

polychaetes.

A negative control or reference sediment is used to give a measure of acceptability of

the test. 

A similar method using the test species Polydora cornuta is described in

Environment Canada’s method EPS/1/RM/41

Data

Uses/Application

This is a sediment toxicity method used to evaluate the acute effects (reduction in

survival ) of marine and estuarine sediments on polychates. Polychaetes are an

important component of the benthic com munity and are sensitive to both organic and

inorganic chemicals. The results of this acute toxicity test can be used to predict

temporal or spatial distribution of sediment toxicity. The test method may be useful in

assessing sediment contamination, registration of pesticides, assessments of new

and existing chemicals, Superfund site assessment, and assessment and cleanup of

hazardous waste treatm ent, storage, and disposal facilities. 

Advantages Polychaetes are burrowers and deposit-feeders; therefore, contaminant exposure is

likely. 

A 10-day test provides data on the short-term effects that may be useful for

comparisons to other species but does not provide information on delayed effects. 

Limitations Polychaetes are not considered benchmark species (USEPA and USACE, 1998). 

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section.

Reference ASTM. 2001b. ASTM Book of Standards. Volume 11.05. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

171

Fact Sheet No. 2.2.3-12

Method Title Chronic Estuarine Amphipod Sediment Bioassay

Purpose This test measures the growth and reproduction of the estuarine amphipod

crustacean, Leptocheirus plumulosus, after exposure to sediments for 28 days in the

laboratory. 

Method Summary This is a laboratory method for determining the chronic toxicity of contaminants

associated with whole sediments collected from estuarine or marine environm ents

(or estuarine or marine sediment spiked with compounds in the laboratory). The

toxicity method uses an estuarine crustacean, the amphipod, Leptocheirus

plumulosus. The toxicity test is conducted for 28 d in 1-L glass chambers containing

175 mL sediment and about 725 mL of overlying water. Test temperature is 25°± 2°,

and the recommended overlying water salinity is 5 ppt ± 2ppt (for test sediment with

pore water at 1 ppt to 10 ppt) or 20 ppt ± 2 ppt (for test sediment with pore water >10

ppt). 400 mL of overlying water is renewed three times a week, at which times test

organisms are fed. The endpoints in the toxicity test are survival, growth, and

reproduction of amphipods. Performance criteria established for this test are that the

average survival of amphipods in negative control treatment must be $ 80% and

there must be measurable growth and reproduction in all replicates of the negative

control treatm ent.

Data

Uses/Application

This is a sediment toxicity method used to evaluate the sublethal effects (reduction in

growth and reproduction) of marine and estuarine sediments on the marine

amphipod, Leptocheirus plumulosus. The test method may be useful in assessing

sediment contamination, registration of pesticides, assessments of new and existing

chemicals, Superfund site assessment, and assessment and cleanup of hazardous

waste treatm ent, storage, and disposal facilities. 

Advantages The marine amphipod, Leptocheirus plumulosus, is considered a benchmark species

which means that they comprise a substantial database, represent the sensitive

range of a variety of ecosystems, and provide comparative data on the relative

sensitivity of local test species. This organisms is a deposit-feeder and burrower

therefore exposure to sediment contaminants will occur through its feeding regime. 

Limitations The test is applicable for use with sediments from oligohaline to fully marine

environments, with a silt content greater than 5% and a clay content less than 85%.

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 2001e. Method for Assessing the Chronic Toxicity of Marine and Estuarine

Sediment-associated Contam inants with the Amphipod Leptocheirus plumulosus,

EPA 600/R-01/020. Office of Research and Development, U.S. Environmental

Protection Agency, Washington, D.C.

Website http://www.epa.gov/waterscience/cs/guid

ancemanual.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

172

Fact Sheet No. 2.2.3-13

Method Title Chronic Marine Polychaete Sediment Bioassay, ASTM Method E1611-00

Purpose This test measures the growth and reproduction of the marine polychaete, Neanthes

arenaceodentata, after exposure to sediments for 28 days in the laboratory. 

Method Summary This is a laboratory method for determining the chronic toxicity of contaminants

associated with whole sediments collected from estuarine or marine environm ents

(or estuarine or marine sediment spiked with compounds in the laboratory). The

toxicity method uses an estuarine polychaete, Neanthes arenaceodentata. The

toxicity test is conducted for 28 d in 1-L glass chambers containing 2-3 cm of

sediment and aerated overlying water. This is a static renewal toxicity test and the

organisms are fed daily. The endpoints in the toxicity test are survival, growth, and

reproduction of amphipods. Performance criteria established for this test include the

average survival of amphipods in negative control treatment must be $ 80% and

there must be measurable growth and reproduction in all replicates of the negative

control treatm ent.

Puget Sound protocols and guidelines recommend a juvenile polychaete bioassay

with a 20-day exposure time. Endpoints are mortality, total biomass, and average

individual biomass. It is a static renewal assay and organisms are fed. 

Data

Uses/Application

This is a sediment toxicity method used to evaluate the sublethal effects (reduction in

growth and reproduction) of marine and estuarine sediments on the marine

polychates. The test method may be useful in assessing sediment contamination,

registration of pesticides, assessments of new and existing chemicals, Superfund

site assessment, and assessment and cleanup of hazardous waste treatm ent,

storage, and disposal facilities. 

Advantages This type of worm is a deposit-feeder and burrower, therefore exposure to sediment

contaminants is likely through the feeding regime.

Limitations This organism is not considered a benchmark species (USEPA and USACE, 1998).

The protocol may have to be modified for tests at salinities less than 20 ppt.

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference ASTM. 2001b. ASTM Book of Standards. Volume 11.05. American Society for

Testing and Materials, West Conshocken, PA.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

173

Fact Sheet No. 2.2.3-14

Method Title Ames Mutagenicity Assay

Purpose This test can be used to determine if a sample is mutagenic or cancer-causing. It

uses the induced reversion of bacterial mutants to detect DNA-damaging

substances. 

Method Summary Sediment samples are dried with anhydrous sodium sulfate, extracted with

methylene chloride, subjected to gel-permeation chromatography cleanup,

evaporated under nitrogen, and brought to volume in DMSO. 

The Ames assay uses 100uL of cultured test strain (Salmonella typhimurium), 500 uL

of either phosphate buffer, and 100uL of the sediment extract or DMSO mixture

(control). The entire mixture is incubated for 20 to 30 minutes in a dry block heater. 

Following incubation, top agar is added containing trace histidine and biotin and the

mixture is poured into plates. Plates are then incubated and the resulting colonies

are counted at 72 hours.

A positive mutagenic response is indicated when the number of revertants on test

plates are greater than or equal to 2 times the number of colonies in the DMSO

solvent control plate. 

Data

Uses/Application

Short-term bioassays generally identify specific genotoxic contaminants or those in

complex mixtures of contaminants, provide baseline data for monitoring changes in

environmental conditions, and predict potential long range genotoxic health effects. 

Advantages The Ames test is relatively quick and inexpensive. It is useful for establishing

priorities for more definitive chemical analysis or toxicological testing. 

Limitations Some chemicals that are mutagenic/carcinogenic, do not give a positive Ames Test

(i.e., dioxin). Because the strain of salmonella used is histidine negative, the test

may also give false positives if histidine is present in environmental samples.

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 1993b. Assessment and Remediation of Contaminated Sediments (ARCS)

Program: Biological and Chemical Assessment of Contaminated Great Lakes

Sediment, EPA 905-R93-006. Great Lakes National Program Office, U.S.

Environmental Protection Agency, Chicago, IL. 

Website http://www.epa.gov/glnpo/arcs/EPA-905-

R93-006/EPA-905-R93-006.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

174

Fact Sheet No. 2.2.3-15

Method Title Mutatox Genotoxicity Assay

Purpose This test can be used to determine if a sample is mutagenic or cancer-causing. It

uses the induced reversion of bacterial mutants to detect DNA-damaging

substances.

Method Summary Sediment samples are dried with anhydrous sodium sulfate, extracted with

methylene chloride, subjected to gel-permeation chromatography cleanup,

evaporated under nitrogen, and brought to volume in DMSO. 

The Mutatox assay uses rat liver S9 for exogenous metabolic activation of

progenotoxins and a dark mutant strain of the luminescent bacterium Photobacterium

phosphoreium for detection of genotoxins. DNA-damaging substances are detected

by measuring the ability of a test extract or specific chemical to restore the

luminescent state in the bacterial cells. The degree of light increase indicates the

relative genotoxicy of the sample. 

Data

Uses/Application

Short-term bioassays generally identify specific genotoxic contaminants or those in

complex mixtures of contaminants, provide baseline data for monitoring changes in

environmental conditions, and predict potential long range genotoxic health effects. 

Advantages The test is relatively quick and inexpensive. It is useful for establishing priorities for

more definitive chemical analysis. 

Limitations Com parative data may be limited. 

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 1993b. Assessment and Remediation of Contaminated Sediments (ARCS)

Program: Biological and Chemical Assessment of Contaminated Great Lakes

Sediment, EPA 905-R93-006. Great Lakes National Program, U.S. Environmental

Protection Agency, Chicago, IL. 

Website http://www.epa.gov/glnpo/arcs/EPA-905-

R93-006/EPA-905-R93-006.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

175

Fact Sheet No. 2.2.3-16

Method Title V79 Sister Chromatid Exchange Assay, NHEERL-AED SOP 1.03.012

Purpose These methods describes a test used to evaluate mutagenic effects of single

compounds and complex mixtures, including sediment extracts and their fractions. 

Method Summary The Sister Chromatid Exchange (SCE) first requires the preparation of media that is

used for plating, dosing and media change procedures. The media is then divided

and applied to toxicity plates and SCE plates. Cell suspensions are prepared and

then added to appropriate vials in a 1:10 dilution ratio. The number of cells per mL

in the cell suspensions are determined using a hemocytometer. Cell suspension

dilutions are then added to the toxicity plates and the SCE plates. These plates are

then incubated for 24 hours to allow for cell attachment. 

Following incubation, media and solvents are mixed to prepare a dosed medium that

is then added to both the toxicity plates and the SCE plates: the toxicity plates

receive a minimum of 3 mL of dosed media and the SCE plates receive a minimum

of 7 mL of dosed media. The plates are then incubated for 5 hours. Media is renewed

with “clean media” on the toxicity plates and then the plates are returned to the

incubator for 6 days. The media is changed again and the plates are incubated for

20 hours. 

Cells are harvested from both the toxicity plates and the SCE plates and slides are

made. On the toxicity slides, the number of colonies are counted. The SCE slides

are scanned for suitable chromosome spreads. The number of chromosomes and

the number of SCEs per spread are recorded, along with other observations.

Data

Uses/Application

The SCE Assay is commonly used to determine genetic damage and mutational

events by cytogenetic analysis.

The mean colony per dose determined on the toxicity test slides is used to find the

percent survival of a dose as compared to the control or blank for that test. The

number of SCE/number of chromosomes is evaluated statistically to find the mean

per dose which is compared to the control or blank for that test. 

Advantages The assay has been used to evaluate effects of single compounds (i.e., mitocmycin

C, benzo(a)pyrene) and to evaluate whole sediment extracts and sediment fractions

from sites with known contaminated sediment problems.

Limitations The tests need to be monitored daily since contamination will skew the results. Any

contaminated plates should be removed, taped with autoclave masking tape,

wrapped in foil, and autoclaved. 

There may also be problems with chromosome, staining and spreading quality.

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

176

Fact Sheet No. 2.2.3-16 (contd.)

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296, Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.03.012.pdf

Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

177

Fact Sheet No. 2.2.3-17

Method Title Bioaccumulation Test for Marine, Estuarine and Freshwater Sediments, EPA Method

100.3

Purpose This test measures bioaccumulation in the freshwater Lumbriculus variegatus after

exposure to sediments for 28 days in the laboratory. 

Method Summary The Lumbriculus variegatus is exposed to sediments for 28-days in a 4 to 6 L

container containing 1 to 2 L of sediment and 1 to 4 L of overlying water. Test

temperature is 23 +/- 1 °C. 1 L of overlying water will be renewed every 12 hours.

No supplemental food will be added during the experiment. The endpoint is

bioaccumulation.

In some cases, body burdens will not approach steady-state body burdens in a 28-d

test (e.g., organic compounds with a log Kow >5, be metabolically refractory, or have

low depuaration rates). Depending on the goals of the study and the adaptability of

the test species to long-term testing, it may be necessary to conduct an exposure

longer than 28-d (or a kinetic study) to obtain a sufficiently accurate estimate of

steady-state tissue residues of these compounds. Use of long-term tests or

toxicokinetic approaches is recommended specifically for slowly accumulated

compounds and for a greater than 80 percent accuracy in test species achieving

steady state.

ASTM also provides guidance for bioaccumulation tests with marine test species

such as the polychaetes, Nereis diversicolor, Neanthes virens and the bivalve

Macoma nasuta, Macoma balthica, and Yoldia limatula (ASTM Method E1688;

ASTM 2001b), and freshwater test species such as the Diproeia spp and

Lumbriculus variegates. They recommend selecting at least one species

representing filter-feeding, deposit-feeding and burrowing species.

Data

Uses/Application**

This is a laboratory method for determining the bioaccumulation of contaminants

associated with sediments collected from freshwater environments. The data are

sometimes multiplied by an adjustment factor to accommodate for different steadystate rates with different contaminant mixtures. 

Data from bioaccumulation tests are used to derive bioaccumulation factors (BAFS)

and to determine biota-sediment accumulation factors (BSAFS) in equilibrium

partitioning models.

Bioaccumulation data is needed in ecological or human health risk assessments,

therefore the procedures are designed to generate quantitative estimates of steadystate tissue residues. These tests are also used to assist in the development of

sediment quality criteria and to assess the potential impacts of disposal of dredged

materials. Biota-sediment accumulation factors (BSAFS) are often compared

between laboratory-exposed and field-exposed organisms to determine the validity of

laboratory experiments and to better predict contaminant-specific lipid partitioning

tendencies. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

178

Fact Sheet No. 2.2.3-17 (contd.)

Advantages Previous bioaccumulation tests used 10-day exposures. A 28-day exposure is a

practical compromise between cost, data accuracy and data utility. Observed

steady-state tissue levels were reached in 28-days in 69% of the tests. The data

should be sufficiently accurate for quantitative risk analysis in most cases. In cases

in which more accurate estimates are required, either a long-term exposure or an

alternative approach can be used. 

Limitations Additional research is needed on the standardization of bioaccumulation procedures

with sediment. Steady-state is reached at different times with different contaminants,

thus 28-day is an approximate period of time that may prove inaccurate with the

wrong mixture of contaminants. 

Specifically, the 28-day time period appears to underestimate steady-state of DDT

and dieldrin, so it may be necessary to use adjustment factors for sites with

significant DDT and dieldrin concentrations. 

Please note the list of general limitations for all solid-phase toxicity tests in the

introduction to this section. 

Reference USEPA. 2000a. Bioaccumulation Testing and Interpretation for the Purpose of

Sediment Quality Assessment: Status and Needs, EPA 823-R-00-001. Office of

Water and Office of Solid Waste, U.S. Environmental Protection Agency,

Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitl

eOW.html

Last Accessed: 2/13/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

179

2.3 Biota

The health and community structure of endemic organisms are often evaluated in order to assess

the impacts of chemical contamination on the environment. At Superfund sites containing

contaminated sediments, phytoplankton, zooplankton, benthic invertebrates and fish can all be

monitored. These organisms can potentially be exposed to contaminant stressors via ingestion

pathways or direct contact/absorption from the water column. Therefore, they are collected either

for identification and enumeration analyses to determine community structure or they are collected

for chemical tissue analyses to investigate contaminant uptake, bioaccumulation and potential

biomagnification in the food chain.

There are three common approaches to evaluating environmental risk to receptors: 1) the use of

literature screening values; 2) a “desk-top” risk assessment that can model existing site-specific

contaminant data to ecological receptors for subsequent comparison to literature toxicity values; or

3) field investigation/laboratory analysis that involves a site investigation and laboratory analysis of

contaminant levels in media and/or experimentation using bioassay procedures (USEPA, 1997a).

The methods provided in the following section provide a summary of those methods that would be

used in the third approach: field investigation and laboratory analyses. These fact sheets intend to

provide Superfund managers with a summary of the existing methods that may be applicable to

their site, the method’s relative strengths, and the method’s relative weaknesses. These analyses

will help to determine the relationship between the exposure of a contaminant and the response it

elicits. 

2.3.1 Chemical and Physical Analyses

Section 2.3.1 presents field sample collection and processing methods for biota. Biota are

collected at Superfund sites for chemical residue studies, population/community studies, and

toxicity testing/bioassays; all directed at assessing exposure-response relationships at the site

(USEPA, 1997a). Methods are provided for sample collection and processing of phytoplankton,

zooplankton, periphyton, benthic invertebrates and fish. Biota methods were predominantly

gathered from the following sources:

• The USEPA’s Office of Water

• The USEPA’s EMAP program

• The USEPA’s Great Lakes Program
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

180

Fact Sheet No. 2.3.1-1

Method Title Phytoplankton Sample Collection and Preservation in the Great Lakes, LMMB 023t

Purpose This method describes the collection and preservation of phytoplankton samples for

com munity analyses

Method Summary Water samples are collected using a Rosette sampler (see Fact Sheet 2.1.1-6). 1L

aliquots from each discrete sampling depth are composited, and approximately 1 L of

the com posite sample is transferred to a sample bottle. The sample is preserved

with Lugol’s Solution for analysis (final concentration 1%). Samples are stored in the

dark and refrigerated. 

Data

Uses/Application

Phytoplankton com munity analyses are useful for bioassessm ents to determine

com munity disturbance as a result of contamination.

Advantages One of few standard methods for water column bioassessment

Limitations This collection method is not suitable for chlorophyll a or productivity measurements.

Populations of phytoplankton are seasonal and highly variable. Use of one or more

reference stations is essential.

Reference USEPA. 1997b. Method LMMB 023: Standard Operating Procedure for

Phytoplankton Sample Collection and Preservation. Lake Michigan Mass Balance

Study Methods Compendium, Volume 1: Sample Collection Techniques, EPA 905-R97-012c. Great Lakes National Program Office, U.S. Environmental Protection

Agency, Chicago, IL. 

Website http://www.epa.gov/glnpo/lmmb/m ethods/phyt

ocol.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

181

Fact Sheet No. 2.3.1-2

Method Title Chlorophyll-a Sampling Method and Preservation: Field Procedure in the Great

Lakes, LMMB 015

Purpose This method is used to filter chlorophyll-a samples.

Method Summary Water samples are collected using Niskin bottles or other suitable sampling device.

The water samples are transferred from the Niskin bottles to opaque sample bottles

for storage. The water sample for chlorophyll-a analysis is vacuum filtered through a

47 mm diameter glass fiber filter (see Fact Sheet 2.3.1-3). The entire procedure is

conducted in subdued (green) light to prevent photodecomposition. During filtration,

the samples are treated with a solution of MgCO3to prevent acid induced

transformation of chlorophyll to its degradation product, Phaeophytin. Sample filters

are stored in aluminum foil pouches and frozen until analysis. 

Data

Uses/Application

Often used as a surrogate for productivity or standing crop measurements,

chlorophyll a measurem ents are also used to monitor plankton blooms. Also used to

calibrate SeaWiFS or other remote sensing images.

Advantages Sim ple collection, extraction, and analysis methods allow economical spatial and

temporal variations to be monitored.

Limitations Filters must be extracted and analyzed within 28 days.

Reference USEPA. 1997b. Method LMMB 015: Standard Operating Procedure for Chlorophylla Sampling Method: Field Procedure. Lake Michigan Mass Balance Study Methods

Compendium, Volume 1: Sample Collection Techniques, EPA 905-R-97-012c. Great

Lakes National Program Office, U.S. Environmental Protection Agency, Chicago, IL. 

Website http://www.epa.gov/glnpo/lmmb/m ethods/chlf

ield.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

182

Fact Sheet No. 2.3.1-3

Method Title Chlorophyll a and Phaeophytin Field Filtering Protocols

Purpose This method describes the methods used for the immediate processing of water

samples analyzed for Chlorophyll a and Phaeophytin. 

Method Summary Chlorophyll a and Phaeophytin samples must be filtered no more than 4 hours after

collection. Samples that cannot be filtered immediately after collection must be held

at 4°C until filtered. Filtering can be accomplished by the use of a vacuum pump or

by using positive pressure, as described below

Vacuum Filtration: Filter the water onto two 47 mm diameter glass fiber filter pads.

Do not exceed a vacuum of 15 psi or a filtration duration of greater than 5 minutes.

Add 1 mL of saturated MgCO3solution during the last few seconds of filtering after

the nutrient filtrate has been removed. 

Record the volume filtered on the data sheet. The filtrate should be saved for

dissolved nutrient analyses. Approximately 40 mL of filtrate will be collected into a

pre-labeled, clean 60 mL Nalgene screw-capped bottle and stored on dry ice. 

Carefully remove the filters using forceps, fold in half, and wrap in aluminum foil.

Label the samples and place package on dry ice.

Positive Pressure: The alternative method is to use positive pressure to push a

sample through the filter. A disposable, graduated 50-cc polypropylene syringe fitted

with a stainless steel or polypropylene filtering assembly is used to filter the site water

through 25 mm diameter glass fiber filter pads; the volume of water must be

documented. If conditions allow, up to 200 mL of site water should be filtered for

each chlorophyll sample. After filtering, add 1 mL of MgCO3

solution to the syringe

and pass through the filter pad. Rem ove the filter, fold and place in aluminum foil.

Again, approximately 40 mL of filtrate will be collected into a pre-labeled, clean 60 mL

Nalgene screw-capped bottle and stored on dry ice.

Methods LMMB 085 and 086 describe similar filtering protocols (USEPA, 1997b).

`Data

Uses/Application

Chlorophyll a measurem ents are indicative of the primary producer’s relative

abundance and composition in the water column sample. 

Advantages Sim ple collection, extraction, and analysis methods allow economical spatial and

temporal variations to be monitored.

Limitations If filtration cannot occur in under 4 hours, the phytoplankton cells can possibly lyse.

The sample on the filter paper may degrade over time, and must be extracted and

analyzed within 28 days.

Reference USEPA. 2000b. Coastal 2000 Northeast Component: Field Operations Manual,

Environmental Monitoring and Assessment Program (EMAP), EPA/620/R-00/002.

Office of Research and Development, U.S. Environmental Protection Agency,

Washington, DC. 

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

183

Fact Sheet No. 2.3.1-4

Method Title Primary Productivity Using 14C: Field Procedure in the Great Lakes, LMMB 016

Purpose This method is used to determine primary productivity and primary productivity

parameters from water.

Method Summary Water samples are collected using Niskin bottles or other suitable samplers. When

there is a thermal stratification in the water column, samples are collected from both

the hypolimnion and the epilimnion. The water samples are transferred from the

Niskin bottles to opaque sample bottles for storage in a light-tight, insulated

container. 

The following procedures are conducted in subdued (green) light to avoid

photodegradation. Water samples are carefully transferred to incubation bottles.

Water samples are inoculated with a known quantity of bircarbonate substrate, which

is labeled with the radiotracer 14C. Samples are incubated at various light intensities

for 2 - 4 hours. After incubation, a 100 mL aliquot of each sample is filtered through a

47 mm cellulose acetate filter (0.45 μm pore size). The filter is placed into a

scintillation vial and 0.5 N HCl is added. The vials sit at room temperature for 1 hour.

20 mL of liquid scintillation cocktail is added to each vial. The vials are stored until

they are analyzed by liquid scintillation counting to determine the quantity of carbon

fixed by the algae into organic matter. 

Data

Uses/Application

Primary productivity is a key measurement in many site assessments, particularly

those affected with nutrient enrichment.

Advantages This method is the standard for productivity determination.

Limitations Use of 14 C requires NRC License. Disposal of 14 C waste is often problematic.

Reference USEPA. 1997b. Method LMMB 016: Standard Operating Procedure for Primary

Productivity Using 14C: Field Procedure. Lake Michigan Mass Balance Study

Methods Compendium, Volume 1: Sample Collection Techniques, EPA 905-R-97-

012c. Great Lakes National Program Office, U.S. Environmental Protection Agency,

Chicago, IL. 

Website http://www.epa.gov/glnpo/lmmb/methods/c14

field.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

184

Fact Sheet No. 2.3.1-5

Method Title Zooplankton Sample Collection and Preservation in the Great Lakes, LMMB 024

Purpose This method describes the collection and preservation of zooplankton samples for

com munity analyses.

Method Summary Water samples are collected using a plankton tow net that is maneuvered using a

winch on the starboard side of the vessel. The tow net has a flow meter and

screened sample bucket attached to the end. The flow meter should be calibrated

every survey season. For sampling, the net is lowered to the desired depth (usually

20 meters from the water surface) and raised at a constant slow speed until the rim is

above the water. In shallower waters, the samples are usually collected from 1 meter

above the bottom to the surface. The net is then lifted out of the water and rinsed

from the outside to free organisms from the side of the net, concentrating them into

the sample bucket. The sample bucket is removed, and the sample is rinsed into a

sample container. The organisms are then narcotized with 20 mL of soda water and

left to sit for 30 minutes. The samples are preserved with 20 mL of formalin solution. 

 

Data

Uses/Application

Tow nets survey the biological com munity by collecting abundance and taxa

composition data from sampling locations. They are often used to gain information

on particular species of larval fish and an overall estimation of fish populations and

comm unities. Fish population and com munity data are used to measure the status

and trends of environmental pollution freshwater, estuarine and marine organisms to

assess water quality criteria and to monitor surface water quality. 

Advantages The tow net is easy to handle and it is sm all enough for use on boats 4 m or larger in

length. The design reduces current vibrations in the water directly in front of the net.

Limitations Zooplankton populations are highly seasonal and within season can vary spatially

depending upon currents and microclimate. Effective use of reference areas

required.

Reference USEPA. 1997b. Method LMMB 024: Standard Operating Procedure for

Zooplankton Sample Collection and Preservation. Lake Michigan Mass Balance

Study Methods Compendium, Volume 1: Sample Collection Techniques, EPA 905-R97-012c. Great Lakes National Program Office, U.S. Environmental Protection

Agency, Chicago, IL. 

Website http://www.epa.gov/glnpo/lmmb/methods/zoo

fld.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

185

Fact Sheet No. 2.3.1-6

Method Title Field-based Periphyton Survey in Wadeable Streams

Purpose To perform a quick semi-quantitative assessment of algal biomass and taxonomic

composition in the field.

Method Summary This protocol describes a field-based rapid survey of periphyton biomass and coarselevel taxonomic composition (e.g., diatoms, filamentous greens, blue-green algae).

