Techniques of Water-Resources Investigations

of the United States Geological Survey

Chapter A3

METHODS FOR THE DETERMINATION OF

ORGANIC SUBSTANCES IN WATER

AND FLUVIAL SEDIMENTS

Edited by R. L. Wershaw, M. J. Fishman,

R. R. Grabbe, and L. E. Lowe

This manual is a revision of "Methods for Analysis of Organic Substances in Water," by

Donald F. Goerlitz and Eugene Brown, Book 5, Chapter A3, published in 1972.

Book 5

LABORATORY ANALYSIS
DEPARTMENT OF THE INTERIOR

DONALD PAUL HODEL, Secretary

U.S . GEOLOGICAL SURVEY

Dallas L. Peck, Director

UNITED STATES GOVERNMENT PRINTING OFFFICE, WASHINGTON : 1987

For sale by the Books and Open-File Reports Section, U.S . Geological Survey,

Federal Center, Box 25425, Denver, CO 80225
PREFACE

This series of manuals on techniques describes methods used by the U.S . Geological

Survey for planning and executing water-resources investigations. The material is

grouped under major subject headings called books and is further subdivided into

sections and chapters. Book 5 is on laboratory analyses. Section A is on water. The unit

of publication, the chapter, is limited to a narrow field of subject matter. "Methods for

the Determination of Organic Substances in Water and Fluvial Sediments" is the third

chapter to be published under Section A of Book 5. The chapter number includes the

letter of the section.

This manual was prepared by many chemists and hydrologists of the U.S . Geological

Survey and provides accurate and precise methods for the analysis of water,

water-suspended-sediment mixtures, and bottom-material samples . Supplements, to be

prepared as the need arises, will be issued as they become available.

Reference to trade names, commercial products, manufacturers, or distributors does

not constitute endorsement by the U.S. Geological Survey or recommendation for use .

This manual is a revision of "Methods for Analysis of Organic Substances in Water"

by D.F. Goerlitz and Eugene Brown (U.S . Geological Survey Techniques of WaterResources Investigations, Book 5, Chapter A3, 1972).
TECHNIQUES OF WATER-RESOURCES INVESTIGATIONS OF THE

U.S . GEOLOGICAL SURVEY

The U.S . Geological Survey publishes a series of manuals describing procedures for planning and conducting

specialized work in water-resources investigations . The manuals published to date are listed below and may be ordered

by mail from the U.S. Geological Survey, Books and Open-File Reports, Federal Center, Building 810, Box 25425, Denver,

Colorado 80225 an authorized agent of the Superintendent of Documents, Government Printing Office) .

Prepayment is required . Remittance should be sent by check or money order payable to U.S . Geological Survey. Prices

are not included in the listing below as they are subject to change. Current prices can be obtained by writing to the USGS,

Books and Open File Reports. Prices include cost of domestic surface transportation . For transmittal outside the U.S .A.

(except to Canada and Mexico) a surcharge of 25 percent of the net bill should be included to cover surface

transportation. When ordering any of these publications, please give the title, book number, chapter number, and "U.S .

Geological Survey Techniques of Water-Resources Investigations."

TWI 1-D1. Water temperature-influential factors., field measurement, and data presentation, by H. H. Stevens, Jr ., J. F. Ficke, and G. F. Smoot,

1975, 65 pages.

TWI 1-D2. Guidelines for collection and field analysis of ground-water samples for selected unstable constituents, by W. W. Wood. 1976. 2 4

pages.

TWI 2-Dl. Application ofsurface geophysics to groundwater investigations, by A. A. R. Zohdy, G. P. Eaton, and D. R. Mabey. 1974 . 11 6 pages.

TWI 2-El . Application of borehole geophysics to water-resources investigations, by W. S. Keys and L. M. MacCary. 1971 . 126 pages.

TWI 3-Al. General field and office procedures for indirect discharge measurement, by M. A. Benson and Tate Dalrymple . 1967 . 30 pages.

TWI 3-A2. Measurement of peak discharge by the slope-area method, by Tate Dalrymple and M. A. Benson. 1967. 1 2 pages.

TWI 3-A3. Measurement of peak discharge at culverts by indirect methods, by G. L. Bodhaine. 1968. 6 0 pages .

TWI 3-A4. Measurement of peak discharge at width contractions by indirect methods, by H. F. Matthai . 1967. 44 pages.

TWI 3-A5. Measurement of peak discharge at dams by indirect methods, by Harry Hulsing. 1967. 29 pages.

TWI 3-A6. General procedure for gaging streams, by R. W. Carter and Jacob Davidian . 1968. 13 pages.

TWI 3-A7. Stage measurements at gaging stations, by T. J . Buchanan and W. P. Somers. 1968 . 28 pages.

TWI 3-A8. Discharge measurements at gaging stations, by T. J. Buchanan and W. P. Somers. 1969 . 65 pages.

TWI 3-A9. Measurement of time of travel and dispersion in streams by dye tracing, by E. P. Hubbard, F. A. Kilpatrick, L. A. Martens, and J .

F. Wilson, Jr. 1982. 44 pages .

TWI 3-All. Measurement of discharge by moving-boat method, by G. F. Smoot and C. C. Novak. 1969. 22 pages.

TWI 3-A12. Fluorometric procedures for dye tracing, Revised, by James F. Wilson, Jr., Ernest D. Cobb, and Frederick A. Kilpatrick . 1986 . 41

pages.

TWI 3-A13. Computation of continuous records of streamflow, by Edward J. Kennedy . 1983 . 53 pages.

TWI 3-A14. Use of flumes in measuring discharge, by F. A. Kilpatrick, and V. R. Schneider. 1983 . 46 pages.

TWI 3-A15. Computation of water-surface profiles in open channels, by Jacob Davidian . 1984. 48 pages.

TWI 3-Bl. Aquifer-test design, observation, and data analysis, by R. W. Stallman . 1971 . 26 pages.

TWI 3-B2. Introduction to ground-water hydraulics, a programmed text for self-instruction, by G. D. Bennett . 1976. 17 2 pages .

TWI 3-B3. Type curves for selected problems of flow to wells in confined aquifers, by J . E. Reed. 1980. 106 p.

TWI 3-Cl. Fluvial sediment concepts, by H. P. Guy. 1970 . 55 pages.

TWI 3-C2. Field methods of measurement of fluvial sediment, by H. P. Guy and V. W. Norman. 1970. 5 9 pages .

TWI 3-C3. Computation of fluvial-sediment discharge, by George Porterfield. 1972. 66 pages.

TWI 4-Al. Some statistical tools in hydrology, by H. C. Riggs. 1968 . 39 pages.

TWI 4-A2. Frequency curves, by H. C. Riggs, 1968. 15 pages.

TWI 4-Bl. Low-flow investigations, by H. C. Riggs . 1972. 1 8 pages.

TWI 4-B2 Storage analyses for water supply, by H. C. Riggs and C. H. Hardison. 1973 . 20 pages .

TWI 4-B3. Regional analyses of streamflow characteristics, by H. C. Riggs. 1973. 1 5 pages .

TWI 4-D1. Computation of rate and volume of stream depletion by wells, by C. T. Jenkins. 1970. 1 7 pages.

TWI 5-Al. Methods for determination of inorganic substances in water and fluvial sediments, by M. W. Skougstad and others, editors. 1979. 626

pages.

TWI 5-A2. Determination of minor elements in water by emission spectroscopy, by P. R. Barnett and E. C. Mallory, Jr . 1971 . 31 pages.

TWI 5-A3. Methods for the determination of organic substances in water and fluvial sediments, edited by R. L. Wershaw, M. J . Fishman, R. R.

Grabbe, and L. E. Lowe. 1987. 80 pages.

TWI 5-A4. Methods for collection and analysis of aquatic biological and microbiological samples, edited by P. E. Greeson, T. A. Ehlke, G. A.

Invin, B. W. Lium, and K. V. Slack . 1977 . 332 pages.

TWI 5-A5. Methods for determination of radioactive substances in water and fluvial sediments, by L. L. Thatcher, V. J . Janzer, and K. W.

Edwards. 1977. 95 pages.

TWI 5-A6. Quality assurance practices for the chemical and biological analyses of water and fluvial sediments, by L. C. Friedman and D. E.

Erdmann . 1982. 181 pages.
TWI 5-Cl. Laboratory theory and methods for sediment analysis, by H. P. Guy. 1969. 5 8 pages.

TWI 7-Cl. Finite difference model for aquifer simulation in two dimensions with results of numerical experiments, by P. C. Trescott, G. F.

Pinder, and S. P. Larson. 1976. 11 6 pages.

TWI 7-C2. Computer model of two-dimensional solute transport and dispersion in ground water, by L. F. Konikow and J . D. Bredehoeft. 1978 .

90 pages.

TWI 7-C3. A model for simulation of flow in singular and interconnected channels, by R. W. Schaffranek, R. A. Baltzer, and D. E. Goldberg.

1981 . 110 pages.

TWI 8-Al . Methods of measuring water levels in deep wells, by M. S. Garber and F. C. Koopman. 1968. 23 pages.

TWI 8-A2. Installation and service manual for U.S . Geological Survey monometers, by J. D. Craig. 1983 . 57 pages .

TWI 8-B2. Calibration and maintenance of vertical-axis type current meters, by G. F. Smoot and C. E. Novak. 1968. 1 5 pages.
Charlene W. Bayer

Michael G. Brooks

Rosalyn Carter

Marvin J. Fishman

Linda C. Friedman

Rolland R. Grabbe

Robert J. Keck

Jerry A. Leenheer

James A. Lewis

Luis E. Lowe

Bernard A. Malo

Douglas B. Manigold

Janice L. O'Byrne

CONTRIBUTORS

Mary C. Olson

Deanna L. Peterson

Janet B. Pruitt

Paul K. Roscio

Michael P. Schroeder

Thomas R. Steinheimer

Marguerite B. Sweny

Ronald S. Torbet

John D. Vaupotic

Dorothy M. Walker

Robert L. Wershaw

William R. White

Marvin L. Yates
Preface ------------------------------

Abstract -----------------------------

Introduction--------------------------

Purpose ----------------------------

Scope------------------------------

Definitions ---------------------------

Significant figures ---------------------

Recovery correction --------------------

Quality control -------------------------

Qualification of the analyst ---------------

Reagents and standards -----------------

Equipment ---------------------

Analytical procedure -------------------

Use and documentation of standard laboratory

procedures ------------------------

Approved methods --------------------

Official methods ---------------------

Provisional methods ------------------

Special methods ----------------------

Sample collection -----------------------

Preparation of the samplers --------------

Sample handling and preservation ----------

Extraction, fractionation, and identification -----

Extraction --------------------------

Batch extraction---------------------

Continuous extraction -----------------

Fractionation ------------------------

Liquid chromatography ----------------

Adsorption -----------------------

Liquid-liquid partition----------------

Ion exchange ---------------------

Gel permeation --------------------

Gas-liquid chromatography --------------

Identification ------------------------

Thermal-conductivity detector -----------

Flame-ionization detector ---------------

Electron-capture detector --------------

Mass spectrometer -------------------

Other detectors ---------------------

Spectrophotometer -------------------

Quantitative analysis ----------------

Qualitative analysis - - - - - - - - - - - - - - - - -

Ancilliary methods of confirmation ----------

Selected references --------------------

Analytical methods ----------------------

Carbon, organic, dissolved, wet oxidation

(0-1100-83) ------------------------

Carbon, organic, total, wet oxidation

(0-3100-83) -------------------------

Carbon, organic, suspended, wet oxidation

(0-7100-83) ------------------------

Carbon, inorganic plus organic, total in bottom

material, dry weight, induction furnace

(0-5101-83) ------------------------

Carbon, inorganic, total in bottom material, modified Van Slyke (0-5102-83) -------------

CONTENTS

Page

III

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Page

Analytical methods-Continued

Carbon, organic, dissolved, fractionation

(0-1103-83) ------------------------

Organochlorine and organophosphorous compounds, total recoverable (0-3104-83) and dissolved (0-1104-83), gas chromatographic ------ 27

Organochlorine and organophosphorous compounds, recoverable from bottom material

(0-5104-83) and recoverable from suspended

sediment (0-7104-83), gas chromatographic - - 31

Organochlorine compounds, recoverable from fish

tissue, gas chromatographic

(0-9104--83) ------------------------

Chlorophenoxy acids, total recoverable

(0-3105-83) and dissolved (0-1105-83), gas

chromatographic --------------------- 40

Chlorophenoxy acids, recoverable from bottom

material (0-5105-83) and recoverable from suspended sediment (0-7105-83), gas chromatographic --------------------------

Triazines, total recoverable, gas chromatographic (0-3106-83) -------------------

Carbamate pesticides, total recoverable, high-performance liquid chromatographic

(0-3107-83) ------------------------ 49

Oil and grease, extractable, extraction-gravimetric

(0-3108--83) ------------------------ 51

Oil and grease, extractable from bottom material,

extraction-gravimetric (0-5108-83) --------- 53

Fuel oils, light, total recoverable, gas chromatographic (0-3109-83) ------------------- 54

Phenols, total recoverable, colorimetric, 4-

aminoantipyrine (0-3110-83) - - - - - - - - - - - - - 55

Methylene blue active substances, total recoverable, colorimetric (0-3111-83) ------------ 57

TNT, RDX, and picric acid, total recoverable,

high-performance liquid chromatographic

(0-3112_83) ------------------------ 58

Polynuclear aromatic hydrocarbons (PNA), total

recoverable, high-performance liquid chromatographic (0-3113-83) -------------------

Ethylene and propane, total recoverable, gas

chromatographic, purge and trap

(0-3114--83) ------------------------

Purgeable organic compounds, total recoverable,

gas chromatographic/mass spectrometric, purge

and trap (0-3115-83) ------------------ 67

Acid extraction compounds, total recoverable, gas

chromatographic/mass spectrometric

(0-3117-83) ------------------------

Base/neutral extractable compounds, total recoverable, gas chromatographic/mass spectrometric

(0-3118-83) ------------------------ 76

23

35

43

46

60

63

71
FIGURES

TABLES

Page

1 . Concentrations of pesticides and PCB's in mixed standard solutions used for gas chromatograph calibration of water and water-suspended sediment --------------------- 29

2. Column fractionation scheme for silica gel column for organochlorine insecticides, PCB's, and

PCN's ----------------------------------------------------------------- 29

3. Column fractionation scheme for alumina and silica columns for insecticides, PCB's, and PCB's

in bottom material and suspended sediment ------------------------------------ 32

4. Concentrations of pesticides and PCB's in mixed standard solutions used for gas chromatograph calibration of bottom material -------------------------------------- 33

5. Column fractionation scheme for alumina and silica columns for organochlorine insecticides,

PCB's, and PCN'sinfish --------------------------------------------------- 36

6. Concentrations of pesticides and PCB's in mixed standard solutions used for gas chromatograph calibration of fish tissue ------------------------------------------ 37

7. Concentration of herbicides in mixed standard solutions used for gas chromatograph calibration

of water and water-suspended sediment --------------------------------------- 41

8. Concentration of herbicides in mixed standard solutions used for gas chromatograph calibration

of bottom material-------------------------------------------------------- 44

9 . Concentration of triazines in mixed standard solutions used for gas chromatograph calibration - 47

10 . Concentration of carbamates in mixed standard solutions used for liquid

chromatograph calibration ------------------------------------------------- 50

Page

1 . Diagram of apparatus for the manometric determination of carbon dioxide ---------------- 20

2. Flow chart showing dissolved organic carbon fractionation analytical scheme -------------- 24

3, 4 . Diagrams of:

3 . Purge vessel --------------------------------------------------------- 64

4 . Stripping and gas chromatographic system in the stripping mode ---------------- 66
METHODS FOR THE DETERMINATION OF

ORGANIC SUBSTANCES IN WATER

AND FLUVIAL SEDIMENTS

Abstract

This manual describes analytical methods used by the U.S.

Geological Survey to determine organic substances in water,

water-suspended-sediment mixtures, and bottom material. Some

of the analytical procedures yield determinations for specific compounds, whereas others provide a measure of the quantity of

groups of compounds present in the sample . Examples of the first

category are procedures for the organochlorine and organophosphate insecticides, chlorophenoxy acid and triazine herbicides,

and specific substituted phenols . Examples of the second category

are the various organic carbon analyses and the polychlorinated

biphenyl methods . The analytical methods are presented in a

standard format; topics covered include conditions for application

of the method, a summary of the method, interferences, required

apparatus and reagents, analytical procedures, calculations, reporting of results, and estimation of precision.

Introduction

The U.S . Department of the Interior has a basic

responsibility for the appraisal, conservation, and

efficient use of the Nation's natural resources-ineluding water as a resource as well as water involved in the use and development of other resources. As one of several Department of Interior

agencies, the U.S . Geological Survey's primary function in relation to water is to assess its availability

and utility as a national resource for all uses. The

Geological Survey's responsibility for water appraisal includes not only assessments of the location,

quantity, and availability of water, but also determinations of water quality. Inherent in this responsibility is a need for extensive water-quality studies reEdited by R .L . Wershaw, M .J . Fishman,

R .R. Grabbe, and L.E. Lowe

lated to the physical, chemical, and biological

adequacy of natural and developed surface- and

ground-water supplies. Included, also, is a need for

supporting research to increase the effectiveness of

these studies.

As part of its mission, the Geological Survey is

responsible for generating a large part of the waterquality data for rivers, lakes, and ground water that

are used by planners, developers, water-quality

managers, and pollution-control agencies. A high

degree of reliability and standardization of these

data is paramount .

This manual is one chapter in a series prepared to

document and make available data-collection and

analysis procedures used by the Geological Survey.

The series describes procedures for planning and

executing specialized work in water-resources investigations. The unit of publication, the chapter, is

limited to a narrow field . This format permits flexibility in revision and publication as the need arises .

For convenience, the chapters on methods of waterquality analysis are grouped into the following categories: inorganic substances, minor elements by

emission spectroscopy, organic substances, aquatic

biological and microbiological samples, radioactive

substances, and quality assurance .

Provisional drafts describing new or revised analytical methods are distributed to field offices of the

Geological Survey for their use . These drafts are

subject to revision based on use or because of advancement in knowledge, techniques, or equipment .

After a method is sufficiently developed and confirmed, it is described in a supplement to the chapter
2

or in a new edition of the chapter, and the publication is available from the Superintendent of Documents, U.S. Government Printing Office, Washington, DC 20402.

Purpose

The purpose of this manual is to record and disseminate methods used by the U.S . Geological Survey to analyze samples of water, water-suspendedsediment mixtures, and bottom material collected in

connection with ongoing water-quality investigations. The manual is an update and enlargement of

Techniques of Water-Resources Investigations of

the U.S . Geological Survey, Book 5, Chapter A3,

"Methods for Analysis of Organic Substances in

Water," by D.F. Goerlitz and Eugene Brown, published in 1972. Of special note is the present manual's inclusion of methods for analyzing samples of

water-suspended-sediment mixtures and of bottom

material collected from streams, lakes, and reservoirs.

Although excellent and authoritative manuals on

water analysis are available, most of them emphasize either municipal, industrial, or agricultural

water use. No single reference or combination of

references is adequate to serve as a comprehensive

guide to the broader phases of water-quality investigations conducted by the U.S . Geological Survey.

These investigations are intended to define the

chemical, physical, and biological characteristics of

the Nation's surface- and ground-water resources,

as well as to indicate the suitability of these resources for various beneficial uses.

Rapid changes in technology are constantly providing new and improved methods of studying

water-quality characteristics. Methods manuals

must be revised more frequently than before so as to

gain the advantages of improved technology and to

obtain water-quality data in the most efficient manner possible, with a high degree of quality control to

ensure nationwide uniformity and standardization of

data.

Scope

This manual describes techniques and procedures

found to be suitable for analyzing representative

samples of water and fluvial sediments for dissolved

and sorbed organic constituents . Because of the typTECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

ically low concentration and complex matrices of

samples, the procedures include pretreatment steps

to increase concentration and to remove interfering

substances. The techniques involving laboratory

equipment represent the current state of technology.

For each method, the following topics are covered :

application, principles of the method, interferences,

apparatus and reagents required, details of the analytical procedure, calculations, reporting of results

(units and significant figures), and analytical precision data, when available. Each method, where applicable, applies to the determination of constituents

in solution (dissolved), the determination of total or

total recoverable constituents (substances both in

solution and adsorbed. on or a part of suspended

sediment), and, finally ;, the determination of total or

recoverable constituents from samples of bottom

material .

Associated with each method is one or more fourdigit identifying numbers preceded by the letter "0."

The letter indicates that the method applies to an

organic substance ; identifying numbers in other

chapters in this series are preceded, for example, by

"P" (for physical characteristic), "I" (for an inorganic substance), "R" (for a radioactive substance), or

"B" (for a biological characteristic or determination) . The first digit of the identifying number indicates the type of determination (or procedure) for

which the method is suitable, as follows :

0 ------Sample preparation .

1 ------Manual method for dissolved constituents.

2 ------Automated method for dissolved constituents.

3 ------Manual method for analyzing watersuspended-sediment mixtures .

4 ------Automated method for analyzing watersuspended-sediment mixtures.

5 ------Manual method for analyzing samples of

bottom material.

6 ------Automated method for analyzing samples

of bottom material.

7 ------Method for suspended constituents.

9 ------Method for fish and other materials.

The second, third, and fourth digits are unique to

each method.

Following each identifying number is a two-digit

number that indicates the year of last approval of

the method. If revisions of a method are issued within the calendar year of last approval, suffixes A, B,

and so forth are added to the year designation to

identify such subsequent revisions. This numbering
DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS 3

system simplifies unequivocal identification of each

method and also simplifies updating of the manual

as new or revised methods are introduced.

Definitions

Reporting the results of analyses of water and

fluvial-sediment samples requires the use of a number of terms that are based on the combination of

physical phases sampled (water or sediments) and

analytical methods used. These terms are defined

below.

Dissolved.-Pertains to the constituents in a representative water sample that pass through a 0.45-

micrometer membrane filter . This is a convenient

operational definition used by Federal agencies

that collect water data. Determinations of "dissolved" constituents are made on subsamples of

the filtrate .

Suspended, recoverable.-Pertains to the constituents extracted from the suspended sediment that

is retained on a 0.45-micrometer membrane filter .

Complete extraction is generally not achieved,

and thus the determination represents something

less than the "total" amount (that is, less than 95

percent) of the constituent present in the suspended phase of the sample. To achieve comparability

of analytical data, all laboratories performing

such analyses would have to use equivalent extraction procedures, because different extraction

procedures are likely to produce different analytical results .

Determinations of "suspended, recoverable"

constituents are made either by analyzing portions of the material collected on the filter or,

more commonly, by computing the difference between (1) dissolved and (2) total recoverable concentrations of the constituent .

Suspended, total.-Pertains to the constituents of

the suspended sediment that is retained on a 0.45-

micrometer membrane filter. This term is used

only when the analytical procedure ensures measurement of at least 95 percent of the constituent

determined. Knowledge of the expected form of

the constituent in the sample, as well as of the

analytical methodology used, is required to determine when the results should be reported as "suspended, total ."

Determinations of "suspended, total" constituents are made either by analyzing portions of the

material collected on the filter or, more commonly, by computing the difference between (1) dissolved and (2) total concentrations of the constituent.

Total, recoverable .-Pertains to the constituents

in solution after a representative watersuspended-sediment sample is digested (usually

using a dilute acid solution) . Complete dissolution

of all particulate matter is often not achieved by

the digestion treatment, and thus the determination represents something less than the "total"

amount (that is, less than 95 percent) of the constituent present in the dissolved and suspended

phases of the sample. To achieve comparability of

analytical data, all laboratories performing such

analyses would have to use equivalent digestion

procedures, because different digestion procedures are likely to produce different analytical

results .

Total, recoverable also pertains to the constituents extracted from a representative watersuspended-sediment sample. Complete extraction

generally is not achieved, and thus the determination represents something less than the "total"

amount (that is, less than 95 percent) of the constituent present in the dissolved and suspended

phases of the sample. To achieve comparability of

analytical data, all laboratories performing such

analyses would have to use equivalent extraction

procedures, because different extraction procedures are likely to produce different analytical

results .

Total.-Pertains to the constituents in a representative water-suspended-sediment sample. This

term is used only when the analytical procedure

ensures measurement of at least 95 percent of the

constituent present in both the dissolved and suspended phases of the sample. Knowledge of the

expected form of the constituent in the sample, as

well as of the analytical methodology used, is required to judge when the results should be reported as "total ." (Note that the word "total" does

double duty here, indicating both that the sample

consists of a water-suspended-sediment mixture

and that the analytical method determines all of

the constituent in the sample.)

Recoverable from bottom material .-Pertains to

the constituents extracted from a representative

sample of bottom material. Complete extraction is

generally not achieved, and thus the determination often represents less than the total amount

(that is, less than 95 percent) of the constituent in

the sample. To achieve comparability of analytical
data, all laboratories performing such analyses

would have to use equivalent extraction procedures, because different extraction procedures are

likely to produce different analytical results .

Total in bottom material .-Pertains to constituents in a representative sample of bottom material . This term is used only when the analytical

procedure ensures measurement of at least 95

percent of the constituent determined. Knowledge

of the expected form of the constituent in the

sample, as well as of the analytical methodology

used, is required to judge when the results should

be reported as "total in bottom material."

In describing an analytical method, it is necessary

to compare the result obtained by the method with

the value that is sought, normally the true concentration of the chemical substance in the sample. Definitions of terms that are used for this purpose are

given below.

Accuracy.-A measure of the degree of conformity

of the values generated by a specific method or

procedure with the true value. The concept of accuracy includes both bias (systematic error) and

precision (random error) .

Bias.-A persistent positive or negative deviation of

the values generated by a specific method or procedure from the true value, expressed as the difference between the true value and the mean

value obtained by repetitive testing of the homogeneous sample.

Limit of detection.-The minimum concentration

of a substance that can be identified, measured,

and reported with 99 percent confidence that the

analyte concentration is greater than zero; determined from analysis of a sample in a given matrix

containing analyte .

Precision.-The degree of agreement of repeated

measurements by a specific method or procedure,

expressed in terms of dispersion of the values

generated about the mean value obtained by repetitive testing of a homogeneous sample.

Definitions of other terms used in this. manual are

given below.

External standard.-A mixture of compounds of

interest (analytes to be determined) prepared in a

suitable organic solvent and diluted to approximate environmental residue concentrations ; used

for calibrating and checking detector response

prior to instrumental analysis. External standards

establish response and retention factors necessary

for quantitative analysis when internal standard

or standard addition methods are not used.

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

Internal standard.-A compound similar in physical and chemical properties to the analyte in the

sample; added to the final extract just prior to

instrumental analysis. Internal standard responses are incorporated into quantitative analysis calculations, thus serving to normalize all data

to a known amount of a common reference . Internal standard materials must be chosen carefully;

they must exhibit proper chromatographic behavior and yet must not occur either naturally or as a

result of pollution . When using mass sensitive detectors, internal standards may be chosen from

stable heavy isotope analogs of analytes of interest. Other types of gas and liquid chromatographic detectors require other kinds of compounds. An

internal standard will correct for the biases associated with the instrumental determinative step in

an analytical procedure.

Spike.-Spikes result from the addition of a known

amount of one or more of the compounds of interest to the sample prior to analysis. Analysis yields

accuracy data (from a synthetic matrix) or recovery data (from an authentic matrix) . Accuracy

reflects the best results that can be expected, and

recovery reflects the degree of influence of matrix

effects on accuracy.

Surrogate .-A compound similar in physical and

chemical properties to the analytes of interest ;

added to the sample upon receipt in the laboratory

(or, ideally, at the time of field sampling) . A surrogate is not used as an internal standard for

quantitative measurement purposes. Surrogates

may be added to every sample to provide quality

control by monitoring for matrix effects and gross

sample-processing errors. They should not occur

naturally or be present in polluted water samples.

Also called "surrogate spike."

Significant figures

The number of significant figures to which the

results of analysis, in milligrams or micrograms per

liter, are reported by the Geological Survey reflects

a compromise between precision of measurement

and a desire for a degree of uniformity in tabulations

of analytical data. One of the commonly used methods, which applies only to the expression of the precision of a determination, is to include all digits

known with certainty and the first (and only the

first) doubtful digit . This method has one obvious

disadvantage: published data so reported may not be
DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

interpreted to mean the same thing by all users of

the data.

Recovery correction

Values reported by Geological Survey laboratories

are not corrected for the percentage of constituent

recovered . Therefore, almost all of the data for determinations of specific compounds such as pesticides, are biased to the low side. Average recoveries

are always less than 100 percent .

Quality Control

Quality control includes the acquisition and documentation of information on personnel, reagents and

standards, equipment, and analytical procedures.

The principles discussed in this section apply to all

analytical procedures described in this manual and

represent the minimum level of analytical quality

control required to produce acceptable data. This

section supplements the practices described in Book

5, Chapter A6 of the Techniques of Water-Resources

Investigations series of the U.S . Geological Survey.

Qualification of the analyst

Before performing any analyses, the analyst must

demonstrate the ability to produce data of acceptable accuracy and precision using the method by

successfully analyzing replicate aliquots of reference

materials over the range of the method. To be considered successful, results obtained must fall within

two standard deviations of the expected values for

each constituent measured.

Reagents and standards

The purity of each reagent must be verified by

analysis employing the analytical method at the lowest detection limit that will be reported. The frequency of verification is a function of the stability of

each reagent . All reagents and adsorbents must be

free of interfering contaminants . The presence of an

interfering contaminant requires remedial action

and reanalysis to verify reagent purity.

Before processing any samples, the analyst must

demonstrate, through analysis of an aliquot of reagent water equivalent in volume to a sample aliquot,

that all glassware and reagent interferences are

Equipment

5

under control . A reagent water blank must be analyzed each time a set of samples is analyzed or there

is a change in reagents.

Stock standard solutions need to be prepared from

materials of highest available purity. All data concerning the preparation must be recorded in a

notebook reserved for standard-solution data. Prior

to use, individual component standards need to be

analyzed to ensure the concentration and component

response. Solutions of individual standards also need

to be analyzed by an independent laboratory, or by a

second analyst in the preparing laboratory, to confirm the results . All differences must be resolved

before a standard solution is used.

As specified in the analytical method, one to seven

standard solutions in the concentration range of the

procedure need to be analyzed with each sample set,

and the results must agree with expected values

before sample results can be reported.

A notebook containing all information on repair,

maintenance, and daily operating conditions should

be maintained and available at each instrument

work station .

Analytical procedure

The analytical procedures described in this manual must be followed exactly . Whenever possible,

method performance must be demonstrated with

each sample by the use of surrogate spikes . Appropriate surrogates are listed for each analytical

method along with applicable acceptance criteria .

For analytical procedures for which appropriate surrogates are unavailable, the laboratory must arrange to receive at least 10 percent of samples in

duplicate. One portion of each of these samples will

be spiked with a mixture of the compounds of interest and analyzed by the analytical method. If the

recovery for any constituent does not fall within

control limits for method performance, the results

reported for that constituent in all samples

processed as part of the same set must be qualified

as "suspect." The laboratory must monitor the frequency of suspect data to ensure that it remains

below 5 percent . For those samples (sediment, soil,

and core material) for which the use of spikes is not

appropriate, an estimate of analytical precision must
6

be obtained and reported by analyzing in duplicate

at least 10 percent of the samples in each set .

Because of the rapid advances in analytical technology, the analyst is permitted certain options to

improve separations or to lower the cost of measurement. Such modification of methods, however, must

conform to the following section, "Use and Documentation of Standard Laboratory Procedures."

Use and Documentation of

Standard Laboratory

Procedures

The Water Resources Division (WRD) of the U.S .

Geological Survey recognizes two types of waterquality analytical methods-approved methods and

special methods. Definitions of the two types of

methods and requirements for approval of analytical

methods are given for the guidance of users of this

manual who provide data to or evaluate data from

WRD programs.

Approved methods

Two categories of approved analytical methods

have been formally established-official methods,

and provisional methods. A description of methods

in each category and requirements for their approval

are given below.

Official methods

Methods in this category are considered the official water-quality analytical methods of the WRD.

They are published in the Techniques of Water-Resources Investigations (TWI) series . Data collected

by these methods may be stored in the national data

file, WATSTORE, and published in the annual basic

data reports of the WRD. Requirements for approval

are as follows :

1 . Submission of documentation of the proposed

method or modification of a presently used

method, in TWI format, to the Chief, Quality of

Water Branch .

2. Submission to and approval by the Quality of

Water Branch of a method-development report

giving information and supporting data on the

following:

a. applicable range, detection limit, and sensitivity

ofthe method;

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

b. known and possible interferences;

c. precision and bias of the method; these data

should, at a minimum, include results of singlelaboratory and multiple-operator tests, including at least 10 replicate analyses each of

pure solutions, natural water, and spiked natural waters at three concentrations covering the

applicable range of the method; and

d. production rates compared with other methods,

when possible.

3. Submission of statements describing hazardous

chemical reactions and (or) reagents that are involved in the method, sample-preservation requirements, and level of skill and (or) special

training needed by personnel using the method.

4 . In addition to the minimum requirements for approval listed above, if it is determined that the

proposed method will be used in the Central Laboratories System, the Quality of Water Branch will

initiate plans to obtain technical reviews of the

method-documentation and method-development

report by at least two colleagues, one of whom

must be outside the author's laboratory. Further,

if the method is developed in one of the Central

Laboratories, the other Central Laboratory will be

expected, if equipment is available, to confirm the

precision and bias of the method and to compare

results with present methodology, if a method

exists . These requirements must be met within 30

days. Approval of the method will depend on obtaining the reviews and additional data. The

Quality of Water Branch may give provisional approval, as outlined below, during the period of

review.

Provisional methods

Provisional methods are those believed to produce

data comparable to data obtainable from official

methods but likely to be used by WRD to such a

limited extent that they have not received extensive

in-house testing by WRD personnel . Many of the

methods published in such reliable compendia as

"Standard Methods," "Environmental Protection

Agency Methods for Chemical Analysis of Water

and Wastes," and "The American Society for Testing and Materials Book of Standards" and used by

in-house, cooperator, or contractor laboratories in

support of WRD programs would be accepted for

approval in this category . These methods will not be

published in the TWI series . Data collected by these

methods may be stored in WATSTORE if a parame-
DETERMINATION OF ORGANIC SUBSTANCES IN WATER ANDFLUVIAL SEDIMENTS

ter code exists, and the data may be published in the

annual basic data reports. Requirements for acceptance of methods in this category are as follows:

1. Submission of a description of the method to the

Chief, Quality of Water Branch, with an explanation of why approval of the method is desired.

2. Provision of the same type of information on precision and bias as is required for "Official Methods" approval, with the exception that data obtained by other than WRD personnel will be

accepted for consideration.

Special methods

In contrast to the two categories of approved analytical methods, there are some methods that have

specialized or limited application and, therefore,

need not be submitted to the Chief, Quality of Water

Branch, for approval. These include methods used in

support of research, experimental or developmental

methods used by a Central Laboratory, and screening methods used in the field or in the laboratory.

Data collected by these methods are not stored in

WATSTORE or published in the annual basic data

reports . However, the data may be published in interpretive reports or project data reports, provided

the method is fully described or an appropriate reference is cited to provide a basis for peer evaluation

of analytical results . Defense of the validity of such

a method is, therefore, the responsibility of the individual reporting the data.

Ordinarily, only approved methods are used in the

Central Laboratories unless a special method is requested and concurrence in its use is obtained from

the individual responsible for analytical technology

transfer in the Office of the Analytical Services Coordinator . Analysts are not to use a modification of

an approved method without satisfying all previously stated requirements for an approved method.

Furthermore, when a laboratory uses an alternate

method to satisfy a specific analytical request because of technical or management considerations,

the analytical method used must have been previously shown, to the satisfaction of the individual

responsible for analytical technology transfer, to

have equivalent or better sensitivity, precision, and

bias compared with the method requested. If these

conditions are not met, the requester must be notified promptly and must approve the change in

method prior to analysis.

Sample Collection

Preparation of the samplers

Collection and preservation of a representative

sample of a natural water body is the first and most

important task in the determination of any substance in water. If the sampling procedure or preservation procedure is faulty, the entire determination

will be of doubtful validity, at best. It is, therefore,

important that each step in the collection and preservation of a sample be carried out with utmost care.

The sampling techniques used will depend on the

compound or group of compounds to be analyzed .

Some organic compounds are associated mainly with

suspended or bottom sediments, others are generally

found as a surface film floating on water, and still

others are dissolved in water.

One must be careful to avoid contamination of a

sample. This is particularly difficult when a sample

is being taken for trace-level organic determinations, in which case it is necessary to prevent the

sample from contacting the myriad of plastics, oils,

and greases that are in common use today. Not only

do these materials introduce contamination into the

sample, but also, in many instances, they sorb the

compounds of interest.

Bed-sediment and suspended-sediment samples

pose special problems which must be dealt with in a

carefully prescribed way.

The sampler should be completely disassembled

and then (1) washed thoroughly with an Alconox

solution and a stiff brush, (2) rinsed with tapwater,

(3) rinsed with acetone, and (4) rinsed with

tapwater. The sampler should then be reassembled

and rinsed again. Immediately prior to its use in the

field, it should be thoroughly rinsed with native

water.

Sample handling and preservation

The samples in general should be collected in specially cleaned sample bottles that have been baked

at 300-350'C overnight. Immediately after the

samples have been collected, they should be chilled

in ice water. After chilling, the sample container

should be placed in an insulated shipping container

with sufficient ice to keep the sample cold until it

reaches the laboratory for analysis. Glass bottles
8

must be packed in such a way that they will not

break during shipping.

There is no single preservative that may be added

to a sample for all forms of organic analysis; each

sample must be treated according to the analytical

procedure to be followed. For the determinations

described in this manual, the following general

methods of sample preservation should be used in

addition to chilling at 4'C.

*Ethylene and propane: Collect samples in vials

containing 1 mL of formalin in a manner that

precludes headspace formation .

*Herbicides: Acidify with concentrated H2SO4 at a

rate of 2 mL per liter of sample in the laboratory.

" Oil and grease : Add concentrated H2SO4 until the

pH of the sample is below 3.0 . Generally, 5 mL

per liter of sample will be sufficient .

* Phenolic material : Acidify the sample to pH 4.0

with H3PO4, and add 1 g of CUS04-5H20 per liter

of sample.

Samples collected for dissolved-constituent determinations must be filtered immediately at the collection site using either pressure or vacuum filtration

apparatus and a 0.45-micrometer silver membrane

filter . Samples collected for specific compound determinations, such as pesticides, should be shipped

immediately to the laboratory, where they are

filtered through an approximately 0.3-micrometer

mean pore size glass fiber filter .

Sample bottles used for collecting purgeable organic compound samples must be filled so that a

meniscus forms at the mouth of the sample bottle .

Then the cap should be attached so that no bubble or

headspace is present . Any sample that has a bubble

or headspace at the collection site must be discarded

and a new sample collected .

Extraction, Fractionation,

and Identification

The determination of a specific organic compound

generally requires that the compound be isolated

from some or all of the other components in the

sample. This is true whether the sample is a natural

water being analyzed for a particular pesticide or a

biological fluid being analyzed for a particular amino

acid. The problem is further complicated if the sample consists of more than one phase, with the desired

component distributed between the phases. This disTECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

tribution of a component between two or more

phases is either a dynamic equilibrium process, in

which the component will redistribute itself between

the phases if the concentration in one phase is

changed, or a nonequilibrium process, in which the

component is irreversibly bound by one or more of

the phases in the system. Consider, for example, a

stream in which the running water is carrying a

suspended-sediment load. Each of the mineral components in the sediment is a separate phase; if a

surface film of an insoluble oil is present, it constitutes another phase; colloidal particles of organic

polymers such as humic material constitute still another phase. Organic analysis in a natural water

system therefore requires that the organic compounds first be isolated from the other components

of the stream (extraction) and then separated one

from another (fractionation) prior to measurement

of the amount of each compound present in the

sample.

Extraction

Two different types of liquid-extraction techniques are commonly used to extract organic solutes

from waters and sediments: batch extraction and

continuous extraction.

Batch extraction

Extraction of an organic compound or a group of

compounds from a water sample is generally done

by shaking the sample with an immiscible solvent in

a separatory funnel . All of the compounds that are

soluble in the two liquid phases (designated phases 1

and 2) will distribute themselves between the

phases. It can be shown thermodynamically that the

concentrations of solutes in the two phases are related to one another at equilibrium by the distribution

coefficient

K=

MA(1 )

MA(2)

where

MP) is the concentration of solute A in phase 1

and

MA(2) is the concentration of solute A in phase 2.

Therefore, if the concentration of solute A in

phase 1 and K are known, the concentration of solute A in phase 2 can be calculated. In an extraction

procedure, the distribution coefficients need to be
DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS

measured for the various possible extraction solvents in order to choose a solvent that will provide

high recovery of the desired solute after a reasonable number of repetitions (generally three or fewer) .

Continuous extraction

In the various continuous-extraction procedures,

fresh solvent, free of solute, is continually introduced into the extraction chamber in which the sample to be extracted is contained . Continuous-extraction equipment has been devised for extraction of

both solid and liquid samples. In some of the continuous-extraction devices for liquid samples, the sample also moves through the device and it is possible

to extract a large volume of liquid with a relatively

small extractor.

Fractionation

Chromatography has been defined by Denny

(1976) as follows: " * * * any separative process in

which a mixture carried in a moving phase (either

liquid or gas) is separated as a result of differential

distribution of the solutes between the mobile phase

and a stationary liquid or solid phase around or over

which the mobile phase is passing . The systems to

which this definition applies include all chromatographic processes, from paper chromatography to

ion exchange and gel permeation."

The stationary phase is a solid, or a solid on which

is adsorbed liquid that is insoluble in the mobile

phase, which may be either a gas or a liquid . If the

mobile phase is a gas, we shall refer to the process

as gas chromatography, and if it is liquid, we shall

refer to it as liquid chromatography.

Liquid chromatography

Liquid chromatography was developed before gas

chromatography. The pioneering work was done

during the first decade of the 20th century in the

United States by David Talbot Day of the U.S . Geological Survey and in Russia by Mikhail Tswett.

At least four different types of interactions have

been found to take place between the solutes and

stationary phases, the type that occurs depending on

the physical and chemical properties of the solutes

and the stationary phases, and these have, in turn,

given rise to four different types of liquid chromatography . These four types of interactions are

Adsorption

" adsorption,

" liquid-liquid partition,

" ion exchange, and

" gel permeation.

In some systems, more than one type of interaction occurs. It is instructive to consider briefly each

type of interaction .

Adsorption of molecules by the surface of an adsorbent occurs because of weak physical forces, such

as London forces or electrostatic forces, between the

molecules and the adsorbent molecules or because of

weak chemical bonds, such as charge transfer or

hydrogen bonds, between the molecules . If a chromatographic fractionation is based on adsorption,

the adsorption must be reversible . Therefore, only

reversible adsorption is considered here; however, it

should be noted that many adsorption processes are

not reversible . Although discussion of irreversible

adsorption is beyond the scope of this paper, it

should be pointed out that irreversible adsorption

often prevents the complete recovery of organic pollutants from sediments .

The most common theory of adsorption of a solute

from a solvent onto an adsorbent surface is an extension of a theory derived for the adsorption of

gases by adsorbent surfaces. Langmuir (1918) derived a model for adsorption that is based on the

following assumptions:

" The adsorbed molecules form a monolayer on the

surface of the adsorbent .

" All of the adsorption sites on the adsorbent surface

are equivalent,

" There is no interaction between the adsorbed molecules.

Because some of the assumptions that have been

made in the derivation of the equations of the

Langmuir model do not hold in all systems, other,

more complex adsorption equations have been derived. These are generally empirical relationships

that have been found to work for some systems.

The behavior of particular solutions in an adsorption system will, in general, be a function of the

surface properties of the adsorbent and the physical/chemical properties of both the solute and the

solvent.

All sorbent surfaces are made up of functional

groups that can interact with other molecules and

bind them. The capacity of a surface to bind molecules depends on the number and distribution of the
10

active functional groups. In general, the particles of

adsorbent that are packed into a chromatographic

column do not provide uniform surface for adsorption. Some of the surface is on the outside of the

particles, and some of the surface is contained in

voids within the particle. If the solute molecules are

too large, they will not enter the voids and therefore

will be excluded from part of the surface of the

particles . In some adsorbents, the active functional

groups are not uniformly distributed over the surface, and in some instances some of the groups may

be sterically hindered so that they cannot interact

with solute molecules . Part of the surface may be

deactivated by the presence of a strongly adsorbed

molecule . For example, water will strongly adhere to

the active sites on the surface of silica gel, effectively removing the affected sites from participating in

other adsorption reactions.

The active functional groups on an adsorbent surface may be classified into two groups: polar and

nonpolar . In general, polar groups are those that can

participate in hydrogen bonding . Nonpolar groups

bind mainly by London dispersion forces.

From the above discussion, it follows that the behavior of a solute in a chromatographic system that

consists of a solute, a solvent, and an adsorbent

surface will be a function of the following :

" the polarity of the solute molecule,

" the size of the solute molecule,

" the distribution of the active sites on the adsorbent,

" the polarity of the adsorbent sites,

" the affinity of the adsorbent surface for the solvent molecules, and

" the polarity of the solvent.

It is therefore possible, by proper manipulation, to

achieve a wide variety of different separations .

Liquid-liquid partition

In partition chromatography, two immiscible liquid phases and a stationary phase are used. One of

the liquid phases is bound to the stationary phase by

adsorption or, in some instances, by chemical bonding, and the other liquid phase moves freely through

the column. We shall call the liquid phase that is free

to move the mobile phase. The solutes that are to be

fractionated are placed in the column dissolved in

the mobile phase. The solutes then distribute themselves between the two liquid phases. After the solutes have been placed on the column, they are eluted

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

by the addition of solvent at the head of the column .

In this way, a continuous partitioning process takes

place on the column and the various solutes are separated one from another . This process was first elucidated by Martin and Synge (1941) . Although other

theories of partition chromatography have been developed since the work of Martin and Synge, their

work contains the basic principles and is the one that

is most widely used. In their work they have developed the concept of "height equivalent to one theoretical plate" (HETP) by drawing an analogy between the mode of operation of a chromatographic

column and a distillation column . They have defined

HETP as "the thickness of the layer such that the

solution issuing from it is in equilibrium with the

mean concentration of solute in the non-mobile

phase throughout the layers." They have assumed

that diffusion of solute from one plate to another is

negligible and that at equilibrium the distribution

ratio of one solute between the two liquid phases is

independent of the concentration of the solute and of

the presence of other solutes . The equations they

derived for partition chromatography may be extended to adsorption chromatography .

Martin and Synge have shown that the movement

of the position of the band of maximum concentration of a solute in a partition chromatographic column will be a function of the partition coefficient of

the solute, K, between the two liquid phases. Because the K values will be different for different

solutes, fractionation of the solutes will occur and

the amount of separation between the bands of maximum concentration of the solutes will be a function

of the total number of plates in the column.

The HETP is a function of the rate of diffusion of

the solute and of the rate of flow of the solvent.

There is an optimum flow rate that minimizes the

effect of diffusion . The smaller the HETP, the more

efficient the column will be for separation. In general, columns made up of smaller particles give smaller

HETP values than do columns composed of larger

particles . However, columns composed of small particles require high pressures to force the solvent

through the column.

Ion exchange

In ion-exchange chromatography, a resin which

consists of either anionic or cationic sites attached to

a cross-linked polymeric backbone is used. The resin,

which is insoluble in water, is formed into beads for

packing into a chromatographic column . When buff-
DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

er is added to the column, the beads swell. Ions can

then migrate in and out of the beads interacting

with the charged sites on the polymer. Ions having

charges opposite the charges of the fixed sites on

the polymer will be bound to the polymer. In ionexchange chromatography, the electrolyte molecules

to be fractionated are added to a buffer solution and

placed on the column of ion-exchange resin. After all

of the solute to be fractionated has migrated onto

the column, more buffer solution is added to the

head of the column to elute the solute fractions. The

solute ions to be fractionated compete for the fixed

charged sites on the polymer with the salt ions in the

buffer. Because, in general, the strengths of the

interactions of the various ions with the fixed

charged sites are different, fractionation of the solute ions will take place on the column .

Gel permeation

Gel-permeation chromatography is normally used

to obtain a molecular size fractionation of a mixture

of similar compounds. In principle, gel-permeation

chromatography is a relatively simple process ; however, as discussed below, the process can be complicated by adsorption and by ion-exchange effects .

In gel-permeation chromatography, the stationary

phase generally consists of porous polymer beads

which are insoluble in the mobile phase. The manufacturer of the beads attempts to make the dimensions of the pores in the beads uniform. Solute molecules that are larger than the largest pores in the

beads do not penetrate the beads but pass through

the column and are eluted from the column first .

Smaller molecules can penetrate into the pores of

the beads and are thereby retarded in their flow

through the column. Within limits, the smaller the

molecule the more retardation takes place and the

longer it takes for the molecule to be eluted from the

column. Thus, molecules are eluted from the column

in decreasing order of molecular size.

In a classical gel-permeation experiment, it is assumed that the stationary phase is completely inert

to the solute molecules, that the process of fractionation is brought about purely by differential penetration of the solute molecules into the pores of the

beads, and that molecular forces between the solute

molecules and the stationary phase are negligible . In

many instances, however, this is not the case and

interactions such as adsorption and ion exchange

can take place.

Gas-liquid chromatography

Identification

In 1952, James and Martin introduced a

gas-liquid "detector" (chromatograph) using a design first suggested by Martin and Synge (1941) .

This separation principle is similar to that discussed

in the section on "Liquid-Liquid Partition ." In the

stationary phase, normally a fairly high boiling polymer is bound to support material by adsorption. The

support material can be almost any inert material

having a large surface area and good structural

strength. Diatomaceous earth is the most commonly

used support, and organosilicone oils are the most

widely used liquid phases. Recently, very high resolution columns have been constructed by coating the

liquid phase directly onto the walls of synthetic

fused silica capillary tubing.

An inert gas such as He or Nz is normally used as

a mobile phase (carrier gas) to move the sample

from an injection port to a detector . As in partition

chromatography, the solutes distribute themselves

between the stationary phase and the mobile phase.

The concept of HETP also applies to gas-liquid

chromatography and gives an indication of the efficiency of a gas chromatographic column . A rather

simple method of calculating the number of theoretical plates is given by McNair and Bonelli (1969) and

is quite useful for measuring the efficiency of different columns of the same length and materials.

The U.S . Geological Survey laboratories perform

all gas chromatographic pesticide determinations on

two columns of dissimilar polarity such as methyl

silicone and 50 percent trifluoropropyl-methyl silicone. An analyte must have the proper retention

time on both columns in order to be considered identified . If peaks with the proper retention time on

both columns are found but the peak areas are different, one usually calculates the quantity of the

analyte based on the smaller peak because almost all

interferent compounds produce positive peak areas.

A large variety of detectors have been used over

the years, the most successful being the thermalconductivity and flame-ionization detectors, both of

which respond to virtually all compounds. More recently, the electron-capture detector has gained

widespread use ; it is somewhat more selective in its

response .
12

Thermal-conductivity detector

A thermal-conductivity detector employs a wire

filament whose electrical resistance varies with temperature . A constant current is passed through the

filament, which is enclosed in a detector block. Carrier gas, usually He, at a constant flow rate will

conduct heat away from the filament at a constant

rate, establishing a baseline. Any change in carrier

gas composition, such as occurs when a sample is

introduced, causes a change in filament temperature, with a corresponding change in resistance.

This is detected by a Wheatstone bridge, and the

resulting signal is recorded on a strip chart recorder .

Flame-ionization detector

A flame-ionization detector uses a hydrogen

flame to burn and ionize organic compounds . A cathode-anode electrode system with a fixed potential

between the electrodes monitors the flame. When a

compound is ionized in the flame, the resulting

change across the electrode gap is measured by an

electrometer and is displayed on a strip chart

recorder.

Electron-capture detector

Electron-capture detectors have been used extensively to detect chlorinated pesticides by gas-liquid

chromatography . This type of detector is probably

the most sensitive available for these compounds; for

example, as little as 1 pg (picogram) of lindane can

be detected. Its use, however, is limited to compounds that have a significant electron-capture

cross section. An electron-capture detector is a relatively simple device; it consists of a two-electrode ion

chamber, an internal radiation source, and associated electronic circuitry for measuring the ion current

between the electrodes in the chamber.

Mass spectrometer

A mass spectrometer as a detector uses an ionization chamber to ionize atoms and molecules eluting

from the GC column . The ions are segregated according to their mass-to-charge ratio, and a plot of

intensity versus mass-to-charge ratio is the mass

spectrum, which usually provides a great deal of

information about the molecular weight and structure of the compound. Special techniques, such as

single or multiple ion monitoring, greatly increase

the sensitivity of the mass spectrometer so that a

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

few picograms of a compound can be detected. If a

data system is used with the gas chromatograph-mass spectrometer system, the system

becomes a powerful qualitative and quantitative tool .

Computerized library searches can be conducted in

an attempt to identify components by comparing

their spectra with thousands of library spectra.

There are software packages to perform automated

quantitation and identification routines .

Other detectors

Other less sensitive detectors such as microcoulometric, flame photometric, photoionization, and

electrolyte conductivity detectors are sometimes

used in gas chromatography because they respond

to only specific elements incorporated into the molecules and this aids in molecular identification .

Spectrophotometer

When electromagnetic radiation impinges upon a

material, the radiation interacts with the molecules

of the material and, in general, some of the radiant

energy is absorbed. The amount of radiation of a

given wavelength (or frequency) that is absorbed is

a function of the atomic or molecular structure of

the material being irradiated. Radiation is absorbed

by a molecule when the frequency of the radiation

corresponds to the energy of a quantum transition

from one electronic vibrational state to another,

from one nuclear vibrational state to another, or

from one molecular vibrational or rotational state to

another. The electronic vibrational transitions cause

absorption in the visible and ultraviolet regions of

the spectrum (200 to 780 nm), nuclear and molecular vibrational transitions in the infrared region

(0 .78 gm to 30 gm), and rotational transitions in the

far infrared and microwave regions (30 gm to 1 m).

Absorption spectrophotometry may be used for

both quantitative and qualitative analysis.

Quantitative analysis

The amount of monochromatic radiation that is

absorbed by the substance is a function of the thickness of the sample and of the concentration of the

substance in the sample. Thus

A2, = log

I~.

I°

= abc,

x
DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

where A. is the absorbance of radiation of wavelength X, I?,is the intensity of the incident radiation

of wavelength X, I, is the intensity of the transmitted radiation of wavelength X, a is the extinction

coefficient at the given wavelength, b is the sample

thickness, and c is the concentration of the substance . The relationship A. = abc is Beer's law.

From it we see that in order to analyze a solution for

a given compound, it is necessary to measure the

path length of the light through the sample and to

evaluate the extinction coefficient for the compound

in the solvent . In practice, this is accomplished by

measuring the absorbances of the sample solution

and a group of suitable standards in a cell of known

path length . A plot of the absorbances of the standards versus concentration is then prepared, and the

concentration of the unknown sample is determined

from the analytical curve.

For most dilute solutions, Beer's law will hold;

however, departures from Beer's law do occur in

which concentration is not directly proportional to

absorbance. These departures are caused by light

scattering, fluorescence, decomposition, saturation,

hydrogen bonding, ion pair formation, molecular aggregation or disaggregation, solvation, and chemical

reactions. Therefore, to avoid inaccurate results it is

imperative in quantitative analysis that standards

that closely approximate the sample in concentration

and substrate composition be used.

Qualitative analysis

In general, the electronic, nuclear, and molecular

energy transitions in any given molecular species

will be different from those in any other molecular

species, and therefore the absorption spectrum of a

pure substance, that is, a plot of absorbance versus

wavelength of the sample, will be unique and will

allow one in theory to identify the substance . Unfortunately, in practice it may not be possible to obtain

sufficient sample to obtain a suitable spectrum.

Within any class of chemical compounds that are

characterized by particular structural elements there

will be a characteristic group of absorption bands in

the absorption spectra. Even if it is not possible to

assign a given spectrum to a particular compound, it

should be possible to determine the class of compounds . Normally, most diagnostic absorption bands

occur in the infrared region of the spectrum ; however, for some species the ultraviolet and visible regions also contain useful information .

Selected references

13

Ancillary methods of confirmation

In addition to the two-column gas chromatographic procedures used for the pesticides methods,

other confirmation procedures may be used, such as

gas chromatography employing (1) a mass spectrometer detector monitoring at least three major

ions or (2) an element specific detector such as the

Hall electrolytic conductivity detector operating in

the halogen, sulfur, or nitrogen mode. These supplemental confirmation techniques are normally used

when the concentrations of pesticides exceed 2 wg/L

in water for a minimum sample size of 1 L or 20

ltg/kg in bed materials for a minimum sample size

of 50 g dry weight equivalent.

Alford, A., 1975, Environmental applications of mass spectrometry : Biomedical Mass Spectrometry v.2, p. 229-253.

Budzikiewicz, A., Djerassi, C., and Williams, D.H ., 1967, Mass

spectrometry of organic compounds: San Francisco, HoldenDay, 690 p .

Burlingame, A.L ., Kimble, B.J ., and Derrik, P.J., 1976, Mass

spectrometry: Analytical Chemistry, v .48, p . 368R-403R .

Cram, S .P., and Juvet, R.S ., Jr., 1976, Gas chromatography :

Analytical Chemistry, v .48, p . 411R-442R.

Denny, R.C ., 1976, A dictionary of chromatography: New York,

Wiley, 191 p.

Guy, H.P ., and Norman, V.W., 1970, Field methods for measurement of fluvial sediment : U.S . Geological Survey Techniques

of Water-Resources Investigations, Book 3, Chapter C2, 59

P .

James, A.T ., and Martin, A.J .P., 1952, Gas-liquid partition chromatography : The separation and microestimation of volatile

fatty acids from formic acid to dodecanoic acid : Biochemical

Journal, v. 50, p. 679-690.

Langmuir, Irving, 1918, The adsorption of gas on plane surfaces

of glass, mica and platinum : Journal of the American Chemical Society, v .40, p . 1361-1402.

Martin, A.J .P., and Synge, R.L .M., 1941, A new form of chromatogram employing two liquid phases. I . A theory of chromatography . II. Application to the microdetermination of the

higher monoamino acids in proteins: Biochemical Journal, v .

35,p .1358-1368.

McFadden, W.H ., ed ., 1973, Techniques of combined gas liquid

chromatography/mass spectrometry : Applications in organic

analysis: New York, Wiley, 463 p.

McLafferty, F.W., 1967, Interpretation of mass spectra: New

York, Benjamin, 229 p .

McNair, H.M., and Bonelli, E.J ., 1969, Basic gas chromatography: Berkeley, Calif., Consolidated Printers, 306 p .

Oswald, E.O ., Albro, P.W., and McKinney, J.D ., 1974, Utilization

of gas-liquid chromatography coupled with chemical ionization of potentially hazardous environmental agents and their

metabolites : Journal of Chromatography, v. 98, p. 363-448 .

Safe, Stephen, and Hutzinger, Otto, 1973, Mass spectrometry of

pesticides and pollutants : Cleveland, Ohio, CRC Press, 220 p.
14

Analytical Methods

Carbon, organic, dissolved, wet oxidation

(0-1100-83)

Parameter Code

Carbon, organic, dissolved, (mg/L as C) ----------- 00681

1 . Application

This method is suitable for the analysis of water,

water-suspended-sediment mixtures, brines, and

waste waters containing at least 0 .1 mg/L of dissolved organic carbon (DOC). The method is not

suitable for the determination of volatile organic

constituents .

2. Summary of method

The sample is acidified, purged to remove inorganic forms of carbon, and oxidized with persulfate

in an autoclave at 116-130'C. The resultant carbon

dioxide is measured by nondispersive infrared spectrometry.

3. Interferences

3.1 Inorganic forms of carbon usually present

in most waters are readily converted to carbon dioxide when acidified but will interfere if the sample is

not purged adequately. Purgeable organic compounds are lost during this step.

3.2 Water samples containing large concentrations of reducing agents will interfere by decomposing the persulfate oxidant .

4. Apparatus

4.1 Ampules, precombusted, 10 mL, glass,

Oceanography International, or equivalent.

4.2 Ampule purging and sealing unit, Oceanography International, or equivalent.

4.3 Autoclave, Oceanography International

0512AU, or equivalent.

4.4 Carbon analyzer, Oceanography International, or equivalent.

5. Reagents

Carbon-free reagent water is required . All reagents should be analyzed to determine carbon content, and any reagent that yields a significant blank

value should be rejected.

5.1 Carbon standard solution, 1.00

mL = 1 .00 mg C (carbon) : Dissolve 2.1254 g potassium hydrogen phthalate (primary standard grade,

dried at 105° C for 1 h) in reagent water and dilute

to 1,000 mL. Store at 4°C.

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

5.2 Phosphoric acid solution, 1.2 N Add 83

mL H3P04 (85 percent) to reagent water and dilute

to 1 L with reagent water. Store in a tightly stoppered bottle .

5.3 Potassium persulfate, reagent grade,

granular : Finely divided forms of this reagent should

be avoided .

6. Procedure

6.1 Add 0.2 g potassium persulfate (a dipper

calibrated to deliver 0.2 g may be used) and 0.5 mL

1 .2 N phosphoric acid solution to precombusted

ampules.

6.2 Pipet (class A pipets are recommended) a

volume of water sample (10 .0 mL maximum) into an

ampule and adjust the volume to 10 mL with rea

gent water. If the sample contains precipitated material, it should be homogenized to ensure accurate

and reproducible subsampling.

6.3 Prepare one reagent blank (10 mL reagent water plus acid and oxidant) for every 15 to 20

water samples .

6.4 Prepare standards to cover the range 0.1

to 40 mg C/L by dilution of the carbon standard

solution (1 .00 mL = 1 .00 mg C.

6.5 Immediately place the filled ampules on

the purging and sealing unit and purge them at 60

mL/min for 6 min with purified oxygen .

6.6 Seal the ampules according to instructions

in the manufacturer's manual .

6.7 Place the sealed samples, blanks, and a

set of standards in ampule racks inside an autoclave

and digest for 4 h at 116-130'C.

6.8 Set the sensitivity range of the carbon

analyzer unit by adjusting the zero and span controls in accordance with instructions in the manufacturer's manual .

6.9 Break the combusted ampules in the cutter assembly of the carbon analyzer, sweep the carbon dioxide into the infrared cell with nitrogen gas,

and record the area of each carbon dioxide peak.

CAUTION: Combusted ampules are under positive

pressure and should be handled with care to prevent

them from exploding.

7. Calculations

7.1 Prepare an analytical curve by plotting

the peak area of each standard versus the concentration (mg/L) of the organic carbon standards .

7.2 The relationship between peak area and

carbon concentration is curvilinear. Operating
DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

curves must be defined each day the samples are

analyzed .

7.3 Determine the concentration of dissolved

organic carbon in each sample by comparing its peak

area with the analytical curve.

7.4 Multiply the result of step 7 .3 by the appropriate dilution factor.

8 Report

Report dissolved organic carbon concentration

(DOC) as follows: less than 10 mg/L, one decimal,

10 mg/L and above, two significant figures.

9 Precision

9.1 Multiple determinations of four different

concentrations of aqueous potassium acid phthalate

samples at 2.00, 5.00, 10 .0, and 40.0 mg/L of car

bon resulted in mean values of 2.2, 5.3, 9 .9, and 38

mg/L and standard deviations of 0.13, 0.15, 0.11,

and 1 .4, respectively.

9.2 The precision may also be expressed in

terms of percent relative standard deviation, as follows:

Selected references

Fredericks, A.D ., 1968, Concentration of organic carbon in the

Gulf of Mexico: Office of Naval Research, Report 68-27T,

65p.

Oceanography International Corporation, 1970, The total carbon

system operating manual: College Station, Tex., 51 p.

Carbon, organic, total, wet oxidation

(0-3100-83)

Parameter Code

Carbon, organic, total (mg/L as C --------------- 00680

1 . Application

This method is suitable for the analysis of water,

water-suspended-sediment mixtures, brines, and

waste waters containing at least 0.1 mg/L of total

organic carbon (TOC). The method is not suitable

for the determination of volatile organic constituents.

2. Summary of method

The sample is acidified, purged to remove inorganic forms of carbon, and oxidized with persulfate

in an autoclave at 116-130'C. The resultant carbon

dioxide is measured by nondispersive infrared spectrometry.

3. Interferences

3.1 Inorganic forms of carbon usually present

in most waters are readily converted to carbon dioxide when acidified but will interfere if the sample is

not purged adequately. Purgeable organic compounds are lost during this step.

3 .2 Water samples containing large concentrations of reducing agents interfere by decomposing the persulfate oxidant .

4. Apparatus

4.1 Ampules, precombusted, 10 mL, glass,

Oceanography International, or equivalent.

4.2 Ampule purging and sealing unit, Oceanography International, or equivalent.

4.3 Autoclave, Oceanography International

0512AU, or equivalent.

4.4 Carbon analyzer, Oceanography International, or equivalent.

4.5 Homogenizer, Willems Polytron PT10ST, Brinkman Instruments, or equivalent.

5. Reagents

Carbon-free reagent water is required. All reagents should be analyzed to determine carbon content, and any reagent that yields a significant blank

value should be rejected.

5.1 Carbon standard solution, 1 .00

mL = 1 .00 mg C (carbon) : Dissolve 2 .1254 g potassium hydrogen phthalate (primary standard grade,

dried at 105'C for 1 h in reagent water and dilute to

1,000 mL. Store at 4°C.

5.2 Phosphoric acid solution, 1 .2 N.- Add 83

mL H3P04 (85 percent) to reagent water and dilute

to 1 L with reagent water. Store in a tightly stoppered glass bottle.

5.3 Potassium persulfate, reagent grade,

granular: Finely divided forms of this reagent should

be avoided .

6. Procedure

15

6.1 Add 0.2 g potassium persulfate (a dipper

calibrated to deliver 0 .2 g may be used) and 0.5 mL

of 1 .2 N phosphoric acid solution to precombusted

ampules.

Number

of Mean

replicates (mg/L)

Relative standard

deviation

(percent)

9 --------- 2 .2 5 .9

10 --------- 5 .3 2 .8

10 --------- 9 .9 1 .1

10 -------- 38 3.7
1 6

6.2 Homogenize the sample until it is uniformly suspended. Rinse the homogenizer with distilled water after each use .

6.3 Pipet (class A pipets are recommended) a

volume of water sample (10 .0 mL maximum) into an

ampule and adjust the volume to 10 mL with reagent water.

6.4 Prepare one reagent blank (10 mL reagent water plus acid and oxidant) for every 15 to 20

water samples.

6.5 Prepare standards to cover the range 0.1

to 40 mg C/L by dilution of the carbon standard

solution (1 .00 mL = 1.00 mg C.

6.6 Immediately place the filled ampules on

the purging and sealing unit and purge them at 60

mL/min for 6 min with purified oxygen.

6.7 Seal the ampules according to instructions

in the manufacturer's manual.

6.8 Place the sealed samples, blanks, and a

set of standards in ampule racks, place the racks in

an autoclave, and digest for 4 h at 116-130'C.

6.9 Set the sensitivity range of the carbon

analyzer by adjusting the zero and span controls in

accordance with instructions in the manufacturer's

manual.

6.10 Break the combusted ampules in the cutter assembly of the carbon analyzer, sweep the carbon dioxide into the infrared cell with nitrogen gas,

and record the area of each carbon dioxide peak.

CAUTION: Combusted ampules are under positive

pressure and should be handled with care to prevent

them from exploding .

7. Calculations

7.1 Prepare an analytical curve by plotting

the peak area of each standard versus the concentration (mg/L) of the organic carbon standards.

7.2 The relationship between peak area and

carbon concentration is curvilinear. Operating

curves must be defined each day the samples are

analyzed.

7.3 Determine the concentration of total organic carbon in each sample by comparing its peak

area with the analytical curve.

7.4 Multiply the result of step 7.3 by the appropriate dilution factor.

8. Report

Report total organic carbon concentration (TOC)

as follows: less than 10 mg/L, one decimal ; 10 mg/L

and above, two significant figures.

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

9. Precision

It is estimated that the percent relative standard

deviation for total organic carbon will be greater

than that reported for dissolved organic carbon

(method 0-1100).

P.

1 . Application

Selected references

Fredericks, A.D ., 1968, Concentration of organic carbon in the

Gulf of Mexico : Office of Naval Research, Report 68-27T,

65

Oceanography International Corporation, 1970, The total carbon

system operating manual: College Station, Tex., 51 p.

Carbon, organic, suspended, wet oxidation

(0-7100-83)

Parameter Code

Carbon, organic, suspended (mg/L as C) ---------- 00689

This method is suitable for the analysis of suspended organic carbon (SOC) constituents found in

natural waters, brines, and waste waters. The

method is not suitable for the determination of volatile organic constituents .

2. Summary of method

The sample is collected on a silver filter, acidified,

purged to remove inorganic carbon, and oxidized

with persulfate in an autoclave at 116-130'C. The

resultant carbon dioxide is measured by nondispersive infrared spectrometry.

3. Interferences

3.1 Inorganic forms of carbon usually present

in most waters are readily converted to carbon dioxide when acidified but will interfere if the sample is

not purged adequately. Purgeable organic compounds are lost during this step.

3.2 Water samples containing large concentrations of reducing agents will interfere by decomposing the persulfate oxidant .

4. Apparatus

4.1 Ampules, precombusted, 10 mL, glass,

Oceanography International, or equivalent.

4.2 Ampule purging and sealing unit, Oceanography International, or equivalent.

4.3 Autoclave, Oceanography International

0512AU, or equivalent.

4.4 Carbon analyzer, Oceanography International, or equivalent.
5. Reagents

Carbon-free reagent water is required. All reagents should be analyzed to determine carbon content, and any reagent that yields a significant blank

value should be rejected.

5.1 Carbon standard solution, 1 .00

mL = 1 .00 mg C (carbon) : Dissolve 2.1254 g potassium hydrogen phthalate (primary standard grade,

dried at 105°C for 1 h in reagent water and dilute to

1,000 mL. Store at 4°C.

5.2 Phosphoric acid, 14.7 N(85 percent), reagent grade.

5.3 Potassium persulfate, reagent grade,

granular : Finely divided forms of this reagent should

be avoided.

6. Procedure

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

6.1 Carefully remove the silver filter from its

container with forceps and coil it into a roll about 'A

inch in diameter using a steel or glass mandril .

6.2 Drop the coiled filter into a precombusted

glass ampule . Add 0 .5 mL of 14.7 N (85 percent)

phosphoric acid and 8 mL reagent water.

6.3 Repeat steps 6 .1 and 6.2 using the blank

filter supplied by field personnel .

6.4 Cover the top of the ampule with aluminum foil and heat on a steam bath for 12 to 24 h.

6.5 Prepare standards to cover the range 0.1

to 40 mg C/L by dilution of the carbon standard

solution (1 .00 mL= 1 .00 mg C. Pipet 10.0 mL of

each standard into precombusted glass ampules containing 0 .5 mL of 14.7 Nphosphoric acid.

6.6 Introduce 0.2 g potassium persulfate (a

dipper calibrated to deliver 0 .2 g may be used) and

rinse down any solid adhering to the neck of the

ampule with 2 mL of reagent water.

6.7 Immediately place the filled ampules on

the purging and sealing unit and purge them at 60

mL/min for 6 min with purified oxygen.

6.8 Seal the ampules according to instructions

in the manufacturer's manual.

6.9 Place the sealed samples, blanks, and a

set of standards in ampule racks inside an autoclave

and digest for 4 h at 116-130'C.

6.10 Set the sensitivity range of the carbon

analyzer unit by adjusting the zero and span controls in accordance with instructions in the manufacturer's manual.

6.11 Break the combusted ampules in the cutter assembly of the carbon analyzer, sweep the carbon dioxide into the infrared cell with nitrogen gas,

and record the area of each carbon dioxide peak.

CAUTION: Combusted ampules are under positive

pressure and should be handled with care to prevent

them from exploding .

7. Calculations

7 .1 Prepare an analytical curve by plotting

the peak area of each standard versus the concentration (mg/L) of the organic carbon standards .

7 .2 The relationship between peak area and

carbon concentration is curvilinear . Operating

curves must be defined each day the samples are

analyzed .