To perform a field-based periphyton survey, the investigator establishes three

transects across the habitat being sampled (preferably riffles or runs in the reach in

which benthic algal accumulation is readily observed and characterized). Three

locations are then selected along each transect. Algae are characterized in each of

the selected locations by immersing a bucket with a 50-dot grid in the water. 

Macroalgal biomass and microalgal cover are then determined in the bucket.

EPA’s EMAP field document describes similar field-based periphyton surveys

(USEPA, 1998, EPA/620/R-94/004F).

Data

Uses/Application

Species relative abundance and taxa richness are data derived from these protocols.

These data parameters provide information pertaining to the status and trends of

environmental pollution and its impacts on freshwater, marine and estuarine

comm unities. 

Biological impairment resulting from pollution is often evaluated using metrics of

biotic integrity derived from the aforementioned data parameters that evaluate

community, population and functional parameters. Examples of metrics based on

species composition include species richness, total number of genera, total number

of divisions, shannon diversity (for diatoms), percent com munity similarity of diatoms,

pollution tolerance index for diatoms, and percent sensitive diatoms. Furthermore,

other metrices infer ecological conditions based on documented preferences. These

metrices include the percent aberrant diatoms, percent motile diatoms, simple

diagnostic metrics, inferred ecological conditions with simple autecological indices

(SAI), inferred ecological conditions with weighted average indices, and impairment

of ecological conditions. 

Advantages The field-based periphyton survey requires less effort than the laboratory methods. It

is able to assess algal biomass over larger spatial scales than substrate sampling

and laboratory analysis. Coarse-level taxonomic characterization of comm unities is

also possible with this technique. 

Limitations The field methods are not as accurate as the laboratory analysis. 

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition,

EPA 841-B-99-002. Office of Water, U.S. Environmental Protection Agency,

Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/ Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

186

Fact Sheet No. 2.3.1-7

Method Title Laboratory-Based Periphyton Survey: Single Habitat Sampling in Wadeable Streams

Purpose To determine the periphyton abundance and composition in area of interests.

Method Summary Single habitat sampling protocol outlines a procedure for collecting periphyton from a

single substrate/habitat combination that characterizes the study reach. A

preliminary, visual habitat assessment should be performed prior to sample collection

to determine the percent cover of each substrate type and the estimated relative

abundance of organisms.

Collection techniques depend on the substrate type and the dominant periphyton.

Several subsamples should be collected from the same substrate/habitat

combination and composited into a single container. Periphyton samples should be

collected during periods of stable stream flow.

If the samples are going to be assayed for chlorophyll a, the samples should not be

preserved until they have been subsampled. Following subsampling and

preservation ( Lugol’s solution, “M3" fixative, buffered 4% formalin, 2%

glutaraldehyde, or other preservative) the samples are transported back to laboratory

on ice. 

EPA’s EMAP field document describes similar protocols for periphyton surveys in

wadeable streams (USEPA, 1998).

Data

Uses/Application

Periphyton can be collected to: 1) determine taxonomic composition and relative

abundance, 2) determine chlorophyll, 3) determine biomass, and 4) determine

acid/alkaline phosphate activity.

Single habitat sampling provides periphyton biomass data.

Advantages Variability in habitat differences between streams may be reduced if periphyton

collection is performed from a single substrate/habitat combination. 

Limitations Spatial variability can lead to samples not being representative of site. Adequate

replication and spatial coverage required.

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition.

EPA 841-B-99-002. Office of Water;\\, U.S. Environmental Protection Agency;

Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/ Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

187

Fact Sheet No. 2.3.1-8

Method Title Laboratory-Based Rapid Periphyton Survey: Multi habitat Sampling in Wadeable

Streams

Purpose Algae sampling methods for subsequent laboratory assessments of species

composition.

Method Summary Multi-habitat sampling is a procedure developed to sample periphyton from wadeable

streams. It should be conducted at the reach scale (30-40 stream widths) to ensure

sampling the diversity of habitats that occur in the stream. The protocol first calls for

visual estimates or quantitative transect-based assessments to determine the

percent cover of each substrate type and the estimated relative abundance of

macrophytes, macroscopic filamentous algae, diatoms and other microscopic algal

accumulations, and other biota. Following preliminary investigations, algae are

collected from all available substrates and habitats roughly in proportion to their areal

coverage in the reach. Periphyton samples should be collected during periods of

stable stream flow. Sm all amounts of subsample (about 5 mL or less) are usually

sufficient. The objective is to collect a single com posite sample that is representative

of the periphyton assemblage present in the reach. Collection techniques depend on

the substrate type and the dominant macroinvertebrates. However, this protocol

recommends that specimens of macroalgae be collected by hand in proportion to

their relative abundance in the reach. Samples are combined into single, water-tight,

unbreakable, wide-mouth containers. After adding the appropriate preservative,

(Lugol’s solution, “M3" fixative, buffered 4% formalin, 2% glutaraldehyde, or other

preservative), the samples are transported back to laboratory on ice. 

EPA’s EMAP field document describes similar protocols for periphyton surveys in

wadeable streams (USEPA, 1998). 

Data

Uses/Application

Multi-habitat sampling will provide information pertaining to species composition.

Changes in species composition among habitat are often evident as changes in color

and texture of the periphyton. These data provide information pertaining to the

effects of pollution on environmental comm unities. 

Multi habitat sampling can also be conducted to collect periphyton for chlorophyll

determination, biomass determination and acid/alkaline phosphate activity.

Advantages The investigators may get a better sense of how habitat changes may impact

different benthic comm unities. 

Limitations There may be variability of data due to differences in habitat between streams. The

results may not be sensitive to subtle water quality changes because of habitat

variability between reaches. 

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition,

EPA 841-B-99-002. U.S. Environmental Protection Agency; Office of Water;

Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/ Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

188

Fact Sheet No. 2.3.1-9

Method Title Artificial Substrate Samplers of Macroinvertebrates in Wadeable Streams

Purpose These methods describe the use of artificial substrate samples that have long been

used in algal investigations and in situations where bottom substrate sampling is not

possible due to physical obstacles. The Rapid Bioassessment Protocols (RBPs) list

artificial substrates as a sampling methodology to collect both periphyton and benthic

macroinvertebrates. 

Method Summary Artificial substrate samplers typically use glass slides as substrate, but also are

deployed with glass rods, ceramic tiles and other substances.

The samplers are positioned in the euphotic zone of good light penetration for

maximum abundance and diversity of macroinvertebrates. Optimum time for

substrate colonization is six weeks. At least two to three samplers should be

installed at each collecting site. 

To retrieve the samplers, they are approached from downstream, lifted quickly and

placed in a polyethylene jug or bag containing 10% formalin or 70-80% ethanol. 

The organisms can be removed in the field by disassembling the sampler in a tub or

bucket partially filled with water and scrubbing the rocks or plates with a soft-bristle

brush to remove clinging organisms. The contents of the bucket are poured through

a No. 30 or 60 sieve and the contents of the sieve are washed into a jar and preserve

with 10% formalin or 70-80% ethanol. 

The use of artificial substrate samplers in macroinvertebrate field and laboratory

studies is also presented in USEPA, 1990b.

Data

Uses/Application

The benthic composition and abundance data from artificial substrate samplers are

used to measure the status and trends of environmental pollution, and effects on

freshwater, estuarine, and marine macroinvertebrates, and to assess surface water

quality.

The RBP protocols caution that artificial substrates should only be used for benthic

macroinvertebrates when other collection devices fail. The substrate used must be

representative of the natural habitat. 

Advantages Artificial substrates allow sample collection in locations that are typically difficult to

sample effectively. As a passive sample collection device, artificial substrates permit

standardized sampling by eliminating subjectivity in sample collection techniques.

Sample collection using artificial substrates may require less skill and training than

direct sampling of natural substrates. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

189

Fact Sheet No. 2.3.1-9 (contd.)

Limitations The limitations commonly encountered when using the artificial substrate sampler

include susceptibility to vandalism, sampling bias for insects, difficulty in anchoring

the device, and the lengthy time from initiation to conclusion of sampling (up to 8

weeks). Furthermore, the material of the substrate will influence the composition

and structure of the community. Orientation and length of exposure of the substrate

will influence the composition and the structure of the community.

Reference Barbour, et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition.

EPA 841-B-99-002. U.S. Environmental Protection Agency; Office of Water;

Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/ Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

190

Fact Sheet No. 2.3.1-10

Method Title Algae and Macroinvertebrate Sampling with Frames

Purpose To delineate the percent coverage of the colonial forms of algae and

macroinvertebrates.

Method Summary A 0.1 m

2

or 1m2square-shaped metal frame can be laid flat along rocky shores,

beaches etc. and be used to delineate the percent coverage of colonial frames. At

least ten frames should be used for counting organisms to characterize species

abundance and distribution adequately. Samples of the algae and

macroinvertebrates should be removed from a measured area for species

identification and weighed for biomass determination. 

Data

Uses/Application

An investigator can gather macrobenthos data using frames from locations where

conventional sampling devices are not practical. Macrobenthos data are used in

benthic com munity analyses to measure the status and trends of environmental

pollution, and its effects on freshwater, estuarine, and marine macroinvertebrates,

and to assess water quality criteria and monitor surface water quality.

Advantages This method is useful for sampling beach infauna.

Limitations Frames delineate organisms present on the surface, however all organisms

burrowed beyond the investigator’s line of site will not be counted. Rely upon

statistics to derive species abundance and distribution for the entire region. 

Reference USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating the

Biological Integrity of Surface Waters, EPA/600/4-90/030. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/clariton/clhtml/pubtitleORD.ht

ml

Last Accessed: 2/13/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

191

Fact Sheet No. 2.3.1-11

Method Title Benthic Organism Collection from a Marine Environm ent, NHEERL-AED SOP

1.02.001

Purpose This procedure describes the methods required to collect infaunal and epibenthic

marine organisms for tissue analyses or for use in toxicological evaluation.

Method Summary Benthic macroinvertebrates: Sediments are frequently collected with grab samplers,

such as the Young-modified Van Veen grab. This particular sampler is constructed

entirely of stainless steel and can be Kynar-coated to make it suitable for collecting

sediment samples for both biological and chemical analyses. The samples should

be numbered and the depth of the sediment at the middle of the sampler should be

recorded on the data sheet. The sampler should be at least half full. The data sheet

should also include a general description of the grab such as the presence or

absence of surface floc, color and sm ell of surface sediments, and visible fauna. 

Worms: Sediments from grab samples are emptied into a tub and then passed

through sieves. The appropriate sieve size could be selected based on sediment

type and organisms to be collected. Worms are picked from the sieves, rinsed free

of sediment, and placed in sample jars.

Quahogs: Quahogs should be collected with the aid of a professional quahog

fisherman. The fisherman should be provided with one extra individual for

assistance, a sampling location chart, and prelabelled and organized sample bags. 

Mytilus: Mussels are collected with a scallop dredge towed at 2-3 knots for 5-10

minutes. The catch is then hauled back, dumped on board, and sorted. 

Oysters: Hand collection in shallow water is recommended.

Soft-shell clams: Soft-shelled clams should be collected by hand at low-tide. After

locating siphon holes on the tidal region of the flat, the clams can be dug out, taking

care to avoid breaking the “soft-shell” during excavation. 

USEPA’s Coastal and Northeast EMAP document describes similar protocols for

benthic organism collection (USEPA, 2000b).

Data

Uses/Application

Benthic organisms can be collected to determine species composition and

abundance in a particular sampling reach, or the organisms may be used for tissue

residue analyses in toxicological evaluation.

Advantages All activities can be performed by hand or from a sm all boat.

Limitations While the use of sediment grab samplers for collecting macroinvertebrates and

worms is almost unanimously recommended, the application of other methods is

highly site-specific.

Reference USEPA and the Naval Construction Battalion Center. 1992. Standard Operating

Procedures and Field Methods Used for Conducting Ecological Risk Assessment

Case Studies. Technical Document 2296. Naval Command, Control and Ocean

Surveillance Center, RDT&E Division, San Diego, CA. 

Website http://www.duxbury.battelle.org/compend

ium/methods/NHEERL-AED-SOP1.02.001.pdf

Last Accessed:

Fact Sheet No. 2.3.1-12
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

192

Method Title Benthic Macroinvertebrate Protocols in a Wadeable Stream: Single Habitat

Approach, 1-Meter Kick Net

Purpose To determine macroinvertebrate diversity and abundance in wadeable streams. 

Method Summary The single habitat approach to benthic macroinvertebrate sampling emphasizes

sampling a single, dominant habitat (i.e., riffles or runs) as a way of standardizing

assessments.

Benthic macroinvertebrate samples are collected system atically using a 1 m kick

net. Once the net is in position on the substrate, a rectangular quadrant that is

approximately 0.5 m

2

is visually defined. The net is held securely while the substrate

is kicked vigorously for 20 seconds. After 20 seconds, the net is removed with a

quick upstream motion to wash the organisms to the bottom of the net. The kicks

collected from different locations in flowing water habitats will be composited to

obtain a single homogeneous sample. Kick net samples collected from pool habitats

are combined into a separate com posite sample. The percentage of habitat type is

recorded along with observations of aquatic flora and fauna and a habitat

assessment will be performed. 

The samples composited from the kick nets will be preserved in 95% ethanol. The

samples are then returned to the laboratory for species enumeration and

identification. 

The USEPA EMAP field document describes similar methods for using a

1-m kick net (USEPA, 1998).

Data

Uses/Application

Species relative abundance and taxa richness are data derived from these protocols.

These data parameters provide information pertaining to the status and trends of

environmental pollution and its impacts on freshwater, marine and Estuarine

comm unities. 

Biological impairment resulting from pollution is often evaluated using metrics of

biotic integrity derived from the aforementioned data parameters that evaluate

community, population and functional parameters.

Advantages The 1-m kick net method provides a rapid, reproducible, and inexpensive method for

the collection of macroinvertebrates from suitable environments.

Limitations Single habitats (i.e., cobble substrates) cannot be solely analyzed in reaches where

the substrate represents less than 30% of the sampling reach.

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition,

EPA 841-B-99-002. Office of Water, U.S. Environmental Protection Agency,

Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/ Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

193

Fact Sheet No. 2.3.1-13

Method Title Benthic Macroinvertebrate Protocols in a Wadeable Stream: Multi-habitat Approach:

D-Frame Dip Net

Purpose To determine macroinvertebrate diversity and abundance in wadeable streams. 

Method Summary This method focuses on a multi-habitat scheme designed to sample major habitats in

proportional representation within a sampling reach. Benthic macroinvertebrates are

collected system atically from all available instream habitats by kicking the substrate

or jabbing with a D-frame dip net. A total of 20 jabs (or kicks) are taken from all

major habitat types in the reach resulting in sampling of approximately 3.1 m

2

of

habitat.

The samples collected from this protocol will then be sent to the laboratory where

species enumeration and identification will be conducted. 

Data

Uses/Application

Species relative abundance and taxa richness are data derived from these protocols.

These data parameters provide information pertaining to the status and trends of

environmental pollution and its impacts on freshwater, marine and estuarine

comm unities. 

Biological impairment resulting from pollution is often evaluated using metrics of

biotic integrity derived from the aforementioned data parameters that evaluate

community, population and functional parameters.

Advantages It is important to use a multi-habitat approach when the stream under investigation

varies in gradient and substrate type. 

Limitations Differences in sampling techniques can lead to variability and difficulty comparing

data among researchers.

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition,

EPA 841-B-99-002. Office of Water, U.S. Environmental Protection Agency,

Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/ Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

194

Fact Sheet No. 2.3.1-14

Method Title Photographic Habitat Documentation of the Benthic Community

Purpose To document a habitat or alterations in a station over time (e.g., increase in canopy

cover, changes in channelization of a stream, and effects of flooding etc.)

Method Summary Photography of aquatic environm ents usually involves SCUBA equipment. The

SCUBA diver will place a photographically identifiable 1.0 m

2

area frame or marker in

the habitat to be photographed and an additional nearby marker on which the

camera is placed each time a photograph is taken to ensure consistency. 

Data

Uses/Application

Photography is a tool used to characterize benthic composition and potential

alterations over time in environm ents with sessile organisms that may change over

time in relation to a new stressor.

Photographic images of the abundance and diversity of sessile organisms over time

is a way in which to monitor the status and trends of environmental pollution, and its

effects on freshwater, estuarine, and marine organisms, and to assess surface water

quality.

Advantages Photographic documentation is a rapid and inexpensive tool to use to support benthic

bioassessments, particularly in areas with significant populations of sessile

organisms. 

Limitations Photography is generally limited to environm ents with suitably clear water that are

inhabited by sessile animals and rooted plants (e.g., estuarine habitats containing

corals, sponges, and attached algal forms). Underwater photography generally

requires trained SCUBA divers.

Reference USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating the

Biological Integrity of Surface Waters, EPA/600/4-90/030. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/clariton/clhtml/pubtitleORD.ht

ml

Last Accessed: 2/13/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

195

Fact Sheet No. 2.3.1-15

Method Title Sediment Profile Cam era

Purpose This method describes use of a sediment profile camera to evaluate the in situ

macroinvertebrate community.

Method Summary Photographs are taken of the benthic com munity prior to any other benthic sampling

to avoid disruption of the surface. The sediment profiling index (SPI) system consists

of a digital camera enclosed in a waterproof, pressure resistant housing, a 45-degree

prism that penetrates the sediment to a depth of 25 cm, and a mirror that reflects an

image of the sediment profile through the camera lens and to the digital camera. The

camera prism is mounted on an assembly that can be moved up and down by

producing tension or slack on the winch wire.

As the camera is lowered, tension on the winch wire keeps the prism in the ‘up’

position until the support frame hits the bottom. At this point the tension on the winch

wire is reduced causing the inner frame to move to the ‘down’ position, penetrating

the undisturbed sediment water interface. The upper 25 cm of the sea floor, as seen

in profile, is then photographed in high resolution with a film or digital camera. An

additional camera mounted on the frame photographs the sediment surface before

the prism penetrates the sediment. After each image is taken, the camera is raised

two or three meters off the bottom and redeployed for taking another image

(‘sample’).

Data

Uses/Application

SPI cameras are used to evaluate macrofauna com munity structure and assess the

benthic habitat. SPI technology can readily quantify over 20 physical, chemical, and

biological parameters including: sediment grain size; prism penetration; surface

pelletal layer; sediment surface relief; mud clasts; redox area; redox contrast; current

apparent redox boundary; relict redox boundaries; methane gas vesicles; apparent

faunal dominants; voids; burrows; surface features (e.g., worm tubes, epifauna,

shell); dredged material; microbial aggregations; and successional stage. SPI data

have been accepted in the United States by the by the U.S. Environmental Protection

Agency and by the U.S. Army Corps of Engineers for describing baseline benthic

habitat conditions at proposed dredged material disposal sites, for monitoring

changes in sediment structure and the benthic com munity from dredged material

disposal, and for monitoring the recovery of disposal sites and their surrounding

environm ent. 

Advantages Rapid photographic evidence of sediment conditions.

Limitations Provides only a partial picture of benthic com munity structure, and little information

on benthic macroinfauna comm unities. Generally, limited to screening in soft bottom

sediments.

Reference USEPA. 1990a. Environmental Monitoring and Assessment Program: Near Coastal

Component, 1990 Demonstration Project, Field Operations Manual. DRAFT.

Contract # 68-C8-0066. Office of Research and Development. Narragansett, Rhode

Island. 

Website http://www.epa.gov/emap/html/pubs/docs/groupd

ocs/estuary/field/90fldman.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

196

Fact Sheet No. 2.3.1-16

Method Title Macroinvertebrate Drift Nets in Wadeable Streams

Purpose To collect macrobenthos inhabiting a wide range of habitat types from shallow

flowing streams or shallow areas in rivers for quantitative evaluations.

Method Summary For synoptic surveys, one net set above each of the major areas of population

concentrations is usually adequate; but for definitive studies a minimum of two drift

nets should be set at each station so that drift from above a pollution source, drift

from the polluted reach and drift from the clean water downstream from the recovery

zone can be compared. 

Use nets with a 929 cm2upstream opening and mesh equivalent to the U.S.

Standard No. 30 screen (0.595 mm pore size). Set drift nets for any specified time

(usually 3 hours). Sampling between dusk and 1 AM is optimum . For definitive

studies, install four nets at each station two about 25 cm from the bottom and tow

about 10 cm below the surface in water not exceeding 3 meters in depth. At the end

of the specified sampling period, remove the net from the water by loosening the

cable clamps and raising the net over the top of the steel rods, taking care not to

disturb the bottom upstream of the net. Concentrate the material in the net in one

corner by swishing up and down in the water and then wash into a bucket half-filled

with water. Then sieve and handle the sample in the regular manner. 

Standard methods 10500 describe similar collection methods (APHA, 1999).

Data

Uses/Application

Drift nets collect macrobenthos in order to characterize the composition and

abundance of macroinvertebrate biota that drift in the water column. A summary of

stream net samplers is presented in Table 2.3.1-1.

Macroinvertebrate data such as these are used to measure the status and trends of

environmental pollution and its effects on freshwater, estuarine, and marine

macroinvertebrates, to assess water quality criteria and to monitor surface water

quality.

Advantages Standard collection method used throughout United States.

Limitations It is unknown where the organisms come from; terrestrial species may make up a

large part of sample in summ er and periods of wind and rain. Drift nets do not collect

non-drifting organisms.

Reference USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating the

Biological Integrity of Surface Waters. EPA/600/4-90/030. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/clariton/clhtml/pubtitleORD.ht

ml

Last Accessed: 2/13/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

197

Table 2.3.1-1. A Summary of Stream Net Samplers Used to Collect Organisms from Flowing Water (USEPA, 1990b1

)

Net Sampler

Habitats and

substrates Effectiveness of Device Advantages Limitations

Surber Stream

Bottom

Sampler

Shallow, flowing

streams, less than 32

cm in depth with good

current; rubble

substrate, mud, sand

and gravel

Performance depends on

current and substrate

Encloses area sampled;

easily transported or

constructed; samples a

unit area

Difficult to set in some

substrate types, that is

large rubble; cannot be

used efficiently in still,

slow-moving streams

Portable

Invertebrate

Box Sampler

Same as Surber Same as Surber Same as Surber except

completely enclosed with

stable platform; can be

used in weed beds.

Same as Surber

Hess Sampler Same as Surber Same as Surber Same as Surber except

completely enclosed with

stable platform; can be

used in weed beds.

Same as Surber

Hess Stream

Bottom

Sampler

Same as Surber Same as Surber Same as Surber except

completely enclosed with

stable platform; can be

used in weed beds.

Same as Surber

Stream-bed

Fauna

Sampler

Same as Surber Same as Surber Same as Surber except

completely enclosed with

stable platform; can be

used in weed beds.

Same as Surber

Drift nets Flowing rivers and

streams; all substrate

types

Effective in collecting all

taxa which drift in the

water column

Low sampling error; less

time, money and effort;

collects

macroinvertebrates from

all substrates, usually

collects more taxa

Unknown where

organisms come from;

terrestrial species may

make up a large part of a

sample in summer and

periods of wind and rain;

does not collect nondrifting organisms

1 USEPA. 1990b. EPA/600/4-90/030.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

198

Fact Sheet No. 2.3.1-17

Method Title Stream-Net Samplers: Surber, Portable Invertebrate Box Sampler, Hess Sampler,

Hess Stream Bottom Sampler, and Stream-Bed Fauna Sampler

Purpose To collect macrobenthos inhabiting a wide range of habitat types from shallow

flowing streams or shallow areas in rivers for quantitative evaluations.

Method Summary The sampler is positioned with its net mouth open, facing upstream. The samplers

are brought down quickly to reduce the escape of rapidly moving organisms. There

should be no gaps under the edges of the frame that would allow for washing of

water under the net and loss of benthic organisms. 

Rem ove the sample after a specified period of time, by inverting the net into the

sample container (wide-mouthed jar) with 10% buffered formalin fixative or 70-80%

ethanol. Examine the net closely for sm all organisms clinging to the mesh, and

remove them (preferably with forceps) for inclusion in the sample. 

Standard method 10500 describe similar collection methods (APHA, 1999).

Data

Uses/Application

Stream-net samplers collect relatively quantitative and qualitative macroinvertebrate

samples from the water column of flowing streams and rivers. The

macroinvertebrates collected in the nets will be taxonomically identified and counted

to determine the macroinvertebrate composition and abundance in that reach of the

river. Composition and abundance data are used to measure the status and trends

of environmental pollution and its effects on freshwater, estuarine, and marine

macroinvertebrates, to assess water quality criteria, and to monitor surface water

quality.

Advantages Rapid, reproducible and inexpensive sampling technique for in-stream fauna. 

Limitations It is difficult to set in some substrate types, such as large rubble. It cannot be used

effectively in still, slow moving streams. Organisms often wash under the bottom

edge of some samplers such as the Surber.

Reference USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating the

Biological Integrity of Surface Waters. EPA/600/4-90/030. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/clariton/clhtml/pubtitleORD.ht

ml

Last Accessed: 2/13/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

199

Fact Sheet No. 2.3.1-18

Method Title Mussel Collection Using Brails

Purpose To sample bivalve mussels in large (non-wadeable) rivers

Method Summary A crowfoot brail will be dragged a measured distance of 100 meters. Each brail

sample is then brought on board the boat, sorted and counted. The area sampled is

calculated in square meters by multiplying the length of the brail by 100 m. Catch

success is expressed in terms of the average catch of mussels per square per drag.

Brail sampling is randomized within fishing areas and by time periods during two

com plete harvest seasons. The crowfoot brails can often be made or rented from a

comm ercial fisherman.

A minimum of six 100 m long hauls (drags) should be accomplished where a single

brail is used. If a significant mussel population is found, then qualitative or

quantitative SCUBA samples should be taken. 

All samples should be identified to species, growth cessation rings counted, and

measured for determination of population age structure. 

Data

Uses/Application

Brail sampling provides both qualitative and quantitative data pertaining to mussel

abundance in a given region. Useful in estuarine areas where mussels comprise a

dominant benthic community.

Advantages Brail sampling is an inexpensive, bioassessment technique for those riverine

environm ents where mussels are an important component of the ecosystem. 

Limitations Mussel fishing with brails is highly dependent on experience of the user; however,

they are very efficient in the hands of experienced users as attested to by almost 100

years of continuous use. 

Reference USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating the

Biological Integrity of Surface Waters. EPA/600/4-90/030. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/clariton/clhtml/pubtitleORD.ht

ml

Last Accessed: 2/13/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

200

Fact Sheet No. 2.3.1-19

Method Title Electrofishing

Purpose Fish collection method for environmental assessments

Method Summary Electrofishing is a method for collecting fish using electricity. Most electrofishing in

freshwater is done with pulsed DC electrical current equipment. In a boat-rigged

shocker (boom shocker), one or two people net the fish and another operates the

boat and equipment. The fish are nearly always driven into cover as a result of

electric stimulus making them difficult to capture. Once driven from cover, the fish

are kept within effective range of the electrical field and are immobilized making it

possible to pick them up with long-handled dip nets.