7.3 Calculate the concentration of suspended

organic carbon in the original water sample from the

equation

S-B

Carbon, suspended organic (mg/L) =

V

17

where

S = apparent carbon concentration of sample, in

mg/L,

B = apparent carbon concentration of blank, in

mg/L,

V = number of liters of water filtered (in field) .

8. Report

Report suspended organic carbon concentrations

(SOC) as follows : less than 10 mg/L, one decimal ;

10 mg/L and above, two significant figures.

9. Precision

It is estimated that the percent relative standard

deviation for suspended organic carbon will be

greater than that reported for dissolved organic carbon (method 0-1100) .

Selected references

Menzel, D.W., and Vacaro, F.F., 1964, The measurement of dissolved organic and particulate carbon in seawater: Limnology and Oceanography, v. 9, p. 138-142 .

Oceanography International Corporation, 1970, The total carbon

system operating manual: College Station, Tex., 51 p.

Carbon, inorganic plus organic, total in bottom

material, dry weight, induction furnace

(0-5101-83)

Parameter Code

Carbon, total in bottom material

dry weight (g/kg as C) ----------------------00693
18

1 . Application

This method is suitable for the determination of

total carbon in bottom material at concentrations of

0.1 g/kg and above.

2. Summary of method

A sample is oxidized, in the presence of oxygen

and a catalyst, in a crucible in an induction furnace .

The carbon dioxide evolved is measured by thermal

conductivity .

3. Interferences

Sulfur, halides, and water vapor can interfere but

are eliminated by traps. Carbon impurities in the

reagents can interfere.

4. Apparatus

4.1 Carbon determinator, Leco WR-12, or

equivalent, consisting of an oxygen purification system, an induction furnace, dust traps, moisture

traps, a halide trap, a sulfur trap, a catalytic furnace, and thermistor cells .

4.2 Combustion crucibles, carbon-free, disposable, Leco 528-018, or equivalent.

4.3 Dryer-balance, consisting of a Mettler LP

15 dryer, a Mettler PC 440 balance, and a Mettler

GC 301 application input device, or equivalents .

4.4 Glass reagent scoops, Leco 501-032, or

equivalent.

4 .5 Grinder, Torsion MG 2 electric mortar

grinder, or equivalent.

4 .6 Sieves, U.S . No. 18 (2-mm opening) and

U.S . No. 60 (0 .25-mm opening) .

5. Reagents

5.1 Carbon standards, 1 g steel rings con5 .2 Catalytic furnace reagent, platinum on

silica, Leco 501-587.

5.3 Combustion accelerators

5 .3 .1 Copper accelerator, Leco 501-263, or

equivalent.

5.3 .2 Iron chip accelerator, Leco 501-077,

or equivalent.

5.4 Oxygen, commercial grade.

5.5 Sodium carbonate, anhydrous, reagent

grade.

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

5.6 Trap reagents

5.6 .1 Anhydrone, Leco 501-171, or equivalent, for moisture trap.

5.6 .2 Antimony, Leco 769-608, or equivalent, for halide trap.

5.6 .3 Ascarite, Leco 183-001, or equivalent, for oxygen purification system.

5.6 .4 Manganese dioxide, Leco 501-587, or

equivalent, for sulfur trap.

6. Procedure

6.1 Sieve the wet sample through a 2-mm

sieve. Place the sieved sample in an aluminum pan

that has been fired at. 550 ° C for 1 h and dry the

material at 40'C overnight.

6.2 Grind the dried material in an electric

grinder, to a powder that passes a No. 60 sieve.

6.3 To determine the percent of dry material,

the initial powder must be weighed before drying.

Dry an accurately weighed amount of the powder

(approximately 1 g) in a combination dryer-balance

at 105'C to a constant weight. Determine the percent of dry material.

6.4 Follow the instructions in the manufacturer's manual to leak-test, blank, and calibrate the

carbon determinator.

6 .5 Place a new combustion crucible on an

analytical balance and add 1 g (maximum) of the

sample (obtained in step 6.2) . Record the sample

weight to three significant figures.

6.6 Add a scoopful of iron chip accelerator,

spreading it evenly over the sample.

6.7 Add a scoopful of copper accelerator in a

similar manner.

6.8 Combust the sample in the determinator

and record the reading of percent carbon for the

sample.

7.1 Calculate the weight of dry bottom material in the sample from the equation

SxD W=-,

100

where

W= weight of dry bottom material in sample, in

91

S = weight of sample, from step 6.5, in g, and

taining known amounts of carbon C. 6.9 Repeat the determination using a smaller

5.1 .1 High standard, ca. 0.8 percent C, sample, if the reading is higher than 5.000 percent

Leco 501-506. C.

5.1 .2 Low standard, ca. 0.1 percent C, Leco

501-502 . 7. Calculations
D =percent of dry material in sample, from step

6.3 .

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

7.2 Calculate the concentration of carbon in

the bottom material from the equation

C

lOxDVMxC,

(g/kg) _ ,

W

where

DVM= direct readout of percent carbon for sample weight as set on compensator,

W= weight of dry bottom material in sample,

obtained in step 7.1, in g, and

CS = compensator setting, in g.

8. Report

Report total carbon concentrations in bottom material as follows: less than 1 g/kg, one decimal; 1

g/kg and above, two significant figures.

9. Precision

Single-operator precision data for carbon steel

standards for each standard (10 replicates each) is

as follows:

Carbon

Steel Mean determined Relative standard

(percent) (percent) deviation(percent)

0.0440---------------- 0.0442 3 .6

0 .166----------------- 0.166 2 .1

0 .941----------------- 0.943 0 .86

1 . Application

Selected reference

Leco Corporation, Instruction manual, WR-12, Carbon determinator, model 761-100, 1976: 3000 Lakeview Ave., St. Joseph, MI 49085.

Carbon, inorganic, total in bottom material,

modified Van Slyke (0-5102-83)

Parameter Code

Carbon, inorganic, total in bottom material,

dry weight (g/kg as C ----------------------00686

This method is suitable for the determination of

inorganic carbon in bottom material at concentrations of 0.1 g/kg and above.

2. Summary of method

A dry bottom-material sample is placed in a modified Van Slyke apparatus and treated with aqueous

hydrochloric acid. The sample is heated, and the

19

amount of carbon dioxide evolved is measured

manometrically.

3. Interferences

Sulfides and sulfites form gaseous hydrogen sulfide and sulfur dioxide, respectively, in the presence

of acid and heat . These gases are evolved from the

sample along with the carbon dioxide and give a

positive error.

4. Apparatus

Numbers in parentheses refer to figure 1.

4.1 Addition tube (1), 25 mm od (outside diameter) x30 cm, with 19/22 ground-glass fitting

and a 120 ° three-way stopcock (see item 4.15), custom-fitted to the condenser (4) .

4.2 Auto-bubbler gas absorption pipet (2),

Burrell, part 40-105-10 or equivalent .

4.3 Bunsen burner (3).

4.4 Condenser (4), custom-made, 200 mm

long with an expansion bulb (23 mm od). The condenser is fitted to the addition tube (1), as shown in

figure 1 .

4.5 Constant-temperature bath and circulator (5), Forma Scientific 2006, or equivalent.

4.6 Dryer-balance, consisting of a Mettler LP

15 dryer, a Mettler PC 440 balance, and a Mettler

GC 301 application input device, or equivalents.

4 .7 Funnel (6), custom-made, 100-mL or

greater capacity, with a 19/22 ground-glass joint.

4 .8 Gas buret (7), 100-mL capacity, Kimax

30025-A, or equivalent, fitted to stopcock (13) (see

item 4.15) .

4 .9 Gas buret cooling jacket (8), 25 mm odx

30 cm, custom-made to encase the buret (7).

4 .10 Grinder, Torsion MG 2 electric mortar

grinder, or equivalent.

4 .11 Leveling bulb reservoirs A (9) and B

(10), 250-mL capacity, Corning 2080, or equivalent.

4 .12 Rubber stopper (11), size 6, one hole .

4 .13 Sample vial (12), custom-made, 25 mL

capacity, 25 mm id (inside diameter) x85 mm roundbottom Pyrex tube, flanged to fit tightly around the

rubber stopper (12) .

4.14 Sieves, metal, U.S. No. 18 (2-mm opening), 15 cm id, and U.S . No. 60 (0 .25-mm opening),

18.7 cm id .

4.15 Stopcocks A (13) and B (14) three-way,

120', narrow bore, Kimax 4160F, or equivalent.

4.16 Tygon tubing (15), '/4 in id, I/16 in wall,

used to make connections in the apparatus.
0 TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

Figure l .-Apparatus for the manometric determination of carbon dioxide .
5 . Reagents

5.1 Hydrochloric acid, 3 .2 N Add 270 mL

concentrated HCl (sp . gr. 1 .19) to 500 mL deionized

water. CAUTION: Add the acid slowly with stirring.

Allow to cool to room temperature and dilute to 1L

with deionized water.

5.2 Sodium carbonate, anhydrous, reagent

grade: Dry the material at 105 ° C for at least 1 h

and store in a desiccator.

5.3 Sodium hydroxide, 10 N Dissolve 400 g

NaOH in 500 mL deionized water. CAUTION: Add

the pellets slowly with stirring . Allow to cool to room

temperature and dilute to 1 L.

5.4 Sulfuric acid, 7.2 N Add 200 mL concentrated H2SO4 (sp . gr. 1.84) to 500 mL deionized

water. CAUTION: Add the acid slowly with stirring.

Allow to cool to room temperature and dilute to 1 L.

Add 5 mg of methyl red indicator.

6. Procedure

DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

6.1 Set up the apparatus as follows:

6.1 .1 Assemble the apparatus as in figure

1.

6.1 .2 Add 7.2 N H,S04 to leveling bulb A

(9) and the gas buret as follows:

6.1 .2 .1 Turn stopcock A (13) to the position that connects the left arm with the gas buret.

6.1 .2 .2 Add 200 mL of 7.2 N H2 SO4 to

leveling bulb A (9) and fill the gas buret by raising

the leveling bulb until the buret is filled just to stopcock A (13).

6.1 .2 .3 Close stopcock A (13) . Lower leveling bulb A (9) and place it in a ring support . The

support must be positioned approximately 50 cm

below stopcock A (13).

6.1 .3 Add 10 N NaOH solution to leveling

bulb B (10) and to the auto-bubbler (2) as follows :

6.1 .3 .1 Turn stopcock A (13) to the position that connects both arms.

6.1 .3 .2 Mark a line about 1 cm above the

check valve (16) of the auto-bubbler (2) .

6.1 .3 .3 Fill the auto-bubbler (2) to the

mark (step 6.1 .3 .2) by adding 200 mL of 10 N

NaOH solution to leveling bulb B (10) and raising

the bulb.

6 .2 Circulate tapwater through the apparatus

until a constant temperature is achieved . Record the

temperature to the nearest 0.1 °C.

6 .3 Record the atmospheric pressure to the

nearest mm of mercury.

(1

21

6.4 Calibrate the system as follows :

6.4 .1 Weigh 50 to 80 mg sodium carbonate

on an analytical balance. Record the weight to the

nearest 0.1 mg.

6.4 .2 Transfer the sodium carbonate to a

sample vial . Add two boiling chips, and secure the

vial to the rubber stopper (11) by applying an upward, twisting motion.

6 .4 .3 Close stopcock B (14) .

6 .4 .4 Turn stopcock A (13) to the position

that connects the left arm with the gas buret.

6.4 .5 Pour approximately 20 mL of 3.2 N

HCl into the funnel (6) .

6.4 .6 Open stopcock B (14) to the position

that connects the addition tube (1) with the sample

vial . Allow 15 mL of 3.2 NHCl to flow into the vial .

CAUTION: Add the acid slowly . Do not allow the

acid level in the addition tube to reach stopcock B

4) .

6.4 .7 Heat the bottom of the vial for at

least 2 min until no more gas is evolved . This can be

determined by watching the liquid level in the buret.

6.4 .8 Turn off the burner and fill the addition tube (1) and the funnel (6) with water.

6.4 .9 Open stopcock B (14) to the position

that connects the addition tube (1) with the sample

vial and flood the path from the sample vial to stopcock A (13) . Close stopcock A (13) .

6 .4 .10 Move leveling bulb A (9) next to the

buret and position it so that the liquid in the bulb

and in the buret are at the same level .

6 .4 .11 Read and record the volume of gas

in the buret, to the nearest 0.1 mL.

6 .4 .12 Raise leveling bulb A (9) and turn

stopcock A (13) to the position that connects the

buret with the auto-bubbler (2) . Push the gas from

the buret into the auto-bubbler by raising leveling

bulb A (9) until the H2 SO4 in the buret reaches stopcock A (13).

6.4 .13 Lower leveling bulb A (9) until the

NaOH level in the auto-bubbler (2) reaches its original level (step 6.1 .3 .2) .

6.4 .14 Raise and lower leveling bulb A (9)

as in steps 6.4 .12 and 6.4 .13, two more times. Finish

with the NaOH at its original level .

6.4 .15 Close stopcock A (13) and position

leveling bulb A (9) as in step 6.4 .10.

6.4 .16 Read and record the volume of gas

in the buret, to the nearest 0.1 mL.

6.4 .17 Calculate (see steps 7.1-7.3) the

concentration of carbon in the Na2 CO3 . If recovery is
22

at least 95 percent, proceed to step 6.5 . If recovery

is less than 95 percent, check for leaks and ensure

that the pH of the NaOH is no less than 14. Correct

any problems and repeat the analysis (steps

6.4 .1-6.4 .16) until a recovery of at least 95 percent

is attained .

6.5 Prepare for the next determination by returning leveling bulb A (9) to a position above stopcock A (13), removing the vial, opening stopcock A

(13) and allowing the liquid above the vial to empty

into a waste container.

6 .6 Close stopcock A (13) when the H2SO4

level reaches the stopcock .

6.7 Prepare the sample for analysis as follows:

6.7 .1 Sieve the wet sample through a 2-

mm sieve. Place the sample in an aluminum pan that

has been fired at 550° C for 1 h. Dry the material at

40'C overnight .

6.7 .2 Grind the dried material in an electric

grinder to a powder that passes a No. 60 sieve .

6.7 .3 Dry an accurately weighed amount of

the powder (approximately 1 g) in a combination

dryer-balance at 105'C to a constant weight. Determine the percent of dry material .

6.8 Analyze the sample as follows :

6.8 .1 Weigh approximately 2 g of sample

obtained in step 6.7 .2, to the nearest mg.

6.8 .2 Transfer the sample to a sample vial

using a small funnel . Add two boiling chips, and

secure the vial to the rubber stopper by applying a

slight upward, twisting motion.

6 .8 .3 Place leveling bulb A (9) in a position

approximately 50 cm below stopcock A (13) and

repeat steps 6.4 .3-6 .4 .16, 6.5, and 6 .6 .

6.8 .4 Repeat the analysis with a smaller

sample if the volume of evolved carbon dioxide exceeds the volume measurable in the system (approximately 20 mL).

7. Calculations

7.1 Calculate the weight of dry bottom material in the sample from the equation

_ SxD

W 100

where

W= weight of dry bottom material in sample, in

91

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

S = weight of sample, from step 6.8 .1, in g, and

D = percent of dry material in sample, from step

6.7 .3 .

7.2 Calculate the volume of carbon dioxide

evolved from the sample from the equation

1%'= vi - Vf

where

V = volume of carbon dioxide evolved from sample, in mL,

Vi = volume reading from step 6.4 .11, in mL, and

Vf= volume reading from step 6.4 .16, in mL.

7.3 Calculate the inorganic carbon concentration in the original sample from the equation

V 273°C

C (g/kg) = WX

273° C + t

Patm - Pwat er

x

760 mm X 22 .4

where

V=volume of carbon dioxide evolved from

sample, from step 7.2, in mL,

W= weight of dry bottom material in sample,

from step 7 .1, in g,

t = temperature of carbon dioxide, from step

6.2, in ° C

Patm = atmospheric pressure determined in step

6.3, in rnm of mercury, and

Pwater = vapor pressure of water at temperature

t, from a reference source, in mm of

mercury.

8. Report

12.0

Report inorganic carbon in bottom material as

follows: less than 10 g/kg, one decimal ; 10 g/kg and

above, two significant figures.

9. Precision

Precision data are not available.

Selected references

Rader, L.F., and Grimaldi, F.S ., 1961, Chemical analyses for

selected minor elements in Pierre shale: U.S . Geological Survey Professional Paper :391-A, 45 p.

Van Slyke, D.D ., and Folch, J., 1940, Manometric carbon determination : Journal of Biological Chemistry, v. 136, p . 509-

541.
1 . Application

3. Interferences

DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

Carbon, organic, dissolved, fractionation

(0-1103-83)

Parameter Code

Carbon, organic, dissolved, fractionation --- None assigned .

Dissolved organic carbon fractionation analysis

can be applied to water samples whose DOC concentrations range between 5 and 25 mg/L and whose

specific conductance is less than 2,000 ltmhos/cm at

25 ° C. Water samples whose DOC is less than 5

mg/L can be freeze-concentrated to the specific conductance limit . DOC concentrations greater than 25

mg/L need to be diluted with organic carbon-free

reagent water to approximately 25 mg/L DOC prior

to analysis . DOC that is sufficiently volatile to be

lost during a 6 min gas purge of an acidified sample

is not included in the fractionation.

2. Summary of method

A flow chart of the analytical scheme of DOC

fractionation analysis is given in figure 2.

2.1 Dissolved organic carbon is first fractionated and classified as hydrophobic or hydrophilic organic solute on the basis of the solute's capability for

physical adsorption. Hydrophobic organic solutes are

separated from hydrophilic organic solutes by physical adsorption of hydrophobic solutes upon Amberlite-XAD-8 resin. Both the hydrophobic and hydrophilic organic solute classes are secondarily

fractionated into acid, base, and neutral compound

classes, thus giving a total of six characteristic DOC

fractions. The procedure requires 2 days to complete .

2 .2 Hydrophobic acids and bases are selectively desorbed from XAD-8 resin with aqueous alkali and acid, respectively. Hydrophobic neutral sol

utes are not desorbed with aqueous solvents. After

removal of the hydrophobic solutes from solution by

adsorption upon XAD-8 resin, the hydrophilic solutes are fractionated by selectively adsorbing hydrophilic bases as cations on a cation-exchange resin,

followed by selective adsorption of the anionic hydrophilic acids upon an anion exchange resin. Hydrophilic neutral organic solutes are not adsorbed by

any of the adsorbents. The fractionation is based on

an organic carbon materials balance using DOC as

the quantifying parameter.

3 .1 Water samples whose specific conductance exceed 2,000 gmhos/cm contain inorganic

23

ionic salts in concentrations that exceed the capacity

of the ion-exchange resins. These samples can be

analyzed if the DOC exceeds 5 mg/L after the sample is diluted with reagent water to a specific conductance of 2,000 lamhos/cm .

3.2 A few samples will form organic precipitates when they are acidified to pH 2 in the analytical fractionation scheme . Care should be taken to

suspend these precipitates by stirring, so that they

are incorporated into the column containing XAD-8

resin.

3 .3 Colloidal clay will foul the resin adsorbents. All samples should be field-filtered prior to

analysis to remove particulate and colloidal material.

3.4 All reagents must be tested for contamination by running reagent blanks .

4 Apparatus

4.1 Carbon analyzers

4.1 .1 Beckman 915, or equivalent.

4.1 .2 Oceanography International, or

equivalent.

4.2 Clamps: Size 18 pinch clamps with compression screw for ball-and-socket joints .

4.3 Columns: All columns are custom-prepared from 11 mm od (outside diameter) x 7 mm id

(inside diameter) Pyrex glass tubing, and are con

nected with 18/7 ball-and-socket ground-glass

joints .

4.3 .1 Anion-exchange columns: Two columns are needed. They are 18 cm long between the

base of the socket joint at the top of the column and

the 6 mm odx 2 mm id glass nipple at the bottom of

the column. Column capacity is approximately 6 mL.

4.3 .2 Cation-exchange column: The column

is 10 cm long between the base of the socket joint on

the top and the four indentations used to hold the

glass-wool plug above the ball joint on the bottom.

Column capacity is approximately 2.5 mL. Indentations are 1 cm from the ball joint .

4.3 .3 XAD-8 column: The column is 8 cm

long between the four indentations at the ends of

the column. Ball joints are fused to both ends of the

column above and below the indentations . Column

capacity is approximately 3 mL. Indentations are 1

cm from the ball joints .

4.3 .4 Tubing-column adaptors : Two socket-joint and two balljoint fittings fused to 6 mm

od x2 mm id glass nipples are adaptors, which en

able connection of the Teflon tubing to the columns.
24 TECHNIQUES OFWATER- RESOURCES INVESTIGATIONS

DOC 1

Adjust sample pH to 2

with HCI

DOC 6

Figure 2. -Dissolved organic carbon fractionation analytical scheme .

3 mL AG-50 BIO RAD

H+ saturated cationexchange resin

6 mL AG-MP-1 BIO RAD

OH - saturated anionexchange resin

Step 1 Step 2 Step 3 Step 4

200 mL filtered sample Elute with Sample at Elute with

at pH 7 0.1 N HCI pH 2 0.1 N NaOH
ty.

5. Reagents

DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

4 .4 Extraction apparatus, Soxhlet: 145 mL

thimble capacity, 300 mL flask capacity .

4.5 Graduate cylinders : One 200 mL capacity and five 25 mL capacity, with ground-glass stoppers.

4.6 Pump: Cole Palmer Masterflex with silicone-rubber tubing (1-5 mL/min), or equivalent.

4.7 Tubing, Teflon : 4 in od x .085 in id (32

mm od x 22 mmid) .

4.8 Silver membrane filter: 0.45 gm porosi5.1 Acetonitrile : Reagent grade.

5 .2 Diethyl ether: Reagent grade.

5 .3 Glass wool: Fine fiber . Clean by Soxhlet

extraction with methanol for 24 h.

5.4 Hydrochloric acid, 1 .0 N and 0.1 N Prepared by diluting 82.9 and 8.29 mL of concentrated

HCl (sp. gr. 1 .19) to 1 L, respectively, in carbon-free

distilled water.

5.5 Methanol: Reagent grade.

5.6 Resin adsorbents: All adsorbents must be

extensively cleaned before use .

5.6 .1 Anion-exchange resin: BioRad AGMP-1, 20-50 mesh, chloride-saturated . Clean by

Soxhlet extraction with methanol for 24 h. Store in

methanol .

5.6 .2 Cation-exchange resin: BioRad AGMP-50, 20-50 mesh, hydrogen-saturated. Clean by

Soxhlet extraction with methanol for 24 h. Store in

methanol .

5.6 .3 XAD-8 resin: Available from Rohm

and Hass, 20-50 mesh. Clean the XAD-8 resin by

first slurrying with 0.1 NNaOH, and then decanting

off the fines, and store in 0.1 N NaOH for 24 h.

Decant the sodium hydroxide, slurry in methanol,

and decant off the fines with the methanol . Perform

sequential 24-h Soxhlet extractions with methanol,

acetonitrile, and diethylether. Store resin in methanol.

5.7 Sodium hydroxide, 2 N, 1 N, 0.1 NPrepared by dissolving 80 g, 40 g, and 4 g, respectively, of analytical reagent-grade sodium hydroxide

in 1 L of carbon-free distilled water. Purify by passing through OH-saturated AG-MP-1 anion-exchange resin .

5.8 Water, carbonfree: Prepare by double

distillation of tapwater in a glass still . Prepare bulk

quantities of 50 L or more per batch and store in

25

clean glass containers. Water blank should be <0.2

mg/L DOC.

6 Procedure

6.1 Samples need to be collected in organicfree glass containers. Filter 200 mL of sample onsite through a silver membrane filter of 0 .45gm po

rosity. Chill on ice (recommended method of sample

preservation) .

6.2 Initial parameters: Before preparing the

columns, take two 10-mL aliquots of sample for

DOC analysis (DOC 1 in fig. 2) . Take a third 10-mL

aliquot and determine pH and specific conductance .

If the pH is less than 6.5, carefully adjust the pH of

the sample to 7.0 by dropwise addition of 1.0 N

NaOH. Once the DOC and specific conductance parameters are known, dilute or concentrate the sample as specified in the section on "Application."

6 .3 Column packing and final resin purification : The final tasks in the resin purification procedure are done after the columns are packed with

the resin adsorbents. All three columns should be

packed at the same time so the reagent solutions

used to prepare the columns can be pumped through

the columns simultaneously. All resin adsorbents are

used only once.

6.3 .1 Anion-exchange column: Place a

small glass-wool plug in the bottom of the anionexchange column. Pack a 6-mL bed of purified

AG-MP-1 resin by pouring the resin slurried in

methanol into the column. Do not let the column run

dry during the packing procedure. Always keep

methanol or water above the resin bed in the column. Pack two columns. Connect the packed columns in series to the transfer tubing and pump; then

pass 100 mL of reagent water, 100 mL of 1 .0 N

HCI, 100 mL of 2 N NaOH, and 50 mL of reagent

water through the two columns at 4 mL/min. Disconnect the columns and discard the resin in the first

column of the series. The first column serves as a

pre-column to adsorb reagent contaminants which

are usually present in the sodium hydroxide . The

second column is used in the analysis. For best results, use the prepared anion-exchange column immediately, as the hydroxide-saturated resin is unstable over time and the blank organic carbon values

increase during storage .

6.3 .2 Cation exchange column: Place a

glass-wool plug at the four indentations at the bottom of the cation-exchange column. Pack a 3-mL
26

bed of purified AG-MP-50 resin using the procedure specified in step 6.3 .1 . Connect the packed column to the transfer tubing and pump; then pass 100

mL of reagent water, 100 mL of 1 .0 N NaOH, 100

mL of 1 .0 N HCl, and 50 mL of reagent water

through the column at 4 mL/min. Do not let the

column run dry after it is prepared.

6.3 .3 . XAD-8 column: Place a glass-wool

plug at the four indentations at one end of the column. Pack a 3-mL bed of purified XAD-8 resin

using the procedure specified in step 6.3 .1, and place

a second glass-wool plug at the four indentations

above the resin bed. Connect the packed column to

the transfer tubing and pump; then pass 500 mL of

reagent water, 50 mL of 0.1 NHCI, 50 mL of methanol, and 100 mL of reagent water through the

column at 4 mL/min. Do not let the column run dry

after it is prepared.

6.4 After the three columns are packed and

prepared, clamp the three columns together in the

following series : XAD-8 column first, cation-ex

change column second, anion-exchange column

third. Connect the column series to the transfer tubing and pump, and pass reagent water through the

columns at 4 mL/min until the DOC in the effluent is

1 .0 mg/L or less, as monitored by the Beckman 915

organic carbon analyzer. The volume of water

needed to rinse the residual methanol from the columns varies between 1 and 2 L. This is the best

point in the 2-day analytical procedure for the overnight pause.

6.5 Just before the sample is analyzed, disconnect the XAD-8 column and pump 50 mL of

0.1 N NaOH, followed by 100 mL of reagent water,

at 4 mL/min through the XAD-8 column, to desorb

any hydrophobic acid contaminants that may have

been previously adsorbed from the reagent purification solvents.

6.6 Pump the water sample through the

XAD-8 column at 2 mL/min and collect precisely

160 mL of eluent in a 200-mL glass-stoppered grad

uate cylinder. Follow the sample with a 20-mL wash

of reagent water so that a total of 180 mL are

collected .

6.7 Pump 0.1 NHCI through the XAD-8 column at 1 mL/min until 23 mL are collected . Collect

23 mL of 0.1 N HCI in a 25-mL graduate cylinder

having a glass stopper . This fraction is used to determine DOC 2 (see fig. 2) .

6 .8 Carefully adjust the pH of the sample

eluent from 6 .6 to 2.0 by dropwise addition of concentrated HCI, while stirring the sample.

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

6.9 Reconnect the XAD-8 column as given in

step 6.4, pump the 180 mL of acidified sample

through the column series, and take aliquots for

DOC determination in the following sequence:

6.9 .1 Discard the first 50 mL of sample eluate. This volume is diluted with reagent water from

the dead volume in the three columns.

6.9 .2 Collect the next 23 mL of eluate in

25-mL glass-stoppered graduate cylinder. This fraction is used to determine DOC 6 (see fig. 2) .

6.9 .3 Disconnect the anion-exchange column and collect 23 mL of eluate from the cationexchange column in a 25-mL glass-stoppered gradu

ate cylinder. This fraction is used to determine DOC

5 (fig . 2).

6.9 .4 Disconnect the cation-exchange column and collect 23 mL of eluate from the XAD-8

column in a 25-mL glass-stoppered cylinder. This

fraction is used to determine DOC 4 (fig. 2) .

6 .9 .5 Pump the remainder of the sample

through the XAD-8 column. Desorb the hydrophobic

acids by pumping 0.1 NNaOH through the column

at 1 mL/min. Collect 23 mL of eluate in a glassstoppered 25-mL graduate cylinder. This fraction is

used to determine DOC 3 (fig. 2) .

6.10 Thoroughly shake and mix each of the

collected fractions in the graduate cylinders before

taking aliquots for DOC determination. For samples

with low DOC values, the Oceanography International system of carbon determination must be used.

The Beckman 915 analyzer can be satisfactorily

used if sample DOC is 15-25 mg/L. Analyze each

fraction for DOC using the above methodology specified .

6.11 Run a complete DOC fractionation of reagent water blank for each set of samples, and correct each DOC fraction value with the respective

blank value obtained.

7. Calculations

Refer to figure 2 and the procedural description

for definition of terms and sample fractions . All

DOC fractions need to be reported in mg/L units

calculated for the concentration in the original water

sample prior to dilution or concentration. Following

is the list of parameters and the computation formulas :

7.1 Total hydrophobic DOC (mg/L) = mg/L

DOC 1- (1 .125 x mg/L DOC 4) (1 .125 is a dilution

coefficient equal to total volume divided by sample

volume) .
7.2 Total hydrophilic DOC (mg/L) = 1.125 x

mg/L DOC 4.

7.3 Hydrophobic base DOC (mg/L) = (mg/L

DOC 2 x 0.023)/0 .160 (0 .023 is the fraction volume, in liters ; 0 .160 is the sample volume, in liters) .

7 .4 Hydrophobic acid DOC (mg/L) = (mg/L

DOC 3 x 0.023)/0 .160.

7 .5 Hydrophobic neutral DOC (mg/L) = total

hydrophobic mg/L DOC-hydrophobic base mg/L

DOC-hydrophobic acid mg/I, DOC.

7.6 Hydrophilic base DOC (mg/L) = 1.125 x

(mg/L DOC 4-mg/L DOC 5).

7.7 Hydrophilic acid DOC (mg/L) = 1.125 x

(mg/L DOC 5-mg/L DOC 6).

7.8 Hydrophilic neutral DOC (mg/L) = 1.125

x mg/L DOC 6.

Subtract the respective blank DOC values from

the equivalent sample DOC fractions .

8. Report

Report all fraction DOC concentrations to two significant figures, in mg/L.

9. Precision

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

9.1 Two factors influence precision: the variability of the reagent blank DOC that elutes from the

columns, and the variability of the DOC determina

tion . The maximum average deviation for duplicate

DOC determinations is about 5 percent of the DOC

mean and, therefore, also 5 percent of the DOC

value of each fraction. The total precision is the sum

of the two factors.

9.2 The average deviations in the reaction

blank for dissolved organic carbon fractionation

analysis are as follows:

Averagedeviation

Fraction (mg/L)

Total hydrophobic DOC -------------- 0 .2

Total hydrophilic DOC --------------- .2

Hydrophobic base DOC -------------- .1

Hydrophobic acid DOC -------------- .1

Hydrophobic neutral DOC ------------ .4

Hydrophilic acid DOC --------------- .5

Hydrophilic base DOC --------------- .5

Hydrophilic neutral DOC ------------- .3

Selected references

Leenheer, J .A., and Huffman, E.W.D ., Jr., 1976, Classification of

organic solutes in water by using macroreticular resins : U.S.

Geological Survey Journal of Research, v. 4, no . 6, p .

737-751

Stuber, H.A ., and Leenheer, J.A ., 1978, Fractionation of organic

solutes in oil shale retort waters for sorption studies on

processed shale : Preprints, American Chemical Society, Division Fuel Chemistry, v. 23, no. 2, p. 105-174 .

Thurman, E.M., Aiken, G.R ., and Malcolm, R.L., 1978, The use of

macroreticular nonionic resins to preconcentrate trace organic acids from water : Joint Conference on Sensing of En

vironmental Pollutants, American Chemical Society, 4th,

1977, New Orleans, Proceedings, p. 630-634.

Thurman, E.M ., Malcolm, R.L ., and Aiken, G.R., 1978, Prediction

of capacity factors for aqueous organic solutes adsorbed on a

porous acrylic resin : Analytical Chemistry, v. 50, no. 6, p.

775-779.

Organochlorine and organophosphorous

compounds, total recoverable (0-3104-83)

and dissolved (0-1104-83), gas

chromatographic

Code

Parameter Totalrecoverable Dissolved

1 . Application

27

This method is suitable for the determination of

organochlorine insecticides, polychlorinated biphenyls (PCB's), polychlorinated naphthalenes (PCB's),

and organophosphorous insecticides in water and

water-suspended-sediment mixtures containing at

least 0.01 pg/L of the analyte.

2. Summary of method

Organochlorine and organophosphorous compounds are extracted from water and waterAldrin--------------------- 39330 39331

Chlordane------------------ 39350 39352

DDD ---------------------- 39360 39361

DDE ---------------------- 39365 39366

DDT ---------------------- 39370 39371

Diazinon------------------- 39570 39572

Dieldrin-------------------- 39380 39381

Endosulfan----------------- 39388 82354

Endrin -------------------- 39390 39391

Ethion--------------------- 39398 82346

Polychlorinated biphenyls ------ 39516 39517

Polychlorinated naphthalenes --- 39250 82360

Heptachlor ----------------- 39410 39411

Heptachlor epoxide----------- 39420 39421

Lindane-------------------- 39340 39341

Malathion ------------------ 39530 39532

Methoxychlor --------------- 39480 82350

Methyl parathion ------------ 39600 39602

Methyl trithion -------------- 39790 82344

Mirex --------------------- 39755 39756

Parathion ------------------ 39540 39542

Perthane------------------- 39034 82348

Toxaphene ----------------- 39400 39401

Trithion ------------------- 39786 82342
28

suspended-sediment mixtures with hexane . Organophosphorous compounds are determined on a gas

chromatograph with flame-photometric detectors.

The extracts are then purified using adsorption

chromatography on an alumina column . If PCB's,

PCB's, and toxaphene are present, the extracts are

further purified using a silica gel column. The organochlorine compounds are then determined by gas

chromatography using electron-capture detectors.

3. Interferences

Compounds having chemical and physical properties similar to the compound of interest may cause

interference. Sulfur and organosulfur compounds

will interfere, but these substances can be removed

by treating the final extracts with mercury; however, the mercury treatment will also remove organophosphorous compounds.

4. Apparatus

4.1 Alumina column: Plug a disposable pipet

with glass wool. Fill to a depth of 3 cm with alumina; then add 0.5 cm anhydrous sodium sulfate .

4.2 Boiling chips, micro, granules, Hengar

H-1366C, or equivalent: Rinse with hexane, air dry,

and heat overnight at 300'C.

4.3 Concentrator, Kuderna-Danish (K-D),

250-mL flask, 5.0-mL receiver, and one-ball Snyder

column .

4.4 Evaporative concentrator, Organomation

N-Evap, or equivalent.

4.5 Gas chromatograph, Tracor 560, or

equivalent.

4 .5 .1 The following conditions are recommended for organochlorine compounds:

Columns, borosilicate glass, 1 .8 m x 2

mm id (inside diameter) operated at 200'C: Column

packing materials are (1) 3 percent SP 2100 on

100/120 mesh Supelcoport, or equivalent; and (2)

1 .5 percent SP 2250 -h 1.95 percent SP 2401 on

100/120 mesh Supelcoport, or equivalent.

Detectors, electron capture, operated at

345'C.

Injection port temperature, 220'C.

Carrier gas, nitrogen or 5 percent methane in argon, flow rate 30 mL/min.

4.5 .2 The following conditions are recommended for organophosphorous pesticides:

Columns, borosilicate glass, 1 .8 m x 2 mm

id operated at 175°C: Column packing materials are

(1) 5 percent SP 2100 on 100/120 mesh SupelcoTECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

port, or equivalent; and (2) 2 percent SE-30 -1- 3

percent OV-210 on 100/120 mesh chromosorb HP,

or equivalent.

Detectors, flame photometric, Melpar, or

equivalent, operated at 210'C.

Injection port temperature, 210°C.

Carrier gas, helium or nitrogen, flow rate

30 mL/min.

4.6 Glass filters, 142 mm, 0.3 gm mean pore

size, Gelman, or equivalent: Prepare the filters by

rinsing with acetone and hexane, evaporating the

solvent, and heating overnight at 300 ° C.

4.7 Silica column; to a 130 mm x 10 mm id

glass tube having a coarse-porosity fritted disc, add

1 cm anhydrous sodium sulfate, 10 cm silica, and 1

cm anhydrous sodium sulfate .

5. Reagents

5.1 Alumina adsorbent, Woelm neutral aluminum oxide, or equivalent: Prepare deactivated adsorbent by adding 8 g deionized water to 92 g alumi

na and shake for at least 2 h on a wrist-action

shaker. The alumina is tested for required deactivation by attempting to elute the organochlorine compounds of interest from a test column with 10 mL

hexane. If the test compounds do not elute with 10

mL hexane, further deactivation is required .

5.2 Mercury, metallic, reagent grade.

5.3 Pesticide mixed standards, analytical

reference grade, EPA analytical reference standards, or equivalent: Prepare individual stock solu

tions by weighing about 10 mg of each compound to

at least three significant figures and quantitatively

transfer each compound to a 25.0-mL volumetric

flask. Dilute to volume with benzene and mix thoroughly. Aliquots are removed and diluted to volume

with iso-octane to obtain the final concentrations

listed in table 1 .

5.4 Silica adsorbent, Woelm silica, 70-150

mesh, or equivalent: Prepare deactivated adsorbent

by adding 0.2 g deionized water to 99.8 g silica and

shake for at least 2 h on a wrist-action shaker. The

deactivation is evaluated by attempting to reproduce

the elution scheme in table 2. If the test compounds

do not elute with 25 mL of hexane from the first

silica fraction, additional deactivation is required .

5 .5 Sodium sulfate, granular, anhydrous:

Heat overnight at 300°C and store in a covered

beaker at 130°C.

5.6 Solvents, benzene, hexane, and iso-octane, pesticide residue quality, distilled in glass, Burdick and Jackson, or equivalent.
DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

Table 1 . Concentrations of pesticides and PCB's in mixed

standard solutions used for gas chromatograph calibration of water and water-suspended sediment

[Picograms permicroliter]

6. Procedure

All glassware must be washed in warm detergent

solution, rinsed with organic-free water, and heated

at 300 ° C overnight . Immediately before use, all

glassware is rinsed with the solvent. Do not use

stopcock grease on any ground-glass joints. For the

determination of dissolved components, filter the

sample through a glass fiber filter. Pour the filtrate

into the original sample bottle and continue with the

procedure .

6.1 A blank must accompany each group of

samples . For each sample, rinse a 1,000-mL separatory funnel and a 125-mL Erlenmeyer flask with

hexane .

6.2 Weigh the sample bottle plus the sample

and record the weight to three significant figures.

6.3 Pour the sample into the separatory funnel and allow the bottle to drain completely. Weigh

the empty bottle and cap, and record the weight to

Table 2 . Column fractionation scheme for silica gel column

for organochlorine insecticides, PCB's, and PCB's

Fraction 2

29

Fraction 1

(20 mL hexane eluate) (30mL benzene eluate)

Heptachlor 70 Heptachlor 30

percent percent

DDE85 percent DDE 15 percent

Endosulfan Lindane

Mirex Perthane

PCB's DDD

PCB's DDT

Methoxychlor

Heptachlor epoxide

Endrin

Dieldrin

Chlordane

three significant figures. Calculate and record the

sample weight.

6.4 Add 25 mL hexane to the sample bottle,

rinse the sides thoroughly, and pour the solvent into

the separatory funnel . The Teflon-lined cap is not

rinsed because of the potential for contamination

from solvent that has contacted the threads and surface beneath the Teflon liner. Shake the funnel vigorously for at least 1 min, venting often. Allow the

layers to separate, and drain the aqueous layer. Pour

the hexane extract into the Erlenmeyer flask . Extract the sample twice more, using 25 mL hexane

each time, and collect the extracts in the Erlenmeyer

flask. Add about 0.5 g anhydrous sodium sulfate to

the flask, cover with foil, and set aside for at least 2

h or refrigerate until analysis can continue.

6.5 Quantitatively transfer the extract with

hexane to the K-D apparatus, add a boiling chip, and

attach a Snyder column. Concentrate the extract to

about 5 mL on a water bath maintained at about

90 ° C. Remove the K-D apparatus from the water

bath, dry the joints with a towel, rinse the lower

joint with hexane as the receiver is disconnected,

and place the receiver on an evaporative concentrator to reduce the volume to about 0.5 mL. Rinse the

walls of the receiver two or three times with a few

drops of hexane during the final concentration . Dilute the extract to 1 .0 mL and analyze the extract by

gas chromatography using flame-photometric detectors for the determination of organophosphorous insecticides .

6.6 Prepare gas chromatograph calibration

curves daily with the mixed standards (table 1) . Operating conditions must be identical to those used for

sample analysis. Calibrate both analytical columns.

Record the volume of the standard injected and the

retention time and integrated peak area of each

component in the standard.

Mixture

number Compound

High standard

concentration

Low standard

concentration

1 ----Lindane 40 20

Heptachlor 40 20

Aldrin 40 20

DDE 40 20

DDD 40 20

Mirex 40 20

Methoxychlor 70 35

2 ----Aldrin 40 20

Heptachlor epoxide 40 20

Dieldrin 40 20

Endrin 40 20

3 ----Chlordane 200 100

4 ----Aldrin 40 20

Endosulfan 40 20

Perthane 40 20

DDT 40 20

5 ----Toxaphene 600 300

6 ----p,p-DDE 40 20

o,p-DDD 40 20

o,p-DDT 40 20

7 ----Aroclor 1016 (a PCB) 300 150

8 ----Aroclor 1254 (a PCB) 400 200

9 ----Aroclor 1260 (a PCB) 300 150

10 ----Diazinon 100 50

Malathion 130 65

Dursban 100 50

DEF 100 50

Ethion 100 50

11 ----Methyl parathion 100 50

Ethyl parathion 100 50

Methyl trithion 200 100

Trithion 200 100
30

6 .7 Inject an aliquot of sample extract into

the gas chromatograph . Record the volume injected.

Identify peaks by retention time. The identification

must be made on both analytical columns. Record

the retention time and integrated area of any identified peak. Dilute any extract containing an identifiable component above the highest standard (see table 1) to bring it within that range. Identified

compounds may be quantitated using the calculations described below.

6.8 Following analysis for organophosphorous insecticides, transfer the extract to an alumina

column . Elute with hexane and collect 10 mL. Add

1 .0 mL iso-octane to the eluate and reduce the volume to 1 .0 mL on an evaporative concentrator, rinsing the sides of the receiver two or three times with

iso-octane during the concentration . Analyze the

concentrated eluate by gas chromatography as described in steps 6.6 and 6.7 using electron-capture

detectors for organochlorine compounds.

6.9 If the extract contains multiple component mixtures such as PCB's, PCB's, toxaphene, or

interferences, it might be necessary to perform the

silica gel cleanup to obtain the fractionation shown

in table 2.

7. Calculations

7.1 Calculate the response factor of each

identified component in the calibration standard using the equation

A,

RF =

Cs X V, ,

where

RF= response factor of identified component in

calibration standard, in area/pg,

CS = concentration of standard, in pg/gL,

V = volume of standard injected, in gL, and

A, = integrated peak area of identified component in calibration standard.

7.2 Calculate the concentration of each identified component in the original water sample from

the equation

Concentration (gg/L) =

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

A2x V2

V3 XWXRF ,

where

RF= response factor of identified calibration

standard component, in area/pg,

A2 = integrated peak area of identified sample

component,

V2 = final volume of sample extract, in mL,

V3 =volume of sample extract injected, in gL,

and

W=weight of sample in g, expressed in mL

(1 .000 mL = 1 .000 g)

8. Report

8.1 Report concentrations of organochlorine

compounds (except chlordane, perthane, toxaphene,

PCB's, and PCB's) and organophosphorous insecti

cides as follows: less than 0.01 gg/L, as "less than

0.01 gg/L"; 0.01 to 0.10 gg/L, one significant figure ; 0.10 gg/L and above, two significant figures.

8.2 Report concentrations of chlordane,

perthane, PCB's, and PCB's as follows: less than 0.1

Fig/L, as "less than 0.1 Fig/L" ; 0.1 gg/L and above,

two significant figures.

8.3 Report concentrations of toxaphene as

follows : less than 1.0 gg/L, as "less than 1 .0 gg/L";

1.0 gg/L and above, two significant figures .

9. Precision

9 .1 Precision for dissolved insecticides and

PCB's (Aroclors 1248 and 1254) in distilled water

for seven replicates at each concentration are as

follows:

Compound

Concentration

spiked

(wg/L)

Mean

concentration

determined

(,tg1L)

Relative standard

deviation

(percent)

Chlordane ---------- 0 .13 0 .09 13

.25 .15 30

.50 .39 13

Aroclor 1248 -------- .20 .20 13

.41 .28 23

.82 .55 8 .8

Aroclor 1254 -------- .12 .07 28

.24 .20 15

.49 .24 4 .1

Lindane------------ .014 .012 12

.027 .035 11

.054 .064 7 .3

Heptachlor---------- .010 .007 15

.020 .015 16

.040 .042 4 .3

Aldrin ------------- .009 .008 17

.017 .016 14

.035 .042 5.0

Perthane ----------- .12 .038 9.4

.23 .17 16

.47 .14 4 .0

Endosulfan --------- .021 .015 8 .9

.042 .047 5 .2

.084 .077 6.7
DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

Mean

Concentration concetration Relative standard

spiked determined deviation

Compound (W9/L) (991L) (Percent)

9.2 It is estimated that the percent relative

standard deviation for total recoverable insecticides

and PCB's will be greater than that reported for

dissolved insecticides and PCB's.

Selected references

Goerlitz, D.F., and Brown, Eugene, 1972, Methods for analysis of

organic substances in water: U.S . Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter

A3, 40 p.

Goerlitz, D.F., and Law, L.M., 1971, Note on removal of sulfur

interferences from sediment extracts for pesticide analysis:

Bulletin of Environmental Contamination and Toxicology, v.

6, p . 9-10.

1972, Chlorinated naphthalenes in pesticide analysis: Bulletin of Environmental Contamination and Toxicology, v. 7,

p. 243-251.

1974, Determination of chlorinated insecticides in suspended sediment and bottom material: Journal of the Association of Official Analytical Chemists, v . 57, p . 176-181.

Organochlorine and organophosphorous

compounds, recoverable from bottom material

(0-5104-83) and recoverable from suspended

sediment (0-7104-83), gas chromatographic

Codes

Parameter bottom mlate'al R~ om

suspended sediment

Aldrin----------------------- 39333 39332

Chlordane--------------------39351 39353

Parameter

DDD------------------------ 39368 39362

1 . Application

This method is suitable for the determination of

recoverable organochlorine insecticides, polychlorinated biphenyls (PCB's), polychlorinated naphthalenes (PCB's), and organophosphorous insecticdes in

bottom material and suspended sediment containing

at least 0.1 leg/kg ofthe analyte.

2. Summary of method

3. Interferences

31

Codes

Recoverablefrom Recoverablefrom

bottom material suspended sediment

Organochlorine and organophosphorous insecticides, PCB's, and PCB's are extracted from suspended sediment and bottom material with acetone

and hexane. The organophosphorous insecticides are

determined by gas chromatography using flamephotometric detectors . The extracts are then purified using adsorption chromatography on an alumina column. If PCB's, PCB's, and toxaphenes are

present, the extracts are further purified using a

silica gel column. The organochlorine compounds are

determined by gas chromatography using electroncapture detectors.

Compounds having chemical and physical properties similar to the compound of interest may cause

interference . Sulfur and organosulfur compounds

will interfere, but these substances can be removed

by treating the final extracts with mercury; however, the mercury treatment will also remove organophosphorous compounds.

p,p-DDE ----------- .020 .025 19 DDE------------------------ 39368 39367

.040 .051 11 DDT------------------------39373 39372

.080 .15 3.3 Diazinon--------------------- 39571 39573

p,p-DDD ----------- .030 .020 13 Dieldrin --------------------- 39383 39382

.060 .062 9 .5 Endosulfan------------------- 39389 82355

.12 .13 6 .2 Endrin ----------------------39393 39392

p,p-DDT ----------- .053 .033 19 Ethion ----------------------39399 82347

.11 .10 7 .7 Polychlorinated biphenyls-------- 39519 39518

.21 .23 7 .0 Polychlorinated naphthalenes----- 39251 82361

Methoxychlor ------- .022 .016 8.5 Heptachlor-------------------39413 39412

.044 .041 7.9 Heptachlor epoxide------------- 39423 39422

.087 .079 5.0 Lindane ---------------------39343 39342

Mirex-------------- .020 .012 34 Malathion-------------------- 39531 39533

.041 .028 21 Methoxychlor----------------- 39481 82351

.082 .072 4.9 Methyl parathion -------------- 39601 39603

Diazinon ----------- .23 .15 20 Methyl trithion ---------------- 39791 82345

Malathion ---------- .26 .13 32 Mirex -----------------------39758 39757

Methyl parathion----- .22 .16 9.2 Parathion--------------------39541 39543

Parathion----------- .15 .12 6 .3 Perthane--------------------- 81886 82349

Methyl trithion------- .15 .07 12 Toxaphene -------------------39403 39402

Ethion ------------- .15 .12 7 .4 Trithion ---------------------39787 82343

Trithion ------------ .25 .18 7 .6
32

4. Apparatus

4.1 Alumina column: To a 130-mm x 10-mm

id (inside diameter) glass tube having a coarseporosity fritted disc, add 1 cm anhydrous sodium

sulfate, 10 em alumina, and 1 cm anhydrous sodium

sulfate.

4.2 Boiling chips, granular, micro, Hengar

H-1366C, or equivalent: Rinse with hexane, air dry,

and heat at 300'C overnight.

4.3 Centrifuge tube, 30 mL, Pyrex, graduated.

4.4 Concentrator, Kuderna-Danish (K-D),

500-mL flask, 5.0-mL volumetric receiver, and oneball Snyder column .

4.5 Gas chromatograph, Tracor model 550,

or equivalent.

4.5 .1 The following conditions are recommended for organochlorine compounds:

Columns, borosilicate glass, 1 .8 m x 2 mm

id operated at 200 ° C: Column packing materials are

(1) 3 percent SP 2100 on 100/120 mesh Supelco

port, or equivalent; and (2) 1.5 percent SP

2250 + 1.95 percent SP 2401 on 100/120 mesh

Supelcoport, or equivalent.

Detectors, electron capture, operated at

345°C.

Injection port temperature, 220'C.

Carrier gas, nitrogen or 5 percent methane in argon, flow rate 30 mL/min.

4.5 .2 The following conditions are recomTECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

4.9 Silica column: To a 130-mm x 10-mm id

glass tube having a coarse-porosity fritted disc, add

1 cm anhydrous sodium sulfate, 10 cm silica, and 1

cm anhydrous sodium sulfate .

5. Reagents

5.1 Alumina adsorbent, Woelm neutral aluminum oxide, or equivalent: Prepare deactivated adsorbent by adding 8 g deionized water to 92 g alumi

na and shake for at least 2 h on a wrist-action

shaker. The alumina is tested for required deactivation by attempting to elute the compounds of interest from a test column according to the column fractionation scheme (table 3) . If the test compounds do

not elute with 20 mL hexane from the first alumina

fraction, further deactivation is required .

5.2 Mercury, metallic, reagent grade.

5.3 Pesticide mixed standards, analytical

reference grade, EPA analytical reference standards, or equivalent: Prepare individual stock solu

tions by weighing about 10 mg of each compound to

at least three significant figures and quantitatively

transfer each compound to a 25-mL volumetric

flask. Dilute to volume with benzene and mix thoroughly. Aliquots are removed and diluted to volume

with iso-octane to obtain final concentrations listed

in table 4.

Table 3. Column fractionation scheme for alumina and silica columns for insecticides, PCB's, and PCB's in bottom

material and suspended sediment

mended for organophosphorous pesticides: ALIIMINA COLUMN

Columns, borosilicate glass, 1 .8 m x 2 mm Fraction 1

(20 mL hexane eluate)

Fraction 2

(25 mI, hexane eluate)

Fraction 3

(20 mL benzene eluate)

id operated at 175'C: Column packing materials are Aldrin Dieldrin

(1) 5 percent SP 2100 on 100/120 mesh Supelco- Chlordane Endri:n Diazinon

port, or equivalent; and (2) 2 percent SE-30 -I- 3

p,p-DDD Heptachlor epoxide Ethion

percent OV-210 on 100/120 mesh chromosorb HP, p,p-DDE

p,p-DDT

Endosulfan Parathion

Methyl parathion

or equivalent. Heptachlor Malathion

Detectors, flame photometric, Melpar, or Perthane Trithion

equivalent, operated at 210'C.

Lindane Methoxychlor

Mirex

Injection port temperature, 210 ° C. PCB's

Carrier gas, helium or nitrogen, flow rate PCN's

30 mL/min.

Toxaphene

SILICA COLUMN

4.6 Glassfilters, 142 mm, 0.3 gm mean pore

size, Gelman or equivalent: Prepare the filters by

Fraction 1

(25 mLhexane eluate)

Fraction 2

(30 mL benzene eluate)

rinsing with acetone and hexane, evaporating the Aldrin

Mirex

Chlordane

p,p-DDD

solvent, and heating overnight at 300° C. PCB's p,p-DDE

4.7 Glass wool, fine, rinsed with hexane, air PCN's p,p-DDT

dried, and heated at 300° C overnight.

p,p-DDE Perthane

Heptachlor

4.8 Shaker, wrist-action, Burrell or equiva- Lindane

lent .

Toxaphene
DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

5 .4 Silica adsorbent, Woelm silica, 70-150

mesh, or equivalent: Prepare deactivated adsorbent

by adding 0 .2 g deionized water to 99.8 g silica and

shake for at least 2 h on a wrist-action shaker . The

silica is tested for required deactivation by attempting to reproduce the elution scheme in table 3. If the

test compounds do not elute with 25 mL hexane

from the first silica fraction, further deactivation is

required.

5.5 Sodium sulfate, granular, anhydrous,

heat overnight at 300 ° C and store in a covered

beaker at 130'C.

5 .6 Solvents, acetone, benzene, hexane, and

iso-octane, distilled in glass, pesticide analysis quality, Burdick and Jackson, or equivalent.

5.7 Water, deionized, organic-free .

6. Procedure

All glassware must be washed in warm detergent

solution, rinsed with organic-free water, and heated

Table 4 Concentrations of pesticides and PCB's in mixed

standard solutions used for gas chromatograph calibration of bottom material

[Picograms per microliter]

33

at 300'C overnight. Prior to use, all glassware is

rinsed with the solvent. Do not use stopcock grease

on any ground-glass joints. For bottom-material

samples, begin at step 6 .1 . For suspended-sediment

samples, first determine the weight of the

water-suspended-sediment mixture, then filter the

sample to isolate the suspended sediment. Use the

filter and the retained sediment and begin at step

6 .2 .

6 .1 Subsampling for determination of moisture:

6.1 .1 Decant excess water from the bottom

material . Use a spatula to thoroughly mix the moist

solid. Weigh 10 g of solid into a tared weighing dish.

Record the weight to three significant figures.

6.1 .2 Place the tared dish containing the

sample in an oven at 130'C overnight. Remove from

oven, allow to cool, weigh, and record the weight to

three significant figures.

6.2 Add either the filter from the suspendedsediment filtration or the calculated amount of bottom material (not more than 100 g) to a 500-mL

Erlenmeyer flask with a ground-glass joint . Stir the

sample and slowly add deionized water until the

mixture has the consistency of paste or until water

begins to separate from the solid .

6.3 Add 20 mL acetone to the Erlenmeyer

flask containing the sample and stopper securely .

Mix the contents of the flask for 20 min using the

wrist-action shaker. Add 80 mL hexane and shake

again for 10 min. Decant the extract into a 1-L

separatory funnel containing approximately 600 mL

deionized water.

6.4 Add another 20 mL acetone to the

Erlenmeyer flask and mix for 20 min. Add 80 mL

hexane, mix 10 min, and decant the extract into the

separatory funnel . Repeat the process as in the second extraction one more time, and collect the acetone-hexane extract in the separatory funnel containing the deionized water.

6.5 Gently mix the contents of the separatory

funnel for about 1 min, and allow the layers to separate. Discard the aqueous layer and collect the ex

tract in a 500-mL Erlenmeyer flask. Add about 1 g

anhydrous sodium sulfate to the flask, cover with

foil, and allow to stand for at least 2 h or store in a

refrigerator until the analysis can continue.

6.6 Quantitatively transfer the extract with

hexane to a K-D flask fitted with a 5-mL volumetric

receiver. Add a boiling chip, attach a one-ball Sny

der column, and concentrate the extract to about 5

Mixture

number Compound

High standard

concentration

Low standard

concentration

1 ----Lindane 40 20

Heptachlor 40 20

Aldrin 40 20

p,p-DDE 40 20

p,p-DDD 40 20

Mirex 40 20

Methoxychlor 70 35

2 ----Aldrin 40 20

Heptachlor epoxide 40 20

Dieldrin 40 20

Endrin 40 20

3 ----Chlordane 200 100

4 ----Aldrin 40 20

Endosulfan 40 20

Perthane 40 20

p,p-DDT 40 20

5 ----Toxaphene 600 300

6 ----o,p-DDE 40 20

o,p-DDD 40 20

o,p-DDT 40 20

7 ----Aroclor 1016 (a PCB) 300 150

S ----Aroclor 1254 (a PCB) 400 200

9 ----Aroclor 1260 (a PCB) 300 150

10 ----Diazinon 100 50

Malathion 130 65

Ethion 100 50

11 ----Methyl parathion 100 50

Parathion 100 50

Methyl trithion 200 100

Trithion 200 100
34

mL on a water bath at about 90° C. Remove the K-D

apparatus from the water bath, allow to cool, dry the

joints with a towel, and rinse the joints into the

receiver with hexane . Disconnect the receiver and

concentrate on an evaporative concentrator to 2-4

mL, rinsing down the sides of the receiver during

concentration with small amounts of hexane . Adjust

the volume of extract in the receiver to 5.0 mL with

hexane .

6.7 Prepare an alumina column for adsorption

chromatography cleanup, referring to table 3 for the

fractionation scheme. Elute the column with 30 mL

hexane to remove contaminants. Discard the eluate.

Quantitatively transfer the extract obtained in step

6 .6 to the top of the column and elute using 45 mL

hexane (the column void volume is about 5 mL).

Collect the first 20 mL (fraction 1) and the second

20 mL (fraction 2) in graduated centrifuge tubes.

Change the elution solvent to benzene and collect 30

mL of eluate (fraction 3) . Reduce the second and

third fractions to 1 .0 mL each on an evaporative

concentrator and analyze by gas chromatography .

Treat the first fraction as described in step 6.8 .

6.8 Prepare a silica column. Elute the column

with 30 mL hexane and discard the eluate. Reduce

the volume of the first alumina fraction on an evapo

rative concentrator to about 0.5 mL and quantitatively transfer it to the top of the silica column . Add

hexane to the top of the column and collect 25 mL of

eluate (fraction 1) in a graduated centrifuge tube.

As the last of the hexane enters the top sodium

sulfate layer, add benzene to the top of the column

and collect 30 mL of eluate (fraction 2) in a graduated centrifuge tube. Reduce the volume of each of

these fractions to 1.0 mL on an evaporative concentrator and analyze each by gas chromatography .

Sulfur can be removed from the first alumina or first

silica fraction by adding several drops of mercury

and shaking for at least 1 min. The addition of mercury is continued until no further reaction occurs, as

evidenced by blackening of the mercury.

6.9 Prepare gas chromatograph calibration

curves daily with the mixed standards listed in table

4. Operating conditions must be identical to those

used for sample analysis. Calibrate both analytical

columns. Record the volume of the standard injected

and the retention time and integrated peak area of

each component in the standard.

6.10 Inject an aliquot of sample extract into

the gas chromatograph . Record the volume injected .

Identify peaks by retention time. The identification

must be made on both analytical columns. Record

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

the retention time and integrated area of any identified peak. Dilute any extract containing an identifiable component above the highest standard (see table 4).

7. Calculations

7.1 Calculate the wet weight required for a

dry weight equivalent of 50 g:

Wet weight (in g) =

W

YV2

x 50 g,

where

wet weight = weight of sample used for extraction, in g,

W = wet weight of sample, in g, and

W2 = dry weight of sample, in g.

7.2 Calculate the response factor of each

identified component in the calibration standard:

A,

RF' =

Cx V, ,

where

RF= response factor of identified component in

calibration standard, in area/pg,

CS = concentration of standard, in pg/gL,

V, = volume of standard injected, in gL, and

A, = integrated peak area of identified component in calibration standard.

7.3 Calculate the concentration of each identified component in the original bottom-material sample from the equation

A2 X V2

Concentration (gg/kg) =

V3xWxRF

where

RF= response factor of identified calibration

standard component, in area/pg,

A2 = integrated peak area of identified sample

component,

V2 = final volume of sample extract, in mL,

V3 = volume of sample extract injected, in gL,

and

W= dry weight equivalent of sample, in g.

7.4 Calculate the concentration of each identified component in the original suspended sediment

from the following equation :

A2 XV2

Concentration ("o-/L) =

V

X

V4 XRF
8. Report

9. Precision

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

where

RF= response factor of identified calibration

standard component, in area/pg,

AZ =integrated peak area of identified sample

component,

V2 = final volume of sample extract, in mL,

V3 = volume of sample extract injected, in gL,

and

V4 =weight of sample in g expressed in mL

(1 .000 mL = 1 .000g) .

8.1 Bottom material

8.1 .1 Report concentrations of organochlorine compounds (except chlordane, perthane, toxaphene, PCB's, and PCB's) and organophosphorous

insecticides in bottom material as follows : less than

0.1 pg/kg, as "less than 0.1 Ftg/kg" 0.1 to 1 .0

lag/kg, one significant figure; 1.0 Fig/kg and above,

two significant figures .

8.1 .2 Report concentrations of chlordane,

perthane, PCB's, and PCB's in bottom materials as

follows : less than 1.0 gg/kg, as "less than 1.0

gg/kg"; 1.0 lag/kg and above, two significant

figures .

8.1 .3 Report concentrations of toxaphene

in bottom materials as follows: less than 10 Fig/kg,

as "less than 10 gg/kg"; 10 lag/kg and above, two

significant figures .

8.2 Suspended sediment

8.2 .1 Report concentrations of organochlorine compounds (except chlordane, perthane, toxaphene, PCB's, and PCB's) and organophosphorous

insecticides in suspended sediment as follows: less

than 0.01 gg/L as "less than 0.01 lag/L" ; 0.01 to

0.10 Fig/L, one significant figure ; 0.1 Fig/L and

above, two significant figures.

8.2 .2 Report concentrations of chlordane,

perthane, PCB's, and PCB's in suspended sediment

as follows: less than 0.1 pg/L, as "less than 0.1

lag/L" ; 0.1 wg/L and above, two significant figures.

8.2 .3 Report concentrations of toxaphene

in suspended sediment as follows: less than 1 .0

lag/L, as "less than 1 .0 Fig/L" ; 1 .0 gg/L and above,

two significant figures.

It is estimated that the percent relative standard

deviation for recoverable insecticides and PCB's

from bottom material and suspended sediment will

35

be greater than that reported for dissolved insecticides and PCB's (method 0-1104-83).

Selected references

Goerlitz, D.F ., and Brown, Eugene, 1972, Methods for analysis of

organic substances in water: U.S . Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter

A3, 40 p .

Goerlitz, D.F ., and Law, L.M., 1971, Note on removal of sulfur

interferences from sediment extracts for pesticide analysis :

Bulletin of Environmental Contamination and Toxicology, v .

6, p . 9-10.

1972, Chlorinated naphthalenes in pesticide analysis: Bulletin of Environmental Contamination and Toxicology, v 7,

p . 243-251.

1974, Determination of chlorinated insecticides in suspended sediment and bottom material: Journal of the Association of Official Analytical Chemists, v. 57, p. 176-181 .

Organochlorine compounds, recoverable from

fish tissue, gas chromatographic (0-9104-83)

Parameter Code

Aldrin ----------------------------- None assigned .

Chlordane

DDD

DDE

DDT

Dieldrin

Endosulfan

Endrin

Polychlorinated biphenyls

Polychlorinated naphthalenes

Heptachlor

Heptachlor epoxide

Lindane

Methoxychlor

Mirex

Perthane

Toxaphene

1 . Application

This method is suitable for the determination of

organochlorine compounds in fish containing at least

0.1 gg/kg of the analyte .

2. Summary of method

A homogenized sample of whole fish or fish fillet

is extracted with petroleum ether to isolate the fat.

The organochlorine compounds are extracted from

the fat with acetonitrile . The acetonitrile extract is

diluted with water and extracted with petroleum

ether to partition the organochlorine compounds.
36

The petroleum ether extract is concentrated and purified using adsorption chromatography . Organochlorine compounds are identified and quantified by

gas chromatography using electron-capture detectors.

3. Interferences

Sulfur and organosulfur compounds will interfere,

but these substances can be removed by treating the

final extracts with mercury.

4. Apparatus

4.1 Alumina column: To a 130-mmx10-mm

id glass tube having a coarse-porosity fritted disc,

add 1 cm anhydrous sodium sulfate, 10 cm alumina,

and 1 cm anhydrous sodium sulfate .

4.2 Blender, 3 .8-L-capacity, Waring, threespeed with stainless steel container, cover, and blade

assembly, or equivalent.

4 .3 Blender, 1 .2 L-capacity, Waring, with explosion-resistant motor base, borosilicate glass containers, vinyl/plastic cover, and stainless steel blade

assembly, or equivalent.

4 .4 Boiling chips, micro, granular, Hengar

H-1366C, or equivalent. Rinse with hexane, air dry,

and heat overnight at 300'C.

4.5 Concentrator, Kuderna-Danish (K-D),

500 mL, with 5-mL and 10-mL volumetric receivers

and a one-ball Snyder column.

4.6 Evaporative concentrator, Organomation

N-Evap, or equivalent.

4.7 Fish tissue preparation equipment, consisting of a nonporous ceramic or stainless steel cutting board, a heavy-duty stainless steel knife, and a

stainless steel spatula having a 25-cm blade.

4.8 Gas chromatograph, Tracor 560, or

equivalent.

4.8 .1 The following conditions are recommended:

Columns, borosilicate glass, 1 .8 mx2 mm

id operated at 200'C: Column packing materials are

(1) 3 percent SP 2100 on 100/120 mesh Supelco

port, or equivalent; and (2) 1.5 percent SP

2250 + 1.95 percent SP 2401 on 100/120 mesh

Supelcoport, or equivalent.

Detectors, electron capture, operated at

345°C.

Injection port temperature, 220'C.

Carrier gas, nitrogen or 5 percent methane in argon, flow rate 30 mL/min.

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

4.9 Glassfilter, 142 mm, 0.3 gm mean pore

size, Gelman, or equivalent: Prepare the filters by

rinsing with acetone and hexane, evaporating the

solvent, and heating overnight at 300°C.

4.10 Silica column: To a 10-mmx 130-mm id

glass tube having a sealed-in, coarse-porosity fritted

disc, add 1 cm anhydrous sodium sulfate, 10 cm

silica, and 1 cm anhydrous sodium sulfate.

5. Reagents

5.1 Acetonitrile, petroleum ether saturated :

Place 800 mL acetonitrile in a 1-L separatory funnel. Add sufficient petroleum ether (about 50 mL) so

that after vigorously mixing, an excess of petroleum

ether is visible above the acetonitrile . Draw off and

retain the lower layer.

5.2 Alumina adsorbent, Woelm neutral aluminum oxide, or equivalent: Prepare deactivated adsorbent by adding 8 g deionized water to 92 g alumi

na and shake for at least 2 h on a wrist-action

shaker. The alumina is tested for required deactivation by attempting to elute the organochlorine compounds of interest from a test column according to

the column fractionation scheme (table 5) .

5.3 Mercury, metallic, reagent grade.

Table 5. Column fractionation scheme for alumina and silica columns for organochlorine insecticides, PCB's, and

PCB's in fish

ALUMINA COLUMN

Fraction 1 Fraction 2

20 mL hexane eluate 20 mL hexane eluate

Aldrin Dieldrin

Chlordane Endrin

Heptachlor Heptachlor epoxide

p,p-DDD Endosulfan

p,p-DDE

p,p-DDT

Lindane

Mirex

Perthane

PCB's

PCB's

Toxaphene

SILICA COLUMN

Fraction 1 Fraction 2

(25 mL hexane eluate) (30 mL bezene eluate)

Aldrin Chlordane

Mirex p,p-DDD

PCB's p,p-DDE

PCB's p,p-DDT

p,p-DDE Perthane

Heptachlor

Lindane

Toxaphene
DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS 37

5.4 Silica adsorbent, Woelm silica, 70-150

mesh, or equivalent: Prepare deactivated adsorbent

by adding 0 .2 g deionized water to 99.8 g silica, and

shake for at least 2 h on a wrist-action shaker. The

silica is tested for required deactivation by attempting to reproduce the elution scheme in table 5.

5 .5 Sodium chloride, granular, reagent

grade: Heat at 300°C overnight before use .

5.6 Sodium chloride solution, saturated : Dissolve 360 g sodium chloride in 1,000 mL deionized

water. Add additional sodium chloride in about 5 g

increments, stirring well after each addition, until an

excess of the salt is observed.

5.7 Sodium hypochlorite solution, (5 percent), Clorox bleach, or equivalent.

5 .8 Sodium sulfate, granular, anhydrous:

Heat overnight at 300°C and store in a covered

beaker at 130'C.

5.9 Solvents, acetonitrile, benzene, hexane,

iso-octane, petroleum ether, distilled in glass, pesticide analysis quality, Burdick and Jackson, or equivalent.

5.10 Pesticide mixed standards, analytical

reference grade, EPA analytical reference stanTable 6. Concentrations of pesticides and PCB's in mixed

standard solutions used for gas chromatograph calibration of fish tissue

[Picograms per microliter]

dards, or equivalent: Prepare individual stock solutions by weighing about 10 mg of each compound to

at least three significant figures, and quantitatively

transfer each compound to a 25-mL volumetric

flask. Dilute to volume with benzene and mix thoroughly. Aliquots are removed and diluted to volume

with iso-octane to obtain final concentrations listed

in table 6.

6. Procedure

All glassware must be washed in warm detergent

solution, rinsed with organic-free water, and heated

at 300'C overnight . Rinse all glassware, fish-preparation equipment, and blender containers with petroleum ether before use, and air dry . After use with

fish samples, the glassware should be washed immediately in a warm solution of detergent and bleach to

aid in the removal of residual oils and odor. Do not

use stopcock grease on ground-glass joints .

6.1 Sample preparation :

6.1 .1 Scrape the scales from whole fish

samples and cut off tough fins . Discard scales and

fins. Chop whole fish into small pieces. If sample is a

fillet, slice into small pieces.

6.1 .2 Add fish pieces to a stainless steel

blender container, cover, and blend until a homogeneous sample is obtained. Weigh 50 g of blended

sample and place in a tared, glass blender container.

Record the weight of the sample in the glass blender

container to three significant figures. (Use excess

sample for a duplicate, or store it frozen if a rerun is

needed.)

6.2 Extraction of fat from fish samples:

6.2 .1 Add 100 g anhydrous sodium sulfate

to the weighed fish in the glass blender container

and blend until thoroughly mixed. Scrape the sides

of the container with a spatula during mixing to

ensure homogeneity.

6.2 .2 Add 150 mL petroleum ether to the

sample in the blender container, cover, and blend at

high speed for 2 min using the explosion-resistant

base. (Start blending at low speed to avoid splashing

and gradually increase to high). CAUTION: Petroleum ether is extremely flammable . Provide additional

air circulation around the blender base to prevent

solvent fumes from contacting the motor housing .