Other USEPA documents and APHA Standard Method describe similar use of

electrofishing for sample collection methods (Barbour et al, 1999; USEPA, 1998;

APHA, 1999). 

Data

Uses/Application

Electrofishing is a technique used to survey the biological community. As a result,

fish species will be identified and counted to determine the organism abundance and

composition in that region. Abundance and composition data are used to measure

the status and trends of environmental pollution and its effects on marine, estuarine

and freshwater organisms, to assess water quality criteria, and to monitor surface

water quality. 

Advantages Efficient method that can be used to obtain reliable information on fish abundance,

length-weight relationships, and age and growth of fish in most streams of order 6 or

less. Usually results in more consistent success under varying conditions than

ordinary seining. It allows greater standardization of catch per unit effort, it requires

less time and manpower than use of ichthyocides, and it is less selective than seining

(although it is selective towards size and species). If properly used, adverse effects

on fish are minimized, and it is appropriate in a variety of habitats.

Limitations Individuals involved in electrofishing must have completed a certified course in

electrofishing or have been trained by someone certified and experienced in

electrofishing. If target assemblage is a comm on species, then seining may be just

as effective. Cannot be used in water with high turbidity. Need very specific

conditions and equipment. Sampling efficiency is affected by turbidity, conductivity,

aquatic vegetation, depth etc; although it is less selective than seining, electrofishing

also is size and species specific. Effects of elctrofishing increase with body size.

Species specific behavioral and anatomical differences also determine vulnerability to

electroshocking. Electrofishing is a hazardous operation that can injure field

personnel.

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters, EPA/600/R-92-111. Office of Research and

Development, Washington, D.C. 

Website http://www.epa.gov/bioindicators/htm l/fish_m eth

ods.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

201

Fact Sheet No. 2.3.1-20

Method Title Chemical Fishing

Purpose Fish collection method for environmental assessments

Method Summary Fish toxicants are used for sampling fish populations in impounded waters and

streams throughout the United States. Only registered fish chemical toxicants, such

as rotenone, cresol, copper sulfate, antimycin A and sodium cyanide, can be used to

collect fish in the U.S. The ideal icthyocide is nonselective; easily, rapidly, and safely

used; readily detoxified; and not detected and avoided by fish. Chemical sampling is

usually employed on a spot basis (e.g., a short reach of river or an embayment of a

lake or reservoir). A concentration of 0.5 ppm active ingredient will provide good

recovery of most species of fish in acidic or slightly alkaline water. Emulsion

products are applied via manual pumps , spraying equipment, power-driven pumps,

or a drip spout coming from a flowing system.

Standard Method 10600 describes similar uses of chemical fishing for fish sample

collection (APHA, 1999). 

Data

Uses/Application

Chemical fishing surveys are often used to gain information on particular species of

fish and an overall estimation of fish populations and comm unities. Fish population

and com munity data are used to measure the status and trends of environmental

pollution and its effects on freshwater, estuarine and marine organisms, to assess

water quality criteria, and to monitor surface water quality. Rotenoning provides

greater standardization of unit of effort than seining. Rotenoning has the potential, if

used effectively, to provide more com plete censuring of the fish population than

seining or electrofishing.

Advantages Advantages of rotenone: The effective use of rotenone is independent of habitat

complexity.

Limitations Disadvantages of rotenone: Use of rotenone is prohibited in many states, application

and detoxification can be time and manpower intensive. Effective use of rotenone is

affected by temperature, light, dissolved oxygen, alkalinity, and turbidity. Rotenoning

typically has a high environmental impact; concentration miscalculations can produce

substantial fish kills downstream of the study site.

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters, EPA/600/R-92-111. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/bioindicators/html/fish_method

s.htm l

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

202

Fact Sheet No. 2.3.1-21

Method Title Fish Collection Using Seine Nets

Purpose Fish collection method for environmental assessments

Method Summary A strip of strong netting is hung between a stout float line and a strong, heavily

weighted lead line at the bottom. In deepwater, one end of the hauling lines is

anchored on shore and the boat plays out the line until it reaches the end. The boat

then lays out the net parallel to the beach. When all of the net is in the water, the

boat brings the end of the second hauling line ashore. The net is then beached as

rapidly as possible. In shallow waters, a person can lay out the one end of the

hauling line and replicate the role of the boat. There are many different types of

seines; selecting the appropriate seine depends on the study design, sampling

methods and habitat type. 

Other USEPA documents and APHA Standard Methods describe the similar use of

seine nets for fish collection (Barbour et al, 1999; USEPA, 1998; Standard Method

10600, APHA, 1999).

Data

Uses/Application

Seine nets survey the biological com munity by collecting abundance and taxa

composition data from sampling locations. Organism abundance and composition

data are used to measure the status and trends of environmental pollution and its

effects on freshwater, estuarine and marine organisms, to assess water quality

criteria, and to monitor surface water quality. The Rapid Bioassessment Protocols

(RBPs) list seining as a viable way to collect fish samples. 

Advantages Seines are lightweight and easily transported and stored. Seine repair and

maintenance are minimal and can be accomplished onsite. Seine use is not

restricted by water quality parameters. Effects on the fish population are minimal

because fish are collected alive and are generally unharmed.

Limitations Not effective in deep water. Not effective in areas that have snags, large rocks and

boulders, or sunken debris that may tear or foul the net. Quantitative seining is very

difficult. Previous experience and skill, knowledge of fish habitats and behavior and

sampling effort are probably more important in seining than in the use of any other

approaches. Seining sample effort and results are more variable than sampling with

electrofishing and rotenoning. Seine use is generally restricted to slower water with

smooth bottoms, and is most effective in sm all streams or pools without litter cover or

debris. Standardization of unit of effort to ensure data com parability is difficult

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters, EPA/600/R-92-111. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/bioindicators/htm l/fish_m eth

ods.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

203

Fact Sheet No. 2.3.1-22

Method Title Entanglement Nets

Purpose Fish collection method for environmental assessments

Method Summary Entanglement nets, including gill nets and trammel nets, are used to sample fish

populations in estuaries, lakes, reservoirs, and larger rivers. Gill nets are the more

commonly used entanglement nets. They are usually set as an upright, vertical

fence of netting and can have either a variable or uniform mesh size. Gill nets

selectively capture particular species of fish since the mesh size determines the size

range of the fish to be sampled. They can be set at the surface, mid-depth, or on the

bottom depending on the objectives of the study and target species within the fish

community. Tramm el nets are used in all types of riverine habitat. If a river channel

is to be fished, the net is floated or drifted downstream. They are very efficient for

taking fish like carp, buffalo, shovelnose sturgeon and freshwater drum. 

Data

Uses/Application

Entanglement nets survey the biological com munity by collecting abundance and

taxa composition data from sampling locations. They are often used to gain

information on particular species of fish and an overall estimation of fish populations

and comm unities. Fish population and com munity data are used to measure the

status and trends of environmental pollution and its effects on freshwater, estuarine

and marine organisms, to assess water quality criteria, and to monitor surface water

quality. 

Advantages This method describes an effective way to sample fish populations. The results are

expressed as the number or weight of fish taken per length of net per day (catch per

unit effort).

Limitations Entanglement nets need to be monitored for by-catch. Non-target species may be

caught and will not survive long in the net. Tidal currents, predation, optimum fishing

time and types of anchors, floats and line must be considered when setting

entanglement nets in estuaries.

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters, EPA/600/R-92-111. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/bioindicators/html/fish_metho

ds.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

204

Fact Sheet No. 2.3.1-23

Method Title Entrapment Devices

Purpose Fish collection method for environmental assessments

Method Summary With entrapment devices, fish enter an enclosed area (which may be baited) through

a series of one or more funnels. They are set in structurally complex areas where

fish movement and density are anticipated to be highest in order to maximize net

catches. Common entrapment devices include the hoop net, fyke net, and minnow

trap.

Data

Uses/Application

Entrapment devices survey the biological com munity by collection abundance and

taxa composition data from sampling locations. They are often used to gain

information on particular species of fish and an overall estimation of fish populations

and comm unities. Fish population and com munity data are used to measure the

status and trends of environmental pollution and its effects on freshwater, estuarine

and marine organisms, to assess water quality criteria, and to monitor surface water

quality. 

Advantages They are used to sample reservoirs and wide river channels with slow velocity

conditions. The catch is recorded as numbers of weight per unit of effort, usually fish

per net day.

Limitations Entrapment devices are generally deployed overnight, requiring separate filed activity

for deployment and retrieval. Traps are often vandalized in unsecured areas.

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters, EPA/600/R-92-111. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/bioindicators/html/fish_method

s.htm l

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

205

Fact Sheet No. 2.3.1-24

Method Title Pop Nets

Purpose Fish collection method for environmental assessments

Method Summary Pop nets are rectangular devices, constructed of mesh netting used for the collection

of fish. They are designed to be deployed from the surface and released with a

mechanical device. They are set and received by two individuals and are easily

dissembled for transport.

Data

Uses/Application

Pop nets survey the biological com munity by collecting abundance and taxa

composition data from sampling locations. They are often used to gain information

on particular species of fish and an overall estimation of fish populations and

comm unities. Fish population and com munity data are used to measure the status

and trends of environmental pollution and its effects on freshwater, estuarine and

marine organisms, to assess water quality criteria, and to monitor surface water

quality. 

Advantages Useful for sampling fish in shallow, riverine waters in heavily vegetated areas and

nonvegetated areas where seining or electroshocking may be difficult. 

Limitations Pop nets will sample a relatively sm all area/volume of surface water. Thus,

representativeness may be a concern if used for population studies.

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters. EPA/600/R-92-111. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/bioindicators/html/fish_method

s.htm l

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

206

Fact Sheet No. 2.3.1-25

Method Title Trawls

Purpose Fish collection method for environmental assessments

Method Summary Trawls are designed like seines; however, they are much larger and intended to be

towed behind boats in large, open water areas. There are four types of trawls

available: beam trawls, used to capture bottom fish; otter trawls, used to capture

near-bottom and bottom fish; mid-water trawls, used to collect schooling fish at

various depths; and surface tow nets, used to collect fish at or near the surface. The

trawls are deployed behind the boat, often with power winches and large motors. 

Under the Coastal EMAP Program, fish are specifically collected with a high rise

sampling trawl with a 13.5 meter footrope with a chain sweep. Tow duration is 10

minutes with a towing speed of 2-3 knots against the prevailing current. Speed over

the bottom should be 1-3 knots (USEPA, 2000b). 

The trawl is retrieved used hydraulics and the contents of the net are often emptied

onto sorting tables. Fish are sorted, enumerated and examined for gross

pathological examinations. Selected specimens are retained and properly processed

for tissue chemical analyses.

The APHA Standard Method describe similar trawl methods to collect fish (Standard

Method 10600 (APHA, 1999).

Data

Uses/Application

Trawl nets survey the biological com munity by collecting samples that will be

analyzed for species composition, relative abundance, chemical analysis, and

pathological examination. They are often used to gain information on particular

species of fish and an overall estimation of fish populations and comm unities. Fish

population and com munity data are used to measure the status and trends of

environmental pollution and its effects on freshwater, estuarine and marine

organisms to assess water quality criteria, and to monitor surface water quality. 

Advantages Trawl nets are very effective in large, open water areas and can effectively sample

selected bottom, mid-water, and surface oriented species at specific life history

stages. 

Limitations Not effective in deep water. Not effective in areas that have snags, large rocks and

boulders and sunken debris that may tear or foul the net. Quantitative trawling is very

difficult. Previous experience and skill, knowledge of fish habitats and behavior and

sampling effort are probably more important in seining than in the use of any other

approaches. Trawling sample effort and results are more variable than sampling with

electrofishing and rotenoning. Trawl use is generally restricted to slower water with

smooth bottoms, and is most effective in sm all streams or pools without litter cover or

debris. Standardization of unit of effort to ensure data com parability is difficult

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters. EPA/600/R-92-111. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C. 

Website http://www.epa.gov/bioindicators/html/fish_method

s.htm l

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

207

Fact Sheet No. 2.3.1-26

Method Title Fish Processing Method in the Great Lakes, LMMB 025

Purpose This method describes the aging, compositing, and grinding method for fish.

Method Summary Whole fish are collected (intact, with all body fluids and no incisions), wrapped in

aluminum foil, placed in 4 mil thick polyethylene bags, tagged, and frozen as soon as

possible on board the vessel. Fish should be transferred in coolers and stored at -

20°C. To age the fish, scales were removed from the fish, and the annual rings on

the scales are read to determine the age. Some fish that have been stocked may

contain coded wire tags (CWT) or clipped fins. Stocking or tagging records may

provide useful information in aging such fish. 

For homogenization, fish are removed from the freezer and allowed to thaw in their

bags over an 8 - 12 hour period. The contents of the bags are weighed and

recorded. Fish may be composited based on species, location, size and season

sampled. Fish are measured (millimeters) on a measuring board and weighed to the

nearest gram. The measuring board, scalpel, and balance are cleaned between

each group. Each com posite sample is homogenized, using various size vertical

cutters or Robot Coupe cutter. Subsamples of the homogenized tissue are placed in

clean sample containers and frozen (-20°C) until analyzed. 

Standard Method 10600D.3 also discusses the field processing of fish, including

length, weight, and age measurem ents (APHA, 1999). 

Data

Uses/Application

Fish are often collected for chemical analyses of tissue to determine whether

contaminants are accumulating in biological populations.

Advantages These are standard methods and are performed fairly consistently to prepare tissue

for chemical analyses which enhances data consistency in resultant data.

Limitations Study design should consider advantages/limitations of homogenates of whole body,

fillet, or offal.

Reference USEPA. 1997b. Method LMMB 025: Fish Processing Method. Lake Michigan Mass

Balance Study Methods Compendium, Volume 1: Sample Collection Techniques,

EPA 905-R-97-012c. Great Lakes National Program Office, Chicago, IL.

Website http://www.epa.gov/glnpo/lmmb/methods/fpml

mm b.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

208

Fact Sheet No. 2.3.1-27

Method Title Fish Processing

Purpose To determine fish diversity and abundance

Method Summary Once a catch is brought on deck, fish are identified to species, measured, counted,

examined for external pathology and processed for chemical analysis. 

The fish are measured with a measuring board, the fork length to the nearest

millimeter. Coastal EMAP procedures specify that the first 30-50 individuals of each

species should be measured. The remaining fish will be identified to species and

counted (USEPA, 2000b, EPA/620/R-00/002). 

All individuals that are measured, will be examined for evidence of gross external

pathology. The examination is intended to be a rapid scan of the surface of

individual. 

Fish should be cut the entire length of the abdominal cavity. The later musculature is

removed from one side of the animal’s visceral cavity to facilitate the fixation of the

internal organs. The opercula is removed and immersed in fixative. The sample

(whole fish or head, visceral cavity and abnormalities excised) is placed in a plastic

bag with many perforations. This bag is then placed in the fixative. Specimen’s

should be fixed in Dietrich’s fixative for one or two days . Samples may be

transferred to another preservative, such as ethyl alcohol (70-75%) or isopropanol

(40-45%), for storage. 

Samples for fish tissue contaminant analysis or electrophoresis must be iced, placed

in dry ice, or liquid N2

for temporary storage or shipping. Special preservation

techniques must be used for histological, histochemical, or biomarker analyses. 

Samples are then identified to the species level. Data recorded include species

composition and diversity, population density and biomass, and physiological

condition of indigenous comm unities of aquatic organisms. 

Data

Uses/Application

Species relative abundance and taxa richness are data derived from these protocols.

These data parameters provide information pertaining to the status and trends of

environmental pollution and its impacts on freshwater, marine and estuarine

comm unities. 

Biological impairment resulting from pollution is often evaluated using metrics of

biotic integrity derived from the aforementioned data parameters that evaluate

community, population and functional parameters. Fish metrics commonly

determined from this data include species richness, trophic composition, and fish

abundance and condition. Metrices are collectively evaluated in indices such as the

Index of Biotic Integrity (IBI) which aggregates 12 biological metrics to assess fish

assemblage data. 

Advantages These are considered basic and standard methods for fish processing.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

209

Fact Sheet No. 2.3.1-27 (contd.)

Limitations As with any tissue processing, care must be taken to avoid laboratory and crosscontamination.

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition,

EPA 841-B-99-002. Office of Water, U.S. Environmental Protection Agency,

Washington, D.C.

Website http://www.epa.gov/owow/monitoring/rbp/ Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

210

Fact Sheet No. 2.3.1-28

Method Title Swallows: Sampling Procedures

Purpose This method describes the use of tree swallows at contaminated sites to quantify

population level impacts and population level chemistry data. 

Method Summary Approximately 60 swallow boxes, 30 at each of 2 sites, will be attached to posts or

other suitable structures in suitable habitat. Boxes will be placed approximately 20-

30 meters apart, but this can vary depending on the structure of the habitat. Each

nest box will be visited approximately once per week until egg laying begins. After

that time, nests may be visited more often to collect egg or just hatched young

samples. After the eggs have been hatched, boxes will be visited at least once per

week until the young reach 12 days of age. Whether eggs or young are present in

the nest box, the number of eggs and young present will be recorded on a data

sheet. A sample of 2-3 eggs and/or just-hatched eggs (pippers) and a sibling 12-dayold tree swallow nestling will be collected from a minimum of 5-10 boxes at each site.

Food samples from the stomachs of tree swallow nestlings will be removed at the

time that they are collected and dissected. A pooled food sample from each site,

along with the pipper and nestling samples may be analyzed for organochlorine

chemicals, including total PCBs and a full dioxin scan if sufficient mass is available. 

Nestling tree swallows may be ligatured to obtain additional food samples for insect

species identification and for chemical analysis of food. Ligatures, black electrical zip

ties, will be placed on all nestlings in a nest box and left in place for 1 hour. Care will

be taken that the zip ties are loose enough to allow normal breathing. After 1 hour,

the food boli will be removed from the throats of the nestlings using a pair of bluntnosed forceps and the ligatures removed. 

Data

Uses/Application

Swallows are collected for determining population impacts (i.e., reproductive

successes, deformities etc) and for chemical analyses.

Advantages Evaluation of population level impacts is important to determine potential exposures

and effects over the long term. 

Chemical analyses of swallow tissue indicate whether or not bioaccum ulative

chemicals are present and whether they are being transferred up the food chain.

Limitations Sampling bird species is less precise since their mobility allows them to forage in a

great range of areas. Therefore, population level and tissue contaminant level

results may not always be indicative of the conditions at the site of concern. 

Single species tests sometimes skew results since species will react differently to

contaminant exposures due to various biochemical or physiological traits. 

Reference USGS. 1998. Tree Swallow Sample Collection and Processing Procedures,

Technical Operating Procedure WE-410.0. Upper Mississippi Science Center, U.S.

Geological Survey, LaCrosse, W I.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

211

Fact Sheet No. 2.3.1-29

Method Title Sample Processing of Swallows

Purpose This method describes the use of tree swallows at contaminated sites to quantify

population level impacts and population level chemistry data. 

Method Summary Pippers or 12-day old nestlings will be removed from the nest box and weighed.

Pippers and nestlings will be visually examined for gross abnormalities. Nestlings will

be decapitated with a pair of sharp scissors, contents in the upper gastrointestinal

tract removed with forceps after an incision is made along the length of the stomach,

and the carcass remainder placed in a chemically clean jar, which has been

purchased in that condition. The above will be done within 2 hours after removal

from the nest box. The carcass remainders and food samples will be maintained

frozen until transported to the storage in a freezer. Samples will be shipped to an

analytical laboratory for processing.

Data

Uses/Application

Swallows are collected for determining population impacts (i.e., reproductive

successes, deformities etc) and for chemical analyses used in ecological risk

assessments. Evaluation of population level impacts is important to determine

potential exposures and effects over the long term. Chemical analyses of swallow

tissue indicate whether or not bioaccum ulatitve chemicals are present and whether

they are being transferred up the food chain.

Advantages Swallows are often used in ecological risk assessments at freshwater ponds

because of their exclusive insect diet and comparative ease of collection. 

Limitations Sampling bird species is less precise since their mobility allows them to forage in a

great range of areas. Therefore, population level and tissue contaminant level

results may not always be indicative of the conditions at the site of concern. 

Single species tests sometimes skew results since species will react differently to

contaminant exposures due to various biochemical or physiological traits. 

Reference Custer, C.M., T.W . Custer, P.D. Allen, K.L. Stromborg, and M.J. Melancon. 1998.

Reproduction and environmental contamination in tree swallows nesting in the Fox

River drainage in Green Bay, Wisconsin, USA. Environmental Toxicology and

Chemistry. 17:1786-1798.

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

212

2.3.2 Chemical and Physical Analysis

Section 2.3.2 contains methods for sample preparation and the chemical analysis of biota. These

methods characterize the chemical composition of biota samples collected by methods described

in Section 2.3.1. Samples are often analyzed for the presence of inorganic and organic

contaminants that may pose a threat to human or ecological health. Analyzing biological tissue

provides a direct measure of the uptake and bioaccumulation of pollutants from the environment

and can be used to correlate environmental concentrations of contaminants with body residues. 

Many of the methods described have been developed over time to optimize the detection,

identification, and quantification of potential chemicals of concern. Several are performance-based

and may be further modified to enhance the accuracy and precision of the method. Special

preparation and clean up procedures are utilized when analyzing biological tissue, due to the often

high lipid content of the samples. Lipids tend to concentrate environmental pollutants, but they can

also interfere with the analysis of these contaminants.

The chemical methods for the analysis of biological tissue are less routinely performed than for the

analysis of water and sediment. NOAA’s National Status and Trends Program (1998) developed

many methods for the chemical analysis of biota and was a main source of information in the

preparation this compendium. Other sources of information presented in Section 2.3.2 included: 

• The USEPA’s Office of Water

• The USEPA’s Lake Michigan Mass Balance Study Methods Compendium, 1997v

• The USEPA’s Fish Field and Laboratory Methods for Evaluating the Biological Integrity

of Surface Waters, 1993

• The USEPA’s Test Methods for Evaluating Solid Waste, Physical/Chemical Methods

(SW846 Methods)

• Standard Methods for Examination of Water and Wastewater, 1999

• ASTM
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

213

Fact Sheet No. 2.3.2-1

Method Title Sample Preparation for Metal Contam inants in Tissue

Purpose This method describes how fish tissue is processed and prepared for metal

contaminant analyses in the laboratory. 

Method Summary Prior to use, utensils and bottles should be thoroughly cleaned with a detergent-free

solution, rinsed with tap water, soaked in acid, and then rinsed with metal-free water.

Sample size requirements vary with tissue type and detection limit requirements.

When filleting the fish, special care should be taken to avoid contaminating target

tissues (especially muscle) with slime and/or adhering sediment from the fish skin.

The procedure previously outlined for the preparation of fillet samples should

generally be followed. Unless specifically sought as a sample, the dark muscle

tissue that may exist in the vicinity should not be separated from the light muscle

tissue. 

Samples should be frozen after resection and kept at -20°C. Samples may or may

not be homogenized before acid digestion and subsequent preparation before

analysis.

ASTM Method D4309 also describes the preparation of biological samples for

inorganic chemical analysis (ASTM, 2001a).

Data

Uses/Application

This method is followed by investigators preparing fish tissue for trace metal analysis.

Advantages Control of metal contamination is addressed in this method.

Limitations The major difficulty in trace metal analyses of tissue samples is controlling

contamination of the sample after collection. In the field, sources of contamination

include sampling gear, grease from winches or cables, engine exhaust, dust or ice

used for cooling. Sample resection and any subsampling of the organisms should be

carried out in a controlled environment (i.e., a Class 100 clean room).

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters, EPA-600-R-92-111. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/bioindicators/html/fish_metho

ds.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

214

Fact Sheet No. 2.3.2-2

Method Title Total Mercury in Tissue, Sludge, Sediment, and Soil by Acid Digestion and BrCl

Oxidation, Appendix to Method 1631

Purpose These procedures may be used in conjunction with EPA Method 1631B for

determination of mercury in tissue, sludge, sediment, soil, industrial samples, and

certified reference materials.

Method Summary Digestion I—This procedure is preferred for matrices containing organic materials,

such as sludge and plant and animal tissues, because the organic matter is

completely destroyed. In this procedure, a 0.2 - 1.5 g sample is digested with

HNO3/H2SO4. The digestate is diluted with BrCl solution to destroy the remaining

organic material.

Digestion II—This procedure is preferred for geological materials because of rapid

and com plete dissolution of cinnabar (HgS), which is otherwise more slowly

attacked by the BrCl in Digestion I. In this procedure, a 0.5 - 1.5 g sample is

digested with aqua regia (HCl/HNO3) to solubilize inorganic materials.

The Hg concentration in the digestate is determined using EPA Method 1631B.

These procedures, in conjunction with Method 1631B, allow determination of Hg

at concentrations ranging from 1.0 to 5000 ng/g in solid and semisolid matrices.

The method detection limit for Hg has been determined to be in the range of 0.24

to 0.48 ng/g when no interferences are present. The minimum level of

quantization (ML) has been established as 1.0 ng/g. These levels assume a

sample size of 0.5 g.

Using Method LMMB 053 (USEPA, 1997d) to measure total mercury in fish, the

fish tissue is digested in nitric acid for 30 minutes at room temperature then in a

bomb at 190°C for 15 minutes. The digestate is diluted to 25 mL using Milli-Q

water and sample aliquots are analyzed using the CVAFS purge and trap method.

Specific procedures for the measurement of mercury in plankton are described in

Method LMMB 051 (USEPA, 1997d).

Data

Uses/Application

The extent of mercury bioaccumulation is an important parameter to support

human health, ecological risk assessments and bioaccumulation models.

Advantages The dual amalgam trap system and fluorescence detector provide greater

sensitivity and specificity in the presence of interferences, and this system must

be used to overcome interferences, if present, and to achieve the sensitivity

required, if necessary.

Limitations This method does not measure methyl mercury, which may need to be monitored

separately in contaminated sediments.

Reference USEPA. 2001d. Appendix to Method 1631: Total Mercury in Sludge, Sediment,

Soil, and Tissue by Acid Digestion and BrCl Oxidation, EPA-821-R-01-013. Office

of Water, U.S. Environmental Protection Agency, Washington, DC.

Website http://www.brooksrand.com/FileLib/163

1e.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

215

Fact Sheet No. 2.3.2-3

Method Title Versatile Combustion-Amalgamation Technique for the Photometric Determination of

Mercury in Fish and Environmental Samples, LMMB 052

Purpose For the direct detection of total mercury in fish and environmental samples.