6.2 .3 Slowly decant the petroleum ether

extract into a Buchner funnel fitted with filter paper. Slowly filter the extract into the filtration flask

using a water aspirator. (Care must be taken to

prevent solids from entering the filtrate .)

Mixture

number Compound

High standard

concentration

Low standard

concentration

1------Lindane 40 20

Heptachlor 40 20

Aldrin 40 20

p,p-DDE 40 20

p,p-DDD 40 20

Mirex 40 20

Methoxychlor 70 35

2------ Aldrin 40 20

Heptachlor epoxide 40 20

Dieldrin 40 20

Endrin 40 20

3 ------Chlordane 200 100

4------ Aldrin 40 20

Endosulfan 40 20

Perthane 40 20

p,p-DDT 40 20

5------ Toxaphene 600 300

6------ o,p-DDE 40 20

o,p-DDD 40 20

o,p-DDT 40 20

7------Aroclor 1016 (a PCB) 300 150

8------Aroclor 1254 (a PCB) 400 200

9------Aroclor 1260 (a PCB) 300 150
38

6 .2 .4 Scrape down the walls of the blender

container and break up any caked material with a

spatula. Add 100 mL petroleum ether, cover, and

repeat the extraction (step 6.2 .2), blending for 1

min. Scrape down the walls of the blender container

and blend for another minute.

6.2 .5 Slowly decant the petroleum ether

extract into the Buchner funnel and filter .

6.2 .6 Repeat steps 6.2 .4 and 6.2 .5 . Quantitatively transfer the remaining fish residue from

the blender to the Buchner funnel using the spatula

and small amounts of petroleum ether to aid in the

transfer . Continue aspirating until the residue appears dry .

6 .2 .7 Quantitatively transfer the filtrate to

a 500-mL Erlenmeyer flask . Add 2 g anhydrous sodium sulfate, cover the flask, and allow to stand for

at least 1 h.

6.2 .8 Weigh a 10-mL receiver for the concentration step and record its weight to two significant figures . Quantitatively transfer the extract into

a K-D flask fitted with the weighed receiver. Add a

boiling chip, fit the K-D flask with a Snyder column,

and concentrate the extract to about 5 mL on a

water bath maintained at 90°C.

6.2 .9 Remove the K-D apparatus from the

water bath, disconnect the receiver, and place it on

an evaporative concentrator to evaporate any re

maining petroleum ether. The receiver contains the

fat extract .

6.2 .10 Weigh the receiver, record the

weight, and calculate the weight of the extract. If

the extracted fat weighs more than 3 g, take 3.0 g

for the liquid-liquid partitioning (step 6.3) and use

the formula in calculation 7.1 to determine the final

sample weight. If the weight of the extracted fat is 3

g or less, the analysis is based on the weight of the

original sample (step 6 .1 .1) .

6.3 Liquid-liquid partitioning:

6.3 .1 Quantitatively transfer the extract (3

g or less) to a 125-mL separatory funnel using small

amounts of petroleum ether to bring the total volume to about 15 mL.

6.3 .2 Add 30-mL petroleum-ether-saturated acetonitrile to the separatory funnel . Stopper and

shake vigorously for at least 1 min, venting often.

Allow the layers to separate.

6.3 .3 Drain the acetonitrile layer (bottom)

into a 1,000-mL saturated sodium chloride solution,

and add 100 mL petroleum ether. Retain the fat

extract in the 125-mL funnel.

TECHNIQUES OFWATER- RESOURCES INVESTIGATIONS

6.3 .4 Repeat steps 6.3 .2 and 6 .3 .3 twice.

Combine all acetonitrile extracts in the 1,000-mL

separatory funnel .

6.3 .5 Stopper the 1,000-mL separatory

funnel and shake vigorously for 1 min, venting often. Allow the layers to separate and drain the aque

ous layer into a 1-L glass bottle . Pour the petroleum

ether extract from the funnel into a 250-mL

Erlenmeyer flask.

6 .3 .6 Return the aqueous layer to the separatory funnel, add 100 mL petroleum ether to the

bottle, swirl to rinse sides, and pour into separatory

funnel.

6.3 .7 Repeat the extraction (step 6.3 .5) .

Allow the layers to separate, discard the aqueous

layer, and add the extract to the Erlenmeyer flask

containing the first extract (step 6.3 .5) . If the extract is highly colored, wash it twice by extracting it

with 100-mL portions of deionized water. Discard

the aqueous layers after each extraction and transfer the final extract to the 250-mL Erlenmeyer

flask . Add 2 g sodium sulfate to the flask, cover, and

allow the extract to stand over sodium sulfate for at

least 1 h.

6.4 Concentration and cleanup :

6.4 .1 Quantitatively transfer the extract

with hexane to a K-D flask fitted with a 5-mL volumetric receiver. Add a boiling chip, attach a Snyder

column, and concentrate the extract to about 5 mL

on a 95'C water bath. Remove the K-D apparatus

from the water bath, allow to cool, wipe the joints

with a towel, disconnect the receiver, rinse the lower

joint with hexane into the receiver, and concentrate

the extract on an evaporative concentrator to 2-4

mL. Rinse down the sides of the receiver during

concentration with small amounts of hexane . Adjust

the volume of extract in the receiver to 5 .00 mL with

hexane.

6 .4 .2 Prepare an alumina column for the

alumina fractionation, referring to table 5 for the

fractionation scheme. Elute the column with 30 mL

hexane to remove contaminants . Discard the eluate .

Quantitatively transfer the extract from step 6.4 .1

to the top of the column and elute using 45 mL

hexane (the column holdup is about 5 mL). Collect

the first 20 mL (fraction 1) and the second 20 mL

(fraction 2) in graduated centrifuge tubes. Reduce

the second alumina fraction to 1 .0 mL on the evaporative concentrator and analyze by gas chromatography. Treat the first alumina fraction as described

in step 6.4 .3 .
7 Calculations

DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS

6.4 .3 Prepare a silica column for the silica

fractionation, referring to table 5. Elute the column

with 30 mL hexane and discard the eluate . Reduce

the volume of the first alumina fraction to about 0.5

mL on the evaporative concentrator and quantitatively transfer it to the top of the silica column. Add

hexane to the top of the column and collect 25 mL of

eluate (fraction 1) in a graduated centrifuge tube.

As the last of the hexane enters the top sodium

sulfate layer, add benzene to the top of the column

and collect 30 mL of eluate (fraction 2) in a graduated centrifuge tube. Reduce the volume of each of

these two fractions to 1 .0 mL on the evaporative

concentrator, and analyze each by gas chromatography-Removal

of sulfur from the first alumina

and first silica fractions is accomplished by adding

several drops of mercury and shaking for at least 1

min. The addition of mercury is continued until no

further reaction occurs, as evidenced by blackening

of the mercury.

6.5 Sample analysis:

6.5 .1 Prepare gas chromatograph calibration curves daily with the mixed standards listed in

table 6. Operating conditions must be identical to

those used for sample analysis. Calibrate both analytical columns. Record the volume of the standard

injected and the retention time and integrated peak

area of each component in the standard .

6.5 .2 Inject an aliquot of sample extract

(from steps 6.4 .2 and 6.4 .3) into the gas chromatograph. Record the volume injected . Identify

peaks by retention time. The identification must be

made on both analytical columns. Record the retention time and integrated area of any identified peak.

Dilute any extract containing an identifiable component above the highest standard.

7.1 Determine the equivalent subsample

weight from the following equation:

WXW3

W= W

,

2

(W = W2 when W2 is equal to or less than 3 g)

where

W= equivalent subsample weight, in g,

W = weight of fat taken for cleanup, in g,

W2 = total weight of extracted fat, in g, and

W3 = weight of original fish subsample, in g.

7.2 Calculate the response factor of each

identified component in the calibration standard :

where

RF=response factor of identified component in

calibration standard, in area/pg,

CS = concentration of standard, in pg/gL (step

5.10),

V = volume of standard injected, in gL (step

6.5 .1), and

A, = integrated peak area of identified component in calibration standard (step 6.5 .1) .

7 .3 Calculate the concentration of each identified component (see step 6.5 .2) in the original fish

sample from the equation

where

RF= response factor of identified calibration

standard component, in area/pg (step

7 .2),

A2 = integrated peak area of identified sample

component (step 6.5 .2),

V2 = final volume of sample extract, in mL (step

6.4 .3),

V3 = volume of sample extract injected, in AIL

(step 6.5 .2), and

W=equivalent subsample weight of fish, in g

(calculation 7 .1) .

8. Report

8.1 Report concentrations of organochlorine

compounds (except chlordane, perthane, and toxaphene) in fish samples as follows: less than 0.1

pg/kg, as "less than 0.1 wg/kg" ; 0.1 to 1.0 gg/kg,

one decimal ; 1 .0 gg/kg and above, two significant

figures.

8.2 Report concentrations of chlordane,

perthane, PCB's, and PCB's in fish samples as follows: less than 1 .0 gg/kg, as "less than 1 .0 igg/kg" ;

1.0 leg/kg and above, two significant figures.

8.3 Report concentrations of toxaphene in

fish samples as follows: less than 10 igg/kg, as "less

than 10 gg/kg"; 10 leg/kg and above, two significant figures.

9. Precision

A, RF =

CS x V

2xV

Concentration (gg/L) =

A

V3 x WXRF

Precision data are not available.

39
40

Selected references

Goerlitz, D.F ., and Law, L.M., 1971, Note on removal of sulfur

interferences from sediment extracts for pesticide analysis :

Bulletin of Environmental Contamination and Toxicology, v .

6, p 1-10 .

1972, Chlorinated naphthalenes in pesticide analysis : Bulletin of Environmental Contamination and Toxicology, v . 7,

p. 243-251.

1974, Determination of chlorinated insecticides in suspended sediment and bottom material : Journal of the Association of Official Analytical Chemists, v. 57, p. 176-181 .

Lamar, W.L ., Goerlitz, D.F., and Law, L.M., 1965, Identification

and measurement of chlorinated organic pesticides in water

by electron-capture gas chromatography : U.S . Geological

Survey Water-Supply Paper 1817-B, p. 12.

1966, Determination of organic insecticides in water by

electron-capture gas chromatography, in American Chemical Society, Advances in Chemistry Series, Number 60, Organic Pesticides in the Environment, p . 187-199.

Law, L.M., and Goerlitz, D.F ., 1970, Microcolumn chromatographic cleanup for the analysis of pesticides in water: Journal of the Association Official Analytical Chemists, v . 53, no .

6,p.1276-1286.

Thompson, J .F ., ed., 1974, Analysis of pesticide residues in

human and environmental samples : Research Triangle Park,

N.C., U.S . Environmental Protection Agency, Environmental

Toxicology Division, Health Effects Research Laboratory,

Sec . 5,A(1), p . 1-14.

(U .S .) Federal Water Pollution Control Administration, 1969,

FWPCA method for chlorinated hydrocarbon pesticides in

water and wastewater: Federal Water Pollution Control Administration, Cincinnati, Ohio, p. 29 .

Chlorophenoxy acids, total recoverable

(0-3105-83) and dissolved (0-1105-83), gas

chromatographic

Code

Parameter Total recoverable Dissolved

2,4-D ---------------------39730 39732

2,4-DP --------------------82183 82356

Silvex---------------------39760 39762

2,4,5-T--------------------39740 39742

1 . Application

This method is suitable for the determination of

chlorophenoxy acid herbicides, and their esters and

salts, in water and water-suspended-sediment mixtures containing at least 0.01 pg/L of the analyte .

2. Summary of method

Chlorophenoxy acid herbicides and their esters

are extracted with either diethyl or methyl t-butyl

ether from an acidified water sample. The extracted

herbicides are hydrolyzed to the free acids which are

converted to their methyl esters with boron trifluoTECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

ride-methanol and purified using adsorption chromatography. The methyl esters are determined by

gas chromatography using electron capture detectors.

3. Interferences

Halogenated organic acids, and their salts and

esters, may cause interference .

4 Apparatus

4.1 Boiling chips, granules, micro, Hengar,

H-1366C, or equivalent: Rinse with hexane, air dry,

and heat at 300°C overnight .

4 .2 Centrifuge tube, 5 mL, Pyrex, graduated,

with ground-glass stopper.

4.3 Concentrator, Kuderna-Danish (K-D),

125-mL flask and 5.0-mL receiver, one-ball Snyder

column, and modified micro-Snyder column, Kontes

569251, or equivalent.

4 .4 Florisil column, a disposable glass pipet

with glass-wool plug: Fill to a depth of 1 .5 cm with

florisil adsorbent, followed by 2 cm sodium sulfate.

4 .5 Gas chromatogroph, Tracor Model 550,

or equivalent.

4 .5 .1 The following conditions are recommended:

Columns, borosilicate glass, 1 .8 mx 2 mm

id (inside diameter), operated at 180'C: Column

packing materials are (1) 3 percent SP 2100 on

100/120 mesh Supelcoport ; and (2) 3 percent SP

2250 on 100/120 mesh Supelcoport, or equivalent.

Detector, dual electron capture operated at

350°C.

Injection port temperature, 200°C.

Carrier gas, nitrogen, flow rate 20

mL/min.

4 .6 Glassfilters, 142 mm, 0.3 gm mean pore

size, Gelman, or equivalent: Prepare the filters by

rinsing with acetone and hexane, evaporating the

solvent, and heating overnight at 300'C.

4.7 Glass wool, fine, rinsed with hexane, air

dried, and heated at 300 ° C overnight.

4.8 Sandbath, Tecam, or equivalent.

5. Reagents

5.1 Boron trifuoride-methanol esterification

reagent, 14 percent BF3; (weight/volume ; w/v) in

methanol, Applied Science Labs, or equivalent.

5.2 Florisil adsorbent, commercially activated at 650 ° C, washed with hexane, allowed to air

dry, and stored at 130°C in a glass-stoppered flask:

Prior to use, the florisil is deactivated by adding 10

percent water by weight and shaking for at least 2 h
DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS

on a wrist-action shaker. The florisil is then tested

for activity by attempting to elute the herbicides of

interest with benzene from a test column. If the test

compounds do not elute within 2.0 mL, further deactivation is required until the desired results are obtained .

5.3 Potassium hydroxide solution, 37 percent (w/v) : Dissolve 78 g KOH reagent-grade pellets

in 200 mL deionized water. Iteflux for 8 h.

5.4 Sodium sulfate, acidified: Prepare a slurrv of sodium sulfate with enough diethyl ether to

cover the crystals, and acidify to pH 2 or less by

adding a few milliliters of concentrated sulfuric acid.

Determine the pH by transferring a small portion of

the slurry to a beaker and removing the ether by

evaporation . Add a few drops of deionized water to

the crystals and measure the pH with pH paper.

Allow to air dry overnight . Store in a covered Pyrex

beaker or flask at 130'C.

5.5 Sodium sulfate, granular, anhydrous:

Heat overnight at 300°C and store covered at

130'C.

5.6 Sodium sulfate solution, 5 percent (w/v):

Dissolve 50 g neutral sodium sulfate in deionized

water and dilute to 1 L.

5.7 Solvents, benzene, unpreserved diethyl

ether or methyl t-butyl ether, and iso-octane, distilled in glass, pesticide analysis quality, Burdick and

Jackson, or equivalent: Diethyl ether preserved with

ethanol cannot be used in this procedure because it

results in the formation of extraneous ethyl esters.

5.8 Standards, methyl esters of ehlorophenoxy acid herbicides, EPA analytical reference

grade or equivalent: Prepare a stock solution by

weighing about 10 mg of compound to at least three

significant figures and transfer to a 25-mL volumetric flask. Dilute to volume with benzene and mix

thoroughly. Prepare a series of mixed-compound

standards by volumetric dilution with iso-octane, as

described in table 7.

5.9 Sulfuric acid, concentrated (sp . gr. 1 .84),

Mallinckrodt analytical reagent, A.C.S . grade, or

equivalent.

5.10 Sulfuric acid, (1 -1- 3) : Prepare by adding 1 part concentrated sulfuric acid to 3 parts

deionized water. Store in a refrigerator at VCC.

5 .11 Water, deionized, organic-free .

6 Procedure

All glassware must be washed in warm detergent

solution, rinsed with organic-free water, and heated

Table 7. Concentration of herbicides in mixed standard solutions used for gas chromatograph calibration of water

and water-suspended sediment

[Picograms per microliter]

4 1

at 300'C overnight . Prior to use, all glassware is

rinsed with the solvent it will contact. Stopcock

grease should not be used on ground-glassjoints .

For the determination of dissolved components,

filter the sample through a glass filter to remove the

suspended material . Pour the filtrate into the original sample bottle and continue with the procedure .

6.1 Immediately upon receipt of a sample in

the laboratory, it must be acidified to pH 2 or lower

with concentrated sulfuric acid and stored at VCC.

6 .2 A blank must accompany each group of

samples . For each sample, rinse a 1,000-mL separatory funnel and a 250-mL Erlenmeyer flask with

ether.

6.3 Weigh the sample bottle plus sample and

record the weight to three significant figures.

6 .4 Pour the sample into the separatory funnel and allow the bottle to drain completely. Weigh

the empty bottle and record the weight to three

significant figures. Calculate and record the sample

weight.

6 .5 Add 150 mL ether to the sample bottle,

rinse the sides thoroughly, and pour the solvent into

the separatory funnel . The Teflon-lined cap is not

rinsed because of the potential for contamination

from solvent that has contacted the threads and the

surface beneath the Teflon liner . Shake the funnel

vigorously for at least 1 min, venting often. Allow

the layers to separate and drain the aqueous layer.

Pour the ether extract into the Erlenmeyer flask .

Extract the sample twice more, using 50 mL ether

each time, and collect the extracts in the Erlenmeyer

flask.

6.6 Add 15 mL distilled water, 2.0 mL 37

percent KOH, and a boiling chip to the extract . Fit

the flask with a Snyder column and heat the assem

bly on a steam bath for a total of 90 min, during

which time the ether will evaporate and the herbicide esters are hydrolyzed.

Herbicide High standard

concentration

Lowstandard

concentration

2,4-DP ----------------- 100 50

2,4-D------------------ 100 50

2,4,5-T ---------------- 40 20

Silvex------------------- 40 20
42

6.7 Remove the assembly from the water

bath, allow to cool, and quantitatively transfer the

water to a 125-mL separatory funnel . Extract the

basic solution with 20 mL ether and discard the

ether layer; repeat twice with 10 mL ether and discard the ether layers. The herbicide potassium salts

remain in the aqueous phase. Add 2 mL sulfuric acid

(1 -i- 3) to the contents of the funnel to bring the pH

to 2 or below, and extract the aqueous phase with 20

mL ether; repeat twice with 10 mL ether to extract

the herbicides in their acid forms. Collect the ether

extracts in a 125-mL Erlenmeyer flask containing

about 0.5 g acidified sodium sulfate . Cover the flask

with foil and set aside for at least 1 h, or store in a

refrigerator until analysis can continue.

6.8 Quantitatively transfer the ether extract

into a K-D apparatus fitted with a 5-mL volumetric

receiver. add 1 mL benzene and a boiling chip. Con

centrate the extract to about 0.5 mL on a fluidized

sandbath heated to 60-70°C . Under no circumstances should the extract be allowed to evaporate

completely to dryness. Clear sand from the glass

joint before opening. (Use a water bath at 80'C for

methyl t-butyl ether extracts .) Rinse the bottom

joint with benzene into the receiver.

6.9 After the benzene solution in the receiver

has cooled, add 0.5 mL boron trifluoride-methanol

reagent . The modified Snyder column is used as an

air-cooled condenser, and the contents of the receiver are held at 50°C for 30 min in a sandbath . Cool

the reaction mixture to room temperature and add

sodium sulfate solution until the benzene-aqueous

solution interface is observed in the restricted neck

ofthe receiver. Stopper the receiver, shake vigorously for 1 min, and allow to stand for at least 1 h for

phase separation. Loosen the stopper after shaking .

6.10 Transfer the benzene layer from the receiver to a florisil column. Elute with benzene until a

total of 2.0 mL of benzene has been collected in a

graduated centrifuge tube. Analyze the eluate by

gas chromatography .

6.11 Prepare gas chromatograph calibration

curves daily with the mixed standards listed in table

7. Operating conditions must be identical to those

used for sample analysis. Calibrate both analytical

columns. Record the volume of the standard injected

and the retention time and integrated peak area of

each component in the standard.

6.12 Inject an aliquot of sample extract into

the gas chromatograph . Record the volume injected .

Identify peaks by retention time. The identification

must be made on both analytical columns. Record

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

the retention time and integrated area of any identified peak. Dilute any extract containing an identifiable component above the highest standard .

7. Calculations

7.1 Calculate the response factor of each

identified component in the calibration standard:

8. Report

RF =

A,

CXV,

where

RF=response factor of identified component in

calibration standard, in area/pg,

CS = concentration of standard, in pg/ gL, (step

5 .8,

V, = volume of standard injected, in p.L (step

6.11), and

A, = integrated peak area of an identified component in calibration standard (step

6.11) .

7.2 Calculate the concentration of each identified component in the original sample from the

equation

Concentration (gg/L) =

A2X V2

V3 xWxRF

where

RF=response factor of identified component in

sample, in area/pg,

A2 =integrated peak area of identified component,

V2 = final volume of sample extract, in mL,

V3 = volume of sample extract injected, in gL,

and

W= weight of sample in g, expressed in mL

(1 .000 mL= 1 .000 g).

7.3 Calculate the free chlorophenoxy acid

concentration :

Concentration of acid (wg/L) = Cxf,

where

C = concentration of methyl ester (calculation

7.2), and

_ molecular weight of acid

f molecular weight of methyl ester

Report chlorophenoxy acid herbicide concentrations as follows : less than 0.01 Fig/L, as "less than
9. Precision

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS 43

0.01 gg/L"; 0.01 gg/L to 0.10 pg/L, one significant

figure; 0.10 lag/L and above, two significant figures.

9.1 Precision for dissolved chlorophenoxy

acids in deionized water for 35 replicates using

diethyl ether is as follows:

Mean Relative

Spiked concentration standard

concentration determined deviation

Compound 4L9/L) 4t91L) (Percent)

2,4-D -------------------- 0 .10 0 .075 10 .0

Silvex -------------------- .048 .036 11 .6

2,4,5-T------------------- .058 .045 12 .2

9.2 Precision for dissolved chlorophenoxy

acids in deionized water for 35 replicates using

methyl t-butyl ether is as follows:

Mean Relative

Spiked concentration standard

concentration determined deviation

Compound (AgIL) (W9lL) (percent)

2,4-D -------------------- 0 .10 0.083 10.1

Silvex -------------------- .048 .040 10.4

2,4,5-T------------------- .056 .049 10.0

9.3 It is estimated that the percent relative

standard deviation for total recoverable chlorophenoxy acids will be greater than that reported for

dissolved chlorophenoxy acids.

Selected references

Goerlitz, D.F., and Brown, Eugene, 1972, Methods for analysis of

organic substances in water: U.S . Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter

A3, 40 p.

Goerlitz, D.F., and Lamar, W.L., 1967, Determination of phenoxy

acid herbicides in water by electron-capture and

microcoulometric gas chromatography: U.S . Geological Survey Water-Supply Paper 1817-C, 21 p .

Chlorophenoxy acids, recoverable from bottom

material (0-5105-83) and recoverable from

suspended sediment (0-7105-83), gas

chromatographic

Code

Recoverablefrom Recoverablefrom

Parameter bottom material suspendedsediment

2,4-D------------------ 39731 39733

2,4-DP ----------------- 34609 34608

Silvex ----------------- 39761 39763

2,4,5-T ---------------- 39741 39743

1 . Application

This method is suitable for the determination of

chlorophenoxy acid herbicides, and their esters and

salts, in bottom material and in suspended sediment

isolated from water containing at least 0.1 gg/kg

and 0.01 gg/L of the analyte, respectively.

2. Summary of method

Chlorophenoxy acid herbicides and their esters

are extracted with either diethyl or methyl t-butyl

ether from an acidified slurry of bottom material or

suspended sediment and water. The extracted herbicides are hydrolyzed to the free acids, which are

converted to their methyl esters with boron trifluoride-methanol and purified using adsorption chromatography. The methyl esters are determined by

gas chromatography using electron-capture detectors.

3. Interferences

Halogenated organic acids, and their salts and

esters, may cause interference.

4. Apparatus

4 .1 Boiling chips, granules, micro, Hengar

H-1366C, or equivalent: Rinse with hexane, air dry,

and heat at 300'C overnight .

4 .2 Centrifuge tube, 10-mL, Pyrex, graduated, with ground-glass stopper.

4 .3 Concentrator, Kuderna-Danish (K-D),

125-mL flask, 5.0-mL volumetric receiver, one-ball

Snyder column, and modified micro-Snyder column,

Kontes 569251 or equivalent.

4.4 Florisil column: To a 130 mmx 10 mm id

(inside diameter) glass tube having a coarse-porosity

fritted disc, add 3 cm florisil and 1 cm anhydrous

sodium sulfate .

4.5 Gas chromatograph, Tracor model 550,

or equivalent.

4.5 .1 The following conditions are recommended:

Columns, borosilicate glass, 1 .8 mx 2 mm

id operated at 180°C: Column packing materials are

(1) 3 percent SP 2100 on 100/120 mesh Supelco

port; and (2) 3 percent SP 2250 on 100/120 mesh

Supelcoport, or equivalent.

Detector, dual electron capture operated at

350°C.

Injection port temperature, 200° C.

Carrier gas, nitrogen, flow rate 20

mL/min.

4 .6 Glassfilters, 142 mm, 0.3 gm mean pore

size, Gelman, or equivalent: Prepare the filters by

rinsing with acetone and hexane, evaporating the

solvent, and heating overnight at 300'C.
44

4.7 Glass wool, fine, rinsed with hexane, air

dried, and heated at 300 ° C overnight .

4.8 Oven, Precision model 18EGE, or, equivalent, capable of maintaining 130°C.

4.9 Sandbath, Tecam, or equivalent.

4.10 Shaker, wrist-action, Burrell, or equivalent.

5. Reagents

5.1 Boron trifuoride-methanol esterification

reagent, 14 percent BF3 (weight/volume ; w/v) in

methanol, Applied Science Labs, or equivalent.

5.2 Florisil adsorbent, commercially activated at 650 ° C, washed with hexane, allowed to air

dry, and stored at 130°C in a glass-stoppered flask:

Prior to use, the florisil is deactivated by adding 10

percent water by weight and shaking for at least 2 h

on a wrist-action shaker. The florisil is then tested

for activity by attempting to elute the herbicides of

interest with benzene from a test column . If the test

compounds do not elute within 10.0 mL, further

deactivation is required.

5.3 Hydrochloric acid, concentrated (sp. gr.

1.19), analytical reagent, A.C.S. grade.

5.4 Postassium hydroxide solution, 37', percent (w/v): Dissolve 78 g KOH reagent-grade pellets

in 200 mL deionized water. Reflux for 8 h.

5.5 Sodium sulfate, acidified: Prepare a'slurry of sodium sulfate with enough diethyl ether to

cover the crystals and acidify to pH 2 or less by

adding a few mL of concentrated sulfuric acid. Determine the pH by transferring a small portion of the

slurry to a beaker, removing the ether by evaporation, adding a few drops of deionized water to the

crystals, and measuring the pH with pH paper. Allow to air dry overnight . Store in a covered Pyrex

beaker or flask at 130°C.

5.6 Sodium sulfate, granular, anhydrous:

Heat overnight at 300°C and store, covered at

130°C.

5.7 Sodium sulfate solution, 5 percent (w/v) :

Dissolve 50 g neutral sodium sulfate in deionized

water and dilute to 1 L.

5.8 Solvents, acetone, benzene, unpreserved

diethyl ether or methyl t-butyl ether, and iso-octane:

Diethyl ether preserved with ethanol cannot be used

in this procedure because it results in the formation

of extraneous ethyl esters.

5.9 Standards, methyl esters of chlorophenoxy acid herbicides, EPA analytical reference

grade, or equivalent: Prepare a stock solution by

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

weighing about 10 mg of compound to at least three

significant figures and transfer to a 25 mL volumetric flask . Dilute to volume with benzene and mix

thoroughly. Prepare a series of mixed-compound

standards by volumetric dilution with iso-octane as

described in table 8.

Table 8. Concentration of herbicides in mixed standard solutions used for gas chromatograph calibration of bottom

material

[Picograms permicroliter]

5.10 Sulfuric acid, concentrated (sp. gr.

1 .84), Mallinckrodt analytical reagent, A.C .S . grade,

or equivalent.

5.11 Sulfuric acid, (1 + 3) : Prepare by adding one part concentrated sulfuric acid to three parts

deionized water. Store in a refrigerator at 4'C.

5.12 Water, deionized, organic-free .

6. Procedure

All glassware must be washed in warm detergent

solution, rinsed with organic-free water, and heated

at 300° C overnight. Prior to use, all glassware is

rinsed with the solvent it will contact . Stopcock

grease should not be used on ground-glass joints .

For bottom-material samples, begin at step 6.1 .

For water-suspended-sediment mixtures, first determine the weight of the water, then filter the sample using a glass fiber filter to isolate the suspended

sediment. Use the filter and the retained sediment to

begin the procedure at step 6.2 .

6;1 Moisture determination:

6.1 .1 Decant excess water from the bottom

material. Use a spatula to thoroughly mix the moist

solid . Weigh 10 g of solid into a tared weighing dish.

Record the weight to three significant figures.

6.1 .2 Place the tared dish containing the

sample in an oven at 130'C overnight . Remove from

oven, allow to cool, weigh, and record the weight to

three significant figures.

6.2 Add either the filter from the suspendedsediment filtration or the calculated amount of bottom material (not more than 100 g) to a 500-mL

Erlenmeyer flask with a ground-glass stopper. Stir

the sample and slowly add deionized water until the

Herbicide High standard

concentration

Low standard

concentration

2,4-DP ----------------- 100 50

2,4-D ------------------ 100 50

2,4,5-T --------------- 40 20

Silvex------------------ 40 20
DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS 45

mixture has the consistency of paste or until water

begins to separate from the solid . Acidify the slurry

to pH 2 or below by the dropwise addition of concentrated hydrochloric acid. Use pH paper to determine

the pH. Periodically check the pH, adding more acid,

if necessary, to maintain the pH at 2 or below.

6.3 Measure 20 mL acetone into the

Erlenmeyer flask containing the acidified sample

and stopper securely . Mix the contents of the flask

for 20 min using the wrist-action shaker. Add 80 mL

ether and shake again for 10 min. Decant the extract into a 1-L separatory funnel containing 400

mL of 5 percent sodium sulfate solution. Add 20 mL

acetone to the Erlenmeyer flask and shake 20 min.

Again, add 80 mL ether, shake 10 min, and decant

the acetone-ether extract into the same separatory

funnel . Repeat the process as in the second extraction one more time, and collect the acetone-ether

extract in the separatory funnel containing the sodium sulfate solution.

6.4 Gently mix the contents of the separatory

funnel for about 1 min. Allow the layers to separate.

Discard the aqueous layer and collect the extract in

a 500-mL Erlenmeyer flask.

6.5 Add 2 mL of 37 percent KOH and 30 mL

distilled water to the extract in the 500-mL

Erlenmeyer flask. Add a boiling chip and fit the

flask with a one-ball Snyder column. Evaporate the

ether on a water bath at 80°C in a hood, and continue the heating for a total of 90 min.

6.6 Remove the assembly from the water

bath and allow it to cool. Remove the Snyder column. Quantitatively transfer the water to a 125-mL

separatory funnel. Extract the basic solution once

with 40 mL ether and discard the ether layer; repeat

twice with 20 mL ether and discard the ether layers.

The herbicides remain in the aqueous phase as their

potassium salts . Add 5 mL (1 + 3) sulfuric acid to

the contents of the funnel to lower the pH to 2 or

below (measure with pH paper), and extract the

aqueous phase with 40 mL ether; repeat twice with

20 mL ether. Collect the ether extracts in a 125-mL

Erlenmeyer flask containing about 0.5 g acidified

anhydrous sodium sulfate . Stopper the flask and set

aside for at least 2 h or store in a refrigerator until

analysis can continue.

6.7 Quantitatively transfer the ether extract

into the K-D apparatus fitted with a 5-mL volumetric receiver. Add 1 .0 mL benzene and a boiling chip.

Concentrate the extract to about 0.5 mL on a fluidized sandbath heated to 60 ° -70°C. Under no circumstances allow the extract to evaporate completely to dryness . Clear sand from the glass joint before

opening. Rinse the bottom joint with benzene into

the receiver.

6.8 After the benzene solution in the receiver

has cooled, add 0.5 mL boron trifluoride-methanol

reagent . The modified Snyder column is used as an

air-cooled condenser, and the contents of the receiver are held at 50'C for 30 min in a sandbath . Cool

the reaction mixture to room temperature and add

sodium sulfate solution until the benzene-aqueous

solution interface is observed in the restricted neck

of the receiver. Stopper the receiver, shake vigorously for 1 min, and allow to stand for at least 1 h for

phase separation. Loosen the stopper after shaking.

6.9 Transfer the benzene layer from the receiver to a florisil column. Elute with benzene and

collect 10.0 mL in a graduated receiver. Analyze the

eluate by gas chromatography .

6.10 Prepare gas chromatograph calibration

curves daily with the mixed standards shown in table 8. Operating conditions must be identical to

those used for sample analysis. Calibrate both analytical columns. Record the volume of the standard

injected and the retention time and integrated peak

area of each component in the standard.

6.11 Inject an aliquot of sample extract into

the gas chromatograph . Record the volume injected .

Identify peaks by retention time. The identification

must be made on both analytical columns. Record

the retention time and integrated area of any identified peak. Dilute any extract containing an identifiable component above the highest standard.

7. Calculations

7.1 Calculate the weight required for a dry

weight equivalent of 50 g:

Wet weight =

W1

x 50 g,

W2

where

wet weight = amount of sample to be taken for

extraction, in g,

W, = wet weight of sample, in g, and

W2 = dry weight of sample, in g.

7.2 Calculate the response factor of each

identified component in the calibration standard:

A,

RF =

CS X

V,

,
46

where

RF= response factor of identified component in

calibration standard, in area/pg,

C$ = concentration of standard, in pg/gL,

Vi = volume of standard injected, in gL, and

A1 =integrated peak area of identified compo

nent in calibration standard .

7.3 Calculate the concentration of each identified component in the original bottom-material sample from the equation

Concentration (gg/kg) =

Azx Vz

V3xWxRF

where

RF= response factor of identified calibration

standard component, in area/pg,

Az = integrated peak area of identified sample

component,

Vz = final volume of sample extract, in mL,

V3 = volume of sample extract injected, in gL,

and

W= dry weight equivalent of sample, in g.

7.4 Calculate the concentration of each identified component in the original suspended sediment

from the following equation:

Concentration (gg/L) =

where

RF= response factor of identified calibration

standard component, in area/pg,

Az = integrated peak area of identified sample

component,

Vz = final volume of sample extract, in mL,

V3 = volume of sample extract injected, in gL,

and

V4 =weight of sample in g, expressed in mL

(1 .000 mL = 1 .000 g) .

7.5 Calculate the free phenoxy acid concentrations using the following equation:

Concentration of acid (wg/L or gg/kg) = Cxf,

where

C = concentration of methyl ester determined in

calculation 7.3 or 7.4, and

_ molecular weight of acid

f molecular weight of methyl ester

8. Report

8.1 Report chlorophenoxy acid herbicide concentrations in bottom materials as follows: less than

TECHNIQUES OFWATER - RESOURCES INVESTIGATIONS

Azx Vz

V3 xV4 xRF

0.10 gg/kg, as "less than 0.10 gg/kg" ; 0.10 pg/kg

and above, two significant figures .

8.2 Report chlorophenoxy acid herbicide concentrations in suspended materials as follows : less

than 0.01 gg/L, as "less than 0.01 gg/L"; 0.01 to

0.10 gg/L, two decimals; 0.10 gg/L and above, two

significant figures.

9. Precision

It is estimated that the percent relative standard

deviation for recoverable chlorophenoxy acids from

bottom material and suspended sediment will be

greater than that reported for dissolved chlorophenoxy acids (method 0-1105).