Method Summary 0.05 - 0.1 g fish tissue is heated in a stream of O2for 3.5 minutes in an induction

furnace. The released mercury vapor passes through a series of traps, and the

mercury is collected in a 10 mm diameter column of 24-gauge gold wires. This

amalgam is heated in the induction furnace and volatilized mercury is measured with

a mercury vapor meter.

The detection limit is less than 0.002 μg.

Data

Uses/Application

The extent of mercury bioaccumulation is an important parameter to support human

health, ecological risk assessments and bioaccumulation models.

Advantages This analytical system is easily converted to handle biological materials, water, and

sediments. Total analysis time is about 8 minutes per sample and a single analyst

can make up to 40 determinations in eight hours. The method has high sensitivity,

precision, and accuracy. Also, a sm all sample size is required.

Limitations For analysis of samples outside the range of 0.02 - 5.0 ppm mercury, a change of

operating procedure is required. Frequent changes in attenuation of the mercury

vapor meter are required. The sample must be well homogenized, due to the very

sm all sample size analyzed with this method. Detection limits may not be sufficiently

low for ecological or human health risk assessments.

Reference USEPA. 1997d. Method LMMB 052: Versatile Combustion-Amalgamation

Technique for the Photometric Determination of Mercury in Fish and Environmental

Samples, Lake Michigan Mass Balance Study Methods Compendium, Volume 2:

Organic and Mercury Sample Analysis Techniques, EPA 905-R-97-012c. Great

Lakes National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/glnpo/lmmb/methods/s

can.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

216

Fact Sheet No. 2.3.2-4

Method Title Trace Element Quantification Techniques

Purpose To determine major and trace elements in sediment and biological tissue samples

utilizing atomic absorption and neutron activation techniques.

Method Summary Tissue samples are homogenized and freeze dried, and a dry aliquot is

homogenized and transferred to a TeflonTM bomb. Samples are digested by adding 3

mL HNO3 and leaving the bombs at room temperature overnight. The bombs are

then placed in a 130°C oven for approximately 20 hours. After cooling, 18 mL of

quartz distilled water are added, and the solution volume is determined, and a 20-fold

dilution is made for FAAS analysis of Al, Fe, Mn, Si, and Zn. For analysis of Hg,

sediment samples are digested using a modified version of EPA method 245.6.

Samples were analyzed using the following instrumentation:

Analyte Method

Hg Cold vapor atomic absorption (CVAA)

Cu, Fe, Zn Flame atomic absorption (FAA)

Ag, As, Cd, Cr, Cu, Ni, Pb, Se, Sn Graphite furnace atomic absorption 

 (GFAA)

Ar, Cr, Fe, Se, Ag, Zn Instrumental neutron activation 

 analysis (INAA)

ASTM Method D1971 describes the digestion of samples for determination of metals

by Flame Atomic Absorption (ASTM, 2001a). 

Standard Method 3030K describes the microwave digestion method (APHA, 1999).

Several Standard Methods describe the analysis of metals using various methods:

SM 3112 B for CVAA; SM 3111 for FAA; SM3113B for GFAA. 

Data

Uses/Application

Methods provide low detection limits needed for measuring ambient concentrations

at uncontaminated sites, or reference sites.

Advantages Tissue sample digestion in a TeflonTM bomb is a standard method for “clean”

digestion for metals analysis. The instrumental suite employed in this method takes

advantage of the know strengths of each instrument for trace analysis. For example,

GFAA is much more sensitive than FAA, requiring only a sm all volume of sample for

trace analysis.

Limitations Requires multiple instrument analyses for results on a com plete suite of elements.

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and Trends

Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo NOS ORCA

130. National Oceanic and Atm ospheric Administration, Silver Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications/tm

130.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

217

Fact Sheet No. 2.3.2-5

Method Title Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

Purpose To determine the concentration of 17 metals in sediment and biological tissue

samples utilizing atomic absorption, inductively coupled plasma mass

spectrometry (ICP-MS), and energy dispersive X-Ray fluorescence (XRF).

Method Summary Tissue samples are weighed and freeze-dried. The dried sample is ground in a

mill. 0.5 g aliquots are used directly for XRF analysis or are further digested for

AA or ICP-MS analysis. 

500 mg of dried tissue is placed in a TeflonTM bomb, to which 5 mL of HCl and

3.5 mL of HNO3 are added. The bombs are heated in a 60°C water bath for 3-4

hours. After cooling, the bombs are heated in a 130°C oven for 16 hours. After

cooling, the digestates are diluted to approximately 20 mL with deionized water.

Solution volumes are calculated, and the digestates are analyzed directly by

GFAA and CVAA or diluted 10:1 for ICP-MS analysis.

Analyte Method

Hg Cold vapor/gold foil amalgam

Al, Cd, Cr, Ni, Ag, Pb Graphite furnace atomic absorption (GFAA)

Ag, Al, Cr, Cd, Ni, Pb, Sb, Sn ICP-MS

As, Cu, Fe, Mn, Se, Si, Zn XRF

Several Standard Methods exist for the analysis of metals by a variety of

methods: 3112B for CVAA; 3113B for GFAA; 3120B for ICP-MS (APHA, 1999).

 

Data Uses/Application Metals contaminant data are used in both ecological and human health risk

assessments.

Advantages XRF analysis does not require digestion of the sample. Se and As, which can

be difficult to analyze in tissue by ICP-MS, are easier to analyze by XRF. The

digestion solution of HCl/HNO3 provide better recoveries for Ag. ICP-MS has

the advantage of simultaneous analysis of many elements with detection limits

much lower than the XRF and similar to those of GFAA. ICP-MS is particularly

sensitive for Al, Cr, Ni, Ag, Cd, Sn, Sb, P, and Tl. CVAA is very sensitive and

reliable for Hg analysis.

Limitations Leakage at high pressure can cause loss of Hg from the sample during

digestion. Analysis of GFAA requires the use of matrix modifiers and

standardization of the instrument by method of addition to the sample matrix to

provide accurate results.

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and

Trends Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo

NOS ORCA 130. National Oceanic and Atm ospheric Administration, Silver

Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications/t

m130.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

218

Fact Sheet No. 2.3.2-6

Method Title Chemical Speciation of Arsenic in Water and Tissue by Hydride Generation Quartz

Furnace Atomic Absorption Spectrometry, EPA Method 1632, Revision A

Purpose This method is for determination of inorganic arsenic (IA), arsenite (As +3 ), arsenate

(As +5 ), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA) in tissue

by hydride generation and quartz furnace atomic absorption detection. This method

is designed for measurement of As species in tissue in the range 0.10-500 μg/g dry

weight.

Method Summary A 10- to 50-g wet weight sample is collected into a sample bottle. The tissue sample

is either freeze-dried and stored at room temperature or stored frozen at less than -

18°C. Prior to analysis, tissue samples are digested in HCl or NaOH at 80°C for 16

hours. An aliquot of tissue digestate is placed in a specially designed reaction

vessel, and 6M HCl is added. NaBH4solution is added to convert IA, MMA, and DMA

to volatile arsines. Arsines are purged from the sample onto a cooled glass trap

packed with 15% OV-3 on Chrom osorb ® W AW-DMCS, or equivalent. The trapped

arsines are thermally desorbed, in order of increasing boiling points and carried into

the quartz furnace of an atomic absorption spectrophotometer for detection. To

determine the concentration of As +3, another aliquot of water sample or tissue

digestate is placed in the reaction vessel and Tris-buffer is added. The procedure is

repeated to quantify only the arsine produced from As +3. The concentration of As

+5 is the concentration of As +3 subtracted from the concentration of IA.

Analyte MDL ML

IA (As +3 +As +5 ) 0.03μg/g 0.10 μg/g

Arsenite (As +3 ) 0.02 μg/g 0.10 μg/g

MMA 0.01 μg/g 0.05 μg/g 

DMA 0.04 μg/g 0.10 μg/g 

Data

Uses/Application

The method is for use site characterizations and risk assessments.

Advantages The relative amounts of carcinogenic arsenite (As +3) to total arsenic varies with

surface water body and varies with pH. This method directly quantifies arsenite.

Limitations This method is far more costly than total arsenic determination and may only be

needed where speciation is required.

Reference USEPA. 2001c. Method 1632, Revision A: Chemical Speciation of Arsenic in Water

and Tissue by Hydride Generation Quartz Furnace Atomic Absorption Spectrometry,

EPA 821-R-01-006. Office of Water, U.S. Environmental Protection Agency,

Washington, DC.

Website http://www.brooksrand.com/methods/1632a.pdf Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

219

Fact Sheet No. 2.3.2-7

Method Title Extraction and Lipid Separation of Fish Samples for Contaminant Analysis and Lipid

Determination, LMMB 043

Purpose To prepare tissue samples for the measurement of organic contaminants, such as

polychlorinated biphenyls, polynuclear aromatic hydrocarbons, and chlorinated

pesticides.

Method Summary Using Method LMMB 043 (USEPA, 1997d), 10 g of tissue are combined with sodium

sulfate. The tissue mixture is transferred to a rinsed chromatography column that is

plugged with glass wool. The tissue is extracted twice by eluting the column with 50

mL of a 90/10 petroleum ether/ethyl acetate mixture. The elutant is concentrated

using a Turbovap. A GPC column is used to remove lipids from the extract.

Samples are solvent exchanged into iso-octane and cleaned up using a silica gel

column. Extracts are eluted using a 5/95 ethyl acetate/hexane solvent mixture.

Samples are concentrated for analysis.

Using NS&T procedures (NOAA, 1998), a 0.5 - 15 gram (wet weight) aliquot of the

homogenized tissue sample is spiked with surrogate standards and extracted three

times with dichloromethane in the presence of sodium sulfate by maceration with a

TissumizerTM. The extract is filtered through glass wool and sodium sulfate after

each extraction. The extract is concentrated using the Kuderna-Danish technique

and solvent changed to hexane. The sample is cleaned-up using alumina/silica gel

column chromatography before instrumental analysis. Tissue samples require

further purification by gel permeation chromatography (GPC) prior to instrumental

analysis for pesticides and PCBs. The sample is concentrated to 1 mL in hexane for

analysis.

US EPA’s Fish Field and Laboratory Methods for Evaluating the Biological Integrity of

Surface Waters (USEPA, 1993c) also describe sample preparation methods for

organic contaminants in tissue.

Data

Uses/Application

Tissue contaminant data are used to delineate the spatial and temporal extent of

contamination and used in ecological and human health risk assessments.

Advantages This method provides quantitative extraction of most organic contaminants from

tissue samples, including those with high lipid contents.

Limitations These extraction methods have been validated for non-polar persistent organic

contaminants, such as PCBs, chlorinated pesticides, and PAHs. Methods may not

be applicable for more polar compounds or more reactive compounds.

Reference USEPA. 1997d. Method LMMB 043: Extraction and Lipid Separation of Fish

Samples for Contaminant Analysis and Lipid Determination, Lake Michigan Mass

Balance Study Methods Compendium, Volume 2: Organic and Mercury Sample

Analysis Techniques. EPA 905-R-97-012c. Great Lakes National Program Office,

U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/glnpo/lmmb/methods/hc521a.pdf Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

220

Fact Sheet No. 2.3.2-8

Method Title Purification of Biological Tissue Samples by Gel Permeation Chromatography of

Organic Analyses

Purpose To purify tissue extract samples by separating out lipids and high molecular

weight com ponents from target compounds.

Method Summary The GPC/HPLC is calibrated to verify the instrument performance based on

retention time and area of each of the calibration standards. Sample extracts are

processed through a guard column and two size exclusion columns connected in

series and the desired fraction is collected with a fraction collector. The collected

fraction is then concentrated and analyzed for polycyclic aromatic hydrocarbons,

pesticides, and polychlorinated biphenyls.

Data Uses/Application This method is a clean-up step used in the processing of organic contaminant

sample extracts for GC/MS or GC/ECD analysis.

Advantages A large amount of neutral lipids and high molecular weight com ponents from

tissue samples can be eluded from an alumina/silica gel column

chromatography clean up step. Size exclusion chromatography can separate

the target analytes from these other components. Upon calibration, this method

is also suitable for the isolation of other classes or organic contaminants.

Limitations This method isolates organic contaminants from lipid matrix but does not

separate or isolate individual fractions of organic contaminants.

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and

Trends Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo

NOS ORCA 130. National Oceanic and Atm ospheric Administration, Silver

Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications/t

m130.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

221

Fact Sheet No. 2.3.2-9

Method Title Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS) - Selected Ion Monitoring (SIM)

Mode

Purpose To determine low concentrations of polycyclic aromatic hydrocarbons (PAHs)

and their alkylated homologues in extracts of water, sediments and biological

tissues.

Method Summary Just prior to analysis, an aliquot of internal standard solution is added to the

sample extract producing a final internal standard concentration of

approximately 40 ng/mL. The analytical system includes a temperature

programmable gas chromatography with a fused silica capillary column.

Helium is used as the carrier gas, and the samples are handled by an auto

sampler capable of making 1 - 4 μl injections. A five point calibration curve is

established to dem onstrate the linear range of the detector. The effluent from

the GC capillary column is routed directly into the ion source of the mass

spectrometer (MS). The MS is operated in the SIM mode using appropriate

windows to include the quantization and confirmation masses for target PAHs.

For all compounds detected at a concentration above the MDL, a confirmation

ion is checked to confirm its presence. The response factors of the surrogate

relative to each of the calibration standards are calculated, followed by the

calculation of the sample extract concentration. The sample concentration for

each compound is calculated by dividing the sample extract concentration by

the sample amount.

Data Uses/Application PAH data obtained from this analysis are used for site characterization and site

assessment.

Advantages GC/MS in the SIM mode provides unambiguous and sensitive detection for

PAHs. The PAH quantization method is very rigorous because PAHs have

very strong molecular ion peaks under the mass spectrometric conditions

used. Also, the availability of labeled surrogates internal standards of many of

the analytes makes very accurate determinations of analyte concentrations

possible. Analysis of alkylated PAH homologues can provide site-specific

information that can be used in source identification or product identification.

Limitations GC/MS in SIM mode cannot be used for simultaneous screening for other

organic contaminants of similar polarity or volatility and cannot be used to

identify tentatively identified compounds (TICs).

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and

Trends Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo

NOS ORCA 130. National Oceanic and Atm ospheric Administration, Silver

Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications/t

m130.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

222

Fact Sheet No. 2.3.2-10

Method Title Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

Purpose To quantify chlorinated hydrocarbons (i.e., chlorinated pesticides and PCBs) in

sample extracts.

Method Summary This method is based on high resolution, capillary gas chromatography using

electron capture detection (GC/ECD). Extracts normally have a holding time of

40 days. The instrument’s detector is calibrated before the sample in injected.

Pesticide/PCB calibration is done also as part of the analytical run. If the

response for any peak exceeds the highest calibration solution, the extract is

diluted, a known amount of surrogate and TCMX solution added, and the sample

reanalyzed for those analytes that exceeded the calibration range.

Concentrations in the samples are calculated based on the internal standard

method. Data is reported as ng/g dry weight.

Other methods describing the analysis of PCBs and pesticides by GC/ECD are

NS&T methods, ASTM Methods D5317 and D3534, and SW846 Methods 8081A

and 8082 (NOAA, 1998; ASTM, 2001c).

Data Uses/Application Data are used in site characterization and in risk analysis.

Advantages The ECD is very sensitive and allows for detection of the chlorinated

hydrocarbons at trace concentrations (ppb).

Limitations The detector does not have a linear response over a wide concentration

range and must be used by sufficiently trained personnel. Second column

analysis must be performed to provide unequivocal compound identification.

These methods do not measure the 12 World Health Organization congeners,

which may be desired data in some risk assessments.

Reference USEPA. 1997d. Method LMMB 041: Analysis of Polychlorinated Biphenyls and

Chlorinated Pesticides by Gas Chromatography with Electron Capture Detection,

Lake Michigan Mass Balance Study Methods Compendium, Volume 2: Organic

and Mercury Sample Analysis Techniques, EPA 905-R-97-012c. Great Lakes

National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

Website http://www.epa.gov/glnpo/lmmb/methods/sop501.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

223

Fact Sheet No. 2.3.2-11

Method Title Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA Method 1613

Purpose This method is for determination of tetra- through octa-chlorinated dibenzo-pdioxins (CDDs) and dibenzo furans (CDFs) in tissue.

Method Summary This method is "performance-based.” The sample is extracted by one of two

procedures:

1.Soxhlet or SDS extraction—A 20 g aliquot of sample is homogenized, and

a 10 g aliquot is spiked with the labeled compounds. The sample is mixed with

sodium sulfate, allowed to dry for 12-24 hours, and extracted for 18-24 hours

using methylene chloride:hexane (1:1) in a Soxhlet extractor. The

extract is evaporated to dryness, and the lipid content is determined.

2.HCl digestion—A 20 g aliquot is homogenized, and a 10 g aliquot is placed in

a bottle and spiked with the labeled compounds. After equilibration, 200 mL of

hydrochloric acid and 200 mL of methylene chloride:hexane (1:1) are added,

and the bottle is agitated for 12-24 hours. The extract is evaporated to dryness,

and the lipid content is determined. After extraction, 37Cl4-labeled 2,3,7,8-TCDD

is added to each extract to measure the efficiency of the cleanup process.

Sample cleanups may include back-extraction with acid and/or base, and gel

permeation, alumina, silica gel, Florisil and activated carbon chromatography.

High-performance liquid chromatography (HPLC) can be used for further

isolation of the 2,3,7,8-isomers or other specific isomers or congeners. After

cleanup, the extract is concentrated to near dryness. Immediately prior to

injection, internal standards are added to each extract, and an aliquot of the

extract is injected into the instrument. The analytes are separated by the GC

and detected by a high-resolution ($10,000) mass spectrometer.

CDD/CDF ML (pg/μL) CDD/CDF ML(pg/μL)

2,3,7,8-TCDF 0.5 1,2,3,4,7,8-HxCDD 2.5

2,3,7,8-TCDD 0.5 1,2,3,6,7,8-HxCDD 2.5

1,2,3,7,8-PeCDF 2.5 1,2,3,7,8,9-HxCDD 2.5

2,3,4,7,8-PeCDF 2.5 1,2,3,4,6,7,8-HpCDF 2.5

1,2,3,7,8-PeCDD 2.5 1,2,3,4,7,8,9-HpCDF 2.5

1,2,3,4,7,8-HxCDF 2.5 1,2,3,4,6,7,8-HpCDD 2.5

1,2,3,6,7,8-HxCDF 2.5 OCDF 5.0

1,2,3,7,8,9-HxCDF 2.5 OCDD 5.0

2,3,4,6,7,8-HxCDF 2.5

This method is also described in SW846 Method 8290

Data Uses/Application The method is for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery

Act, the Com prehensive Environmental Response, Compensation and Liability

Act, and the Safe Drinking Water Act.

Advantages Method 1613 is able to meet detection limits required for human health and

ecological risk assessments.

Limitations The GC/MS portions of this method are for use only by analysts experienced

with HRGC/HRMS or under the close supervision of such qualified persons.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

224

Fact Sheet No. 2.3.2-11 (contd.)

Reference USEPA. 1994c. Method 1613: Tetra- through Octa-Chlorinated Dioxins and Furans

by Isotope Dilution HRGC/HRMS, EPA 821-B-94-005. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/waterscience/methods/16

13.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

225

Fact Sheet No. 2.3.2-12

Method Title Toxic Polychlorinated Biphenyls by Isotope Dilution High Resolution

Gas Chromatography/High Resolution Mass Spectrometry, EPA Method 1668

Purpose This method is for determination of the toxic polychlorinated biphenyls (PCBs) in

solids (not tissue).

Method Summary This method is performance-based. A 20-g aliquot of sample is homogenized,

and a 10-g aliquot is spiked with the labeled compounds. The sample is mixed

with sodium sulfate, allowed to dry for 12- 24 hours, and extracted for 18-24

hours using methylene chloride: n-hexane (1:1) in a Soxhlet extractor. The

extract is evaporated to dryness, and the lipid content is determined. After

extraction, samples are cleaned up using back-extraction with sulfuric acid

and/or base, and gel permeation, silica gel, Florisil and activated carbon

chromatography. High-performance liquid chromatography (HPLC) can be used

for further isolation of specific isomers or congeners. After cleanup, the extract

is concentrated to near dryness. Immediately prior to injection, internal

standards are added to each extract, and an aliquot of the extract is injected

into the gas chromatography. The analytes are separated by the GC and

detected by a high-resolution ($10,000) mass spectrometer.

Extract

IUPAC EMDL (ng/kg) EML (ng/kg) EML

(pg/μL)

77 0.5 2 1

123 4 10 5

126 10 4 5

118/167/156/157/169/180/170/189 6 20 10

114 60 200 100

105 40 100 50

EMD: = Estimated Method Detection Limit; EML = Estimated Minimum Level

Data Uses/Application The method is for use in EPA's data gathering and monitoring programs

associated with the Clean Water Act, the Resource Conservation and Recovery

Act, the Com prehensive Environmental Response, Compensation and Liability

Act, and the Safe Drinking Water Act.

Advantages Method 1668 provides data for most, but not all, of the “dioxin-like” PCBs,

including those with the highest TEFs, as determined by the World Health

Organization. This method provides detection limits frequently required in risk

assessments.

Limitations The GC/MS portions of this method are for use only by analysts experienced

with HRGC/HRMS or under the close supervision of such qualified persons.

Method 1668 does not provide data for all of the “dioxin-like” PCBs, as does

Method 1668A.

Reference USEPA. 1997e Method 1668: Toxic Polychlorinated Biphenyls by Isotope

Dilution HRGC/HRMS, EPA-821-R-97-001. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/clariton/clhtml/pubtitl

eOW.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

226

Fact Sheet No. 2.3.2-13

Method Title Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and Tissue by

HRGC/HRMS, EPA Method 1668 Revision A

Purpose This method is for congener-specific determination of more than 150

chlorinated biphenyl (CB) congeners in solids (not tissue).

Method Summary This method is performance-based. A 20-g aliquot of sample is homogenized,

and a 10-g aliquot is spiked with the labeled compounds. The sample is mixed

with anhydrous sodium sulfate, allowed to dry for 12 - 24 hours, and extracted

for 18- 24 hours using methylene chloride:hexane (1:1) in a Soxhlet extractor.

The extract is evaporated to dryness, and the lipid content is determined. After

extraction, a labeled cleanup standard is spiked into the extract which is then

cleaned up using back-extraction with sulfuric acid and/or base, and gel

permeation, silica gel, or Florisil chromatography. Activated carbon and highperformance liquid chromatography (HPLC) can be used for further isolation of

specific congener groups. Prior to the cleanup procedures cited above, tissue

extracts are cleaned up using an anthropogenic isolation column. After

cleanup, the extract is concentrated to 20 μL. Immediately prior to injection,

labeled injection internal standards are added to each extract and an aliquot of

the extract is injected into the gas chromatography (GC). The analytes are

separated by the GC and detected by a high-resolution ($10,000) mass

spectrometer. Without interferences, EMDLs and EMLs will be, respectively,

0.5 and 1.0 ng/kg for soil, tissue, and mixed-phase samples, and EMLs for

extracts will be 0.5 pg/uL. EMD: = Estimated Method Detection Limit; EML =

Estimated Minimum Level

Data Uses/Application This method is for use in data gathering and monitoring associated with the

Clean Water Act, the Resource Conservation and Recovery Act, the

Com prehensive Environmental Response, Compensation and Liability Act,

and the Safe Drinking Water Act.

Advantages Method 1668A provides congener data that can be used for source

identification. Listed PCBs include the 12 World Health Organization “dioxinlike” PCBs. The HRMS method provides lower EMDLs compared to ECD or

low resolution MS analyses and provides unequivocal congener identification.

Limitations The GC/MS portions of this method are for use only by analysts experienced

with HRGC/HRMS or under the close supervision of such qualified persons.

Solvents, reagents, glassware, and other sample processing hardware may

yield artifacts, elevated baselines, and/or lock- mass suppression causing

misinterpretation of chromatograms. The natural lipid content of tissue can

interfere in the analysis of tissue samples for the CBs.

Reference USEPA. 1999c. Method 1668, Revision A: Chlorinated Biphenyl Congeners in

Water, Soil, Sediment, and Tissue by HRGC/HRMS, EPA-821-R-00-002.

Office of Water, U.S. Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/region08/water/waste

water/biohome/biosolidsdown/methods/1

668a5.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

227

Fact Sheet No. 2.3.2-14

Method Title Determination of Percent Dry Weight for Tissues

Purpose To determine the percentages of dry weight and moisture in tissue samples.

Method Summary A 0.5-1 g aliquot of homogenized sample is placed into a pre-weighed beaker

and weighed. The samples are dried for 24 hours in a drying oven set at 63-

65°C. Samples are placed in a desiccator and allowed to cool to room

temperature for at least 30 minutes. The samples are weighed. The samples

are put back in the oven for at least 2 hr after which they are removed from the

oven and allowed to cool for at least 30 min in a desiccator. The sample is

reweighed. If the difference between the first and second weighting is less than

± 0.02 g, the dry weight percent is calculated based on the last weighing. The

difference between the weight of the dried sample and that of the wet sample is

used to calculate the percent dry weight.

 [Vial wt. + Dry sample wt.] - [Vial wt.]

Percent dry weight = [Vial wt. + W et sample wt.] - [Vial wt.] X 100

Data Uses/Application Some exposure assessment models require concentration data on dry weight

basis.

Advantages National database of fish and contaminant data reported on dry weight basis

following this procedure.

Limitations None.

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and

Trends Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo

NOS ORCA 130. National Oceanic and Atm ospheric Administration, Silver

Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications/t

m130.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

228

Fact Sheet No. 2.3.2-15

Method Title Determination of Percent Lipid in Tissue

Purpose To determine the percent lipid (weight/weight basis) of a tissue.

Method Summary An appropriate amount of sodium sulfate-dried tissue sample is extracted three

times with dichloromethane (100 mL each time). An aliquot of 20 mL of the

extract is quantitatively removed for lipid determination. This aliquot is filtered,

further dried with sodium sulfate, and brought to a final volume of 1 mL in

dichloromethane. An aliquot of 100 μL was taken and evaporated to constant

weight. The residual weight of this dried 100 μL portion is used to calculate the

percent lipids of the sample based on the dry weight.

Data Uses/Application Lipid content has been found to be correlated to contaminant concentrations

for specific tissues and whole organisms.

Advantages National database of fish and bivalve contaminant data reported on lipid basis

following this procedure.

Limitations The Bligh and Dyer (1959) Method, using a different solvent system, provides

slightly different lipid values.

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and

Trends Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo

NOS ORCA 130. National Oceanic and Atm ospheric Administration, Silver

Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications/tm

130.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

229

Fact Sheet No. 2.3.2-16

Method Title Microwave Extraction of Marine Tissue for Semivolatile Organic Analytes, AED

LOP 2.03.030, Revision 0

Purpose Microwave-assisted extraction of semi-volatile organic compounds from marine

tissue samples.

Method Summary Homogenize the entire tissue sample using a tissue homogenizer. Determine dry

weight to wet weight ratio. Weigh approximately 1.0 g of homogenized sample into

preweighed aluminum pan. Place pan in drying oven and record weight at 24 and

48 hours. Using the dry/wet ratio, back-calculate the wet weight needed for each

sample, setting the dry weight constant between 0.8 and 1.0 g target dry weight.

Use the sample with the lowest dry/wet ratio (highest percent moisture) and back

calculate the wet weight for that sample (see A). Since the moisture content is not

the same for all the samples, the wet weight will also be different (see B). Adjust

the wet weight of all samples to be equivalent to the standardization samples by

adding hexane rinsed DI water (see C).

A.: Target dry weight/(dry/wet ratio sample A) = grams wet sample A

B: Target dry weight/(dry/wet ratio sample B) = grams wet sample B

C: Grams wet sample A - Grams wet sample B = grams H20 added to

 sample B.

Assemble and prepare extraction vessels according to operation manual (AED

uses a CEM MES-1000 microwave extraction system). Weigh samples directly

into the bottom of the liners. Standardize the wet weight for all samples by adding

hexane rinsed DI water. Add internal standards (IS) as required. For samples < 1

g, grind sample with 5 g of sodium sulfate and transfer to extraction vessel. Add 30

mL of 20/80 hexane/acetone solvent mixture, stir gently with a Teflon spatula, and

insert the liner into a clean, dry, particle-free vessel body. Program the microwave

at 70% power, 200 psi, 30 minutes runtime; 15 minutes at pressure, and 115B C.

After the extraction, pour the top solvent layer from the extraction vessel into a presolvent rinsed 250 mL separatory funnel containing 80 mL of hexane rinsed DI

water. Back extract the DI/acetone; hexane phase in the separatory funnel 3X with

hexane, using 10mL hexane for the first extraction and 5 mL each for the second

and third extractions. Combine the extracts and treat with sodium sulfate to

remove water.

Transfer the extract into a clean rinsed 200 mL Turbo-Vap® tube. Place the flask

into the Turbo-Vap® apparatus and turn on the unit. Adjust the associated

nitrogen pressure regulator to read approximately 5 psi. Reduce the sample

volume to approximately 1 mL. Adjust the final volume to 1.0 mL with hexane.

Rem ove 0.1 mL into a preweighed aluminum pan for lipid weight determination.

Allow it to dry at room temperature for a minimum of 24 hours. Record the weight

of the pan plus sample.

Fractionate the sample using column chromatography with silicic acid.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

230

Fact Sheet No. 2.3.2-16 (contd.)

Data

Uses/Application

Extracting semivolatile organic compounds from the tissue of aquatic fauna.

Extracts can be further processed by separation on silicic acid chromatography

procedures prior to analysis by gas chromatography and/or

gas/chromatography/mass selective detector.

Advantages More time efficient and requires less solvent than other methods of semivolatile

organic compound extraction from tissue, such as sonication or maceration in

solvent.

Limitations This procedure was written to meet the specific needs of the research program at

the U.S. EPS-Atlantic Ecology Division. It is not a U.S. EPA Standard Method and

must not be referred to as such. Mention of trade names or comm ercial products

does not constitute endorsement or recommendation for use.

Reference Unpublished laboratory SOP, EPA NHEERL-AED, Narragansett RI

Website N/A Last Accessed:
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

231

2.3.3 Biological Analysis Methods

Section 2.3.3 provides a compendium of biota-related biological analyses. Biota data like these

are often used to measure the status and trends of environmental pollution on freshwater,

estuarine, and marine macroinvertebrates to assess water quality criteria and monitor surface

water quality. Thus, many of these methods pertain to the analysis of samples collected in the field

to determine species abundance and taxa richness. Biological impairments resulting from pollution

are often evaluated using indices derived from the sampling data that evaluate matrices such as

community, population and functionality parameters. Other fact sheets pertaining to

histopathological analyses are often included. 

The sources of information for the biological analyses fact sheets come from the following

agencies and offices:

• The USEPA’s EMAP Program

• The USEPA’s Environmental Research Laboratory-Narragansett,

• The USEPA’s Great Lakes Program Office

• NOAA’s Status and Trends Program

• The USEPA’s Office of Water
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

232

Fact Sheet No. 2.3.3-1

Method Title Laboratory Identification, Enumeration and Biomass Measurements of Periphyton in

Wadeable Streams

Purpose To determine periphyton species composition and/or biomass in the laboratory

Method Summary The standard laboratory-based method provides the option of sampling natural

substrates in a few different ways. Regardless of the sampling strategy, the samples

are returned to the laboratory where they are homogenized, sorted, identified and

counted in order to derive relative abundance and taxa richness data. “Soft” (nondiatom) samples are homogenized with a tissue homogenizer or a blender. These

thoroughly mixed samples are placed in Palmer counting cells. Approximately 300

algal “cell units” are counted and identified to the lowest possible taxonomic level at

400x magnification. Relative abundances of soft algae are determined by dividing the

number of cells counted for each taxon by the total number of cells counted. 

Diatom samples are subsampled and oxidized. The diatoms are then mounted on a

high refractive index medium to make permanent slides. Diatom valves are counted

and identified to the lowest possible taxonomic level, which should be species and

perhaps variety level, under oil immersion at 1000X magnification. At minimum, count

600 valves (300 cells) and at least until 10 valves of 10 species have been observed. 

Relative abundances of diatoms have to be corrected for the number of live diatoms

observed in the count of all algae. To determine the relative abundance of diatom

species in the algae assemblage, divide the number of valves counted for each

species by the total number of valves counted; then multiply the relative abundance of

each diatom taxon in the diatom count by the relative abundance of live diatoms in the

count of all algae. 

USEPA’s EMAP document describes a similar method for enumerating and measuring

periphyton from wadeable streams (USEPA, 1998).

Data

Uses/Application

Species relative abundance and taxa richness are data derived from these protocols.

These data parameters provide information pertaining to the status and trends of

environmental pollution and its impacts on freshwater, marine and estuarine

comm unities. 

Biological impairment resulting from pollution is often evaluated using metrics of biotic

integrity derived from the aforementioned data parameters that evaluate community,

population and functional parameters. Examples of metrics based on species

composition include species richness, total number of genera, total number of

divisions, shannon diversity (for diatoms), percent com munity similarity of diatoms,

pollution tolerance index for diatoms, and percent sensitive diatoms. Furthermore,

other metrices infer ecological conditions based on documented preferences. These

metrices include the percent aberrant diatoms, percent motile diatoms, simple

diagnostic metrics, inferred ecological conditions with simple autecological indices

(SAI), inferred ecological conditions with weighted average indices, and impairment of

ecological conditions. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

233

Fact Sheet No. 2.3.3-1 (contd.)

Advantages The laboratory-based survey is more accurate in assessing biotic integrity and in

diagnosing causes of impairment than the field-based survey.

Limitations The laboratory-based methods require more time and effort than the field evaluation.

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition,

EPA 841-B-99-002. Office of Water, U.S. Environmental Protection Agency,

Washington, D.C.

Website http://www.epa.gov/owowwtr1/monitoring/rbp/inde

x.htm l

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

234

Fact Sheet No. 2.3.3-2

Method Title Laboratory Periphyton Biomass Determination

Purpose To determine periphyton biomass in the laboratory.

Method Summary To quantify algal biomass, the area of the substrate sampled must be

determined. Periphyton biomass can be estimated with chlorophyll a, ash-free

dry mass, cell densities, and biovolume, usually per cm2. Each of these

measures estimates a different component of periphyton biomass. 

Chlorophyll a: Extract in acetone and measure chl concentration in the extract

with a spectrophotometer or fluorometer. Calculate the chlorophyll a density on

substrates by determining the proportion of original sample that was assessed

for chlorophyll a. 

Ash-Free Dry Mass: A measurement of the organic matter in samples. At

detailed description of the process is beyond the scope of this fact sheet, but

standard methods are readily available (APHA, 1999, USEPA 1995). It is a

fairly simple analysis. It is recommended over dry mass measurem ents

because silt can account for a substantial proportion of dry mass in some

samples. 

Area-Specific Cell Densities and Biovolumes: Cell densities are determined by

dividing the numbers of cells counted by the proportion of sample counted and

the area from which the samples were collected. Cell biovolumes are

determined by summ ing the products of cell density and biovolume of each

species counted and dividing that sum by the proportion of sample counted and

the area from which the samples were collected. 

USEPA EMAP provides similar guidance for laboratory periphyton biomass

determination (USEPA, 1998).

Data Uses/Application Biomass may be especially important in studies that address nutrient

enrichment or toxicity. 

Advantages Periphyton biomass provides information on standing crop, which is useful for

assessing the biological integrity of streams.

Limitations In many cases, sampling benthic algae misses peak biomass, which may best

indicate nutrient problems and potential for nuisance algae growths. 

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second

Edition. EPA 841-B-99-002. USEPA. Office of Water. Washington DC. 

Website http://www.epa.gov/owowwtr1/monitoring/rbp/ind

ex.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

235

Fact Sheet No. 2.3.3-3

Method Title Laboratory Analysis of Benthic Macroinvertebrates in Wadeable Streams

Purpose To determine macroinvertebrate diversity and abundance.

Method Summary Following sample collection, the sediment is sieved and preserved in a 10%

buffered formalin solution, however different mixtures should be used for softbodied organisms (i.e., leeches, aquatic oligochaete, and other soft-bodied

organisms. 

Samples are sorted by hand in the laboratory using a low power (2x) scanning

lens or a stereomicroscope. Approximately one or two tablespoonfuls of the

sample are placed in a white enamel pan filled about one-third full of water.

Ethanol-preserved organisms should float to the top and be removed from the

dish as a sub-sampling procedure. Various staining methods may also be used

for sub-sampling. 

Microscope slide mounts are then prepared for all or parts of organisms for

identification purposes. These slides are then identified to a specific taxonomic

level. For water quality and pollution analyses, it is important that organisms are

identified to the species level. As organisms are identified, the individuals in

each taxonomic category are counted and the numbers are recorded in

laboratory bench sheets. 

Subsampling of benthic samples is not a requirement and is often discouraged

by certain scientists. However, Rapid Bioassessment Protocols recommend a

fixed-count approach to subsampling and sorting the organisms based on the

sample matrix of detritus, sand and mud. This approach calls for sieving

samples as described above and then removing all material in four randomly

selected grids contained the sieved sample. The organisms in these four grids

are enumerated and identified to the lowest possible taxonomic level. 

Several USEPA EMAP documents describe similar protocols to determine

macroinvertebrate diversity and abundance (USEPA, 1990b; USEPA, 2000b;

USEPA, 1995).

Data Uses/Application Macroinvertebrate data such as these are used to measure the status and

trends of environmental pollution. Biological impairment resulting from pollution

is often evaluated using indices derived from the sampling data that evaluate

matrices such as community, population and functional parameters. 

Advantages Examples of indices often derived from the data include the Hilsenhoff’s Family

Biotic Index (HBI), Invertebrate Community Index (ICI), Community Similarity

Indices, Community Loss Index, and the Ohio EPA Invertebrate Community

Index.

Limitations Laboratory analysis of benthic macroinvertebrate samples is costly and requires

a high degree of taxonomic expertise.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

236

Fact Sheet No. 2.3.3-3 (contd.)

Reference Barbour et al. 1999. Rapid Bioassessment Protocols for Use in Streams and

Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second

Edition, EPA 841-B-99-002. Office of Water, U.S. Environmental Protection

Agency, Washington, D.C.

Website http://www.epa.gov/owowwtr1/monitoring/rbp/ind

ex.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

237

Fact Sheet No. 2.3.3-4

Method Title Laboratory Analysis of Water Column Organisms

Purpose To determine composition and abundance from drift net or stream-net samplers.

Method Summary The organism’s collected by drift nets or stream-net samplers are emptied directly

into a white enamel pan or sm all bucket. The organisms can then be hand-picked

into a sample container and filled three-fourths full of preservative (70-80% ethyl

alcohol), however different mixtures should be used for soft-bodied organisms

(i.e., leeches, aquatic oligochaete, and other soft-bodied organisms). Sample

containers should be large enough so that they are not over one-half full of the

washed sample before the preservative is added. 

Samples should be sorted by hand in the laboratory using a low power (2x)

scanning lens or a stereomicroscope). Approximately one or two tablespoonfuls

of the sample will be placed in a white enamel pan filled about one-third full of

water. Ethanol-preserved organisms should float to the top and be removed from

the dish as a sub-sampling procedure. Various staining methods have also been

used for sub-sampling. 

Microscope slide mounts are then prepared for all or parts of organisms for

identification purposes. These slides are then identified to a specific taxonomic

level depending on the needs, experience and available resources. For water

quality and pollution analyses, it is important that organisms are identified to the

species level. As organisms are identified, the individuals in each taxonomic

category are counted and the numbers are recorded in laboratory bench sheets. 

Data

Uses/Application

Macroinvertebrate data such as these are used to measure the status and trends

of environmental pollution on freshwater, estuarine, and marine

macroinvertebrates, to assess water quality criteria, and monitor surface water

quality. Biological impairment resulting from pollution is often evaluated using

indices derived from the sampling data that evaluate matrices such as

community, population and functional parameters. Examples of indices often

applied include the Hilsenhoff Biotic Index (HBI), Invertebrate Community Index

(ICI), Community Similarity Indices, Community Loss Index, and the Ohio EPA

Invertebrate Community Index . 

Advantages Macroinvertebrate biomass (weight of organisms per unit area) is a useful

measure of standing crop which is useful in assessing the biological integrity of

surface waters. The results of this analysis feed into the commonly accepted

incdices used to evaluate com munity health.

Limitations Taxonomic analysis requires a team of highly experienced technicians and/or

scientists. 

Reference USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating

the Biological Integrity of Surface Waters, EPA/600/4-90/030. Office of Research

and Development, Washington, D.C. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

238

Fact Sheet No. 2.3.3-4 (contd.)

Website http://www.epa.gov/clariton/clhtml/pubtitleOR

D.html

Last Accessed: 2/13/03
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

239

Fact Sheet No. 2.3.3-5

Method Title SOP-2: Lab Analysis of Lake Trout Stomachs and Data Entry; Appendix B.

Standard Operating Procedure for Lab Analysis of Coho Salmon Stomachs and

Data Entry, LMMB 026 - Appendix 2 & LMMB 027 - Appendix B

Purpose To examine and quantify the contents of lake trout and coho salmon stomachs.

Method Summary Lake trout stomachs: Prey fish in the stomachs are identified, measured (nearest

mm) and weighed (nearest 0.1 kg). The percent digested state is recorded.

Measures of length include: maximum total length, standard length, vertebral

column length, and length of a multiple of 5 vertebrae. Fish or parts of fish that

cannot be positively identified are recorded as unidentified remains.

Invertebrates are identified, grouped by taxa, and weighed as a taxon group.

The number of individuals in each group is enumerated. Stomach contents are

repackaged and frozen. Using the weight and length of intact prey, conversion

equations are developed to reconstruct total prey length and weight from partial

length measures.

Coho salmon stomachs: The stomach is rinsed with rinse water to remove

excess formalin. Prey fish in the stomachs are identified, measured (nearest

mm) and weighed (nearest 0.1 g (large items and 0.02 g for sm all items). The

percent digested state is recorded. Measures of length include: maximum total

length, standard length, vertebral column length, and length of as many

vertebrae as possible. Invertebrates are identified, grouped by taxa, and

weighed as a taxon group. The number of individuals in each group is

enumerated. The average length and digested state of each taxon group is

recorded. If identification of a prey item is uncertain, the item is examined by a

second identifier and compared to a reference collection of diet items. During

the analysis, examples of each species of prey fish and taxonomic group of

invertebrate is set aside and preserved in 5% formalin. Stomach contents are

repackaged and preserved. Using the weight and length of intact prey,

conversion equations are developed to reconstruct total prey length and weight

from partial length measures.

Standard Method 10600D.2 also discusses diet analysis of fish (APHA, 1999).

Data Uses/Application Stomach content data of upper trophic level fish are important to understand

exposure pathways.

Advantages Standardized method enhances uniformity of data.

Limitations N/A

Reference USEPA. 1997b. Methods LMMB 026 - Appendix 2 & LMMB 027 - Appendix B.

Lake Michigan Mass Balance Study Methods Compendium, Volume 1: Sample

Collection Techniques, EPA 905-R-97-012c. Great Lakes National Program

Office, U.S. Environmental Protection Agency, Chicago, IL

Website

http://www.epa.gov/glnpo/lmmb/methods/qappf

ish.pdf 

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

240

Fact Sheet No. 2.3.3-6

Method Title Gonadal Analysis

Purpose A method to determine the reproductive stage of oysters, mussels, and zebra

mussels.

Method Summary A semi-quantitative histological approach is used to rank reproductive stage. For

oysters and mytilid mussels, a dorsal-ventral slice of tissue is taken and fixed in

Davidson’s fixative (48 hours for oysters and 1 week for mussels). Zebra

mussels are fixed whole in Davidson’s fixative for one week, decalcified with

acetic acid, and embedded whole. Tissue samples are embedded in paraffin,

sectioned at a 5-μm thickness, and stained using a pentachrome staining

protocol. Unstained sections may be used for histopathological analysis.

Stained sections are examined under a compound microscope. Sex and state of

gonadal development are determined. The stage of the gametogenic cycle is

assigned a numerical value. For mytilids and zebra mussels, a mean gonadal

index, ranging from 0 to 5, can be calculated by summ ing the individual stage

numbers. For oysters, the number of individuals with substantial gonadal

development are compared to those having little gonadal volume using an

egg/eggless ratio.

Data Uses/Application This method helps to assess the physiological state of bivalve populations.

Analysis of reproductive stage is important in identifying differences in tissue

composition which might affect between site and interannual comparisons of

contaminant data.

Advantages The pentachrome staining procedure yields better differentiation of tissue types

and mucins. This analysis provides an assessment of sexual stage in the

gametogenic cycle, and if desired, allows for a concomitant histopathological

analysis, with a single sample preparation protocol.

Limitations The procedure cannot be performed on pooled samples, as is commonly done

for chemical parameters, but only on individuals.

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and

Trends Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo

NOS ORCA 130. National Oceanic and Atm ospheric Administration, Silver

Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/publications/t

m130.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

241

Fact Sheet No. 2.3.3-7

Method Title Histopathological Evaluations of Target and Non Target Fish Species

Purpose To determine fish quality in Estuaries

Method Summary Specimens for histopathology analysis are unpacked, logged in, and placed in

70% ethyl alcohol for at least 48 hours prior to examination. A careful visual

inspection is made of the fins and body surfaces. Any abnormalities are noted.

Thorough examinations of the eyes, branchial chambers, buccal cavity, visceral

organs is performed. Representative tissue samples are removed from either

fish or shellfish (USEPA and the Naval Construction Battalion Center, 1992) and

slides are prepared. These slides are then examined using a compound

research microscope to diagnose pathological conditions.

Data Uses/Application Histopathological evaluations provide data that can be used as a com posite

index of the incidence of diseases and contaminant body burdens in selected

resident species. Microscopic examination can determine the presence or

absence of pathological changes and evaluate the health of the animal or its

exposure to contaminated material or infectious agents. Changes include

morphological alterations, variations in the normal staining characteristics, or a

change in the rate of occurrence of features (i.e., mitotic figures) 

Advantages Histopathological investigations provide information on the relationship between

incidence of external abnormalities and internal histopathological abnormalities. 

Limitations It is difficult to prove in a legal context that both external and internal

abnormalities are indicators of degraded environmental systems.

Reference USEPA. 1995. Environmental Monitoring and Assessment Program (EMAP),

Laboratory Methods Manual, Estuaries, Volume 1-Biological and Chemical

Analyses, EPA/620/R-95/008. Office of Research and Development, U.S.

Environmental Protection Agency, Washington, DC.

Website http://www.epa.gov/emap/html/pubs/docs/group

docs/estuary/field/lab_man.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

242

Fact Sheet No. 2.3.3-8

Method Title Histopathology Analysis

Purpose A quantitative or semi-quantitative method to determine the prevalence and

density of parasites, pathologies, and diseases afflicting oysters, mussels, and

zebra mussels.

Method Summary Analyses are conducted on paraffin-embedded tissues sectioned at a 5-μm

thickness and stained using a pentachrome staining procedure. Prepared slides

are examined individually under the microscope using a 10X ocular and a 10X

objective. Conditions evaluated are scored for intensity using either a

quantitative or semi-quantitative scale. Conditions scored quantitatively include

parasites, the number of ceroid bodies, incidences of tissue inflammation,

rickettsial bodies, incidences of tissue edema, and suspected neoplasms and

tumors. A running count of incidences of the condition is kept as the slide is

scanned, to avoid re-examining each slide multiple times for each separate

malady. Evaluation of conditions scored semi-quantitatively related to the

intensity of the effect or the extensiveness of pathologies affecting large tissue

areas. Semi-quantitative measurement may require re-scanning portions of the

tissue for each malady type to completely assess the degree of tissue

involvement. Infection intensity of parasites, the occurrence and extensiveness

of tissue pathologies, and the intensity of diseases are recorded using semiquantitative or quantitative measures.

Data Uses/Application Histopathology is used to help assess the influence of contaminant exposure on

population health.

Advantages Infection intensity quantified by counts or semi-quantitative methods consistently

provides a more robust data set for statistical analysis.

Limitations Prevalence rarely provides an adequate description of the population dynamics

of disease and often yields ambiguous results.

Reference NOAA. 1998. Sampling and Analytical Methods of the National Status and

Trends Program, Mussel Watch Project: 1996 Update, NOAA Technical Memo

NOS ORCA 130. National Oceanic and Atm ospheric Administration, Silver

Spring, MD. 233 pp.

Website http://ccma.nos.noaa.gov/puslications/tm

130.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

243

Fact Sheet No. 2.3.3-9

Method Title Index of Biotic Integrity (IBI)

Purpose The IBI is used to identify and assess degraded and undegraded streams.

Method Summary The steps in developing IBIs are the same for both fish and benthic

macroinvertebrates. Criteria for both reference and degraded sites were

determined based on water chemistry, physical habitat, and land use.

Ecologically-relevant geographic strata were determined using cluster analysis

and nonmetric multidimensional scaling. Candidate metrics were evaluated for

1) their ability to discriminate (based on classification efficiency) between

reference and degraded sites, and 2) for redundancy. The final suite of metrics

used in the IBIs contained those ecologically significant metrics with the best

classification efficiency. Both IBIs were validated using an independent data set

and overall classification efficiencies were calculated.

An example of the metrices developed for the Fish Index of Biotic Integrity are

provided below:

• Number of native species

• Number of benthic species

• % tolerant individuals

• % abundance of dominant species

• % generalists, omnivores and insectivores

• Number of individuals/square meter

• Biomass (grams/square meter) (used for coastal plain streams only)

• % lithophilic spawners

• % insectivores (used for non-coastal plain streams only)

Data Uses/Application IBIs are used to determine biological integrity based on characteristics of the

fish and benthic assemblage at a site. The results of these assessments are

used for watershed managem ent decisions concerning strategies that will

control and minimize point and non-point sources of water pollution. 

Advantages IBIs use multiple attributes to quantitatively assess stream health. It is a

system atic way in which to interpret data for managem ent decisions.

Limitations Different states have developed similar, yet different IBIs. This makes it difficult

to compare the results from one state to another. 

Reference USEPA. 1997f. State of the Streams: 1995-1997 Maryland Biological Stream

Survey Results, Mid-Atlantic Integrated Assessment. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C.

Website http://www.dnr.state.md.us/streams/pubs/ea-99-

6.pdf

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

244

Fact Sheet No. 2.3.3-10

Method Title Fish Bioassessment I and II

Purpose These bioassessment strategies use existing and new information in a

system atic manner to determine the health of the local fish community.

Method Summary Fish Bioassessment I uses a questionnaire to serve as a screening tool to pool

the existing knowledge regarding fishery populations and health from the local

fish community. 

The questionnaire polls state fish biologists and university ichthyologists believed

to be knowledgeable about the fish assemblages in stream reaches of concern.

Potential respondents are contacted initially by telephone to identify appropriate

respondents. Then the questionnaire is mailed to all respondents for completion

followed by follow-up mailings and telephone contact. 

Questionnaire responses should provide information pertaining to the integrity of

the fish community, the frequency of limiting factors and causes, the frequency

and occurrence of particular fish com munity condition characterizations, the

geographic patterns of these variables, the temporal trends in the variables, the

effects of water body type and size on the spatial and temporal trends, the

likelihood of improvement and degradation and the major limiting factors. The

data are then analyzed and results are reported as histograms, pie graphs, or

box plots. 

Based on the results of the Bioassessment I survey, Bioassessment II pursues

standardized field collection, species identification and enumeration, and

com munity analyses using biological indices or quantification of the biomass and

numbers of key species. 

Data Uses/Application The questionnaire provides a qualitative assessment of a large number of water

bodies quickly an inexpensively. Its quality depends on the survey design, the

questions presented, and the knowledge and cooperation of the respondents.

The fish Bioassessment II survey yields an objective, discrete measure of the

health of the fish community. Data provided in this survey can be used to

develop biological criteria, prioritize sites for further evaluation, provide a

reproducible impact assessment, and monitor trends in fish com munity status. 

Advantages Questionnaires can provide information that field surveys cannot such as

historical trends and conditions. Field surveys can be oriented towards gathering

data in areas where data were missing historically.

Limitations Questionnaires are sometimes inaccurate due to hasty responses, they often

report conditions better than they are in reality, and the respondents have

insufficient knowledge to answer the questions.

Reference USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological

Integrity of Surface Waters. EPA/600/R-92. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, D.C.

Website http://www.epa.gov/bioindicators/htm l/fish_m eth

ods.html

Last Accessed: 1/31/2003
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

245

3.0 REFERENCES

APHA. 1999. Standard Methods for the Examination of Water and Wastewater, 20th Edition,

L.S. Clesceri and A.D. Eaton, Eds., American Public Health Association, Washington D.C.

ASTM. 2001a. ASTM Book of Standards. Volume 11.01. American Society for Testing and Materials, West

Conshocken, PA.

ASTM. 2001b. ASTM Book of Standards. Volume 11.05. American Society for Testing and Materials, West

Conshocken, PA.

ASTM. 2001c. ASTM Book of Standards. Volume 11.02. American Society for Testing and Materials, West

Conshocken, PA.

ASTM. 2001d. ASTM Book of Standards. Volume 03.06. American Society for Testing and Materials, West

Conshocken, PA.

ASTM. 2001e. ASTM Book of Standards. Volume 04.08. American Society for Testing and Materials, West

Conshocken, PA.