1 . Application

Selected references

American Public Health Association, 1981, Standard methods for

the examination of water and wastewater (15th ed .) : Washington, American Public Health Association, Inc ., 1,134 p.

American Society for Testing and Materials, 1983, Annual Book

of ASTM Standards, Sect. 11, v. 11 .01 : Philadelphia, American Society for Testing and Materials, 752 p.

Goerlitz, D.F., and Brown, Eugene, 1972, Methods for analysis of

organic substances in water : U.S. Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter

A3,40 p.

Goerlitz, D.F., and Lamar, W.L ., 1967, Determination of phenoxy

acid herbicides in water by electron-capture and

microcoulometric gas chromatography: U.S . Geological Survey Water-Supply Paper 1817-C, 21 p.

Triazines, total recoverable, gas

chromatographic (0-3106-83)

Parameter Code

Ametryn---------------------------- 82184

Atrazine---------------------------- 39630

Cyanazine -------------------------- 81757

Prometon --------------------------- 39056

Prometryn -------------------------- 39057

Propazine--------------------------- 39024

Simazine --------------------------- 39055

Simetryn --------------------------- 39054

Alachlor ---------------------------- None assigned .

Trifluralin--------------------------- None assigned .

This method is suitable for the determination of

triazine herbicides, alachlor, and trifluralin in water

and water-suspended-sediment mixtures containing

at least 0.1 pg/L of each constituent.
2. Summary of method

4. Apparatus

DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS 47

Triazine herbicides are extracted from water with

methylene chloride following adjustment to pH 7 to

9. Optional adsorption chromatography on alumina

is used for the elimination of most nonpesticide interferences . Identification is made by selective gas

chromatographic separation through the use of two

or more dissimilar column packing materials using a

nitrogen specific detector.

3. Interferences

Solvents, reagents, glassware, and other sampleprocessing hardware may yield discrete artifacts or

elevated baselines which may cause misinterpretation of gas chromatograms . All of these materials

must be demonstrated to be free of interferences

under the conditions of the analysis. Specific selection of reagents and purification of solvents in an

all-glass system is required. Glassware should be

cleaned by washing with hot detergent solution,

rinsing with organic-free water, and heating overnight at 300'C.

4.1 Alumina column: To a 15-cm-long disposable Pasteur pipet, add a glass-wool plug, anhydrous sodium sulfate to a depth of 10 em, 3 cm

alumina, and 0.5 em anhydrous sodium sulfate . Tap

the column gently to promote settling to a uniform

bed.

4.2 Boiling chips, granular, micro, Hengar

H-1366C, or equivalent: Rinse with hexane, air dry,

and heat at 300'C overnight .

4.3 Concentrator apparatus, Kuderna-Danish (K-D), with a 500-mL flask, a 10.0-mL receiver,

and a three-ball Snyder column .

4.4 Evaporative concentrator, Organomation

N-Evap, or equivalent.

4.5 Gas chromatograph, Hewlett-Packard

model 5880A/Tracor model 560, or equivalent: Instrument must incorporate a glass-lined injection

port and a glass column,

4.5 .1 The following conditions are recommended:

Columns, Borosilicate glass, 1 .8 m x 2 mm

id (inside diameter) rendered inert by treatment with

silanizing agent: Column packing materials are (1) 1

percent OV-101 on 100/120 mesh U1traBond 20 M

or equivalent, operated at an oven temperature of

175°C; and (2) 100/120 mesh UltraBond PEGS, or

equivalent, operated at an oven temperature of

200°C.

Detector, alkali flame ionization (N/P), operated at 300'C.

Injection port temperature, 200'C.

Carrier gas, helium, flow rate 35 mL/min.

Detector flows, hydrogen at 3.2 mL/min

and air at 100 mL/min.

5. Reagents

5.1 Alumina, Woelm W 200, neutral, activity

I : Prepare activity V (16 percent deactivation) by

mixing 100 g activity I with 19 mL water. Mix on

wrist-action shaker for 2 h and let stand overnight

in a sealed container . Prepare fresh weekly.

5.2 Borosilicate glass wool, filtering grade,

prewashed with hexane and heated overnight at

300°C.

5.3 Potassium hydroxide, 37 percent

(weight/volume; w/v) aqueous solution prepared

from reagent-grade KOH and reagent water.

5 .4 Sodium sulfate, granular, anhydrous,

heated overnight at 300'C and stored at 130'C.

5.5 Solvents, benzene, ethyl ether, hexane,

and methylene chloride, pesticide residue quality,

distilled in glass.

5 .6 Sulfuric acid, 25 percent (volume/volume; v/v) prepared from high-purity concentrated H2SO4 (sp . gr. 1 .84) and reagent water.

5.7 Triazine standards, EPA analytical reference grade or highest purity available : Dissolve 5

mg of standard in benzene in a 50-mL volumetric

flask, dilute to volume, and mix. Dilute stock standard with hexane to working concentration shown in

table 9.

6. Procedure

6.1 Rinse all glassware with methylene chloride before using. Do not use stopcock grease on

ground-glass joints .

6.2 Weigh the sample and capped bottle to

the nearest 0 .1 g and record the weight.

Table 9. Concentration of triazines in mixed standard solutions used for gas chromatograph calibration

[Picograms per microliter]

Mixture

number Triazines Lowstandard

Medium

standard

High

standard

1 ------- Atrazine 500 1000 5000

Propazine 500 1000 5000

Simazine 500 1000 5000

2 ------- Ametryn 500 1000 5000

Prometon 500 1000 5000

Prome- 500 1000 5000

tryn

Simetryn 500 1000 5000
48

6.3 Transfer the sample to a 2,000-mL separatory funnel using a stainless steel powder funnel .

6.4 Weigh the empty sample bottle with cap

to the nearest 0.1 g and calculate the sample weight.

6.5 Dissolve 5 g sodium chloride in sample

and adjust the pH to 7 to 9 using the potassium

hydroxide solution or sulfuric acid solution as necessary.

6.6 Add 75 mL methylene chloride to sample

bottle, swirl, and transfer to separatory funnel . Allow to drain, rinsing the walls ofthe bottle.

6.7 Insert the glass stopper and shake the

funnel vigorously for 1 min. Vent the system several

times duringthis initial shakeout.

6.8 Let stand undisturbed while layers clarify,

and draw off the methylene chloride into a 250-mL

glass-stoppered Erlenmeyer flask.

NOTE: Most water-suspended-sediment

mixtures will require special treatment. If emulsions

occur, add sufficient volume of hexane (50-75 mL)

to float the extract. Shake the separatory funnel

again to mix the solvents, and allow the layers to

separate . Draw off the sample into the sample bottle . The remaining emulsion can be eliminated by

vigorous shaking (CAUTION: Vent often) . Decant

the extract into a 250-mL glass-stoppered

Erlenmeyer flask and return the sample to the separatory funnel. Proceed to step 6.9 .

6.9 Repeat the extraction with two additional

50-mL portions of methylene chloride, collecting the

organic layers in the flask .

6.10 Add 5 gm anhydrous sodium sulfate,

stopper, and shake.

6.11 Quantitatively transfer the combined extracts to the K-D apparatus, add approximately 3

mL hexane as a keeper, add a boiling chip, and

concentrate to 3-5 mL in a water bath maintained at

80°C.

6.12 Continue evaporation to approximately

0.5 mL in a warm water bath under nitrogen or

helium stream using the N-Evap apparatus . Wash

the walls of the tube with 5 mL hexane. Concentrate

to 0.5 mL and wash the walls again with 5 mL

hexane. Concentrate to 0.5 mL.

6.13 Bring the volume to 1 mL with hexane,

and proceed to alumina column cleanup, if necessary.

6.13.1 Alumina column cleanup:

6.13.1 .1 Quantitatively transfer the extract to the column using hexane. Elute the column

with hexane until 6 mL has been eluted, and discard.

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

6.13.1 .2 When the last of the hexane has

just entered the sodium sulfate, elute the column

with hexane-ethyl ether (2 :1) until a 4-mL fraction

has been collected .

6.13.1 .3 Reduce the volume to 1.0 mL

under a nitrogen or helium stream and proceed to

gas chromatographic analysis.

6.14 Frequently use standard triazine mixtures to demonstrate the effectiveness of the alumina in characterizing the eluate composition and providing quantitative recovery.

6.15 Prepare gas chromatograph calibration

curves daily with the mixed standards in table 9.

Operating conditions must be identical to those used

for sample analysis. Calibrate both analytical columns. Record the volume of the standard injected

and the retention time and peak area of each component in the standard .

6.16 Chromatograph and identify the triazines by comparing retention time with standards

(3-percent window for identification) using at least

two ofthe dissimilar column packings described .

7. Calculations

7.1 Calculate the response factor of each

identified component in the calibration standard :

RF =

A,

C3 x V,

where

RF=response factor of identified component in

calibration standard, in area/pg,

C3 = concentration of standard, in pg/gL,

V, =volume of standard injected, in gL, and

A, =integrated peak area of identified component in calibration standard .

7.2 Calculate the concentration of each identified component in the original water sample from

the equation

A2 X

V2

Concentration (pg/L) =

V3 xWxRF

where

RF=response factor of identified calibration

standard component, in area/pg,

A2 =integrated peak area of identified sample

component,

V2 = final volume of sample extract, in mL,

V3 = volume of sample extract injected, in gL,

and
W= weight of sample in g, expressed in mL

(1 .000 mL = 1 .000 g)

8. Report

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

Report concentrations of total recoverable triazines, alachlor, and trifluralin as follows: less than

0.1 gg/L, as "less than 0.1 gg/L"; 0.1 to 1 .0 Fig/L,

one decimal ; 1 .0 lug/L and above, two significant

figures.

9. Precision

9 .1 Recovery and precision (seven replicates)

for total recoverable triazines in tapwater are as

follows :

9.2 Recovery and precision (10 replicates) for

total recoverable triazines from a natural surface

water are as follows:

Compound

Ametryn ----------

Simetryn----------

Carbamate pesticides, total recoverable,

high-performance liquid chromatographic

(0-3107-83)

Parameter Code

Aldicarb ---------------------------- None assigned.

Carbaryl---------------------------- 39750

Carbofuran-------------------------- None assigned.

3-Hydroxycarbofuran------------------ None assigned.

Methomyl--------------------------- 39051

1-Naphthol-------------------------- None assigned .

Propham---------------------------- 39052

1 . Application

This method is suitable for the determination of

carbamate pesticides in water or water-suspendedsediment samples containing at least 2 Fig/L of the

analyte .

2. Summary of method

3. Interferences

49

The carbamates are extracted from water or

water-suspended-sediment mixtures with methylene chloride . The extract is concentrated and analyzed by high-performance liquid chromatography

(HPLC) using a C18 reverse phase column and a

dual-channel variable-wavelength ultraviolet detector .

Compounds that exhibit chemical and physical

properties similar to the compounds of interest can

interfere .

4. Apparatus

4.1 Concentrator, Kuderna-Danish (K-D),

500-mL capacity with a three-ball Snyder column

and a 10-mL graduated receiver tube.

4.2 Evaporative concentrator, Organomation

N-Evap, or equivalent.

Amount

Mean

percent Standard

Relative standard

deviation

Compound spiked (Fig) recovery deviation (percent)

Prometon --------- 0 .5 85 .5 10 .9 12 .8

1 .0 94 .4 6 .60 6 .99

5 .0 87 .0 5 .92 6 .81

Propazine --------- .5 82 .3 11 .6 14 .1

1 .0 97 .0 4 .90 5 .05

5 .0 91 .7 5 .74 6.26

Atrazine ---------- .5 85.1 11 .8 13.9

1 .0 97.3 4.35 4.47

5 .0 87.1 5.26 6.04

Prometryn -------- .5 87 .3 9.98 11 .4

1 .0 98 .3 3.97 4 .04

5 .0 81 .5 4 .50 5 .53

Simazine---------- .5 81 .0 12 .0 14 .8

1 .0 96 .6 8 .68 8 .99

5 .0 61 .8 9 .41 15 .2

Ametryn ---------- .5 89.6 10 .5 11 .7

1 .0 97.2 4 .52 4 .65

5 .0 91.4 5 .70 6.24

Simetryn---------- .5 87.7 1 .01 11 .5

1 .0 92.2 6.42 6 .97

5.0 88 .3 7 .38 8 .36

Amount

spiked (Fig)

Mean

percent

recovery

Standard

deviation

Relative standard

deviation

(percent)

.5 101 .1 8.82 8 .73

1 .0 101 .5 3.83 3 .77

5.0 87 .8 4 .03 4 .58

.5 98 .7 7 .17 7 .27

1 .0 94 .7 2 .08 2 .19

5 .0 86 .4 4 .23 4 .90

Compound

Amount

spiked (Fig)

Mean

percent

recovery

Standard

deviation

Relative standard

deviation

(percent)

Prometon --------- 0.5 95.1 5 .83 6.13

1 .0 96.9 2 .08 2.15

5.0 84.6 4 .58 5 .41

Propazine --------- .5 96 .2 5.46 5 .68

1 .0 100 .9 2 .06 2 .05

5.0 87 .6 4 .62 5 .28

Atrazine ---------- .5 96 .0 7 .34 7 .65

1 .0 98 .1 2 .11 2 .15

5 .0 85 .5 5 .14 6 .01

Prometryn -------- .5 98 .0 5 .09 5 .20

1 .0 77 .9 1 .67 2 .15

5 .0 83 .5 3 .82 4 .57

Simazine---------- .5 96.9 4 .90 5 .05

1 .0 98.8 4 .91 4.97

5 .0 82.8 7 .69 9.29
50

4.3 Filters, 0.5 gm millipore FHUP, catalog

no. 04700, or equivalent, and 0.45 gm millipore

HAWP, catalog no. 04700, or equivalent.

4.4 Liquid chromatograph, Waters Associates ALC/GPC 204 liquid chromatograph equipped

with a dual-channel variable-wavelength detector, a

model 6000A solvent-delivery system, a model 660

solvent flow programmer, a model WISP 710A

microprocessor, and a data module, or equivalent.

4.4 .1 The following conditions are recommended:

Columns, Waters Associates Radial Compression Module with a', Radial PAK reverse-phase

cartridge (octadecylsilane permanently bonded to

unmodified silica), 10 gm particle size, or equivalent.

Wavelengths, 254 and 280 nm.

Solvent, 45 percent water and 55 percent

methanol at a flow rate of 1.0 mL/min isocratic.

4,5 Solvent clarification kit, Waters Associates 85113, or equivalent.

5. Reagents

5.1 Carbamate and metabolite standards,

EPA analytical reference grade or highest purity

available: Prepare by dissolving 5 mg of standard in

acetonitrile in a 50-mL volumetric flask, dilute to

volume, and mix. Dilute this stock solution with acetone to the working concentrations listed in table 10.

Table 10 . Concentration of carbamates in mixed standard

solutions used for liquid chromatograph calibration

[Nanograms per microliter]

5.2 Phosphate buffer, pH 7.5 : Mix 50 mL of

0.1 M KH2 PO4 with 41 mL of 0.1 M NaOH and dilute to 100 mL with deionized water.

5.3 Potassium dihydrogen phosphate, 0.1 MDissolve 13.6 g KH2PO4 in deionized water and dilute to 1 L.

5.4 Sodium hydroxide, 0.1 M Dissolve 4 g

NaOH pellets in deionized water and dilute to 1 L.

5.5 Sodium sulfate, granular, anhydrous:

Heat overnight at 300°C and store at 130°C.

5.6 Solvents, HPLC quality, acetonitrile,

methanol, tetrahydrofuran, and pesticide grade

methylene chloride.

5.7 Water, organic-free .

TECHNIQUESOF WATER - RESOURCES INVESTIGATIONS

6. Procedure

Glassware should be cleaned by washing with hot

detergent solution, rinsing with deionized water, and

heating overnight at 300°C. Just prior to use the

glassware is rinsed with methylene chloride. Stopcock grease should not be used on the ground-glass

joints .

6 .1 Weigh the sample bottle plus the sample

and record the weight. Pour the sample into a 1,000

mL-separatory funnel, add 10 mL phosphate buffer,

and shake until well mixed. Weigh the empty sample

bottle . Calculate the net sample weight and record

the value obtained to three significant figures .

6.2 Add 75 mL methylene chloride to the

sample bottle, swirl to rinse the sides of the bottle,

and transfer the solvent to the separatory funnel .

The Teflon-lined cap is not rinsed because of the

potential for contamination from solvent that has

contacted the threads and surface beneath the

Teflon liner. Shake the separatory funnel vigorously

for 1 min. Vent often. Allow the layer to separate

and draw off the methylene chloride layer into a

250-mL Erlenmeyer flask that contains 1 g anhydrous sodium sulfate.

6.3 Repeat the extraction ofthe water sample

two more times, using 50 mL methylene chloride

each time. Combine all the organic extracts in the

250-mL Erlenmeyer flask containing the first extract .

6.4 Transfer the extract to a 500-mL K-D

apparatus fitted with a three-ball Snyder column

and a 10-mL receiver containing a micro boiling chip

and 4 mL acetonitrile .

6.5 Place the apparatus on a hot-water bath

(75-85°C) until the volume is reduced to about 4

mL. Remove from the heat and allow to cool. Wipe

the joints with a towel. Rinse the bottom joint with

acetonitrile into the receiver.

6.6 Further reduce the volume of solvent to

about 1 mL on an evaporative concentrator with the

water bath at 35°C. Rinse down the sides of the

tube with 1 mL acetonitrile and concentrate to a

final volume of 1 .0 mL. Stopper until chromatographic analysis can begin.

6.7 Prepare the solvents for the mobile phase

by filtering, using the solvent clarification kit and

HAWP filters for water and FHUP filters for the

organic solvents.

6.8 Prepare liquid chromatograph calibration

curves daily by injecting the reference standards

listed in table 10. Operating conditions need to be

Carbamate LOS'

standard

Medium

standard

High

standard

Propham ---------- 9 18 36

Methomyl ---------- 2 4 8

Carbatyl------------ 2 4 8
identical to those used for sample analysis in step

6.9 . Record the volume of the standard injected and

the retention time and integrated peak area of each

component in the standard .

6.9 Inject an aliquot of sample extract into

the calibrated liquid chromatograph . Record the volume injected . Identify the peaks by retention time.

Confirmation is obtained by measuring the peak

area at the two different wavelenghths (254 and

280 nm) and comparing the ratio of the peak areas

to that of the standard . Record the retention time

and integrated area of any identified peak. Dilute

any extract containing an identifiable component

above the highest standard .

7. Calculations

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

7.1 Calculate the response factor of each

identified component in the calibration standard :

A, RF =

Cs x V

,

where

RF= response factor of identified component in

calibration standard, in area/ng,

CS = concentration of standard component, in

ng/EtL,

V = volume of standard injected, in pL, and

A, = integrated peak area of identified component in calibration standard .

7.2 Calculate the concentration of each identified component in the orginial sample from the

equation

Concentration (Ffg/L) =

A2x V2 x 1,000,

RFx V3 x W

where

RF= response factor of identified calibration

standard, in area/ng,

A2 = integrated peak area of identified sample

component,

V2 = final volume of sample extract, in mL,

V3 = volume of sample extract injected, in gL,

and

W= weight of sample in g, expressed in mL

(1 .000 mL = 1.000 g).

8 Report

Report concentrations of carbamates in water or

water-suspended-sediment mixtures as follows: less

than 2 gg/L, as "less than 2.0 gg/L" ; 2.0 gg/L and

above, two significant figures.

9. Precision

Single-operator precision was determined by spiking surface-water samples at four concentrations

and four replicates performed on different days. Results are as follows :

Selected references

Goerlitz, D.F., and Brown, Eugene, 1972, Methods for analysis of

organic substances in water: U.S . Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter

A3, 40 p.

Sparacine, C.M., and Hines, J .W., 1976, High-performance liquid

chromatography of carbamate pesticides : Journal of Chromatography Science, v . 14, p. 549-555.

Oil and grease, extractable,

extraction-gravi metric (0-3108-83)

Parameter Code

Oil and grease, total (mg/L as oil and grease)-------00556

1 . Application

51

This method is suitable for the determination of

oil and grease in water-suspended-sediment mixtures containing at least 1 mg/L of the analyte .

2. Summary of method

2.1 A sample is extracted twice with trichlorotrifluoroethane and the extract is evaporated at

20'C to leave a nonvolatile residue whose weight

represents an estimate of the extractable organic

matter in the sample.

2.2 The procedure approximates the determination of oils and grease in water, and is similar to

Method 2778-70 described by the American Society

for Testing and Materials (1982) and to Method 137,

Oil and Grease, described by the American Public

Mean Relative

Concentration concentration standard

Compound

spiked

(pg1L)

recovered

(vg/L)

deviation

(percent)

Methomyl--------- 2 .51 2 .13 20

5 .02 3 .80 11

10 .0 7 .47 9.4

20.1 15 .5 7.3

Carbaryl---------- 2 .36 2 .33 5 .7

4 .72 4 .58 4 .8

9 .44 9 .18 5 .5

18 .9 18 .6 3 .4

Propham---------- 7 .05 5 .68 13

14 .1 11 .2 8.0

28 .2 22 .2 7.7

56 .4 43 .0 8.2
52

Health Association and others (1981) in "Standard

Methods for the Examination of Water and Wastewater."

3. Interferences

Organic solvents vary considerably in their ability

to dissolve oil substances and other organic matter.

Any method used to obtain an estimate of the

amount of extractable organic matter must, of necessity, be highly empirical . Close attention to all

operations of the analytical procedure is required to

obtain reproducible results .

4. Apparatus

4.1 Dish, aluminum foil, 110 mm in diameter,

100-mL capacity .

4.2 Funnel, separatory, pear-shaped, 2-L catiously, with constant stirring and cooling, add 100

mL concentrated H2SO4 to 100 mL demineralized

water.

5.4 Trichlorotrifluoroethane solvent, 1,1,2-

Trichlorotrifluoroethane, b.p . 47.6°C, reagent

grade.

6. Procedure

6 .1 Collect approximately 900 mL of sample

in a 1-L glass bottle.

6 .2 Transfer the entire contents of the glass

bottle to a 1,000-mL graduated cylinder. Record the

volume . Prepare a 900-mL demineralized water

blank and carry it through the sample-analysis procedure. Subtract the residual weight (blank) from

the sample extract residual weight. If the weight of

the blank exceeds 4.0 mg, a new bottle of solvent

must be obtained to provide a blank of 4.0 mg or

less .

6.3 Transfer the sample from the graduated

cylinder to a 2-L separatory funnel, and add 5 mL

sulfuric acid (1 + 1) . Shake to mix thoroughly.

6.4 Rinse the glass bottle with 25 mL

trichlorotrifluoroethane solvent and transfer the solvent to the graduated cylinder. Rinse the graduate

and transfer the solvent to the separatory funnel.

Shake vigorously for 2 min, stopping to vent the

pressure as necessary .

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

6.5 Allow the layers to separate and then

draw off the solvent and filter it through a small

amount of anhydrous Na2SO4 placed on a small filter

paper (Whatman No. 40, or equivalent) in a funnel.

Collect the filtrate in a tared aluminum-foil dish.

6.6 Repeat steps 6.4 and 6.5, filtering the solvent through the same funnel and adding the filtrate

to that already collected in the aluminum-foil dish.

6.7 Wash the filter paper with three 5-mL

portions of solvent, collecting all washings in the

aluminum-foil dish.

6.8 Evaporate the solvent collected in the aluminum-foil dish at room temperature (20'C) in a

well-ventilated fume hood.

6.9 Rinse the inside of the aluminum-foil dish

with demineralized water to remove traces of sulfuric acid. Dry the dish in a desiccator to remove

waert .

Organic matter, extractable, mg/L =

RX -Rb

mL sample

where

RX = weight of extracted residue, in mg, and

Rb = weight of solvent residue (blank), in mg.

8. Report

9. Precision

x 1,000,

Report organic matter, extractable, watersuspended-sediment, concentrations as follows: less

than 10 mg/L, nearest mg/L; 10 mg/L and above,

two significant figures .

Precision data cannot be given for this determination because of the variable nature of the materials

being extracted .

Selected references

American Public Health Association, 1981, Standard methods for

the examination of water and wastewater (15th ed .) : Washington D.C ., American Public Health Association, Inc .,

p. 461 .

American Society for Testing and Materials, 1982, Annual Book

of ASTM Standards, Part 31, Water: Philadelphia, American

Society for Testing and Materials, p. 627.

pacity

5. Reagents

(Corning 6404, or equivalent).

6.10 Weigh the residue in the dish after the

water has evaporated.

5.1 Sodium sulfate, anhydrous, granular.

7. Calculations

5.2 Suotric acid, concentrated (sp. gr. 1.84) . Determine the mg/L extractable organic matter

5.3 Sulfuric acid, (1 -F 1) : Slowly and cau- in the samples as follows :
1 . Application

2. Summary of method

3. Interferences

4. Apparatus

5. Reagents

6. Procedure

DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS 53

Oil and grease, extractable from bottom

material, extraction-gravi metric (0-5108-83)

Parameter Code

Oil and grease, recoverable from

bottom material (mg/kg as oil and grease) -------00557

1 .1 This method is suitable for the determination of oil and grease in air-dried bottom materials

containing at least 1,000 mg/kg.

1 .2 This method may be used for the determination of oil and grease in wet bottom materials if

the appropriate moisture correction is applied.

A sample is extracted twice with trichlorotrifluoroethane and the extract is evaporated at

20° C to leave a nonvolatile residue whose weight

represents an estimate of the extractable organic

matter in the sample.

Organic solvents vary considerably in their ability

to dissolve oil substances and other organic matter.

Any method used to obtain an estimate of the

amount of extractable organic matter must, of necessity, be highly empirical . Close attention to all

operations of the analytical procedure are required

to obtain reproducible results .

4.1 Dish, aluminum foil, 110 mm in diameter,

100-mL capacity .

4 .2 Funnel, separatory, pear-shaped, 2-L capacity (Corning 6404, or equivalent).

5.1 Sodium sulfate, anhydrous, granular .

5.2 Sulfuric acid, concentrated (sp . gr. 1 .84) .

5.3 Sulfuric acid, (1 + 1) : Slowly and cautiously, with constant stirring and cooling, add 100

mL concentrated H2SO4 to 100 mL demineralized

water.

5.4 Trichlorotrifluoroethane solvent, 1,1,2-

Trichlorotrifluoroethane, b.p . 47.6 ° C, reagent

grade.

6.1 Weigh, to the nearest mg, approximately

1 g of air-dried sample material. Alternatively, a wet

sample may be weighed if a correction is made for

moisture content.

6.2 Quantitatively transfer the weighed sample to a 2-L-capacity separatory funnel . Add approximately 900 mL demineralized water and shake to

mix. Prepare a 900-mL demineralized water blank

and carry it through the sample-analysis procedure .

Subtract the residual weight (blank) from the sample extract residual weight. If the weight of the

blank exceeds 4.0 mg, a new bottle of solvent must

be obtained to provide a blank of 4.0 mg or less .

6.3 Add 5 mL sulfuric acid (1 + 1) . Shake to

mix thoroughly.

6.4 Add 25 mL trichlorotrifluoroethane and

shake vigorously for 2 min, stopping to vent the

pressure as necessary.

6.5 Allow the layers to separate and then

draw off the solvent and filter it through a small

amount of anhydrous Na2SO4 placed on a small filter

paper (Whatman No. 40, or equivalent) in a funnel .

Collect the filtrate in a tared aluminum-foil dish.

6.6 Repeat steps 6.4 and 6 .5, filtering the solvent through the same funnel and adding the filtrate

to that already collected in the aluminum-foil dish.

6.7 Wash the filter paper with three 5-mL

portions of solvent, collecting all washings in the

aluminum-foil dish.

6 .8 Evaporate the solvent collected in the aluminum-foil dish at room temperature (20'C) in a

well-ventilated fume hood.

6.9 Rinse the inside of the aluminum-foil dish

with demineralized water to remove traces of sulfuric acid. Dry the dish in a desiccator to remove

water.

6.10 Weigh the residue remaining in the dish

after the water has evaporated.

7. Calculations

7.1 Determine the mg/kg extractable organic

matter in the air-dried sample as follows:

Organic matter, extractable, mg/kg =

R.-R6

sample weight in g

x 1,000,

where

RX = weight of extracted residue, in mg, and

Rb = weight of solvent residue (blank), in mg.

NOTE: If wet bottom-material sample is used in

preference to air-dried sample, a factor correcting

for moisture content must be applied to above equation.
54

8. Report

8.1 Report organic matter, extractable, airdried bottom material, concentrations as follows :

less than 10,000 mg/kg, nearest 1,000 mg/kg;

10,000 mg/kg and above, two significant figures .

9. Precision

Precision data cannot be given for this determination because of the variable nature of the materials

being extracted .

Fuel oils, light, total recoverable, gas

chromatographic (0-3109-83)

Parameter Code

Fuel oils, light, total recoverable (mg/L) ---- None assigned.

1 . Application

This method is suitable for the determination of

light fuel oils (Clo-C22) in water or watersuspended-sediment mixtures at concentrations of

0.1 mg/L and above.

2. Summary of method

A water or water-suspended-sediment sample

containing light fuel oils (diesel oils) is acidified and

extracted with hexane. The extract is concentrated

and analyzed by temperature-programmed gas chromatography using a flame-ionization detector.

These oils possess a characteristic envelope which

appears during the temperature-programmed run.

This envelope, composed of the various hydrocarbon

peaks, approximates the boiling range profile of the

oil . The area of the envelope is reproducible and is

the basis for quantitation. Characterization is based

on comparison of the peaks in the residue chromatogram with those of a known oil sample.

3. Interferences

Any compound having chemical and physical

properties similar to an analyte of interest may interfere.

4. Apparatus

4.1 Boiling chip, micro, granules, Hengar H1366C, or equivalent.

4.2 Concentrator apparatus, Kuderna-Danish (K-D) type, with 10.0- and 4.0-mL receivers,

200- and 500-mL flasks, and a one-ball Snyder column.

4.3 Evaporative concentrator, Organomation

N-Evap, or equivalent.

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

4.4 Gas chromatograph, Hewlett-Packard

5710, or equivalent.

4.4 .1 The following conditions are recommended:

Columns, borosilicate glass, 1 .8 mx 2

mm id (inside diameter) . Column packing is 5 percent OV-101 on 100/120 mesh Gas Chrom Q, or

equivalent. A capillary column operated under appropriate conditions may also be used.

Temperature program, 3-min postinjection hold, 100 ° C to 230 ° C at 8'C/min, hold for 2

min, end of cycle.

Detector, flame-ionization operated at

250°C.

5. Reagents

Injection port temperature, 2100'C .

Carrier gas, helium, flow rate 30 mL/min.

5.1 Fuel oil standard: A sample of the oil

being determined must be available from the manufacturer or other suitable source . The Environmental

Protection Agency Environmental Monitoring and

Support Laboratory in Cincinnati, Ohio, has several

standard oil references. Weigh about 10 mg of standard to at least three significant figures and quantitatively transfer it to a 25-mL volumetric flask.

Dilute to volume with iso-octane. Prepare standards

in iso-octane at concentrations of 100, 1,000, and

10,000 ng/gL.

5.2 Sodium sulfate, granular, anhydrous :

The sodium sulfate should be heated at 300 ° C overnight and stored in a stoppered, glass container.

5 .3 Solvents, acetone and hexane, pesticide

residue quality, distilled in glass, Burdick and Jackson, or equivalent.

5.4 Sulfuric acid, concentrated (sp . gr. 1.84) .

5.5 Sulfuric acid, 2 N Add 53 mL concentrated H2504 slowly to 500 mL water. Cool and

dilute to 1 L.

5.6 Water, reagent grade, organic-free.

6. Procedure

Glassware must be cleaned by washing with hot

detergent solution, rinsing with deionized water, and

heating overnight at 300°C .Just prior to use, the

glassware is rinsed with methylene chloride. Do not

use stopcock grease on ground-glassjoints .

6.1 Refrigerate the sample until extraction.

The extraction must be performed within 24 to 48 h

after receipt to minimize oil degradation.

6.2 Weigh the bottle containing the sample

and record the weight. Pour the sample into a 1-L
DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS

separatory funnel and weigh the empty sample bottle . Calculate the net sample weight and record the

value obtained to three significant figures . Use 2 N

sulfuric acid to adjust the pH to 1 to 2.

6.3 Add 20 mL hexane to the sample bottle,

swirl, and pour into a separatory funnel . Allow bottle to drain.

6.4 Shake the funnel vigorously for 1 min,

venting often to relieve pressure. Decant the solvent

layer into a 500-mL Erlenmeyer flask.

6 .5 Repeat steps 6 .3 and 6.4 twice, for a total

of three extractions.

6.6 Add about 5 g anhydrous sodium sulfate

to the Erlenmeyer flask containing the extracts and

allow to stand covered for at least 4 h.

6.7 Quantitatively transfer the extract to a

K-D apparatus . Add a boiling chip, and concentrate

on a water bath to about 5 mL. Remove the appara

tus and dry the joints with a towel. Rinse the lower

joint with hexane into the receiver.

6.8 Continue concentration on an evaporative

concentrator until the volume is reduced to about 0.5

mL. Wash the walls of the receiver with small por

tions of hexane during concentration and dilute to

1 .0 mL with hexane .

6.9 Prepare gas chromatograph calibration

curves daily by injecting the series of reference standards. Operating conditions must be identical for

samples and standards . Record the volume of the

standard injected .

6.10 Inject an aliquot of sample extract into

the gas chromatograph. Record the volume injected.

Dilute any extract containing fuel oil in a concentration greater than the highest standard .

6.11 Determine the area under the envelope

of the fuel oil standard . Choose the beginning and

ending peaks so that a reproducible area will be

measured when quantitating different standards and

samples . Use the equations is section 7 to calculate

the concentration of fuel oil in the sample.

7. Calculations

7 .1 Calculate the response factor for the calibration standard:

RF =

A,

V,XC

where

RF= response factor of fuel oil in calibration

standard, in area/ng,

C, = concentration of standard, in ng/gL,

V, = volume of standard injected, in gL, and

A, = integrated peak area of calibration standard.

7.2 Calculate the concentration of fuel oil in

the original water sample from the equation

8. Report

9. Precision

Concentration (mg/L) A2 XV2

V3 xWxRF

55

where

RF = response factor of fuel oil in calibration

standard, in area/ng,

A2 = integrated peak area of sample,

V2 = final volume of sample extract, in mL,

V3 = volume of sample extract injected, in gL,

and

W= weight of sample in g, expressed in mL

(1 .000 mL = 1.000 g).

Report concentrations of fuel oil in water or

water-suspended-sediment mixtures as follows: less

than 0.02 mg/L, as "less than 0.02 mg/L" ; 0.02 to

0.1 mg/L, two decimals, 0.1 mg/L and above, two

significant figures .

Single-operator precision data for No. 2 diesel fuel

spiked into distilled and natural water samples are

as follows:

Number Mean concentration Relative

of determined standard deviation

replicates (mg1L) (percent)

Concentration

ofspikedfueloil Distilled Natural Distilled Natural Distilled Natural

sample (mg/L) water water water water water water

0.085 ------ 5 5 0.064 0.058 28 22

.85 ------- 5 5 .68 .67 4.8 10

8.5 -------- 5 4 7.0 6.8 1.7 7.3

Phenols, total recoverable, colorimetric,

4-aminoantipyrine (0-3110-83)

Parameter Code

Phenols, total recoverable (gg/L as phenol) --------32730

1 . Application

This method may be used to analyze water or

water-suspended-sediment mixtures containing at

least 1.0 lag/L of phenolic material.

2. Summary of method

Steam-distillable phenols react with 4-aminoantipyrine at pH 10.0 ± 0 .2 in the presence of potassium ferricyanide to form a colored antipyrine
dye. This dye is extracted from aqueous solution

with chloroform, and the absorbance is measured at

460 nm. This method is similar in principle to, but

different in detail from ASTM Method D 1783-80

(American Society for Testing and Materials, 1982).

3. Interferences .

Phenol has been selected as the standard for reference. Substituted phenols may produce less color

than phenol. The concentration of phenols determined by this method represents the minimum concentration of phenolic compounds present in the

sample.

Certain bacteria, oxidizing and reducing substances, and highly alkaline waste waters may interfere with this method. Information for removal of

major interferences may be found in ASTM Method

D 1783-80.