Barbour, M.T., J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment Protocols for Use in

Streams and Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish, Second Edition. EPA 841-

B-99-002. Office of Water, U.S. Environmental Protection Agency; Washington, D.C.

Battelle 2001 Natural Recovery of Persistent Organics in Contaminated Sediments at the SangamoWeston/Twelvemile Creek/Lake Hartwell Superfund Site. USEPA National Risk Management Research

Laboratory Cincinnati, OH.

Bligh, E.G. and W.J. Dyer. 1959. A rapid method for total lipid extraction and purification. Canadian Journal of

Biochemistry and Physiology. 37:911.

Brannon, J.M., R.M. Engler, J.R. Rose, P.G. Hunt, and I. Smith. 1976. Selective Analytical Partitioning of

Sediments to Evaluate Potential Mobility of Chemical Constituents During Dredging and Disposal Operations,

W ES Technical Report D-76-7. U.S. Army engineer Waterways Experiment Station, Vicksburg MS.

Carr, R.S. 2001. Marine and Estuarine Sediment Quality Assessment Studies. Proceedings of the U.S.

Geological Survey (USGS) Sediment Workshop, February 4-7, 1997. Available at

http://water.usgs.gov/osw/techniques/workshop/carr.html. September 27, 2001.

Centre for Environm ent, Fisheries and Aquaculture Science (CEFAS) 2002. Guidelines for the conduct of

benthic studies at aggregate dredging sites. Department for Transport, Local Government and the Regions,

London, U.K.

Custer, C.M., T.W . Custer, P.D. Allen, K.L. Stromborg, and M.J. Melancon. 1998. Reproduction and

environmental contamination in tree swallows nesting in the Fox River drainage in Green Bay, Wisconsin,

USA. Environmental Toxicology and Chemistry. 17:1786-1798.

Elonen, Gregory E., RL Spehar, GW Holcombe, RD Johnson, JD Fernandez, RJ Erickson, JE Tietge, and PM

Cook. 1998. Com parative Toxicity of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin to Seven Freshwater Fish Species

During Early Life-Stage Development. Environmental Toxicology and Chemistry, Vol. 17, No. 3, pp 472-483
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

246

Environment Canada. 1990. Acute Lethality Test Using Rainbow Trout. EPS 1/RM/9 Environmental

Protection Service, Environment Canada, Ottawa, Ontario, Canada.

Environment Canada. 1990. Acute Lethality Test Using Daphnia spp.. EPS 1/RM/11 Environmental

Protection Service, Environment Canada, Ottawa, Ontario, Canada.

Environment Canada. 1992. Test of Reproduction and Survival Using the Cladoceran Ceriodaphnia dubia.

EPS 1/RM/21. Environmental Protection Service, Environment Canada, Ottawa, Ontario, Canada.

Environment Canada. 1992. Test of Larval Growth and Survival Using Fathead Minnows. EPS 1/RM/22

Environmental Protection Service, Environment Canada, Ottawa, Ontario, Canada.

Environment Canada. 1992. Growth Inhibition test Using the Freshwater Alga Selenastrum capricornutum.

EPS 1/RM/25. Environmental Protection Service, Environment Canada, Ottawa, Ontario, Canada.

Environment Canada. 1992. Fertilization Assay Using Echinoids. EPS 1/RM/27. Environmental Protection

Service, Environment Canada, Ottawa, Ontario, Canada.

Environment Canada. 1997. Test for Survival and Growth in Sediment Using the Larvae of freshwater Midges

(Chironomus tentans or Chironomus riparius). EPS 1/RM/32. Environmental Protection Service, Environment

Canada, Ottawa, Ontario, Canada.

Environment Canada. 1997. Test for Survival and Growth in Sediment Using the Freshwater Amphipod

Hyalella azteca . EPS 1/RM/33 Environmental Protection Service, Environment Canada, Ottawa, Ontario,

Canada.

Environment Canada. 1998. Toxicity Tests Using Early Life Stages of Salmonid Fish. EPS 1/RM/28, 2

nd

edition. Environmental Protection Service, Environment Canada, Ottawa, Ontario, Canada.

Environment Canada. 1999. Test for Measuring the inhibition of Growth Using the Freshwater Macrophyte,

Lemna minor. EPS 1/RM/37. Environmental Protection Service, Environment Canada, Ottawa, Ontario,

Canada.

Environment Canada. 2001. Test for Survival and Growth in Sediment Using the Spionid Polychaete Worms

(Polydora cornuta). EPS 1/RM/41. Environmental Protection Service, Environment Canada, Ottawa, Ontario,

Canada.

Fitzgerald, S.A., J Val Klump, PW Swarzenski, RA Mackenzie, and KD Richards. 2001. Beryllium-7 as a

Tracer of Short-Term Sediment Deposition and Resuspension in the Fox River, Wisconsin. Environmental

Science and Technology 35/ 300-305.

Folk, R.L. 1974. Petrology of Sedimentary Rocks. Hemphill Publishing Company, Austin TX.

182 pp.

Ho, K.T, A. Kohn, M. Pelletier, H. Mcgee, R. M. Burgess, and J. Serlost. 2000. Sediment Toxicity

Assessment: Comparison of standard and new test designs. Archives of Environmental Contamination and

Toxicology. 39, 462-468.

Ho, K.T., R. A. Kinney, A. Kuhn, M.C. Pelletier and R. M Burgess. 1997. Identification of Acute Toxicants in

New Bedford Harbor Sediments. Environmental Toxicology and Chemistry. 16, 551-558.

Hunt, C.D., D. Redford, H. White, A. Robertson, F. Aikman III, and D. Pabst. Transport, Fate, and Effects of

Sewage Sludge Disposal at the 106-Mile Site: A Summary and Synthesis of Findings. Journal of Marine

Environmental Engineering, 3. 313 - 326.

Ireland, D.S., Burton, G.A. and G.G. Hess. 1996. In Situ Toxicity Evaluations of Turbidity and Photoinduction

of Polycylic Aromatic Hydrocarbons. Environmental Toxicology and Chemistry. Vol. 15: 4. p 574-581. 
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

247

Lipscomb, S.W . 1995. Quality Assurance Plan for Discharge Measurements Using Broadband Acoustic

Doppler Current Profilers. USGS Open-File Report 95-701.

Long E.R., D.D. Mac Donald, S.L. Smith, and F.D. Calder. 1995. Incidence of Adverse Biological Effects with

Ranges of Chemical Concentrations in Marine and Estuarine Waters. Environmental Management 19 (1): 81-

97.

Long, E.R. and L.G. Morgan. 1990. The Potential for Biological Effects of Sediment-Sorbed Contam inants

Tested in the National Status and Trends Program. NOAA Tech. Memo. NOS OMA 62. National Oceanic and

Atm ospheric Administration, Seattle W A.

MacDonald D.D., C.G. Ingersoll, and T.A. Berger. 2000. Development and Evaluation of Consensus-Based

Sediment Quality Guidelines for Freshwater Ecosystems. Archives of Environmental Contamination and

Toxicology 39: 20-31.

Manheim F.T. 1966. A Hydraulic Squeezer for Obtaining Interstitial Water from Consolidated and

Unconsolidated Sediments. Geol. Surv. Prof. Pap. U.S., 550-C: 256 - 261.

Murdoch, Alena and Jose M. Azcue. 1995. Manual of Aquatic Sediment Sampling. Lewis Publishers, Boca

Raton, FL. 219 pp.

NFESC. 2000. Rapid Characterization of Metals in Sediments Using X-Ray Fluorescence (XRF) Technology.

Naval Facilities Engineering Command, Washington DC.

NOAA. 1998. Sampling and Analytical Methods of the National Status and Trends Program, Mussel Watch

Project: 1996 Update, NOAA Technical Memo NOS ORCA 130. National Oceanic and Atm ospheric

Administration, Silver Spring, MD. 233 pp.

PSWQAT. 1997. Recommended Guidelines for Sampling Marine Sediment, Water Column, and Tissue in

Puget Sound, Puget Sound Protocols and Guidelines. Puget Sound Water Quality Action Team, Olympia,

W A.

Sarda, N. and G.A. Burton. 1995. Am monia Variation in Sediments: Spatial, Temporal, and Method-Related

Effects. Environmental Toxicology and Chemistry. Vol 14: 9.

Sasson-Brickson, G. and G.A. Burton, Jr. 1991. In Situ and Laboratory Sediment Toxicity Testing with

Ceriodaphnia Dubia. Environmental Toxicology and Chemistry. Vol 10. P 201-207.

Sediment Management Work Group. 1999. Contaminated Sediment Management Technical Papers.

Sediment Management Work Group, Detroit MI.

USACE/WDNR/WDEC. 2000. Dredged Material Evaluation and Disposal Procedures, A Users Manual for the

Puget Sound Dredged Disposal Analysis (PSDAA) Program U.S. Army Corps of Engineers, Seattle District,

U.S. Environmental Protection Agency Region 10, Washington State Department of Natural Resources,

Washington State Department of Ecology

USACE Engineering and Design.1996. Soil Sampling Engineering Manual. EM 1110-1-1906. Department of

the Army, Washington D.C.

US Army Engineer Waterways Experiment Station. 1994. The Plume Measurement System (PLUMES), A

Commercially Available System, Dredging Research Technical Notes DRP-1-16. Waterways Experiment

Station, U.S. Army Corps of Engineers, Vicksburg, MS.

US Army Engineer Waterways Experiment Station. 1991. Hydrologic Surveys Applicable to Dredging

Operations, Dredging Research Technical Notes DRP-2-03. Waterways Experiment Station, U.S. Army Corps

of Engineers, Vicksburg, MS.

US Army Engineer Waterways Experiment Station. 1989. Monitoring Dredged Material Consolidation and
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

248

Settlement at Aquatic Disposal Sites. Environmental Effects of Dredging Technical Notes EEDP-01-5.

Waterways Experiment Station, U.S. Army Corps of Engineers, Vicksburg, MS.

US Army Engineer Waterways Experiment Station. 1988. Acoustic Tools and Techniques for Physical

Monitoring of Aquatic Dredged Material Disposal Sites. EEDP-01-10. Waterways Experiment Station, U.S.

Army Corps of Engineers, Vicksburg, MS.

USEPA. 2001a. Method 245.7: Mercury in Water by Cold Vapor Atomic Fluorescence Spectrometry, Draft,

EPA 821-R-01-008. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 2001b. Method 1630: Methyl Mercury in Water by Distillation, Aqueous Ethylation, Purge and Trap,

and CVAFS, EPA 821-R-01-020. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 2001c. Method 1632, Revision A: Chemical Speciation of Arsenic in Water and Tissue by Hydride

Generation Quartz Furnace Atomic Absorption Spectrometry, EPA-821-R-01-006. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

USEPA. 2001d. Appendix to Method 1631: Total Mercury in Tissue, Sludge, Sediment, and Soil by Acid

Digestion and BrCl Oxidation, EPA-821-R-01-013. Office of Water, U.S. Environmental Protection Agency,

Washington, DC.

USEPA. 2001e. Method for Assessing the Chronic Toxicity of Marine and Estuarine Sediment-associated

Contam inants with the Amphipod Leptocheirus plumulosus. EPA 600/R-01/020. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 2000a. Bioaccumulation Testing and Interpretation for the Purpose of Sediment Quality Assessment:

Status and Needs, EPA 823-R-00-001. Office of Water and Office of Solid Waste, U.S. Environmental

Protection Agency, Washington, DC.

USEPA. 2000b. Coastal 2000 Northeast Component: Field Operations Manual, Environmental Monitoring and

Assessment Program (EMAP), EPA/620/R-00/002. Office of Research and Development, U.S. Environmental

Protection Agency, Washington, DC.

USEPA. 2000c. Method 6200, Field Portable X-Ray Fluorescence Spectrom etry for the Determination of

Elemental Concentrations in Soil and Sediment. Office of Solid Waste, U.S. Environmental Protection Agency,

Washington DC.

USEPA. 2000d. Methods for Measuring the Toxicity and Bioaccumulation of Sediment-associated

Contam inants with Freshwater Invertebrates, EPA/600/R-99/064. Office of Science and Technology, U.S.

Environmental Protection Agency, Washington, DC.

USEPA. 1999a. National Recommended Water Quality Criteria-Correction, EPA-822-Z-99-001. Office of

Water, U.S. Environmental Protection Agency, Washington D.C.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

249

USEPA. 1999b. Method 1631, Revision B: Mercury in Water by Oxidation, Purge and Trap, and Cold Vapor

Atomic Fluorescence Spectrometry, EPA 821-R-99-005. Office of Water, U.S. Environmental Protection

Agency, Washington, DC.

USEPA. 1999c. Method 1668, Revision A: Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and

Tissue by HRGC/HRMS, EPA-821-R-00-002. Office of Water, U.S. Environmental Protection Agency,

Washington, DC.

USEPA. 1999d. Innovative Technology Verification Report: Aquatic Research Instruments Russian Peat

Borer, EPA/600/R-01/010. Office of Research and Development, U.S. Environmental Protection Agency,

Washington, DC.

USEPA. 1999e. Sediment Sampling Technology: Art’s Manufacturing & Supply, Inc. Split Core Sampler for

Submerged Sediments, Superfund Innovative Technology, Evaluation Program. EPA/600/R-01/009. Office of

Research and Development, U.S. Environmental Protection Agency, Washington, DC. 

USEPA. 1998. EMAP: Surface Waters, Field Operations and Methods for Measuring the Ecological Condition

of Wadeable Streams. EPA/6201/R-94/004F. Office of Research and Development. U.S. Environmental

Protection Agency, Washington, DC.

USEPA and USACE. 1998. Evaluation of Dredged Material Proposed for Discharge in Waters of the U.S.-

Testing Manual, Inland Testing Manual, EPA 823-B-98-004. Office of Water, U.S. Environmental Protection

Agency, Washington, DC.

USEPA. 1997a. Ecological Risk Assessment Guidance for Superfund: Process for designing and Conducting

Ecological Risk Assessments, Interim Final. EPA 540-R-97-006. Office of Solid Waste and Emergency

Response, U.S. Environmental Protection Agency, Washington D.C.

USEPA. 1997b. Lake Michigan Mass Balance Study Methods Compendium, Volume 1: Sample Collection

Techniques, EPA 905-R-97-012c. Great Lakes National Program Office, U.S. Environmental Protection

Agency, Chicago, IL.

USEPA. 1997c.Lake Michigan Mass Balance Study Methods Compendium, Volume 3: Metals,

Conventionals, Radio chemistry, and Biomonitoring Sample Analysis Techniques. EPA 905-R-97-012c. Great

Lakes National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

USEPA. 1997d. Lake Michigan Mass Balance Study Methods Compendium, Volume 2: Organic and Mercury

Sample Analysis Techniques. EPA 905-R-97-012c. Great Lakes National Program Office, U.S. Environmental

Protection Agency, Chicago, IL.

USEPA. 1997e. Method 1668: Toxic Polychlorinated Biphenyls by Isotope Dilution HRGC/HRMS,

EPA-821-R-97-001. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 1997f. State of the Streams: 1995-1997 Maryland Biological Stream Survey Results, Mid-Atlantic

Integrated Assessment. Office of Research and Development, U.S. Environmental Protection Agency,

Washington, D.C.

USEPA. 1996a. Ecological Significance and Selection of Candidate Assessment Endpoints. EPA 540-F-95-

037. ECO Update (January 1996). Office of Solid Waste and Emergency Response, U.S. Environmental

Protection Agency, Washington D.C.

USEPA. 1996b. Method 1639: Determination of Trace Elem ents in Ambient Waters by Stabilized

Tem perature Graphite Furnace Atomic Absorption, EPA 821-R-96-006. Office of Water, U.S. Environmental

Protection Agency, Washington, DC.

USEPA. 1996c. Method 1637: Determination of Trace Elem ents in Ambient Waters by Off-Line Chelation Preconcentration and Stabilized Tem perature Graphite Furnace Atomic Absorption, EPA 821-R-96-004. Office of

Water, U.S. Environmental Protection Agency, Washington, DC.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

250

USEPA. 1996d. Method 1638: Determination of Trace Elem ents in Ambient Waters by Inductively Coupled

Plasma-Mass Spectrometry, EPA 821-R-96-005. Office of Water, U.S. Environmental Protection Agency,

Washington, DC.

USEPA. 1996e. Method 1640: Determination of Trace Elem ents in Ambient Waters by On-Line Chelation Preconcentration and Inductively Coupled Plasma-Mass Spectrometry, EPA 821-R-96-007. Office of Water, U.S.

Environmental Protection Agency, Washington, DC.

USEPA. 1996f. Method 1632: Inorganic Arsenic in Water by Hydride Generation Quartz Furnace Atomic

Absorption. EPA 821-R-96-013. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 1996g. Method 1636: Determination of Hexavalent Chromium by Ion Chromatography. EPA 821-R96-003. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 1996h. Screening of Polychlorinated Biphenyls by Immunoassay, SW846 Method 4020. Office of

Solid Waste, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 1995. Environmental Monitoring and Assessment Program Laboratory Methods Manual, Estuaries,

Volume 1-Biological and Chemical Analyses, EPA/620/R-95/008. Environmental Monitoring and Assessment

Program , U.S. Environmental Protection Agency, Washington DC.

USEPA. 1994a. Field Studies for Ecological Risk Assessment. EPA 540-F-94-014. ECO Update (September

1994). Office of Solid Waste and Emergency Response, U.S. Environmental Protection Agency, Washington

D.C.

USEPA. 1994b. Compendium of Environmental Response Team Standard Operating Procedures. Office of

Solid Waste and Emergency Response, U.S. Environmental Protection Agency, Edison, NJ. 

USEPA. 1994c. Method 1613: Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA 821-B-94-005. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 1994d. Methods for Assessing the Toxicity of Sediment-associated Contam inants with Estuarine and

Marine Amphipods. EPA/600/R-94/025. Office of Research and Development, U.S. Environmental Protection

Agency, Washington, DC.

USEPA. 1993a. Biological and Chemical Assessment of Contaminated Great Lakes Sediment, EPA 905-R93-

006. Great Lakes National Program Office, U.S. Environmental Protection Agency, Chicago, IL.

USEPA. 1993b. Assessment and Remediation of Contaminated Sediments (ARCS) Program: Biological and

Chemical Assessment of Contaminated Great Lakes Sediment, EPA 905-R93-006. Great Lakes National

Program Office, U.S. Environmental Protection Agency, Chicago, IL.

USEPA. 1993c. Fish Field and Laboratory Methods for Evaluating the Biological Integrity of Surface Waters,

EPA/600/R-92-111. Office of Research and Development, U.S. Environmental Protection Agency,

Washington, DC.

USEPA. 1992a. Sediment Classification Methods Compendium. EPA/823-R-92-006. Office of Water, U.S.

Environmental Protection Agency, Washington DC.

USEPA. 1992b. Framework for Ecological Risk Assessment. EPA/630/R-92/001. Risk Assessment Forum,

U.S. Environmental Protection Agency, Washington D.C.

USEPA. 1992c. Monitoring Guidance for the National Estuary Program. EPA 842-B-92-004. Office of Water.

Office of Wetlands, Oceans and Watersheds. Washington. D.C. 

USEPA. 1991. Draft Analytical Method for Determination of Acid Volatile Sulfide in Sediment, EPA-821-R-91-

100. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 1990a. Environmental Monitoring and Assessment Program: Near Coastal Component, 1990
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

251

Demonstration Project, Field Operations Manual. DRAFT. Contract #. 68-C8-0066. Office of Research and

Development. Narragansett, Rhode Island.

USEPA. 1990b. Macroinvertebrate Field and Laboratory Methods for Evaluating the Biological Integrity of

Surface Waters. EPA/600/4-90/030. Office of Research and Development, U.S. Environmental Protection

Agency, Washington, D.C.

USEPA. 1989a. Method 1624, Revision B: Volatile Organic Compounds by Isotope Dilution GC/MS,

EPA440-1-89-100. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA. 1989b. Method 1625, Revision B: Semi-volatile Organic Compounds by Isotope Dilution GC/MS,

EPA440-1-89-100. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

USEPA and the Naval Construction Battalion Center. 1992. Standard Operating Procedures and Field

Methods Used for Conducting Ecological Risk Assessment Case Studies. Technical Document 2296, Naval

Command, Control and Ocean Surveillance Center, RDT&E Division, San Diego, CA. 

USEPA and USACE. 1992. Guidance for Performing Tests on Dredged Material Proposed for Ocean

Disposal. (Draft). U.S. Army Corps of Engineers, New York District and U.S. Environmental Protection

Agency, Region II, New York, New York.

USEPA and USACE. 1991. Evaluation of Dredged Material Proposed for Ocean Disposal, Testing Manual,

EPA 503-8-91-001. Office of Water, U.S. Environmental Protection Agency, Washington D.C.

USGS. 2001. The Krupaseep. Next Generation Seepage Meter.

http://sofia.usgs.gov/sfrsf/entdisplays/krupaseep/. Updated July 23, 2001.

USGS. 1998. Tree Swallow Sample Collection and Processing Procedures, Technical Operating Procedure

WE-410.0. Upper Mississippi Science Center, U.S. Geological Survey, LaCrosse, W I.

Weber, C.I. 1991. Methods for Measuring Acute Toxicity of Effluents in Receiving Waters for Freshwater and

Marine Organisms. 4

th edition. EPA/600/4-901027. Environmental Monitoring Systems Laboratory, Office of

Research and Development, U.S. Environmental Protection Agency, Cincinnati, OH.
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

252

Index

acid digestion

Fact Sheet # 2.2.2 - 1 Total Mercury in Sludge, Settlement, Soil, and Tissue by Acid Digestion

and BrCl Oxidation, Appendix to Method 1631

acid volatile sulfide (AVS)

Fact Sheet # 2.2.2 - 4 Determination of Acid Volatile Sulfide and Selected Sim ultaneously

Extractable Metals in Sediment

acoustic sampling

Fact Sheet # 2.1.1 - 18 Quality Assurance Plan for Discharge Measurements Using Broadband

Acoustic Doppler Current Profilers

Fact Sheet # 2.2.1 – 13 Method No. DRP-2-03: Acoustic Sub-bottom Profiling Systems

algae

Fact Sheet # 2.1.3 – 3 Method No. ERT SOP 2027: Chronic Freshwater Algae Test

Fact Sheet # 2.1.3 16 Method No. NHEERL-AED SOP 1.03.009: Microtox® tests, NHEERL-AED

1.03.009

Fact Sheet # 2.3.1 - 6 Field-based Periphyton Survey in Wadeable Streams

Fact Sheet # 2.3.1 - 7 Laboratory-based Periphyton Survey: Single Habitat Sampling in Wadeable

Streams

Fact Sheet # 2.3.1 - 8 Laboratory-based Rapid Periphyton Survey: Multi-habitat Sampling in

Wadeable Streams

Fact Sheet # 2.3.1 - 10 Algae and Macroinvertebrate Sampling with Frames

alkalinity

Fact Sheet # 2.1.2 - 30 Method No. LMMB 091: Standard Operating Procedure for GLNPO Total

Alkalinity Titration

aluminum

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

antimony

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 4 EPA Method No. 1639: Determination of Trace Elem ents in Ambient

Waters by Stabilized Tem perature Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient

Waters by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

arsenic determination

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 8 EPA Method No. 1632: Inorganic Arsenic in Water by Hydride Generation

Quartz Furnace Atomic Absorption
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

253

Fact Sheet # 2.1.2 - 9 EPA Method No. 1632, Revision A: Chemical Speciation of Arsenic in

Water and Tissue by Hydride Generation Quartz Furnace Atomic

Absorption Spectrom etry

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2-4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 6 Chemical Speciation of Arsenic in Water and Tissue by Hydride Generation

Quartz Furnace Atomic Absorption Spectrometry, EPA Method 1632,

Revision A

assimilation efficiency

Fact Sheet # 2.1.3 - 15 Method No. NHEERL-AED SOP 1.03.013: Growth and Scope for Growth

Measurements with Mytilus edulis

Atterberg limits

Fact Sheet # 2.2.2 - 23 Standard Test Method for Liquid Limit, Plastic Limit, and Plasticity Index of

Soils, ASTM Method D4318

avian samples

Fact Sheet # 2.3.1 - 28 Swallows: Sampling Procedures

Fact Sheet # 2.3.1 - 29 Sample Processing of Swallows

benthic community characterization

Fact Sheet # 2.3.1 - 11 Benthic Organism Collection from a Marine Environm ent, NHEERL-AED

SOP 1.02.001

Fact Sheet # 2.3.1 - 12 Benthic Macroinvertebrate Protocols in a Wadeable Stream: Single Habitat

Approach, 1-Meter Kick Net

Fact Sheet # 2.3.1 - 13 Benthic Macroinvertebrate Protocols in a Wadeable Stream: Multi-habitat

Approach: D-Frame Dip Net

Fact Sheet # 2.3.1 - 14 Photographic Habitat Documentation of the Benthic Community

Fact Sheet # 2.3.1 - 15 Sediment Profile Cam era

Fact Sheet # 2.3.1 - 17 Stream-net Samplers: Surber, Portable Invertebrate Box Sampler, Hess

Sampler, Hess Stream Bottom Sampler, and Stream-bed Fauna Sampler

Fact Sheet #2.3.3 - 3 Laboratory Analysis of Benthic Macroinvertebrates in Wadeable Streams

beryllium -7

Fact Sheet #2.2.2 - 26 Method Title: Beryllium-7 as a Tracer of Short Term Sediment Deposition

bioaccumulation

Fact Sheet # 2.1.1 - 20 Caged Bivalve Deployment

Fact Sheet # 2.2.3 - 17 Bioaccumulation Test for Marine, Estuarine, and Freshwater Sediments,

EPA Method 100.3

Fact Sheet # 2.3.1 - 28 Swallows: Sampling Procedures

Fact Sheet # 2.3.1 - 29 Sample Processing of Swallows

bioassay- algae

Fact Sheet # 2.1.3 - 3 Method No. ERT SOP 2027: Chronic Freshwater Algae Test

bioassay – crustacean

Fact Sheet # 2.1.3 - 1 Method No. ERT SOP 2024: Acute Freshwater Crustacean Bioassay: 48

Hours
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

254

Fact Sheet # 2.1.3 – 4 Method No. ERT SOP 2025: Chronic Freshwater Crustacean Bioassay

(7day)

Fact Sheet # 2.1.3 - 5 Method No. ERT SOP 2028: Chronic Freshwater Crustaceans Bioassay

(10 days)

Fact Sheet # 2.1.3 - 8 Method No. NHEERL-AED SOP 1.03.003: Acute Marine Crustacean

Bioassay

Fact Sheet # 2.1.3 - 10 Method No. NHEERL-AED SOP 1.03.005: Chronic Estuarine Survival,

Growth, and Fecundity Test

Fact Sheet # 2.1.3 - 13 Toxicity Evaluations of Photoinduction of Polycyclic Aromatic Hydrocarbons

(PAH): In Situ Analysis

Fact Sheet # 2.1.3 - 14 Toxicity Evaluations of Photoinduction of Polycyclic Aromatic

Hydrocarbons: Laboratory Analysis of Storm water

bioassay – echinoderm

Fact Sheet # 2.1.3 - 11 Method No. NHEERL-AED SOP 1.03.006: Chronic Echinoderm

Fertilization Test

bioassay - estuarine environments

Fact Sheet # 2.1.3 - 10 Method No. NHEERL-AED SOP 1.03.005: Chronic Estuarine Survival,

Growth, and Fecundity Test

Fact Sheet # 2.1.3 - 11 Method No. NHEERL-AED SOP 1.03.006: Chronic Echinoderm

Fertilization Test

Fact Sheet # 2.2.3 - 12 Chronic Estuarine Amphipod Sediment Bioassay

bioassay – fish

Fact Sheet # 2.1.3 - 2 Method No. ERT SOP 2022: Acute Freshwater fish Bioassay

Fact Sheet # 2.1.3 - 6 Method No. ERT SOP 2026: Chronic Freshwater Fish Bioassay, ERT SOP

2026

Fact Sheet # 2.1.3 - 9 Method No. NHEERL-AED SOP 1.03.003: Acute Marine Fish Bioassay

Fact Sheet # 2.1.3 - 12 Method No. NHEERL-AED SOP 1.03.004: Chronic Marine Fish Bioassay

bioassay - freshwater environments

Fact Sheet # 2.1.3 - 1 Method No. ERT SOP 2024: Acute Freshwater Crustacean Bioassay: 48

Hours

Fact Sheet # 2.1.3 - 2 Method No. ERT SOP 2022: Acute Freshwater fish Bioassay

Fact Sheet # 2.1.3 - 3 Method No. ERT SOP 2027: Chronic Freshwater Algae Test

Fact Sheet # 2.1.3 – 4 Method No. ERT SOP 2025: Chronic Freshwater Crustacean Bioassay

(7day)

Fact Sheet # 2.1.3 - 5 Method No. ERT SOP 2028: Chronic Freshwater Crustaceans Bioassay (10

days)

Fact Sheet # 2.1.3 - 6 Method No. ERT SOP 2026: Chronic Freshwater Fish Bioassay, ERT SOP

2026

Fact Sheet # 2.2.3 - 1 Acute Freshwater Crustacean Sediment Bioassay: Flow-through

Fact Sheet # 2.2.3 - 2 Acute Freshwater Crustacean Sediment Bioassay: In Situ Exposures

Fact Sheet # 2.2.3 - 3 Acute Freshwater Crustacean Sediment Bioassay: Static Laboratory

Exposures

Fact Sheet # 2.2.3 - 4 Acute Freshwater Amphipod and Freshwater Insect Larvae Sediment

Bioassay, EPA Method 100.1

Fact Sheet # 2.2.3 - 5 Chronic Freshwater Amphipod Sediment Bioassay, EPA Method 100.4

Fact Sheet # 2.2.3 - 6 Life-Cycle Freshwater Midge Sediment Bioassay, EPA Method 100.5

bioassay - marine environments

Fact Sheet # 2.1.3 - 7 Method No. NHEERL-AED SOP 1.03.001: Chronic Marine Macroalgae,

Champia parvula, Sexual Reproduction test
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

255

Fact Sheet # 2.1.3 - 8 Method No. NHEERL-AED SOP 1.03.003: Acute Marine Crustacean

Bioassay

Fact Sheet # 2.1.3 - 9 Method No. NHEERL-AED SOP 1.03.003: Acute Marine Fish Bioassay

Fact Sheet # 2.1.3 - 12 Method No. NHEERL-AED SOP 1.03.004: Chronic Marine Fish Bioassay

Fact Sheet # 2.2.3 - 7 Acute Larval Bivalve Sediment Bioassay

Fact Sheet # 2.2.3 - 8 Acute Echinoderm Sediment Bioassay

Fact Sheet # 2.2.3 - 9 Acute Marine Crustacean Sediment Bioassay

Fact Sheet # 2.2.3 - 10 Acute Marine Amphipod Crustacean Sediment Bioassay, EPA Method

100.4

Fact Sheet # 2.2.3 - 11 Acute Marine Polychaete Sediment Bioassay, ASTM Method E1611-00

Fact Sheet # 2.2.3 - 12 Chronic Estuarine Amphipod Sediment Bioassay

Fact Sheet # 2.2.3 - 13 Chronic Marine Polychaete Sediment Bioassay, ASTM Method E1611-00

bioassay - marine/estuarine environments

Fact Sheet # 2.1.3 - 11 Method No. NHEERL-AED SOP 1.03.006: Chronic Echinoderm Fertilization

Test

bioassay – sediment

Fact Sheet # 2.2.1 - 8 Sediment Processing for Chem istry and Toxicity Testing

Fact Sheet # 2.2.3 - 1 Acute Freshwater Crustacean Sediment Bioassay: Flow-through

Fact Sheet # 2.2.3 - 2 Acute Freshwater Crustacean Sediment Bioassay: In Situ Exposures

Fact Sheet # 2.2.3 - 3 Acute Freshwater Crustacean Sediment Bioassay: Static Laboratory

Exposures

Fact Sheet # 2.2.3 - 4 Acute Freshwater Amphipod and Freshwater Insect Larvae Sediment

Bioassay, EPA Method 100.1

Fact Sheet # 2.2.3 - 5 Chronic Freshwater Amphipod Sediment Bioassay, EPA Method 100.4

Fact Sheet # 2.2.3 - 6 Life-Cycle Freshwater Midge Sediment Bioassay, EPA Method 100.5

Fact Sheet # 2.2.3 - 7 Acute Larval Bivalve Sediment Bioassay

Fact Sheet # 2.2.3 - 8 Acute Echinoderm Sediment Bioassay

Fact Sheet # 2.2.3 - 9 Acute Marine Crustacean Sediment Bioassay

Fact Sheet # 2.2.3 - 10 Acute Marine Amphipod Crustacean Sediment Bioassay, EPA Method

100.4

Fact Sheet # 2.2.3 - 11 Acute Marine Polychaete Sediment Bioassay, ASTM Method E1611-00

Fact Sheet # 2.2.3 - 12 Chronic Estuarine Amphipod Sediment Bioassay

Fact Sheet # 2.2.3 - 13 Chronic Marine Polychaete Sediment Bioassay, ASTM Method E1611-00

biomass

Fact Sheet # 2.3.1 - 6 Field-based Periphyton Survey in Wadeable Streams

Fact Sheet # 2.3.1 - 7 Laboratory-based Periphyton Survey: Single Habitat Sampling in Wadeable

Streams

Fact Sheet # 2.3.1 - 8 Laboratory-based Rapid Periphyton Survey: Multi-habitat Sampling in

Wadeable Streams

Fact Sheet # 2.3.3 - 1 Laboratory Identification, Enumeration and Biomass Measurements of

Periphyton in Wadeable Streams

Fact Sheet # 2.3.3 - 2 Laboratory Periphyton Biomass Determination

brail sampling

Fact Sheet # 2.3.1 - 18 Mussel Collection Using Brails

butylins

Fact Sheet # 2.2.2 - 13 Butyltin in Sediments
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

256

cadmium

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 4 EPA Method No. 1639: Determination of Trace Elem ents in Ambient Waters

by Stabilized Tem perature Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.1.2 - 5 EPA Method No. 1637: Determination of Trace Elem ents in Ambient Waters

by Off-Line Chelation Pre-concentration and Stabilized Tem perature

Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 7 EPA Method No. 1640: Determination of Trace Elem ents in Ambient Waters

by On-Line Chelation Pre-concentration and Inductively Coupled PlasmaMass Spectrom etry

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

calcium

Fact Sheet # 2.1.2 - 34 Method No. LMMB 095: Total Hardness Titration

carbon-14

Fact Sheet # 2.3.1 - 4 Primary Productivity Using 14C: Field Procedure in the Great Lakes, LMMB

016

cesium-137

Fact Sheet # 2.2.2 - 25 Method Title: Sediment Age Dating Using Cesium-137

chemical fishing techniques

Fact Sheet # 2.3.1 - 20 Chemical Fishing

chloride

Fact Sheet # 2.1.2 - 27 Method No. ESS Method 140.4: Chloride - Automated Flow Injection

Analysis

Fact Sheet # 2.1.2 - 28 Method No. ESS Method 200.5: Determination of Inorganic Anions in Water

by Ion Chromatography

chlorophyll-a

Fact Sheet # 2.3.1 - 2 Chlorophyll-a Sampling Method and Preservation: Field Procedure in the

Great Lakes, LMMB 015

Fact Sheet # 2.3.1 - 3 Chlorophyll-a and Phaeophytin Field Filtering Protocols

Fact Sheet # 2.3.3 - 2 Laboratory Periphyton Biomass Determination

chromium

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 4 EPA Method No. 1639: Determination of Trace Elem ents in Ambient Waters

by Stabilized Tem perature Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 -10 EPA Method No. 1636, Method Title: Determination of Hexavalent

Chromium by Ion Chromatography

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

257

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

clearance rate

Fact Sheet # 2.1.3 - 15 Method No. NHEERL-AED SOP 1.03.013: Growth and Scope for Growth

Measurements with Mytilus edulis

combustion analysis

Fact Sheet # 2.2.2 - 14 Procedures for Sediment Total Organic Carbon (TOC) Determination

conductivity

Fact Sheet # 2.1.2 - 31 Method No. LMMB 094: Standard Operating Procedure for GLNPO Specific

Conductance: Conductivity Bridge

conductivity, temperature, density (CTD) measurements

Fact Sheet # 2.1.1-1 In Situ sampling with the Hydrolab Datasonde3® Unit

consolidation (see sediment consolidation)

copper

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.1.2 - 7 EPA Method No. 1640: Determination of Trace Elem ents in Ambient Waters

by On-Line Chelation Pre-concentration and Inductively Coupled PlasmaMass Spectrom etry

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

dioxin analysis

Fact Sheet # 2.1.2 - 17 EPA Method No. 1613: Tetra- through Octa-Chlorinated Dioxins and Furans

by Isotope Dilution HRGC/HRMS

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.2.2 - 10 Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA Method 1613

Fact Sheet # 2.3.2 - 11 Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA Method 1613

dissolved organic carbon (DOC)

Fact Sheet # 2.1.2 - 24 Standard Method No. 5310: Total Organic Carbon

Fact Sheet # 2.1.2 - 25 Method No. LMMB 096: Standard Operating Procedure for the Analysis of

Dissolved-Phase Organic Carbon in Great Lakes Waters
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

258

dissolved oxygen

Fact Sheet # 2.1.1 - 1 Method Title: In Situ sampling with the Hydrolab Datasonde3® Unit

Fact Sheet # 2.1.1 - 2 In Situ Dissolved Oxygen sampling with a YSI Model 58 Dissolved Oxygen

Meter and probe DO meter

Fact Sheet # 2.1.1 - 9 Sample and Preservation of Water Specific Parameters

dry weight determination

Fact Sheet # 2.3.2 - 14 Determination of Percent Dry Weight for Tissues

Fact Sheet # 2.2.2 - 17 Procedures for Water Content Determination

electrofishing

Fact Sheet # 2.3.1 - 19 Electrofishing

entrapment devices

Fact Sheet # 2.3.1 - 23 Entrapment Devices

field sampling - in situ measurements

Fact Sheet # 2.1.1-1 In Situ sampling with the Hydrolab Datasonde3® Unit

Fact Sheet # 2.1.1-2 In Situ Dissolved Oxygen sampling with a YSI Model 58 Dissolved Oxygen

Meter and probe DO meter

Fact Sheet # 2.1.1-3 In Situ sampling of Irradiance

Fact Sheet # 2.1.1-4 In Situ Transparency Sampling

Fact Sheet # 2.2.2 - 24 Field Portable X-Ray Fluorescence Spectrom etry for the Determination of

Elemental Concentrations in Soil and Sediment

Fact Sheet # 2.3.1 - 6 Field-based Periphyton Survey in Wadeable Streams

field sampling - discrete sample collection

Fact Sheet # 2.1.1-5 Sample Collection Procedures for Marine water

Fact Sheet # 2.1.1-6 Method No. LMMB 013: Field Sampling Using a Rosette Sampler

Fact Sheet # 2.1.1 - 7 Method No. ERT SOP #2013: Water Sample Collection with the Kemmerer

Bottle and the Bacon Bomb Sampler

Fact Sheet # 2.1.1 -8 Method No. ERT SOP # 2013: Dip Sampler

Fact Sheet # 2.1.1 - 9 Sample and Preservation of Water Specific Parameters

Fact Sheet # 2.1.1 - 10 Method No. LMMB 014: Sampling of Particulate-Phase and DissolvedPhase Organic Carbon in Great Lakes Waters

Fact Sheet # 2.1.1 - 17 LMMB 017: USGS Field Operation Plan: Tributary Monitoring

Fact Sheet # 2.1.1 - 19 Seepage Meters

Fact Sheet # 2.3.1 - 1 Phytoplankton Sample Collection and Preservation in the Great Lakes,

LMMB 023t

Fact Sheet # 2.3.1 - 2 Chlorophyll-a Sampling Method and Preservation: Field Procedure in the

Great Lakes, LMMB 015

Fact Sheet # 2.3.1 - 3 Chlorophyll-a and Phaeophytin Field Filtering Protocols

fish collection

Fact Sheet # 2.3.1 - 19 Electrofishing

Fact Sheet # 2.3.1 - 20 Chemical Fishing

Fact Sheet # 2.3.1 - 21 Fish Collection Using Seine Nets

Fact Sheet # 2.3.1 - 22 Entanglement Nets

Fact Sheet # 2.3.1 - 23 Entrapment Devices

Fact Sheet # 2.3.1 - 24 Pop Nets

Fact Sheet # 2.3.1 - 25 Trawls

fish community assessment

Fact Sheet # 2.3.3 - 10 Fish Bioassessment I and II
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

259

fish age dating

Fact Sheet # 2.3.1 - 26 Fish Processing Method in the Great Lakes, LMMB 025

fish processing

Fact Sheet # 2.3.1 - 26 Fish Processing Method in the Great Lakes, LMMB 025

Fact Sheet # 2.3.1 - 27 Fish Processing

Fact Sheet # 2.3.3 - 5 SOP-2: Lab Analysis of Lake Trout Stomachs and Data Entry; Appendix B.

Standard Operating Procedure for Lab Analysis of Coho Salmon Stomachs

and Data Entry, LMMB 026 - Appendix 2 & LMMB 027 - Appendix B

fish tissue analysis

Fact Sheet # 2.3.2 - 1 Sample Preparation for Metal Contam inants in Tissue

Fact Sheet # 2.3.2 - 7 Extraction and Lipid Separation of Fish Samples for Contaminant Analysis

and Lipid Determination, LMMB 043

furans (see dioxins)

gas chromatography

Fact Sheet # 2.1.2 - 13 EPA Method No. 1625: Semi-volatile Organic Compounds by Isotope

Dilution GC/MS

Fact Sheet # 2.1.2 - 14 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS) - Selected Ion Monitoring

(SIM) Mode

Fact Sheet # 2.1.2 - 15 Method No. LMMB 041: Analysis of Polychlorinated Biphenyls and

Chlorinated Pesticides by Gas Chromatography with Electron Capture

Detection

Fact Sheet # 2.1.2 - 16 Method No. LMMB: PCBs and Pesticides in Surface Water by XAD-2 Resin

Extraction

Fact Sheet # 2.1.2 - 17 EPA Method No. 1613: Tetra- through Octa-Chlorinated Dioxins and Furans

by Isotope Dilution HRGC/HRMS

Fact Sheet # 2.1.2 - 18 EPA Method No. 1668: Toxic Polychlorinated Biphenyls by Isotope Dilution

High Resolution Gas Chromatography/High Resolution Mass Spectrom etry

Fact Sheet # 2.1.2 - 19 EPA Method No. 1668, Revision A: Chlorinated Biphenyl Congeners in

Water, Soil, Sediment, and Tissue by HRGC/HRMS

Fact Sheet # 2.2.2 - 5 Photovac GC Analysis for Soil, Water, and Air/Soil Gas, OSW ER SOP#

2109

Fact Sheet # 2.2.2 - 7 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS)--Selected Ion Monitoring

(SIM) Mode

Fact Sheet # 2.2.2 - 8 Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

Fact Sheet # 2.2.2 - 10 Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA Method 1613

Fact Sheet # 2.2.2 - 11 Toxic Polychlorinated Biphenyls by Isotope Dilution High Resolution Gas

Chromatography/High Resolution Mass Spectrometry, EPA Method 1668

Fact Sheet # 2.2.2 - 12 Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and Tissue by

HRGC/HRMS, EPA Method 1668 Revision A

Fact Sheet # 2.2.2 - 13 Butyltin in Sediments
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

260

Fact Sheet # 2.3.2 - 9 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS)--Selected Ion Monitoring

(SIM) Mode

Fact Sheet # 2.3.2 - 10 Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

Fact Sheet # 2.3.2 - 11 Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA Method 1613

Fact Sheet # 2.3.2 - 12 Toxic Polychlorinated Biphenyls by Isotope Dilution High Resolution Gas

Chromatography/High Resolution Mass Spectrometry, EPA Method 1668

Fact Sheet # 2.3.2 - 13 Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and Tissue by

HRGC/HRMS, EPA Method 1668 Revision A

gonadal analysis

Fact Sheet # 2.3.3 - 6 Gonadal Analysis

grain size analysis

Fact Sheet # 2.2.2 - 16 Sediment Grain Size Analysis, NHEERL-AED SOP 1.01.005

habitat assessment

Fact Sheet # 2.1.1 - 15 Physical Characterization of a stream

Fact Sheet # 2.1.1 - 16 Visual based habitat assessment

hardness

Fact Sheet # 2.1.2 - 34 Method No. LMMB 095: Total Hardness Titration

high resolution gas chromatography

Fact Sheet # 2.3.2 - 11 Tetra- through Octa-Chlorinated Dioxins and Furans by Isotope Dilution

HRGC/HRMS, EPA Method 1613

Fact Sheet # 2.3.2 - 12 Toxic Polychlorinated Biphenyls by Isotope Dilution High Resolution Gas

Chromatography/High Resolution Mass Spectrometry, EPA Method 1668

Fact Sheet # 2.3.2 - 13 Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and Tissue by

HRGC/HRMS, EPA Method 1668 Revision A

histopathology

Fact Sheet # 2.3.3 - 7 Histopathological Evaluations of Target and Non Target Fish Species

Fact Sheet # 2.3.3 - 8 Histopathology Analysis

hydrographic profiles

Fact Sheet # 2.1.1-1 In Situ sampling with the Hydrolab Datasonde3® Unit

Fact Sheet # 2.1.1-2 In Situ Dissolved Oxygen sampling with a YSI Model 58 Dissolved Oxygen

Meter and probe DO meter

Fact Sheet # 2.1.1-3 In Situ sampling of Irradiance

Fact Sheet # 2.1.1-4 In Situ Transparency Sampling

immunoassay screening

Fact Sheet # 2.2.2 - 9 Screening for Polychlorinated Biphenyls by Immunoassay, SW846 Method

4020

Index of Biotic Integrity (IBI)

Fact Sheet # 2.3.3 - 9 Index of Biotic Integrity (IBI)
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

261

Inductively Coupled Plasma Mass Spectrometry (ICPMS)

Fact Sheet # 2.2.2 - 3 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

iron

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

lead

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.1.2 - 5 EPA Method No. 1637: Determination of Trace Elem ents in Ambient Waters

by Off-Line Chelation Pre-concentration and Stabilized Tem perature

Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 7 EPA Method No. 1640: Determination of Trace Elem ents in Ambient Waters

by On-Line Chelation Pre-concentration and Inductively Coupled PlasmaMass Spectrom etry

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

lead-210

Fact Sheet # 2.2.2-15 Method No. LMMB 084, Method Title: Determination of the Activity of Lead210 in Sediments and Soils

light measurements

Fact Sheet # 2.1.1-3 In Situ sampling of Irradiance

macroinvertebrate – analysis

Fact Sheet # 2.3.3 - 3 Laboratory Analysis of Benthic Macroinvertebrates in Wadeable Streams

Fact Sheet # 2.3.3 - 4 Laboratory Analysis of Water Column Organisms

macroinvertebrate - sample collection

Fact Sheet # 2.2.1 - 1 Grab Sampling

Fact Sheet # 2.2.1 - 2 Core Samplers

Fact Sheet # 2.2.1 - 3 Hand Collection

Fact Sheet # 2.2.1 - 4 Hand collection at depth with SCUBA

Fact Sheet # 2.3.1 - 9 Artificial Substrate Samplers of Macroinvertebrates in Wadeable Streams

Fact Sheet # 2.3.1 - 10 Algae and Macroinvertebrate Sampling with Frames

Fact Sheet # 2.3.1 - 11 Benthic Organism Collection from a Marine Environm ent, NHEERL-AED

SOP 1.02.001
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

262

Fact Sheet # 2.3.1 - 12 Benthic Macroinvertebrate Protocols in a Wadeable Stream: Single Habitat

Approach, 1-Meter Kick Net

Fact Sheet # 2.3.1 - 13 Benthic Macroinvertebrate Protocols in a Wadeable Stream: Multi-habitat

Approach: D-Frame Dip Net

Fact Sheet # 2.3.1 - 16 Macroinvertebrate Drift Nets in a Wadeable Stream

Fact Sheet # 2.3.1 - 17 Stream-net Samplers: Surber, Portable Invertebrate Box Sampler, Hess

Sampler, Hess Stream Bottom Sampler, and Stream-bed Fauna Sampler

magnesium

Fact Sheet # 2.1.2 - 34 Method No. LMMB 095: Total Hardness Titration

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

mercury

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 1 EPA Method No. 245.7: Mercury in Water by Cold Vapor Atomic

Fluorescence Spectrom etry

Fact Sheet # 2.1.2 - 2 EPA Method No. 1631, Revision B: Mercury in Water by Oxidation, Purge

and Trap, and Cold Vapor Atomic Fluorescence Spectrom etry

Fact Sheet # 2.1.2 - 3 EPA Method No. 1630: Methyl mercury in water by distillation, Aqueous

Ethylation, Purge and Trap, and CVAFS

Fact Sheet # 2.2.2 - 1 Total Mercury in Sludge, Settlement, Soil, and Tissue by Acid Digestion and

BrCl Oxidation, Appendix to Method 1631

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 2 Total Mercury in Tissue, Sludge, Sediment, and Soil by Acid Digestion and

BrCl Oxidation, Appendix to Method 1631

Fact Sheet # 2.3.2 - 3 Versatile Combustion-Amalgamation Technique for the Photometric

Determination of Mercury in Fish and Environmental Samples, LMMB 052

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

metals analysis methods

Fact Sheet # 2.1.1 - 9 Sample and Preservation of Water Specific Parameters

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 1 EPA Method No. 245.7: Mercury in Water by Cold Vapor Atomic

Fluorescence Spectrom etry

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.1.2 - 7 EPA Method No. 1640: Determination of Trace Elem ents in Ambient Waters

by On-Line Chelation Pre-concentration and Inductively Coupled PlasmaMass Spectrom etry

Fact Sheet # 2.1.2 - 8 EPA Method No. 1632: Inorganic Arsenic in Water by Hydride Generation

Quartz Furnace Atomic Absorption
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

263

Fact Sheet # 2.1.2 - 9 EPA Method No. 1632, Revision A: Chemical Speciation of Arsenic in Water

and Tissue by Hydride Generation Quartz Furnace Atomic Absorption

Spectrom etry

Fact Sheet # 2.1.2 - 10 EPA Method No. 1636: Determination of Hexavalent Chromium by Ion

Chromatography

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.3.2 - 1 Sample Preparation for Metal Contam inants in Tissue

Fact Sheet # 2.3.2 - 4 Trace Element Quantification Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 6 Chemical Speciation of Arsenic in Water and Tissue by Hydride Generation

Quartz Furnace Atomic Absorption Spectrometry, EPA Method 1632,

Revision A

metals toxicity

Fact Sheet # 2.1.2 - 4 EPA Method No. 1639: Determination of Trace Elem ents in Ambient Waters

by Stabilized Tem perature Graphite Furnace Atomic Absorption

microbiological testing

Fact Sheet # 2.1.2 - 5 EPA Method No. 1637: Determination of Trace Elem ents in Ambient Waters

by Off-Line Chelation Pre-concentration and Stabilized Tem perature

Graphite Furnace Atomic Absorption

microwave extraction

Fact Sheet # 2.3.2 - 16 Microwave Extraction of Marine Tissue for Semivolatile Organic Analytes,

AED LOP 2.03.030,

Revision 0

mussels

Fact Sheet # 2.1.1 - 20 Caged Bivalve Deployment

Fact Sheet # 2.3.1 - 18 Mussel Collection Using Brails

mutagenicity testing

Fact Sheet # 2.2.3 - 14 Ames Mutagenicity Assay

Fact Sheet # 2.2.3 - 15 Mutatox Genotoxicity Assay

Fact Sheet # 2.2.3 - 16 V79 Sister Chromatid Exchange Assay, NHEERL-AED SOP 1.03.012

net tow surveys

Fact Sheet # 2.3.1 - 5 Zooplankton Sample Collection and Preservation in the Great Lakes, LMMB

024

Fact Sheet # 2.3.1 - 13 Benthic Macroinvertebrate Protocols in a Wadeable Stream: Multi-habitat

Approach: D-Frame Dip Net

Fact Sheet # 2.3.1 - 16 Macroinvertebrate Drift Nets in a Wadeable Stream

Fact Sheet # 2.3.1 - 21 Fish Collection Using Seine Nets

Fact Sheet # 2.3.1 - 22 Entanglement Nets

Fact Sheet # 2.3.1 - 24 Pop Nets
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

264

nickel

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 4 EPA Method No. 1639: Determination of Trace Elem ents in Ambient Waters

by Stabilized Tem perature Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.1.2 - 7 EPA Method No. 1640: Determination of Trace Elem ents in Ambient Waters

by On-Line Chelation Pre-concentration and Inductively Coupled PlasmaMass Spectrom etry

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

nitrogen determination

Fact Sheet # 2.1.2 - 20 Method No. ESS Method 220.3: Am monia Nitrogen and Nitrate+Nitrite

Nitrogen, Automated Flow Injection Analysis Method

Fact Sheet # 2.1.2 - 21 Method No. ESS Method 230.1: Total Phosphorus and Total Kjeldahl