4. Apparatus

4.1 Distillation apparatus, all glass, consisting of a 1-L Pyrex distilling apparatus and a watercooled condenser (Corning 3360, or equivalent).

4.2 Funnels, Buchner type with fritted-glass

disk (15-mL Corning 36060, or equivalent).

4.3 Photometer, spectrophotometer or filter

photometer operating at 460 nm, and accommodating cells having light paths of 1 .0 and 10 em.

4.4 Funnel, Separatory, pear-shaped, 1,000

mL (Corning 6404, or equivalent).

5. Reagents

All reagents must be prepared with phenol-free

distilled water. Deionized water is usually not satisfactory.

5.1 Aminoantipyrine solution, 2 g/100 mL:

Dissolve 2.0 g of 4-Amnoantipyrine in distilled

water and dilute to 100 mL. This solution is not

stable and must be prepared each day.

5.2 Ammonium chloride solution, 20 g/L:

Dissolve 20 g reagent-grade ammonium chloride in

water and dilute to 1 L.

5.3 Ammonium hydroxide, concentrated (sp .

gr. 0.90), reagent grade.

5.4 Chloroform, spectrophotometric grade.

5.5 Copper sulfate solution, 100 g/L: Dissolve 100 g of CUS04 -5H20 in water and dilute to 1

L.

5.6 Phenol standard solution, 1.00

mL =1.00 mg phenol: Dissolve 1.00 g analytical

reagent phenol in 1,000 mL freshly boiled and

cooled distilled water. Solution is stable for 1 mo.

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

5.7 Phosphoric acid solution: Dilute 10 mL of

85 percent H3P04 to 100 mL with phenol-free water.

5.8 Potassiumferricyanide solution, 8 g/100

mL: Dissolve 8.0 g of K3Fe(CN)6 in water, dilute to

100 mL, and filter. This solution is not stable and

must be prepared each day.

5.9 Sodium sulfate, anhydrous, granular,

ACS reagent grade.

6. Procedure

Samples should be protected from light and analyzed as soon as possible. The analyst is referred to

"Standard Methods for the Examination of Water

and Wastewater," 15th edition (American Public

Health Association 1981) for the analysis of very

alkaline or highly polluted water.

6.1 Measure a volume of sample containing

less than 50 pg phenol (500 mL maximum) into a

beaker. If less than 500 mL of sample is used, dilute

sample with distilled water to 500 mL. Determine

the pH and adjust to below 4.0, if necessary. (Add

5.0 mL copper sulfate solution if it was not added at

sampling.) Transfer the solution to the distillation

apparatus, add boiling stones, and distill . Collect 450

mL distillate and stop. Add 50 mL distilled water to

the residue and proceed with distillation until 500

mL of distillate is collected.

6.2 Prepare a 500-mL distilled-water blank.

Also prepare 500-mL standards containing 5, 10,

20, 30, 40, and 50 gg phenol, using the standard

phenol solution .

6.3 Treat the sample, blank, and standards as

follows: Add 1.0 mL ammonium chloride solution

and 1.0 mL ammonium hydroxide, mix, and adjust

the pH to 10.0 ± 0.2 with concentrated ammonium

hydroxide . Add 3.00 mL aminoantipyrine solution

and mix. Add 3.00 mL potassium ferricyanide solution and again mix. Allow the color to develop for 3

min. A clear to light-yellow solution should result.

6.4 Add 25.0 mL chloroform for 1-and 5-cm

cells and 50.0 mL chloroform for 10-cm cells . Shake

the separatory funnel vigorously for 1 min. Allow

the layers to separate and repeat the shaking to

achieve a higher recovery .

6.5 After the layers have separated, draw off

the lower chloroform layer and filter through a 5-g

layer of anhydrous sodium sulfate, using a sintered

glass funnel, directly into an appropriate absorption

cell. Avoid working in a draft so as to reduce evaporation of the solvent.
Phenolic material (Igg/L) =

8. Report

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

6.6 Measure the absorbance of the sample

and standards against the blank at 460 nm. Prepare

a calibration curve plotting absorbance versus Fig of

phenol.

7. Calculations

Calculate the phenolic content in the sample, in

Fig/L, as follows :

gg phenol in samplex 1,000

mL original sample

Report concentrations of phenolic material as follows: less than 1 lag/L, as "less than 1 leg/L" ; 1 to

100 lag/L, nearest gg/L; 100 gg/L and above, two

significant figures .

9. Precision

Single-operator precision (10 replicates) for phenol spiked into distilled water is as follows:

Selected references

American Public Health Association, 1981, Standard methods for

the examination of water and wastewater (15th ed.) : Washington D.C ., American Public Health Association, Inc .,

1,134 p.

American Society for Testing and Materials, 1982, Annual book

of ASTM standards, Part 31, Water: Philadelphia, American

Society for Testing and Materials, p . 789.

Goerlitz, D.F., and Brown, Eugene, 1972, Methods for analysis of

organic substances in water: U.S . Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter

A3, 40 p.

Methylene blue active substances, total

recoverable, colorimetric (0-3111-83)

Parameter Code

Methylene blue active substances,

total recoverable (mg/L as MBAS) -------------38260

1 . Application

This method is applicable to the analysis of water

or water-suspended-sediment mixtures containing

at least 0.01 mg/L methylene blue active substances

2. Summary of method

3. Interferences

4. Apparatus

5. Reagents

57

(MBAS) relative to linear alkyl sulfonate (LAS)

standard.

Methylene blue reacts with anionic surfactants,

both alkyl benzene sulfonates (ABS) and LAS, to

form a blue-colored dye complex. The complex is

extracted with chloroform, and the methylene blue

active substances are determined spectrophotometrically . This method is similar in substance to the

MBAS method in "Standard Methods for the Examination of Water and Wastewater," 15th edition

(American Public Health Association, 1981).

Phenols, proteins, and inorganic chloride, cyanate,

nitrate, and thiocyanate will complex methylene blue

and give a positive interference . With LAS concentrations from 0.0 to 0 .1 mg/L, tests have shown no

interference from the following individual constituents: 10 mg/L nitrite, 25 mg/L nitrate, 5 mg/L phenol, and 1 mg/L hydrogen sulfide . Organic amines

cause low results .

4.1 Spectrophotometer, for use at 635 nm.

4 .2 Refer to the manufacturer's manual to

optimize instrument.

5.1 Chloroform, spectrophotometric grade.

5.2 Detergent, primary stock standard, 1

mL = 1 mg LAS: Use reference LAS acid. An ampoule of LAS acid may be obtained from the Environmental Monitoring and Support Laboratory, U.S .

Environmental Protection Agency, Cincinnati, OH

45268. The ampoule has a shelf life of 2 yr when

stored unopened in a cool, dark location. The LAS

ampoule is labeled in percent active LAS

(weight/volume) . Weigh an amount of LAS acid

equal to 1,000 g LAS on a 100 percent basis and

dilute with demineralized water to 1,000 mL. To

obtain the amount of active LAS, divide 1 .000 g by

the percent active stated on the ampoule label. For

example, if the LAS solution is stated as 5 .69 percent active, multiply 1 .000 by 100 and divide by

5.69. The result, 17.575g, is the amount of LAS

solution to be weighed out and diluted with demineralized water to 1,000 mL.

5.3 Detergent working standard I, 1 .00

mL = 0.01 mg LAS: Dilute 10 mL of primary stock

Relative standard

Phetwl concentration Meanfound deviation

(RgIL) (WBlL) (Percent)

4 .0 ---------- 4.5 12

40 ------------ 37 5 .5
58

standard to 1,000 mL with demineralized water.

This has a shelflife of 1 mo when refrigerated .

5.4 Detergent working standard II, 1 .00

mL = 0.001 mg LAS: Dilute 100 mL of working

standard I to 1,000 mL with demineralized water.

Prepare fresh daily .

5.5 Methylene blue reagent: Dissolve 0.35 g

methylene blue in 0.01 N H2 SO4 and dilute to 1 L

with 0.01 NH2 SO4 .

5.6 Phenolphthalein solution : Dissolve 2.5 g

phenolphthalein in 250 mL of 95 percent ethyl alcohol and add 250 mL deionized water. Then add

0.02 N NaOH dropwise until a faint pink color appears.

5.7 Sodium hydroxide solution, 0.5 N Dissolve 10 g NaOH in deionized H2 O and dilute to 500

mL.

5.8 Sulfuric acid solution, 5 N Cautiously,

mix with cooling, 138 mL concentrated H2 SO4 (SPgr.

1 .84) in 500 mL deionized water and dilute to

1,000 mL with deionized water.

5.9 Sulfuric acid solution, 0.1 N Cautiously,

mix 3 mL concentrated H2 SO4 (sp . gr. 1 .84) with

deionized water and dilute to 1 L.

6. Procedure

All glassware must be rinsed with dilute HC1 and

deionized water immediately before use .

6.1 Pipet a volume of sample (100 mL maximum) containing less than 0.10 mg MBAS into a

separatory funnel . Prepare a blank and standards in

the same manner.

6.2 Add 2 drops of phenolphthalein solution

to the samples and standards and adjust the pH to

near neutral by the dropwise addition of 0.5 N

NaOH or 0.1 NH2 SO4 . To the blank, standards, and

samples add 1 .0 mL of 5 N H2SO4 , mix, and then

add 5.0 mL methylene blue solution. Mix thoroughly .

6.3 Add 25.0 mL chloroform and shake the

contents of the funnel for 30 s. Allow the layers to

separate.

6.4 Drain off the lower chloroform layer into

a 1 .0-cm cell and measure the absorbance of the

sample and standards against the blank at 635 nm.

7. Calculations

Determine the amount of detergent contained in

the sample, minus the blank, from the analytical

curve. Calculate the amount of MBAS in the sample

using the following equation:

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

mg/L MBAS (as LAS) =

Wx 1000

S

where

W= mg MBAS obtained from calibration curve

minus mg blank MBAS, and

S = mL of sample.

8. Report

Report MBAS concentrations as follows: less than

0.01 mg/L, as "less than 0.01 mg/L"; 0.01 to 1 .0

mg/L, two decimals; 1 .0 mg/L and above, two significant figures.

9. Precision

Deviation of ± 10 percent may be expected in the

range of 1 to 5 mg/L, in surface water.

Selected references

American Public Health Association, 1981, Standard methods for

the examination of water and wastewater (15th ed .): Washington, American Public Health Association, Inc., 1,134 p.

Goerlitz, D.F., and Brown, Eugene, 1972, Methods for analysis of

organic substances in water: U.S . Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter

A3, 40 p.

TNT, RDX, and picric acid, total recoverable,

high-performance liquid chromatographic

(0-3112-83)

Parameter Code

TNT --------------------------------------81360

RDX ---------------------------------- .---81364

Picric acid ----------------------------------82340

1 . Application

This method is suitable for the determination

of 1,3,5-trinitro-1,3,5-triaza-cyclohexane (RDX),

2,4,6-trinitrophenol (picric acid), and 2,4,6-trinitrotoluene (TNT) in water or water-suspended-sediment mixtures containing at least 2 gg/L of analyte .

2. Summary of method

RDX, picric acid, and TNT are extracted from

water or water-suspended-sediment mixtures with

methylene chloride. The extract is concentrated and

subjected to high-performance liquid chromatography (HPLC) analysis using a reverse-phase column

and a dual-wavelength ultraviolet detector.

3. Interferences

Any compounds that exhibit chemical and physical properties similar to the compounds of interest

can interfere .
4. Apparatus

DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

4.1 Concentrator apparatus, Kuderna-Danish (K-D), with concentrator, 500-mL flask, one-ball

and three-ball Snyder columns, and a 10-mL graduated receiver.

4 .2 Evaporative concentrator, Organomation

N-Evap, or equivalent.

4 .3 Filters, 0.5 gm millipore FHUP, catalog

no. 04700, or equivalent, and 0.45 gm millipore

HAWP, catalog no. 04700, or equivalent.

4 .4 Liquid chromatograph, Waters Associates ALC/GPC 204 liquid chromatograph equipped

with a dual-channel, variable-wavelength detector, a

model 6000A solvent-delivery system, a model 660

solvent-flow programmer, a model WISP 710A

microprocessor, and a data module, or equivalent.

4.4 .1 The following conditions are recommended:

Column, 300 mm x 3.9 mm id (inside diameter) stainless steel packed with 10 gm particle

size reverse-phase material : Waters Associates

u-Bonapak C1a packing, or equivalent.

Wavelengths, 254 and 356 nm.

Solvent, 36 percent solution A (step 5.2.1)

and 64 percent solution B (step 5.2 .2) at a flow rate

of 1 .10 mL/min isocratic .

4.5 Solvent clarification kit, Waters Associates 85113, or equivalent.

4.6 Shaker, mechanical, wrist-action.

5. Reagents

5.1 Hydrochloric acid, concentrated (sp. gr.

1 .19), reagent grade.

5.2 Mobile phase solutions : Prepare the ion

pairing, tetrabutylammonium-phosphate solutions

for the mobile phase using Waters Associates PIC

Reagent A, or equivalent.

5.2 .1 Solution A: Add one bottle of PIC

reagent A to 1,000 mL organic-free water. Stir for 5

min and then filter through a 0.45-gm filter, type

HAWP for aqueous solvents.

5.2 .2 Solution B: Add one bottle of PIC

reagent A to 1,000 mL acetonitrile . Stir for 5 min

and then filter through a 0.45-gm filter, type FHUP

for organic solvents.

5.3 RDX, picric acid, and TNT standards,

analytical reference grade or highest purity available: These may be obtained from chemical specialty

suppliers or from military sources. Weigh about 10

mg to three significant figures and dilute to 25 mL

59

with acetonitrile . Prepare standards at 1, 5, and 10

ng/pL in acetonitrile.

5.4 Sodium chloride, anhydrous, granular:

Prepare by heating at 300°C overnight .

5.5 Solvents, acetone, diethylether (unpreserved) methylene chloride, pesticide residue

quality, distilled in glass, Burdick and Jackson, or

equivalent: HPLC-grade acetonitrile .

5.6 Water, organic-free .

6. Procedure

6.1 Pour 500 mL water into a 1-L separatory

funnel . Do not use stopcock grease on ground-glass

joints .

6.2 Add 3 mL concentrated HCl and 85 g

NaCl to the sample contained in the separatory funnel and shake until the salt is dissolved.

6.3 Add 75 mL methylene chloride to the separatory funnel . Stopper and shake for 1 min. Vent

the pressure often. Allow the layers to separate and

draw off the methylene chloride layer into a 250-mL

Erlenmeyer flask.

6.4 Repeat the extraction twice using 50 mL

methylene chloride each time. Combine the extracts

in the 250-mL Erlenmeyer flask.

6.5 Quantitatively transfer the extract to a

500-mL K-D apparatus fitted with a three-ball Snyder column and a 20-mL receiver. Add a micro boiling chip and 4 mL acetonitrile .

6.6 Place the K-D apparatus on a hot-water

bath (approximately 85'C). Reduce the volume of

the extract to about 4 mL. Remove the K-D appara

tus and allow it to cool . Wipe the joints with a towel.

Rinse the lower joint with acetonitrile into the receiver.

6 .7 Use the evaporative concentrator to reduce the volume of solvent to 0.8-0.9 mL by directing a stream of nitrogen onto the surface of the

extract in the receiver in a water bath at 35'C.

6.8 Proceed to HPLC analysis.

6.9 The chromatographic system should be

evaluated each day to determine retention volume

and detector response for RDX, picric acid, and

TNT. Prepare calibration curves daily by injecting

10 gL of a standard mixture at three concentrations

(see step 5.3) . Record the volume of the standard

injected and the retention time and integrated peak

area of each component in the standard. The operating conditions used for calibration must be identical

to those used for sample analysis (step 6.10) .
60

6.10 Add 0.1 mL PIC reagent A to the sample

extract, mix, and make up to a volume of 1.0 mL;

allow the extract to stand 10 min, and then inject 10

gL into the liquid chromatograph . Record the volume injected. Identify the peaks by retention time.

Confirmation is made by measuring the peak area at

two different wavelengths and comparing the ratio

of the peak areas to the peak area ratio of the standard. Dilute any extract containing an identifiable

component above the highest standards.

7. Calculations

7.1 Calculate the response factor of each

identified component in the calibration standard :

RF =

A,

where

RF= response factor of identified component in

calibration standard, in area/ng,

CS = concentration of standard component, in

ng/gL,

V, = volume of standard injected, in gL, and

A, = integrated peak area of identified component in calibration standard .

7.2 Calculate the concentration of each identi-

(1 .000 mL =1 .000 g).

8. Report

Report concentrations of TNT, RDX, and picric

acid in water or water-suspended-sediment mixtures as follows : less than 2 gg/L, as "less than 2

gg/L"; 2.0 gg/L and above, two significant figures.

9. Precision

Single-operator precision for eight replicates, expressed in terms of percent relative standard deviation, is as follows:

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

CS XV

Compound

RDX--------------------

Picric acid ---------------

Goerlitz, D.F., and Law, L.M., 1975, Gas chromatographic

method for the analysis of TNT and RDX explosives contaminating water and soil-core material : U.S . Geological Survey

Open-File Report 75-182, 21 p.

1979b, Direct analyses of RDX and TNT in water by

high-performance liquid chromatograph : U.S . Geological

Survey Open File Report 79-916, 11 p.

Goerlitz, D.F., 1979a, Analysis of picric acid in water by highperformance liquid chromatography : U.S. Geological Survey

Open File Report 79-207, 7 p.

1 . Application

Selected references

This method is suitable for the analysis of water

and water-suspended-sediment mixtures for polynuclear aromatic hydrocarbons (PNA's) containing

at least 1 gg/L of the analyte .

2. Summary of method

PNA's are extracted from water or

water-suspended-sediment mixtures with methylene chloride. The extract is concentrated and subjected to high-performance liquid chromatographic

Concentration

spiked (wglL)

Mean

concentration

recovered

(W91L)

Relative

standard

deviation

(Peru)

2.0 2.2 37

4.0 3.9 20

8.0 7.9 12

16 16 11

2.0 2.2 10

4 .0 4.0 10

8 .0 7.6 4

16 16 5

2.0 1 .7 23

4 .0 3.0 17

8 .0 6 .8 10

16 14 11

fied component in the original water sample from

the equation

Concentration

x

(wg/L)

A2 V2x 1,000

=

Vs x WxRF

Polynuclear aromatic hydrocarbons (PNA),

total recoverable, high-performance liquid

chromatographic (0-3113-83)

Parameter Code

Acenaphthene------------------------------- 34205

where Anthracene--------------------------------- 34220

RF= response factor of identified calibration Benzo(a)anthracene --------------------------34526

standard component, in area/ng, Benzo(g,h,i)perylene-------------------------- 34521

Benzo(a)pyrene------------------------------ 34247

A2 = integrated peak area of identified sample Chrysene----------------------------------- 34320

component, Dibenzo(a,h)anthracene -----------------------34556

V2 = final volume of sample extract, in mL, Fluoranthene -------------------------------34376

V3 = volume of sample extract injected, in gL, Fluorene----------------------------------- 34381

and Naphthalene--------------------------------34696

Phenanthrene------------------------------- 34461

W=weight of sample in g, expressed in mL Pyrene ------------------------------------34469
(HPLC) analysis using a 10-gm reverse-phase column and a dual-channel ultraviolet detector.

3. Interferences

Any compounds that exhibit chemical and (or)

physical properties similar to the compounds of interest can interfere .

4. Apparatus

5. Reagents

DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

4.1 Concentrator apparatus, Kuderna-Danish (K-D), with a 500-mL flask, a three-ball Snyder

column, and a 10-mL graduated receiver tube.

4.2 Evaporative concentrator, Organomation

N-Evap, or equivalent.

4.3 Filtering apparatus, Millipore, or equivalent : The filtering apparatus consists of a 250-mL

reservoir with a glass frit, and a 1,000-mL receiving

reservoir with FH, Millipore no. FHUP 04700 and

HA, Millipore no. HAWP 40700 0.45-gm filters for

the corresponding organic and aqueous solvents, or

equivalent.

4.4 Liquid chromatograph, Waters Associates ALC/GPC 204 liquid chromatograph equipped

with a dual-channel, variable-wavelength detector, a

model 6000A solvent-delivery system, model WISP

710A microprocessor with a model 730 data module

and a model 720 system controller, or equivalent.

4.4 .1 The following conditions are recommended:

Column, reverse-phase, micro-bondapak

C18-10 gm, Waters Associates, or equivalent.

Wavelengths, 254 and 313 nm.

Solvent, 40 to 80 percent acetonitrile/water, linear slope gradient at a flow rate of

1.0 mL/min.

5.1 PNA standards, EPA analytical reference grade or highest purity available : Use methylene chloride as a solvent to prepare stock solutions

in the approximate 100-300 ng/pL concentration

range. Store in the dark at 4'C.

5.2 PNA working-standard solution : Prepare three standard mixtures of 12 PNA's at concentrations of 1, 5, and 10 ng/pL in acetonitrile,

from step 5.1 .

5.3 Sodium sulfate, granular, anhydrous:

Heat overnight at 300'C and store at 130'C.

5.4 Solvents, HPLC-quality acetonitrile, and

methylene chloride: Filter before use with the filtering apparatus described above (step 4.3) .

5.5 Water, organic-free .

6. Procedure

61

Glassware must be cleaned by washing with a hot

detergent solution, rinsing with deionized water, and

heating overnight at 300°C. Just prior to use, the

glassware is rinsed with solvent. Stopcock grease

should not be used on ground-glass joints .

6.1 Weigh the bottle containing the sample

and record the weight. Pour the sample into a 1-L

separatory funnel . Weigh the empty sample bottle .

Calculate the net sample weight and record the

value obtained to three significant figures .

6.2 Add 50 mL methylene chloride to the

sample bottle, swirl to rinse the sides of the bottle,

and transfer the solvent to a separatory funnel . The

Teflon-lined cap is not rinsed because of the potential for contamination from solvent that has contacted the threads and surface beneath the Teflon liner .

Shake the separatory funnel vigorously for 1 min.

Vent often. Allow the layers to separate and draw

off the methylene chloride layer into a 250-mL

Erlenmeyer flask that contains 0 .5 g anhydrous sodium sulfate.

6.3 Repeat the extraction of the water sample

twice using 40 mL methylene chloride each time.

Combine all organic extracts in the 250-mL

Erlenmeyer flask .

6.4 Transfer the extract to a 500-mL K-D

apparatus fitted with a three-ball Snyder column

and a 10-mL receiver containing a micro boiling chip

and 0.5 mL of acetonitrile .

6.5 Place the apparatus on a water bath at

about 80'C and concentrate to about 5 mL. Remove

from the heat and allow to cool. Dry the joints with a

towel. Rinse the lower joint with acetonitrile into the

receiver.

6.6 Further reduce the volume of solvent to

about 1 mL on an evaporative concentrator. Rinse

down the sides of the tube with 1 mL acetonitrile

and concentrate to a final volume of 0.5 mL. Stopper

until chromatographic analysis can begin.

6.7 Optimize the chromatographic conditions.

6.8 Prepare liquid chromatograph calibration

curves daily by injecting the standards described in

step 5 .2 . Operating conditions must be identical to

those used for sample analysis (step 6.9) . Record the

volume of the standard injected and the retention

time and integrated peak area of each component in

the standard . The calibration should be performed at

the beginning and end of a run, and after every

fourth sample.

6.9 Inject an aliquot of sample extract into

the liquid chromatograph . Record the volume inject-
62

ed. Identify the peaks by retention time. Confirmation is made by measuring the peak area at two

different wavelengths and comparing the ratio of

the peak areas to that of the standard. Record the

retention time and integrated area of any identified

peak. Dilute any extract containing an identifiable

component above the highest standard. The 254-nm

detector does not resolve some pairs of compounds.

Determination at another wavelength, 313 nm, is

necessary to distinguish between these pairs . For

example, fluorene and acenaphthene each absorb at

254 nm and are not separated by the column .

Acenaphthene, however, absorbs at 313 nm, whereas fluorene does not . The response ratio of

acenaphthene calculated at 254 nm and 313 nm is

l:l ; therefore, both peaks are analyzed. Chrysene

and benzo(a)anthracene are not clearly distinguishable at 254 nm, but are at 313 nm. When these

compounds are determined at both wavelengths, individual contributions to peak areas can be determined and concentrations calculated.

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

W= weight of sample determined in g expressed

in mL (1 .000 mL = 1.000 g).

8. Report

Report concentrations of individual PNA's in

water or water-suspended-sediment mixturesas follows: less than 1 lag/L, as "less than 1 gg/L"; 1 to

10 gg/L, one significant figure; 10 p.g/L and

greater, two significant figures.

9. Precision

Single-operator precision on seven replicates and

recovery data determined by spiking watersuspended-sediment mixture samples with PNA's

are as follows:

Mean Relative

concentration standard

Concentration recovered deviation

Compound spiked (W9lL) (k9/L) (perce t)

7. Calculations Naphthalene------------- 1 .3 0 .94 6 .9

7.1 Calculate the response factor of each 2 .6 1 .41 20

identified component in the calibration standard :

Fluorene----------------

5 .1

.64

2 .4

.47

24

12

A, 1 .3 .94 7.8

RF= 2.6 1 .6 14

C3 X V Acenaphthene ----------- 2.8 2 .01 8.5

5 .50 3.6 13

where 11 . 6.5 18

RF= response factor of identified component in Phenanthrene------------ .21 .15 8.3

calibration standard, in area/ng,

.42

.84

.36

.66

6.3

8.5

Cs = concentration of standard component, in Anthracene ------------- .052 .038 3.9

ng/gL, .10 .076 5 .1

V = volume of standard injected, in p.L, and .21 .142 7 .4

A, = integrated peak area of identified compoPyrene ----------------- .32

.64

.28

.58

2.8

3 .4 nent in calibration standard . 1 .3 1 .1 5 .6

7.2 Calculate the concentration of each identi- Fluoranthene ------------ 1 .4 1 .1 3 .5

fied component in the original water sample from 2 .8 . 2 .5 5 .5

the equation 5 .5 5 .0 5 .1

Benzo(a)anthracene ------- .43 .39 3 .0

A2 X V2 X 1,000 .85 .80 3 .4

Concentration (pg/L) =

'

1 .7 1 .6 5 .5

VXWXRF Benzo(a)pyrene ---------- .15 .13 4 .8

.29 .29 7 .5

where .58 .55 5 .5

RF= response factor of identified calibration Dibenz(a,h)anthracene----- .13 .12 6 .9

standard component, in area/ng, .26

.51

.28

.50

7 .4

5 .0

A2 = integrated peak area of identified sample Benzo(g,h,i)perylene ----- 1 .6 1 .4 3 .0

component, 3 .2 3 .2 5 .8

V2 =final volume of sample extract, in mL, 6 .4 6 .2 4 .0

V2 =volume of sample extract injected, in gL, Chrysene --------------- .64 .57 5 .1

1 .3 1 .3 5 .2

and 2 .6 2 .3 7 .2
Acheson, M.A ., Harrison, R.M., Perry, R., and Wellings, R.A .,

1976, Factors affecting the extraction and analysis of polynuclear aromatic hydrocarbons in water: Water Research, v.

10,p .207-212.

Giger, Walter, and Schaffner, Christian, 1978, Determination of

polycyclic aromatic hydrocarbons in the environment by

glass capillary gas chromatography : Analytical Chemistry, v.

50,p .243-249.

Goerlitz, D.F., and Brown, Eugene, 1972, Methods for analysis of

organic substances in water: U.S . Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter

A3, 40 p.

Josephson, Julian, 1981, Polynuclear aromatic hydrocarbons: Environmental Science and Technology, v . 15, p. 20-22.

Keith, L.H ., and Telliard, W.A ., 1979, Priority pollutants : Environmental Science and Technology, v. 13, p. 416-423.

Kuratsune, M., and Hirohata, T., 1962, Decomposition of polycyclic aromatic hydrocarbons under laboratory illuminations:

National Cancer Institute Monograph 9, p. 117-125 .

Lee, M.L., and Wright, B.W., 1980, Capillary column gas chromatography of polycyclic aromatic compounds-A review :

Journal of Chromatographic Science, v. 18, p. 345-358.

White, W.R ., and Grabbe, R.R ., 1981, High-performance, liquidchromatographic method for the analysis of polynuclear aromatic hydrocarbons in water: U.S . Geological Survey WaterResources Investigations 81-63, 13 p.

Ethylene and propane, total recoverable, gas

chromatographic, purge and trap (0-3114-83)

Parameter Code

Ethylene-----------------------------------82357

Propane -----------------------------------82358

1 . Application

This method is suitable for the determination of

ethylene and propane in water or water-suspendedsediment mixtures containing at least 0 .1 lag/L of

the analyte . Concentrations higher than 100 gg/L

may be determined by analyzing a smaller aliquot of

the sample.

2. Summary of method

DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

Selected references

A sample is injected into a purge vessel and

sparged with nitrogen. The effluent gas stream is

dried and passed through an alumina trap at

-95'C, where ethylene and propane are adsorbed

by the alumina . The trap is heated to desorb gases,

which are determined by gas chromatography using

a flame ionization detector .

3. Interferences

63

Volatile compounds that have retention times similar to ethylene or propane on the analytical column

can interfere .

4. Apparatus

4.1 Adsorption tube, Pyrex tubing, 1/4 in od

(outside diameter) : Bend Pyrex tubing into a U-tube

approximately 8 in long by 2 in wide. Charge the

tube with 5 g activated alumina held in place with

plugs of glass wool.

4.2 Cold bath, a Dewar flask large enough to

accommodate the lower half of the adsorption tube:

Maintain the bath at -75 to -100°C with either

dry ice/acetone or a refrigerant probe such as the

Neslab CC-100, or equivalent.

4.3 Drying tubes, Pyrex tubing, 1/2 in od by 8

in long: Fill the tubes with Drierite held in place with

glass-wool plugs.

4 .4 Gas chromatograph, Tracor 560, or

equivalent.

4.4 .1 The following conditions are recommended:

Column, borosilicate glass, 1 .8 mx2 mm

id (inside diameter) operated at 90 ° C, packed with

Porapak N, Q, or QS.

Detector, flame-ionization, operated at

250°C.

Injection port temperature, 1100'C .

Carrier gas, helium, flow rate 30 mL/min.

4.5 Gas sampling valve, six port, Valco V-6-

HPa, or equivalent.

4.6 Hot bath, a heating mantle for a 1-L

round-bottom flask filled with sand and powered by

a variable-voltage transformer: The voltage is ad

justed to maintain the sand at approximately 130 ° C.

4.7 Purge vessel : See figure 3.

4.8 Sample vial, glass, 40 mL, screw cap,

fitted with a Teflon-lined septum . Pierce 13075, or

equivalent.

4.9 Syringe needles, 19 gauge, 3 in long.

4.10 Syringes, two syringes, 30 mL and 5

mL, glass, Luer-Lock, equipped with stopcocks, Becton-Dickinson 3152, or equivalent.

4.11 Syringes, gas-tight, syringes of 1 .0-,

2.5-, 10-, and 50-mL capacities, Hamilton, or equivalent, used for delivery of calibration gas and

equipped preferably with sideport needles .

4.12 Teflon tubing, 1A in od, used for the connecting lines .
TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

WE00 P

Ill

about 1 inch outside diameter

glass tubing

Drain

Teflon stopcock

Nitrogen inlet

'/a inch outside diameter

Septum

'/a X/4 inch

Swagelok union

Medium-porosity

glass frit

E- '/a inch outside diameter

about 10 inches
5. Reagents

6. Procedure

-Figure 3.-Purge vessel .

DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS 65

5.1 Alumina, neutral aluminum oxide, activity grade I, Woelm, or equivalent.

5.2 Calibration gas, a certified gas mixture

containing 10 ppm each of ethylene and propane in

nitrogen: The regulator valve must be equipped with

a stainless steel diaphragm. Equip the outlet from

the regulator with an injection septum to facilitate

withdrawal of aliquots of calibration gas with a gastight syringe.

5 .3 Drying agent, indicating Drierite, 8-

mesh: Anhydrous magnesium perchlorate (granular)

is also suitable .

Samples are to be collected in vials containing 1

mL formalin as a preservative and in a manner that

precludes headspace formation. Samples need not be

refrigerated since formalin prevents bacterial decomposition for at least 3 weeks, and probably longer. Rapid loss of ethylene and propane will occur if

the preservative is omitted.

6.1 Adjust the flow of hydrogen and air to the

flame-ionization detector to achieve a linear response from 1 to 100 ng of ethylene and propane .

6 .2 Set the flow of nitrogen through the

purging vessel at 30 to 35 mL/min. This is conveniently measured at the outlet port of the gas-sampling valve (see fig. 4) .

6.3 Place the adsorption tube in the cold bath

and set the gas-sampling valve as shown in figure 4.

6.4 Calibration procedure:

6.4 .1 Inject an aliquot of calibration gas

mixture into the purging vessel using a gas-tight

syringe .

6.4 .2 Start the timer and allow 12 min for

complete adsorption of ethylene and propane by the

alumina trap. NOTE: The minimum time required to

completely strip the gases depends on the dead volume of the system (purging vessel, drying tube, adsorption tube, and connecting lines) and the flow

through it .

6.4 .3 Turn the gas sampling valve to the

position that conducts the flow from the adsorption

tube into the analytical column of the gas chro

matograph. Immediately place the adsorption tube

in the hot bath and begin digital peak integration .

Record integrated peak areas.

6.4 .4 Repeat steps 6 .4 .1 through 6 .4 .3 for

as many other aliquots of calibration gas mixture as

is necessary to cover the expected range of the samples.

6 .5 Sample analysis:

6.5 .1 Open a sample vial and fill a 30-mL

glass syringe by closing the stopcock and pouring

the sample gently into the barrel. Do not attempt to

fill the syringe by suction.

6.5 .2 Introduce 10 mL of sample into the

purging vessel through the injection port.

6.5 .3 Start the timer and allow the sample

to be purged for 12 min (see NOTE above) .

6.5 .4 Turn the gas sampling valve to the

position that conducts the flow from the adsorption

tube into the analytical column of the gas chro

matograph. Immediately place the adsorption tube

in the hot bath and begin digital peak integration .

Record integrated peak areas.

6.5 .5 Drain the sample from the stripping

chamber while gas chromatographic determination

proceeds .

7. Calculations

7.1 Correction for calibration gas:

7.1 .1 The calibration gas is 10 ppm by volume. This must be corrected to give the corresponding concentration by weight.

7.1 .2 For 1 mL of 10 ppm calibration gas,

the volume of ethylene and propane is 1 .0 x 10-5 mL

each.

7.1 .3 Applying the ideal gas law at standard temperature and pressure (STP) where 1 mole

of gas occupies 22.4 L,

N,TIP2 V2

N2 T9P1V1

where

N1 = 1 .0 mole,

N2 = number of moles ethylene or propane,

T1 = 273°K (STP),

P1 = 760 mm Hg (STP),

V1 = 22,400 mL (volume of 1 mole gas at STP),

T2 = actual temperature, in °K,

P2 = actual atmosphere pressure, in mmHg, and

V2 = 1.0 x 10-5 mL (volume of ethylene or propane in 1 .0 mL of calibration gas) .
66 TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

7.2 For either ethylene or propane, calculate

the concentration of ethylene or propane per mL of

calibration gas from the following equation :

N, xMWx 106

V3

where

W= concentration of ethylene or propane in

calibration gas, in lag/mL,

Gas chromatograph

equipped with

flame-ionization detector

Figure 4.-Stripping and gas chromatographic system in the stripping mode.

Six-port

valve (sampling

valve) Vent (outlet port)

V3 = volume of calibration gas, in mL,

N, = number of moles of ethylene or propane,

and

MW=molecular weight of ethylene or propane,

in g/mole.

7.3 Calculate the response factor for the calibration curve from the following equation :

RIP=

A,

WXV4

Strip chart recorder

and integrator
DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

where

RF = response factor of ethylene or propane, inarea/gg,

A, = integrated peak area of standard,

V4 = volume of calibration gas injected, in mL,

and

W = concentration of ethylene or propane calibration gas, in lag/mL .

7.4 The concentration of ethylene or propane

in the sample is calculated from the following equation:

Concentration (gg/L) =

A2 x 1,000

RFx V5

where

A2 = integrated peak area of ethylene or propane,

RF= response factor of ethylene or propane, in

area/gg, and

V5 = volume of sample injected, in mL.