Nitrogen, Semi-Automated Method

Fact Sheet # 2.1.2 - 28 Method No. ESS Method 200.5: Determination of Inorganic Anions in Water

by Ion Chromatography

organic analysis methods

Fact Sheet # 2.1.1 - 9 Sample and Preservation of Water Specific Parameters

Fact Sheet # 2.1.1 - 10 Method No. LMMB 014: Sampling of Particulate-Phase and DissolvedPhase Organic Carbon in Great Lakes Waters

Fact Sheet # 2.1.2 - 11 EPA Method No. 1624b: Volatile Organic Compounds by Isotope Dilution

GC/MS

Fact Sheet # 2.1.2 - 13 EPA Method No. 1625: Semi-volatile Organic Compounds by Isotope

Dilution GC/MS

Fact Sheet # 2.1.2 - 14 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS) - Selected Ion Monitoring

(SIM) Mode

Fact Sheet # 2.1.2 - 15 Method No. LMMB 041: Analysis of Polychlorinated Biphenyls and

Chlorinated Pesticides by Gas Chromatography with Electron Capture

Detection

Fact Sheet # 2.1.2 - 16 Method No. LMMB: PCBs and Pesticides in Surface Water by XAD-2 Resin

Extraction

Fact Sheet # 2.1.2 - 18 EPA Method No. 1668: Toxic Polychlorinated Biphenyls by Isotope Dilution

High Resolution Gas Chromatography/High Resolution Mass Spectrom etry

Fact Sheet # 2.1.2 - 19 EPA Method No. 1668, Revision A: Chlorinated Biphenyl Congeners in

Water, Soil, Sediment, and Tissue by HRGC/HRMS

Fact Sheet # 2.3.2 - 7 Extraction and Lipid Separation of Fish Samples for Contaminant Analysis

and Lipid Determination, LMMB 043

organotins

Fact Sheet # 2.2.2 - 13 Butyltin in Sediments
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

265

oxidation

Fact Sheet # 2.2.2 - 1 Total Mercury in Sludge, Settlement, Soil, and Tissue by Acid Digestion and

BrCl Oxidation, Appendix to Method 1631

oysters

Fact Sheet # 2.3.3 - 6 Gonadal Analysis

PAH analysis methods

Fact Sheet # 2.1.2 - 14 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS) - Selected Ion Monitoring

(SIM) Mode

Fact Sheet # 2.1.3 - 13 Toxicity Evaluations of Photoinduction of Polycyclic Aromatic Hydrocarbons

(PAH): In Situ Analysis

Fact Sheet # 2.1.3 - 14 Toxicity Evaluations of Photoinduction of Polycyclic Aromatic Hydrocarbons:

Laboratory Analysis of Storm water

Fact Sheet # 2.2.2 - 6 Extraction and Clean-up of Sediments for Semi-volatile Organics Following

the Internal Standard Method, LMMB 040

Fact Sheet # 2.2.2 – 7 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectom etry (GC/MS)--Selected Ion Monitoring

(SIM) Mode

Fact Sheet # 2.3.2 - 7 Extraction and Lipid Separation of Fish Samples for Contaminant Analysis

and Lipid Determination, LMMB 043

Fact Sheet # 2.3.2 - 8 Purification of Biological Tissue Samples by Gel Permeation

Chromatography of Organic Analyses

Fact Sheet # 2.3.2 - 9 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS)--Selected Ion Monitoring

(SIM) Mode

particulate organic carbon (POC)

Fact Sheet # 2.1.2 - 24 Standard Method No. 5310: Total Organic Carbon

Fact Sheet # 2.1.2 - 26 Method No. LMMB 097: Standard Operating Procedure for the Analysis of

Particulate-Phase Organic Carbon in Great Lakes Waters

PCB analysis methods

Fact Sheet # 2.1.2 - 15 Method No. LMMB 041: Analysis of Polychlorinated Biphenyls and

Chlorinated Pesticides by Gas Chromatography with Electron Capture

Detection

Fact Sheet # 2.1.2 - 16 Method No. LMMB: PCBs and Pesticides in Surface Water by XAD-2 Resin

Extraction

Fact Sheet # 2.1.2 - 18 EPA Method No. 1668: Toxic Polychlorinated Biphenyls by Isotope Dilution

High Resolution Gas Chromatography/High Resolution Mass Spectrom etry

Fact Sheet # 2.1.2 - 19 EPA Method No. 1668, Revision A: Chlorinated Biphenyl Congeners in

Water, Soil, Sediment, and Tissue by HRGC/HRMS

Fact Sheet # 2.2.2 - 6 Extraction and Clean-up of Sediments for Semi-volatile Organics Following

the Internal Standard Method, LMMB 040

Fact Sheet # 2.2.2 - 8 Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

Fact Sheet # 2.2.2 - 9 Screening for Polychlorinated Biphenyls by Immunoassay, SW846 Method

4020

Fact Sheet # 2.2.2 - 11 Toxic Polychlorinated Biphenyls by Isotope Dilution High Resolution Gas

Chromatography/High Resolution Mass Spectrometry, EPA Method 1668

Fact Sheet # 2.2.2 - 12 Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and Tissue by

HRGC/HRMS, EPA Method 1668 Revision A
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

266

Fact Sheet # 2.3.2 - 7 Extraction and Lipid Separation of Fish Samples for Contaminant Analysis

and Lipid Determination, LMMB 043

Fact Sheet # 2.3.2 - 8 Purification of Biological Tissue Samples by Gel Permeation

Chromatography of Organic Analyses

Fact Sheet # 2.3.2 - 10 Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

Fact Sheet # 2.3.2 - 12 Toxic Polychlorinated Biphenyls by Isotope Dilution High Resolution Gas

Chromatography/High Resolution Mass Spectrometry, EPA Method 1668

Fact Sheet # 2.3.2 - 13 Chlorinated Biphenyl Congeners in Water, Soil, Sediment, and Tissue by

HRGC/HRMS, EPA Method 1668 Revision A

percent dry weight

Fact Sheet # 2.3.2 - 14 Determination of Percent Dry Weight for Tissues

percent lipid

Fact Sheet # 2.3.2 - 15 Determination of Percent Lipid in Tissue

percent moisture

Fact Sheet # 2.3.2 - 14 Determination of Percent Dry Weight for Tissues

periphyton

Fact Sheet # 2.3.1 - 6 Field-based Periphyton Survey in Wadeable Streams

Fact Sheet # 2.3.1 - 7 Laboratory-based Periphyton Survey: Single Habitat Sampling in Wadeable

Streams

Fact Sheet # 2.3.1 - 8 Laboratory-based Rapid Periphyton Survey: Multi-habitat Sampling in

Wadeable Streams

Fact Sheet # 2.3.1 - 9 Artificial Substrate Samplers of Macroinvertebrates in Wadeable Streams

Fact Sheet # 2.3.3 - 1 Laboratory Identification, Enumeration and Biomass Measurements of

Periphyton in Wadeable Streams

Fact Sheet # 2.3.3 - 2 Laboratory Periphyton Biomass Determination

permeability

Fact Sheet # 2.2.2 - 20 Standard Test Method for Permeability of Granular Soils (Constant Head),

ASTM Method D2434

pesticide analysis methods

Fact Sheet # 2.1.2 - 16 Method No. LMMB: PCBs and Pesticides in Surface Water by XAD-2 Resin

Extraction

Fact Sheet # 2.2.2 - 8 Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

Fact Sheet # 2.3.2 - 8 Purification of Biological Tissue Samples by Gel Permeation

Chromatography of Organic Analyses

Fact Sheet # 2.3.2 - 10 Analysis of Polychlorinated Biphenyls and Chlorinated Pesticides by Gas

Chromatography with Electron Capture Detection, LMMB 041

pH

Fact Sheet # 2.1.2 - 29 Method No. LMMB 092: Standard Operating Procedure for Electrometric pH
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

267

phosphorus

Fact Sheet # 2.1.2 - 21 Method No. ESS Method 230.1: Total Phosphorus and Total Kjeldahl

Nitrogen, Semi-Automated Method

Fact Sheet # 2.1.2 - 22 Method No. ESS Method 310.2, LMMB 064: Phosphorus, Total, Low Level

(Persulfate Digestion)

Fact Sheet # 2.1.2 - 23 Method No. ESS Method 310.1, LMMB 063: Ortho-Phosphorus, Dissolved

Automated, Ascorbic Acid

photographic surveys

Fact Sheet # 2.3.1 - 14 Photographic Habitat Documentation of the Benthic Community

Fact Sheet # 2.3.1 - 15 Sediment Profile Cam era

phytoplankton sampling

Fact Sheet # 2.3.1 - 1 Phytoplankton Sample Collection and Preservation in the Great Lakes,

LMMB 023t

plasticity index

Fact Sheet # 2.2.2 - 23 Standard Test Method for Liquid Limit, Plastic Limit, and Plasticity Index of

Soils, ASTM Method D4318

polychlorinated biphenyls (see PCB analysis methods)

polycyclic aromatic hydrocarbons (see PAH analysis methods)

pore water

Fact Sheet # 2.1.1 - 13 In Situ Peepers

Fact Sheet # 2.1.1 - 14 Suction samplers

Fact Sheet # 2.2.1 - 10 Method No. ASTM E 1391-94: Pore Water Extraction through Centrifugation

Fact Sheet # 2.2.1 - 11 Method No. ASTM E 1391-94: Pore Water Extraction from Sediments

through Squeezing

Fact Sheet # 2.2.1 - 12 Method No. ASTM E 1391-94: Pore water extraction from sediment from

Vacuum Filtration, Gas Pressurization, or Displacement

primary productivity

Fact Sheet # 2.3.1 - 4 Primary Productivity Using 14C: Field Procedure in the Great Lakes, LMMB

016

radioisotopes

Fact Sheet # 2.2.2 - 15 Determination of the Activity of Lead-210 in Sediments and Soils, LMMB 084

Fact Sheet # 2.2.2 - 25 Sediment Age Dating Using Cesium-137

Fact Sheet # 2.2.2 - 26 Beryllium-7 as a Tracer of Short Term Sediment Deposition

Fact Sheet # 2.3.1 - 4 Primary Productivity Using 14C: Field Procedure in the Great Lakes, LMMB

016

respiration rate

Fact Sheet # 2.1.3 - 15 Method No. NHEERL-AED SOP 1.03.013: Growth and Scope for Growth

Measurements with Mytilus edulis

scope for growth (SFG) index

Fact Sheet # 2.1.3 - 15 Method No. NHEERL-AED SOP 1.03.013: Growth and Scope for Growth

Measurements with Mytilus edulis
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

268

SCUBA

Fact Sheet # 2.2.1 - 4 Hand collection at depth with SCUBA

Fact Sheet # 2.3.1 - 14 Photographic Habitat Documentation of the Benthic Community

sea floor mapping

Fact Sheet # 2.2.1 - 13 Method No. DRP-2-03: Acoustic Sub-bottom Profiling Systems

Fact Sheet # 2.2.1 - 14 Method No. EEDP-01-10: Side Scan Sonar

sediment consolidation

Fact Sheet # 2.2.1 - 15 Method No. DRP-2-3: Settlement Phases

Fact Sheet # 2.2.2 - 21 Standard Test Method for One-Dimensional Consolidation Properties of

Soil, ASTM Method D2435

sediment cores

Fact Sheet # 2.2.1 - 2 Core Samplers

Fact Sheet # 2.2.1 - 6 Russian Peat Borer

Fact Sheet # 2.2.1 - 7 Split Core Sampler for Submerged Sediments

sediment dating

Fact Sheet # 2.2.2 - 15 Determination of the Activity of Lead-210 in Sediments and Soils, LMMB 084

Fact Sheet # 2.2.2-25 Sediment Age Dating Using Cesium-137

Fact Sheet # 2.2.2-26 Beryllium-7 as a Tracer of Short Term Sediment Deposition

sediment grain size

Fact Sheet # 2.2.2 - 16 Sediment Grain Size Analysis, NHEERL-AED SOP 1.01.005

sediment flux

Fact Sheet # 2.1.1 - 19 Seepage Meters

Fact Sheet # 2.2.1 - 4 Hand collection at depth with SCUBA

sediment sampling

Fact Sheet # 2.2.1 - 1 Grab Sampling

Fact Sheet # 2.2.1 - 2 Core Samplers

Fact Sheet # 2.2.1 - 3 Hand Collection

Fact Sheet # 2.2.1 - 4 Hand collection at depth with SCUBA

Fact Sheet # 2.2.1 - 5 Sediment traps

Fact Sheet # 2.2.1 - 6 Russian Peat Borer

Fact Sheet # 2.2.1 - 7 Split Core Sampler for Submerged Sediments

Fact Sheet # 2.3.1 - 11 Benthic Organism Collection from a Marine Environm ent, NHEERL-AED

SOP 1.02.001

Fact Sheet # 2.3.1 - 12 Benthic Macroinvertebrate Protocols in a Wadeable Stream: Single Habitat

Approach, 1-Meter Kick Net

Fact Sheet # 2.3.1 - 13 Benthic Macroinvertebrate Protocols in a Wadeable Stream: Multi-habitat

Approach: D-Frame Dip Net

sediment toxicity

Fact Sheet # 2.2.3 - 1 Acute Freshwater Crustacean Sediment Bioassay: Flow-through

Fact Sheet # 2.2.3 - 3 Acute Freshwater Crustacean Sediment Bioassay: Static Laboratory

Exposures

Fact Sheet # 2.2.3 - 4 Acute Freshwater Amphipod and Freshwater Insect Larvae Sediment

Bioassay, EPA Method 100.1

Fact Sheet # 2.2.3 - 5 Chronic Freshwater Amphipod Sediment Bioassay, EPA Method 100.4

Fact Sheet # 2.2.3 - 6 Life-Cycle Freshwater Midge Sediment Bioassay, EPA Method 100.5

Fact Sheet # 2.2.3 - 7 Acute Larval Bivalve Sediment Bioassay
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

269

Fact Sheet # 2.2.3 - 8 Acute Echinoderm Sediment Bioassay

Fact Sheet # 2.2.3 - 9 Acute Marine Crustacean Sediment Bioassay

Fact Sheet # 2.2.3 - 10 Acute Marine Amphipod Crustacean Sediment Bioassay, EPA Method

100.4

Fact Sheet # 2.2.3 - 11 Acute Marine Polychaete Sediment Bioassay, ASTM Method E1611-00

Fact Sheet # 2.2.3 - 12 Chronic Estuarine Amphipod Sediment Bioassay

Fact Sheet # 2.2.3 - 13 Chronic Marine Polychaete Sediment Bioassay, ASTM Method E1611-00

Fact Sheet # 2.2.3 - 14 Ames Mutagenicity Assay

Fact Sheet # 2.2.3 - 15 Mutatox Genotoxicity Assay

Fact Sheet # 2.2.3 - 16 V79 Sister Chromatid Exchange Assay, NHEERL-AED SOP 1.03.012

Fact Sheet # 2.2.3 - 17 Bioaccumulation Test for Marine, Estuarine, and Freshwater Sediments,

EPA Method 100.3

sediment water content

Fact Sheet # 2.2.2 - 17 Procedures for Water Content Determination

selected ion monitoring (SIM)

Fact Sheet # 2.2.2 - 7 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS)--Selected Ion Monitoring

(SIM) Mode

Fact Sheet # 2.3.2 - 9 Quantitative Determination of Polynuclear Aromatic Hydrocarbons by Gas

Chromatography/Mass Spectrom etry (GC/MS)--Selected Ion Monitoring

(SIM) Mode

selenium

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 4 EPA Method No. 1639: Determination of Trace Elem ents in Ambient Waters

by Stabilized Tem perature Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

semi-volatile organic compounds

Fact Sheet # 2.1.2 - 13 EPA Method No. 1625: Semi-volatile Organic Compounds by Isotope

Dilution GC/MS

Fact Sheet # 2.2.2 - 6 Extraction and Clean-up of Sediments for Semi-volatile Organics Following

the Internal Standard Method, LMMB 040

Fact Sheet # 2.3.2 - 16 Microwave Extraction of Marine Tissue for Semivolatile Organic Analytes,

AED LOP 2.03.030,

Revision 0

settlement plates

Fact Sheet # 2.2.1 - 15 Method No. DRP-2-3: Settlement Phases

settling particulate matter

Fact Sheet # 2.2.1 - 5 Sediment traps
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

270

shear strength

Fact Sheet # 2.2.2 - 18 Standard Test Method for Field Vane Shear Test in Cohesive Soil, ASTM

Method D2573

side scan sonar

Fact Sheet # 2.2.1 - 14 Method No. EEDP-01-10: Side Scan Sonar

silt

Fact Sheet # 2.2.2 – 16 Sediment Grain Size Analysis, NHEERL-AED SOP 1.01.005

silicon

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

silver

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

simultaneously extracted metals (SEM)

Fact Sheet # 2.2.2 - 4 Determination of Acid Volatile Sulfide and Selected Sim ultaneously

Extractable Metals in Sediment

soil classification

Fact Sheet # 2.2.2 - 18 Standard Test Method for Field Vane Shear Test in Cohesive Soil, ASTM

Method D2573

Fact Sheet # 2.2.2 - 19 Standard Test Method for Specific Gravity of Soil Solids by Water

Pycnometer, ASTM Method D854

Fact Sheet # 2.2.2 - 20 Standard Test Method for Permeability of Granular Soils (Constant Head),

ASTM Method D2434

Fact Sheet # 2.2.2 - 21 Standard Test Method for One-Dimensional Consolidation Properties of

Soil, ASTM Method D2435

Fact Sheet # 2.2.2 - 22 Standard Test Method for Classification of Soils for Engineering Purposes

(Unified Soil Classification System), ASTM Method D2487

Fact Sheet # 2.2.2 - 23 Standard Test Method for Liquid Limit, Plastic Limit, and Plasticity Index of

Soils, ASTM Method D4318

specific gravity

Fact Sheet # 2.2.2 - 19 Standard Test Method for Specific Gravity of Soil Solids by Water

Pycnometer, ASTM Method D854

stream characterization

Fact Sheet # 2.1.1 - 15 Physical Characterization of a stream

Fact Sheet # 2.1.1 - 16 Visual based habitat assessment

Fact Sheet # 2.1.1 - 17 LMMB 017: USGS Field Operation Plan: Tributary Monitoring

sub-bottom profiling

Fact Sheet # 2.2.1 - 13 Method No. DRP-2-03: Acoustic Sub-bottom Profiling Systems
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

271

sulfate

Fact Sheet # 2.1.2 - 28 Method No. ESS Method 200.5: Determination of Inorganic Anions in Water

by Ion Chromatography

thallium

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

tin

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

total organic carbon (TOC)

Fact Sheet # 2.1.2 - 24 Standard Method No. 5310: Total Organic Carbon

Fact Sheet # 2.2.2 - 14 Procedures for Sediment Total Organic Carbon (TOC) Determination

total suspended solids (TSS)

Fact Sheet # 2.1.2 - 32 Method No. LMMB 090: Standard Operating Procedure for GLNPO

Turbidity: Nephelometeric Method

Fact Sheet # 2.1.2 - 33 Method No. LMMB 065: ESS Method 340.2: Total Suspended Solids, Mass

Balance (Dried at 103-105EC) Volatile Suspended Solids (Ignited at 550EC)

toxicity testing

Fact Sheet # 2.1.3 - 1 Method No. ERT SOP 2024: Acute Freshwater Crustacean Bioassay: 48

Hours

Fact Sheet # 2.1.3 - 2 Method No. ERT SOP 2022: Acute Freshwater fish Bioassay

Fact Sheet # 2.1.3 - 3 Method No. ERT SOP 2027: Chronic Freshwater Algae Test

Fact Sheet # 2.1.3 - 4 Method No. ERT SOP 2025: Chronic Freshwater Crustacean Bioassay

(7day)

Fact Sheet # 2.1.3 - 5 Method No. ERT SOP 2028: Chronic Freshwater Crustaceans Bioassay (10

days)

Fact Sheet # 2.1.3 - 6 Method No. ERT SOP 2026: Chronic Freshwater Fish Bioassay, ERT SOP

2026

Fact Sheet # 2.1.3 - 7 Method No. NHEERL-AED SOP 1.03.001: Chronic Marine Macroalgae,

Champia parvula, Sexual Reproduction test

Fact Sheet # 2.1.3 - 8 Method No. NHEERL-AED SOP 1.03.003: Acute Marine Crustacean

Bioassay

Fact Sheet # 2.1.3 - 9 Method No. NHEERL-AED SOP 1.03.003: Acute Marine Fish Bioassay

Fact Sheet # 2.1.3 - 10 Method No. NHEERL-AED SOP 1.03.005: Chronic Estuarine Survival,

Growth, and Fecundity Test

Fact Sheet # 2.1.3 - 11 Method No. NHEERL-AED SOP 1.03.006: Chronic Echinoderm Fertilization

Test

Fact Sheet # 2.1.3 - 12 Method No. NHEERL-AED SOP 1.03.004: Chronic Marine Fish Bioassay

Fact Sheet # 2.1.3 - 13 Toxicity Evaluations of Photoinduction of Polycyclic Aromatic Hydrocarbons

(PAH): In Situ Analysis

Fact Sheet # 2.1.3 - 14 Toxicity Evaluations of Photoinduction of Polycyclic Aromatic Hydrocarbons:

Laboratory Analysis of Storm water
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

272

Fact Sheet # 2.1.3 - 16 Method No. NHEERL-AED SOP 1.03.009: Microtox® tests, NHEERL-AED

1.03.009

Fact Sheet # 2.2.3 - 1 Acute Freshwater Crustacean Sediment Bioassay: Flow-through

Fact Sheet # 2.2.3 - 2 Acute Freshwater Crustacean Sediment Bioassay: In Situ Exposures

toxicology testing - sample prep

Fact Sheet # 2.2.1 - 8 Sediment Processing for Chem istry and Toxicity Testing

Fact Sheet # 2.2.1 - 9 Sediment Processing for Elutriate Toxicity tests

Fact Sheet # 2.2.1 - 10 Method No. ASTM E 1391-94: Pore Water Extraction through Centrifugation

Fact Sheet # 2.2.1 - 11 Method No. ASTM E 1391-94: Pore Water Extraction from Sediments

through Squeezing

Fact Sheet # 2.2.1 - 12 Method No. ASTM E 1391-94: Pore water extraction from sediment from

Vacuum Filtration, Gas Pressurization, or Displacement

trace metals analysis

Fact Sheet # 2.1.2 - 1 EPA Method No. 245.7: Mercury in Water by Cold Vapor Atomic

Fluorescence Spectrom etry

Fact Sheet # 2.1.2 - 2 EPA Method No. 1631, Revision B: Mercury in Water by Oxidation, Purge

and Trap, and Cold Vapor Atomic Fluorescence Spectrom etry

Fact Sheet # 2.1.2 - 3 EPA Method No. 1630: Methyl mercury in water by distillation, Aqueous

Ethylation, Purge and Trap, and CVAFS

Fact Sheet # 2.1.2 - 4 EPA Method No. 1639: Determination of Trace Elem ents in Ambient Waters

by Stabilized Tem perature Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 5 EPA Method No. 1637: Determination of Trace Elem ents in Ambient Waters

by Off-Line Chelation Pre-concentration and Stabilized Tem perature

Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.1.2 - 7 EPA Method No. 1640: Determination of Trace Elem ents in Ambient Waters

by On-Line Chelation Pre-concentration and Inductively Coupled PlasmaMass Spectrom etry

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Trace Element Quantification Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

transparency

Fact Sheet # 2.1.1-4 In Situ Transparency Sampling

trawling

Fact Sheet # 2.3.1 - 25 Trawls

turbidity

Fact Sheet # 2.1.2 - 32 Method No. LMMB 090: Standard Operating Procedure for GLNPO

Turbidity: Nephelometeric Method

volatile organic compounds (VOC)

Fact Sheet # 2.1.2 - 12 Method No. OERR SOP #2109: Photovac GC Analysis for Soil, Water, and

Air/Soil Gas

Fact Sheet # 2.2.2 - 5 Photovac GC Analysis for Soil, Water, and Air/Soil Gas, OSW ER SOP#

2109

water column characterization
A Compendium of Chemical, Physical and Biological Methods 

for Assessing and Monitoring the Remediation of Contaminated Sediment Sites February 17, 2003

273

Fact Sheet # 2.3.3 - 4 Laboratory Analysis of Water Column Organisms

water pycnometer

Fact Sheet # 2.2.2 - 19 Standard Test Method for Specific Gravity of Soil Solids by Water

Pycnometer, ASTM Method D854

water velocity

Fact Sheet # 2.1.1 - 18 Quality Assurance Plan for Discharge Measurements Using Broadband

Acoustic Doppler Current Profilers

x-ray fluorescence

Fact Sheet # 2.2.2 - 3 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

Fact Sheet # 2.2.2 - 24 Field Portable X-Ray Fluorescence Spectrom etry for the Determination of

Elemental Concentrations in Soil and Sediment

zinc

Fact Sheet # 2.1.1 - 11 Method No. EPA Method 1669: Sampling Ambient Water for Trace Metals

at EPA Water Quality Criteria Levels

Fact Sheet # 2.1.2 - 4 EPA Method No. 1639: Determination of Trace Elem ents in Ambient Waters

by Stabilized Tem perature Graphite Furnace Atomic Absorption

Fact Sheet # 2.1.2 - 6 EPA Method No. 1638: Determination of Trace Elem ents in Ambient Waters

by Inductively Coupled Plasma — Mass Spectrom etry

Fact Sheet # 2.2.2 - 2 Trace Element Quantification Techniques

Fact Sheet # 2.2.2 - 3 Method Title: Analysis of Marine Sediment and Bivalve

Tissue by X-Ray Fluorescence, Atomic Absorption and Inductively Coupled

Plasma Mass Spectrom etry

Fact Sheet # 2.3.2 - 4 Method No. NS&T, Method Title: Trace Element Quantification

Techniques

Fact Sheet # 2.3.2 - 5 Analysis of Marine Sediment and Bivalve Tissue by X-Ray Fluorescence,

Atomic Absorption and Inductively Coupled Plasma Mass Spectrom etry

zooplankton sampling

Fact Sheet # 2.3.1 - 5 Zooplankton Sample Collection and Preservation in the Great Lakes, LMMB

024
National Risk Management

Research Laboratory

Cincinnati, OH 45268

Official Business

Penalty for Private Use

$300

EPA/600/R-03/108

April 2004

National Risk Management

Research Laboratory

Cincinnati, OH 45268

Official Business

Penalty for Private Use

$300

EPA/600/R-03/108

April 2004

Please make all necessary changes on the below label,

detach or copy, and return to the address in the upper

left-hand corner.

If you do not wish to receive these reports CHECK HERE ;

detach, or copy this cover, and return to the address in the

upper left-hand corner.

Please make all necessary changes on the below label,

detach or copy, and return to the address in the upper

left-hand corner.

If you do not wish to receive these reports CHECK HERE ;

detach, or copy this cover, and return to the address in the

upper left-hand corner.

Preso

Posta

Per