8. Report

Report concentration of ethylene and (or) propane

as follows: concentrations less than 0.1 Igg/L, as

"less than 0.1 Fig/L" ; 0.1 to 1 .0 gg/L, one decimal ;

1 .0 lag/L and above, two significant figures.

9. Precision

Solutions of ethylene and propane were prepared

in deionized water and stored in sample vials (step

4.8) containing 1 mL formalin as preservative . Replicates were prepared to cover the concentration

range of the method. All of the samples were analyzed within 3 weeks of preparation. Single-operator

precision is as follows:

Selected reference

67

Shultz, D.J., Pankow, J .F ., Tai, D.Y ., Stephens, D.W., and Rathbun, R.E ., 1976, Determination, storage, and preservation of

low molecular weight hydrocarbon gases in aqueous solution:

U.S . Geological Survey Journal of Research, v. 4, p.

247-251.

Purgeable organic compounds, total

recoverable, gas chromatographic/ mass

spectrometric, purge and trap (0-3115-83)

Parameter Code

Chloromethane------------------------------ 34418

Bromomethane------------------------------ 34413

Vinyl chloride -------------------------------39175

Chloroethane -------------------------------34311

Methylene chloride ---------------------------34423

1,1-Dichloroethene ---------------------------34501

1,1-Dichloroethane ---------------------------34496

Trans- 1,2-Dichloroethene---------------------- 34546

Chloroform ---------------------------------32106

1,2-Dichloroethane ---------------------------32103

1,1,1-Trichloroethane -------------------------34506

Carbon tetrachloride--------------------------32102

Bromodichloromethane------------------------32101

1,2-Dichloropropane --------------------------34541

Trans- 1,3-Dichloropropene --------------------- 34699

Trichloroethene------------------------------ 39180

Dibromochloromethane------------------------ 32105

Benzene -----------------------------------34030

1,1,2-Trichloroethane -------------------------34511

Cis-1,3-Dichloropropene -----------------------34704

2-Chloroethylvinyl ether -----------------------34576

Bromoform ---------------------------------32104

1,1,2,2-Tetrachloroethane---------------------- 34516

Tetrachloroethene---------------------------- 34475

Toluene ------------------------------------ 34010

Chlorobenzene ------------------------------34301

Ethylbenzene -------------------------------34371

1 . Application

This method is suitable for the determination of

purgeable organic compounds in water and watersuspended-sediment mixtures containing at least 3

wg/L of a reportable analyte .

2. Summary

A water sample is purged with helium. The purgeable organic compounds are carried with helium and

trapped on a porous polymer trap. The trapped compounds are thermally desorbed into the gas chromatograph. These compounds are separated by gas

chromatography (GC) and detected by mass spectrometry (MS).

Concentration of

ethylene

(Pg1L)

Number of

replicates

Standard

deviation

Relative

standard deviation

(percent)

0 .072----- 7 0 .002 2 .8

.198----- 12 .006 3 .0

1 .69------ 11 .04 2 .4

8 .38------ 12 .10 1 .2

16 .0------- 12 .20 1 .3

35 .1------- 12 .6 1 .7

82 .4------- 11 2 .1 2 .5

132 -------- 11 3 .0 2 .3

Concentration of

propane

(pglL)

Number of

replicates

Standard

deviation

Relative

standard deviation

(percent)

0.099 --- 7 0 .004 4.0

.257 --- 12 .015 5.8

2.52 ---- 11 .05 2.0

12.9 ----- 12 .4 3.1

24.5 ----- 12 .4 1 .6

53.5 ----- 12 .9 1 .7

110 ----- 11 2 .6 2.4

171 ----- 11 4 .0 2.3
68

3 Interferences

3.1 Any purgeable compound that elutes at a

retention time similar to that of the analyte and

produces an ion that is the same as the quantitation

ion of the analyte is a potential interference. Common laboratory solvents such as methylene chloride,

benzene, and chloroform may contaminate the sample and give erroneous results .

3.2 Special handling of samples, such as storage in a dessicator over activated charcoal, may be

required to prevent contamination by common laboratory solvents.

4. Apparatus

4.1 Gas chromatograph/mass spectrometer/data system, Finnigan 3223, or equivalent.

4 .1 .1 Gas chromatographic column, borosilicate glass, 1 .8 m x 2 mm id (inside diameter) that

has been deactivated and packed with 1 percent SP1000 coated on 60/80 mesh Carbopack B, or equivalent .

4.1 .2 Gas chromatographic conditions : GC

conditions need to be optimized for each system. Use

the purgeable standards (see step 5 .3) to adjust con

ditions to obtain good peak separation in a reasonable amount of time. The following conditions should

serve as a starting point for the optimization process:

Injector temperature----- 200oC

Carrier gas flow (He) ---- 20 mL/min

Initial hold temperature -- 45°C

Initial hold time -------- 4 min

Program rate ---------- 8°C/min

Final temperature- - - - - - - 21WC

Final hold time --------- To end of data acquisition

4.1 .3 Mass spectrometer conditions : Analyze the mass range 35-260 amu (atomic mass

units) with a nominal electron energy setting of 70

eV at a scan rate sufficient to obtain a minimum of 5

scans per chromatographic peak.

4.2 Purge and trap device, Chemical Data

Systems model 310, or equivalent: The trap is

packed with the following adsorbents: 1 cm methyl

silicone coated packing (3 percent SP-2100 on 80-

mesh Supelcoport, or equivalent), followed by 15 cm

Tenax, and ending with 8 cm silica gel (Davison

grade 15), or equivalent. The silica gel may be replaced by 5 cm of Ambersorb resin (Rohm and

Haas). New traps are conditioned by heating overnight at 240'C with helium flow (20 mL/min) .

4.3 Syringe, gas-tight, 10 mL, equipped with

a Teflon syringe valve and a 3-in x 19-gauge needle .

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

5. Reagents

5.1 BFB (4-Bromofluorobenzene) solution:

Fill a 50-mL volumetric flask to the mark with

methanol and add a 1 .0-pL capillary pipet filled with

BFB to the volumetric flask. This solution contains

32 ng BFB per gL.

5.2 Methanol, pesticide analysis quality, Burdick and Jackson, or equivalent.

5.3 Purgeable mixed standards, EPA analytical reference grade or highest purity available:

Purgeable standards may be purchased from various

commercial sources or prepared from pure compounds. To prepare purgeable mixed standards, fill a

50-mL volumetric flask to the mark with methanol

and, for each desired component, add a 1 .0-gL capillary pipet filled with the authentic material to the

volumetric flask . Mix thoroughly and store at 4'C.

Calculate the concentration of each analyte from its

density .

5 .4 Surrogates/internal standard solution,

bromochloromethane (EPA, or equivalent), 1-bromo2-chloroethane (EPA, or equivalent), perdeuter

obenzene (Pfaltz and Bauer, or equivalent), and

fluorobenzene (the internal standard, Aldrich Chemical Co., or equivalent) . Add a 1 .0-gL capillary pipet

filled with each component to a 50-mL volumetric

flask filled to the mark with methanol. Mix thoroughly and store at 4°C. Add 5 gL to each sample,

standard, and blank to monitor recovery and to provide an internal standard.

5.5 Water, organic-free .

6. Procedure

6.1 Condition the trap at 220'C for 10 min.

6.2 Mass spectrometer tuning:

6.2 .1 Use perfluorotributylamine to tune

the mass spectrometer in a manner that results in a

satisfactory calibration of mass assignments as well

as agreement with the criteria listed in step 6.2 .2 .

6.2 .2 Set the MS to scan the mass range 35

to 260 amu. Set the GC column temperature to

220-230°C isothermal. Introduce 50 ng of BFB (1 .6

pL of solution, step 5.1) by direct, on-column injection or by purging from 5 mL of reagent water.

Obtain a background corrected mass spectrum of

BFB and verify that all of the following criteria are

met:

Mass Ion abundance criteria

50 -------------------------- 15 to 40 percent of mass 95

75 -------------------------- 30 to 60 percent of mass 95

95 -------------- Base peak, 100 percent relative abundance

96 ---------------------------- 5 to 9 percent of mass 95
DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

Mass Ion abundance criteria

173----------------------------- <2percent ofmass 174

174----------------------------->50 percent of mass 95

175 --------------------------- 5 to 9 percent of mass 174

176 - - - - - - - - - - - - - >95 percent but < 101 percent of mass 174

177--------------------------- 5to9percent ofmass 176

6 .3 Blank analysis:

6.3 .1 Remove the plunger from a 10-mL

syringe and attach a closed syringe valve and needle. Open the sample vial and carefully pour the

water into the barrel of the syringe until it overflows. Replace the syringe plunger, open the syringe

valve, and vent any air present. Adjust the plunger

to the 10.0-mL mark. Add 10 gL of the surrogate/internal standard solution through the valve,

close the syringe valve, and mix.

6.3 .2 Transfer 5.0 mL water into a clean

purge tube. Close the syringe valve and retain the

second 5.0-mL aliquot of water for future analysis,

if needed. The second aliquot is preserved in this

manner since the integrity of the original water

sample is destroyed after it is opened.

6.3 .3 Purge for 11 min.

6.3 .4 Immediately begin the desorb cycle

and data acquisition when purging is complete.

6.3 .5 Begin the temperature program of

the GC oven immediately upon completion of the

desorb cycle.

6.3 .6 Initiate the trap bakeout when data

acquisition has ended.

6.3 .7 Allow the trap to cool to room temperature and return the GC oven temperature to

45'C for the next analysis.

6.3 .8 Analyze the mass spectral data for

the three surrogates and the internal standard . Record the integrated area of the quantitation ion for

each.

6.3 .9 Examine the mass spectral data to

verify that the analytical system is free from contamination.

6.4 Calibration:

6.4 .1 Remove the plunger from a 10-mL

syringe and attach a closed syringe valve and needle. Open the sample vial and carefully pour the

organic-free water into the barrel of the syringe

until it overflows . Replace the syringe plunger, open

the syringe valve, and vent any air present . Adjust

the plunger to the 10.0-mL mark. Add 10 gL of

surrogate/internal standard solution . Also add 10

gL of purgeable standard solution . Close the syringe

valve, and mix.

69

6 .4 .2 Transfer 5.0 mL water into a clean

purge tube. Close the syringe valve and retain the

second 5.0-mL aliquot of sample for future analysis,

if needed. The second aliquot is preserved in this

manner since the integrity of the original water

sample is destroyed after it is opened.

6.4 .3 Perform the analysis as described in

steps 6 .3 .3 through 6 .3 .8 .

6.4 .4 Compare the recovery of the surrogates in the purgeable standard with that observed

in the blank. Deviations of more than ± 30 percent

from theoretical are an indication of a problem (e .g .,

leakage in the purge and trap device) that needs to

be corrected before proceeding further .

6.4 .5 Process the data from the purgeable

standard and record the integrated area of the quantitation ion of each component as well as its retention time.

6.4 .6 Repeat steps 6 .4 .2 through 6.4 .5 with

as many other volumes of the purgeable standard

solution as are necessary to define the working

range ofthe analytical system.

6.5 Sample analysis:

6.5 .1 Allow the water sample to come to

room temperature .

6.5 .2 Remove the plunger from a 10-mL

syringe and attach a closed syringe valve and needle . Open the sample vial and carefully pour the

sample into the barrel of the syringe until it overflows. Replace the syringe plunger, open the syringe

valve, and vent any air present . Adjust the plunger

to the 10.0-mL mark. Add 10 p.L of the surrogate/internal standard solution . Close the syringe

valve, and mix.

6.5 .3 Transfer 5.0 mL of sample into a

clean purge tube. Close the syringe valve and retain

the second 5.0-mL aliquot of sample for future anal

ysis, if needed. The second aliquot is preserved in

this manner since the integrity of the original water

sample is destroyed after it is opened.

6 .5 .4 Perform the analysis as described in

steps 6.3 .3 through 6.3 .8 .

6 .5 .5 Compare the recovery of the surrogates in the sample with that observed in the blank

(step 6 .3 .8) . If the recovery is not in the range of 70

to 130 percent, the sample should be reanalyzed.

6.5 .6 Examine all ofthe mass spectral data

from the sample. Identify analytes by a library

search with a satisfactory match error. Positive

identification is obtained when (1) the retention time

is within 5 percent of the authentic material in the

purgeable standard (step 6.4 .5), and (2) three of the
70 TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

characteristic ions of the analyte maximize

within ± 1 scan of each other.

6 .5 .7 Integrate and record the area under

the quantitation ion for each analyte identified in

step 6.5 .6 . If the areas are greater than the calibra

tion range of the analytical system, the sample

needs to be reanalyzed using a smaller volume of

sample. It may be necessary to add additional internal standard for smaller sample volumes.

6.5 .8 If very high levels of analytes or contaminants are found in a sample, analyze a blank to

demonstrate no carryover.

7. Calculations

7.1 External standard method:

7.1 .1 Calculate a response factor for an

analyte (step 6.4 .5) according to the equation

A

RF= CxV,

where

RF= response factor of analyte, in area/ng,

C = concentration of analyte in purgeable standard, in ng/gL,

V =volume of purgeable standard analyzed, in

gL, and

A = area of quantitation ion of analyte .

7.1 .2 Calculate the concentration of the

analyte in the original water sample from the equation

Concentration (gg/L) =

AS

Vx RF

where

AS = area of quantitation ion of analyte in sample,

VS = volume of original water sample analyzed,

in mL, and

RF= response factor of analyte, in area/ng .

7.1 .3 Calculate the percent recovery of

each surrogate added to the water sample from the

equation

Recovery (Percent) _:

A1

A2

x 100,

where

A1= area of quantitation ion of surrogate in

water sample, and

A2 = area of quantitation ion of surrogate added

to blank.

7 .2 Internal standard method:

7.2 .1 Calculate the response factor of an

analyte (step 6 .4 .5) in the purgeable standard from

the equation

where

RF= response factor of analyte, in area/ng,

AS = area of quantitation ion of analyte, and

CS = amount of analyte in purgeable standard, in

ng.

7.2 .2 Calculate the response factor of the

internal standard (step 6.4 .5) in the purgeable standard from the equation

RFI =

Ali

Ch

where

RFI = response factor of internal standard in

purgeable standard, in area/ng,

AI, = area of quantitation ion of internal standard in. purgeable standard, and

CI, = amount of internal standard in surrogate

and internal standard solution, in ng.

7.2 .3 Calculate a relative response factor

from the equation

RF

RRF =

RFI

where

RRF= relative response factor of analyte,

RF = response factor of analyte determined

(step i' .2 .1), and

RFI=response factor of analyte internal standard determined (step 7.2 .2) .

7.2 .4 Calculate the concentration of an

analyte in the original water sample from the equation

AxCl2

Concentration (pg/L) =

RRFxAI2 x V P

where

A = area of quantitation ion of analyte in analyzed sample,

CI2 = amount of internal standard in surrogate

solution, in ng,

RRF= relative response factor determined (step

7.2 .3),
AI, = area of quantitation ion of internal standard in analyzed sample (step 6.5 .9),

and

V = volume of original water sample analyzed,

in mL.

7 .2 .5 Calculate the percent recovery of

each surrogate added to the water sample from the

equation

where

A, = area of quantitation ion of surrogate added

to water sample, and

A2 = area of quantitation ion of surrogate added

to blank.

8. Report

9. Precision

DETERMINATION OF ORGANIC SUBSTANCES IN WATERAND FLUVIAL SEDIMENTS

Recovery (Percent) =

A1

A2

x 100,

Report concentrations of purgeable organic compounds in water or water-suspended-sediment mixtures as follows: less than 3 wg/L, as "less than 3

gg/L"; 3.0 wg/L and above, two significant figures.

Precision data are not available.

Selected references

Goerlitz, D.F., 1976, Determination of volatile organohalides in

water and treated sewage effluents: U.S. Geological Survey

Open-File Report 76-610, 14 p.

U.S . Environmental Protection Agency, 1979, PurgeablesMethod 624: Federal Register, v. 44, no. 233, p. 69532.

Acid extractable compounds, total recoverable,

gas chromatographic/ mass spectrometric

(0-3117-83)

Parameter Code

4-Chloro-3-methylphenol ----------------------34452

2-Chlorophenol------------------------------ 34586

2,4-Dichlorophenol ---------------------------34601

2,4-Dimethylphenol --------------------------34606

4,6-Dinitro-2-methylphenol --------------------34657

2,4-Dinitrophenol ----------------------------34616

2-Nitrophenol------------------------------- 34591

4-Nitrophenol------------------------------- 34646

Pentachlorophenol ---------------------------39032

Phenol-------------------------------------34694

2,4,6-Trichlorophenol -------------------------34621

8-Methyldecanoic acid-------------------------76992

Undecanoic acid -----------------------------77648

1 . Application

71

This method is suitable for the determination of

acid extractable compounds in water and watersuspended-sediment mixtures containing at least

3-5 pg/L of the analyte .

2. Summary

Acidic organic compounds are extracted from

water and water-suspended-sediment mixtures with

methylene chloride. The extract is concentrated and

subjected to analysis by gas chromatography (GC)

using a flame-ionization detector (FID) or a mass

spectrometric (MS) detector .

3. Interferences

Compounds having chemical and physical properties similar to the compounds of interest may interfere.

4. Apparatus

4.1 Boiling chip, micro, carbon chips: Rinse

with hexane, air dry, and heat at 300 ° C overnight .

Treat with 5 percent aqueous sulfuric acid for 5 min.

Rinse with deionized water until the wastewater is

neutral to pH paper. Heat at 130°C overnight .

4.2 Concentrator, Kuderna-Danish (K-D),

500-mL, all glass, with ground-glass joints, a 10.0-

mL receiver, and a one-ball Snyder column.

4.3 Evaporative concentrator, Organomation N-Evap, or equivalent: Water bath must be

maintained at 50°to 55°C.

4.4 Gas chromatograph/mass spectrometer/data system, Hewlett-Packard 5985B GC/MS

(gas chromatograph/mass spectrometer), or equiva

lent : The gas chromatograph is used with one of the

following options :

4.4 .1 Column, fused silica capillary column, 25 mx 0 .20 mm id (inside diameter), SE-54

bonded column, 0.33-gm film thickness .

Detector, mass spectrometer .

Injection temperature, 260'C.

Carrier gas, 1 mL/min, helium.

Transfer line temperature, 285'C.

Mode, splitless injection .

Program rate, 45' to 300'C, 2.5-min initial hold, 6°C/min, 15-min final hold.

4.4 .2 Columns, two fused silica capillary

columns, 25 m x 0.20 mm id, SE-54 bonded column,

0.33-gm film thickness .

Detectors, mass spectrometer and a flameionization detector, 285 ° C.
72

Injection temperature, 260°C.

Mode, splitless injection .

Carrier gas, 1 mL/min, helium.

Transfer line, 285°C.

Program rate, 45' to 300'C, 2.5-min initial hold, 6 ° C/min, 15-min final hold.

4.5 pHpaper, hydrion, pH range 1 to 14.

5. Reagents

5.1 Acidic organic compound standards,

EPA analytical reference grade or highest purity

available : Weigh about 20 mg of the standard to

three significant figures, quantitatively transfer it to

a 100-mL volumetric flask, and dilute to volume

with methylene chloride. Prepare dilutions in methylene chloride to obtain solutions containing 2, 10,

and 20 ng/gL.

5.2 DFTPP (Deeafuorotriphenylphosphine)

solution, 50 ng/gL: Dilute 20 gL DFTPP (Supelco,

or equivalent, 25 mg/mL solution) to 10 mL in a

volumetric flask with methylene chloride .

5.3 Internal standard, perdeuterophenanthrene (phenanthrene-d,o), Kor Isotopes, Division of

Kor, Inc., or equivalent; perdeuteronaphthalene

(naphthalene-ds), Aldrich Chemical Co., or equivalent ; and perdeuterochrysene (chrysene-d, 2 ), Kor

Isotopes, Division of Kor, Inc., or equivalent: Weigh

about 20 mg perdeuteronaphthalene, perdeuterophenanthrene, and perdeuterochrysene to three

significant figures, quantitatively transfer to a 100-

mL volumetric flask, and dilute to volume with

methylene chloride.

5.4 Sodium hydroxide, pellets, reagent

grade.

5.5 Sodium hydroxide solution, 37 percent

(weight/volume ; w/v): Dissolve 185 g sodium hydroxide pellets in 500 mL organic-free water and

reflux 8 h. Cool and store at 4'C.

5.6 Sodium sulfate, anhydrous, granular,

Mallinckrodt 8024, or equivalent: Heat at 300°C

overnight, slurry with enough diethyl ether to cover

the crystals, and acidify to pH 2 or less by adding a

few millileters of concentrated sulfuric acid. Determine the pH by removing a, portion of the slurry,

evaporating the ether, adding water to the crystals,

and testing the aqueous phase with pH paper. Evaporate the ether by allowing the slurry to stand in an

open container under a fume hood. Store at 130'C

in a glass-stoppered bottle .

5.7 Solvents, diethyl ether, unpreserved, distilled in glass, pesticide analysis quality, Burdick and

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

Jackson, or equivalent; methylene chloride, isopro

panol, glass distilled pesticide quality, Burdick ana

Jackson, or equivalent.

5 .8 Sulfuric acid, concentrated, (sp. gr.

1 .84), reagent grade, Mallinckrodt, or equivalent.

5 .9 Sulfuric acid, (1 + 3): Prepare by adding

one part concentrated sulfuric acid to three parts

organic-free water. Store in a refrigerator at 4'C.

5 .10 Surrogate standards, perdeuterophenol

(phenol-d 6 ), Aldrich Chemical Co., or equivalent;

dibromobenzene, FDA, or equivalent; 2,4,6

tribromophenol, FDA, or equivalent: Weigh 4 mg of

each of the three compounds to three significant

figures, quantitatively transfer to a 100-mL volumetric flask, and dilute to volume with isopropanol .

5 .11 Water;, organic-free .

6. Procedure

All glassware must be washed in warm detergent

solution, rinsed with organic-free water, and heated

at 300°C overnight. All glassware must be rinsed

with (1 + 3) aqueous sulfuric acid, then rinsed with

organic-free water, until the washwater is neutral to

pH paper, and then heated at 130 ° C overnight . Immediately before use, the glassware is rinsed with

methylene chloride . Stopcock grease should not b,

used on ground-glass joints .

6.1 Immediately upon receipt of the sample in

the laboratory, acidify to pH 2 or lower (as indicated

by pH paper) with (1 + 3) sulfuric acid (approxi

mately 3 mL), and store at 4'C. Extraction must

begin within 48 h after receipt of the sample.

NOTE: Extraction may be carried out on the

aqueous phase following extraction of the extractable base/neutral compounds (method 0-3118).

6.2 A blank must accompany each set of samples . For each sample and blank, rinse a 2-L separatory funnel and a 500-mL Erlenmeyer flask with

methylene chloride .

6.3 Weigh the capped sample bottle to three

significant figures and record the weight for subsequent calculations.

6 .4 Pour the sample into the separatory funnel and allow the sample bottle to drain completely.

6.5 Weigh the capped empty sample bottle to

three significant figures and calculate and record

the net sample weight.

6.6 Add 1.0 mL of the surrogate standard

(5 .10) to the sample in the separatory funnel.

6.7 Add 100 mL methylene chloride to the

empty sample bottle and gently swirl to wash tl-
DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

sides of the container with the solvent. The Teflon

liner inside the cap is not rinsed because of the potential for contamination from solvent that has contacted the cap threads and the surface beneath the

liner . Pour the contents of the bottle into the separatory funnel .

6.8 Stopper the separatory funnel and shake

vigorously for at least 1 min, venting often to release the pressure. Allow the layers to separate.

6.9 Drain the organic layer into a 500-mL

Erlenmeyer flask.

6.10 Add 50 mL methylene chloride to the

separatory funnel . Stopper the separatory funnel

and shake for at least 1 min, venting often to release

the pressure . Allow the layers to separate.

6.11 Drain the organic layer into the

Erlenmeyer flask.

6.12 Extract the sample one more time by

repeating steps 6.9 and 6.10.

6.13 Add approximately 30 g sodium sulfate

to the Erlenmeyer flask. Cover the Erlenmeyer flask

with aluminum foil and set aside for no longer than

2 h. (Significant loss of analytes can occur from

sample adsorption onto the sodium sulfate if the

extract is allowed to remain in contact with the sodium sulfate for longer than 2 h.)

6.14 Quantitatively transfer the dried extract

to a K-D apparatus, add a boiling chip, and attach a

Snyder column.

6.15 Concentrate to about 5 mL by heating

the apparatus on a 90'C water bath in a fume hood.

6.16 Allow the K-D flask to cool. Dry the

apparatus with a towel, especially around the

ground-glass joint .

6.17 Separate the Snyder column from the KD flask and rinse the walls of the K-D flask with

approximately 2 mL methylene chloride. Dry the

joints with a towel. Separate the receiver from the

K-D flask and rinse the ground-glass joint of the KD flask into the receiver with methylene chloride.

6.18 Reduce the volume of the extract in the

receiver to less than 0.9 mL on the evaporative concentrator . During the concentration procedure, rinse

the receiver walls two or three times with small

portions of methylene chloride .

6.19 Stopper the receiver with a ground-glass

stopper and store the extract in a freezer until analysis can proceed .

NOTE: The extract obtained from the

base/neutral extraction (method 0-3118) and the

extract from this acidic extraction can be combined

for GC/MS analysis immediately before injection .

73

6 .20 Immediately before analysis, add 0.05

mL of internal standard . Adjust the final volume of

the sample extract in the receiver to 1.0 mL.

6.21 Mass spectrometer tuning:

6.21.1 Use perfluorotributylamine to tune

the mass spectrometer in a manner that results in a

satisfactory calibration of mass assignments as well

as agreement with the criteria listed in step 6.21.2 .

6.21.2 Set the MS to scan the mass range

40-450 amu (atomic mass units) . Temperature program the GC from 45' to 275° C at 20 ° C per min

with an initial hold of 1 .5 min. Inject 50 ng (1 gL of

solution, step 5.2) . Obtain a background corrected

mass spectrum of DFTPP and verify that all of the

following criteria are met:

Mass Ionabundance criteria

51-------------------------- 30-60 percent of mass 198

68------------------------------ <2percent ofmass 69

70------------------------------<2 percent of mass 69

127-------------------------- 40-60 percent ofmass 198

197-----------------------------<1 percent of mass 198

198 -------------- Base peak, 100 percent relative abundance

199---------------------------- 5-9 percent ofmass 198

275 -------------------------- 10-30 percent of mass 198

365----------------------------->1 percent of mass 198

441-------------------------------------- <mass443

422 ---------------------------->40 percent of mass 198

443 -------------------------- 17-23 percent of mass 442

6.22 Analyze the extract by injection of an

aliquot into the GC/MS system optimized as follows :

6.22.1 For systems configured with option

A, analyze the mass range 40 to 450 amu at a scan

rate sufficient to obtain a minimum of 5 scans per

chromatographic peak. Record the total ion current

chromatogram and the mass spectrum of each peak.

6.22.2 For systems configured with option

B, analyze the mass range 40 to 450 amu at a scan

rate sufficient to obtain a minimum of 5 scans per

chromatographic peak. Record the total ion current

chromatogram and the mass spectrum of each peak

as well as the data (retention time and integrated

area of each peak) from the FID.

6.23 Process the data to determine the identity ofthe extractable acidic organic compounds in the

following manner:

6 .23 .1 Identification of the target compound is accomplished by a computerized reverse

search procedure employing a 25-scan retention

time window.
74

6.23.2 Identification of the extractable acid

compounds that are not target compounds is accomplished by a computerized library search versus the

National Bureau of Standards library reference

spectra on each peak. The best computer matches of

mass spectra are reviewed manually .

6.24 Determine the largest characteristic ion

and quantitate the area on this ion for any identified

peak, including the internal standard peak and the

surrogate standards peaks. Alternatively, if the system is configured with option B, the quantitation can

be carried out on the FID trace rather than on the

mass spectrum . The integrated area of an identified

peak is recorded for subsequent calculations. The

better chromatogram (FID or MS) is used for quantitation .

6.25 Confirm the surrogate compounds found

in the sample by injecting an aliquot of the corresponding surrogate standards (step 5.10) into the

gas chromatoggamh and analyze according to steps

6.22 through 6.24. Record the integrated area obtained .

6.26 Confirm any identified extractable acid

compounds found in the sample by injecting an aliquot of the corresponding acidic organic compound

standard (step 5.1) of about the same concentration

into the gas chromatoggamh and analyze according

to steps 6.22 through 6.24. If the concentration of

the compound of interest exceeds the highest standard, dilute the extract and reanalyze it . Record the

integrated area obtained. If an acid standard is not

available, quantitate relative to the internal standard

(see step 7.6) .

7. Calculations

7.1 Calculations of response factors and relative response factors:

7.1 .1 Compute the response factor of the

surrogate standard (step 6.25) and each compound

in the acid standard (step 6.26) using the following

equation:

.452 RF=--~

~c52

where

RF2 = response factor of compound i in standard,

in area/ng

Cs2 = amount of compound i injected, in ng/gL,

and,

A3z = area of compound i peak.

'TECHNIQUES OFWATER- RESOURCES INVESTIGATIONS

7 .1 .2 Calculate the response factor of tb

internal standard using the following equation :

RFI

RRF =

where

RFI=response factor of internal standard, in

area/ng,

CI, = amount of internal standard in acid standard (step 5.1) injected, in ng, and

AI, = area of internal standard peak.

7.1 .3 Calculate a relative response factor

by the following equation:

RF

RFI

where

RRFi = relative response factor,

RF = response factor of compound i, and

RFI = response factor of internal standard .

7.2 Calculations of recoveries of surrogates:

Calculate the percent recovery of each surrogate

standard recovered from the original water

suspended-sediment mixture using the following

equation:

AxCI2 xV3 x100 Percent recovery =

RRFi xAI2 x CS x V4

where

A = area of identified surrogate peak in sample extract,

Cl2 = amount of internal standard injected, in

ng,

RRFi = relative response factor of surrogate i,

A12 = area of internal standard peak in sample

extract,

Cs = concentration of standard, in ng/mL,

V3 =final volume of extract, in mL, and

V4 =volume of sample injected, in pL.

7.3 Calculations of concentrations of analytes : Calculate the concentration of each identified

acid extractable compound in the original water or

water-suspended-sediment mixture using the following equation:

AxC72 x V3x 1,000

Concentration (,pg/L) =

RRFxAI2 x Wx V4

where

A = area of identified peak in sample,

CI2 = amount of internal standard injected, in

ng,
DETERMINATION OF ORGANIC SUBSTANCES IN WATER AND FLUVIAL SEDIMENTS

RRF= relative response factor,

AI2 = area of internal standard peak in sample

extract,

W=weight of sample extracted, expressed in

mL (1 .000g = 1 .000 mL),

V3 = final volume of extract, in mL, and

V4 = volume of sample injected.

7.4 Calculations of concentrations of other

compounds: The concentrations of all other identified extractable acid compounds in the original

water sample, for which there are no standards, are

calculated relative to the concentration of the internal standard and are semiquantitative for the purpose of a general organic scan. The response factor

of the compound is assumed to be exactly equal to

that of the internal standard. Calculate the concentration using the following equation:

A x V3 x 1,000

Concentration (leg/L) =

RFlx Wx V4

where

RFI= response factor of internal standard, in

area/ng,

A = area of identified peak in sample,

W=weight of sample extracted, expressed in

mL (1 .000 g = 1 .000 mL),

V3 = final volume of extract, in mL, and

V4 = volume of sample injected .

8. Report

8.1 Report concentrations of extractable acidic organic compounds (except 4-chloro-3-methyl

phenol, 2,4,6-trichlorophenol, 2,4-dinitrophenol, 4

nitrophenol, 4,6-dinitro-2-methylphenol, and pentachlorophenol) in water and water-suspended-sediment mixtures as follows : less than 6.0 Fig/L, as

"less than 6 .0 gg/L"; 6.0 gg/L and above, two significant figures.

8.2 Report concentrations of 4-chloro-3-

methylphenol, 4-nitrophenol, 4, 6-dinitro-2-methylphenol, and pentachlorophenol as follows : less than

30 gg/L, as "less than 30 ltg/ L"; 30 Igg/L and

above, two significant figures.

8 .3 Report concentrations of 2,4,6-trichlorophenol and 2,4-dinitrophenol as follows: less

than 20 gg/L, as "less than 20 gg/L"; 20 gg/L and

above, two significant figures.

9. Precision

9.1 Surrogate recoveries must be between 30

percent and 130 percent unless a matrix effect can

be demonstrated . Water and water-suspended-sedi75

ment samples were spiked with surrogate standards

and recoveries were determined by two operators in

a single laboratory over an 8-mo period . Results are

as follows:

9.2 Deionized water samples were spiked

with acid extractable compounds and recoveries

were determined by two operators in a single laboratory over a 1-yr period . Results are as follows:

Relative

Number of Average standard

samples recovery deviation

Compound! analyzed (percent) (percent)

Selected references

Glaser, J.A ., Foerst, D.L., McKee, G.D., Quave, S.A., and Budde,

W.L ., 1981, Trace analysis for wastewaters : Environmental

Science and Technology, v . 15, p . 1426-1435.

Realini, P.A ., 1981, Determination of priority pollutant phenols in

water by HPLC: Journal of Chromatography Science, v. 19,

p. 124-129.

Sauter, A.D ., Betowski, L.D ., Smith, T.R ., Strickler, V.A .,

Beimer, R.G ., Colby, B.M., and Wilkison, J .E ., 1981, Fused

silica capillary column GC/MS for the analysis of priority

pollutants : Journal of High Resolution Chromatography and

Chemical Communication, v. 4, p . 366-384.

U.S . Environmental Protection Agency, 1979, Base/neutrals,

acids, and pesticides-Method 625: Federal Register v . 44,

no . 233, p. 69540.

4-Chloro-3-methyphenol ------------- 18 80 27

2-Chlorophenol--------------------- 20 73 25

2,4-Dichlorophenol------------------ 18 84 21

2,4-Dimethylphenol ----------------- 19 74 23

2,4-Dinitrophenol------------------- 15 67 26

2-Methyl-4,6-dinitrophenol ------------ 5 66 33

2-Nitrophenol---------------------- 17 78 32

4-Nitrophenol---------------------- 17 61 44

Pentachlorophenol ------------------ 15 77 31

Phenol---------------------------19 53 44

2,4,6-Trichlorophenol---------------- 17 83 31

Ionused

for

Compound

quantitation

Concentration

range

(ng/pL)

Number

ofsamples analyzedAverage

recovery

(percent)

Relative

standard

deviation

(percent)

PhenoW6 ------------- 99 42-81 45 50 30

2,4-Dibromophenol ------ 252 57-104 46 68 39

2,4,6-Tribromophenol ---- 332 61-75 44 83 34

Ion usedfor

Compound quantitation

Concentration

range (ng/pL)

4-Chloro-3-methyphenol -------------- 142 50-292

2-Chlorophenol---------------------- 128 50-355

2,4-Dichlorophenol------------------- 162 50-316

2,4-Dimethylphenol ------------------ 122 43-319

2,4-Dinitrophenol-------------------- 184 58-436

2-Methyl-4,6-dinitrophenol ------------ 198 50-571

2-Nitrophenol----------------------- 139 50-271

4-Nitrophenol----------------------- 139 50-326

Pentachlorophenol------------------- 266 144-349

Phenol---------------------------- 94 50-331

2,4,6-Trichlorophenol----------------- 196 50-236
76

Base/neutral extractable compounds, total

recoverable, gas chromatographic/mass

spectrometric (0-3118-83)

Parameter Code

1 . Application

This method is suitable for the determination of

methylene chloride extractable base/neutral compounds in water and water-suspended-sediment

mixtures containing at least 5 gg/L of the analyte .

2. Summary of method

Organic base/neutral compounds are extracted

from water and water-suspended-sediment mixtures

TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

with methylene chloride. The extract is concentrate

and analyzed by gas chromatography (GC) using a

flame-ionization detector (FID) or a mass spectrometric (MS) detector.

5. Reagents

5.1 Base/neutral standards, EPA analytical

reference grade or highest purity available: Weigh

20 mg of the compound to three significant figures,

quantitatively transfer to a 100-mL volumetric flask,

and dilute to volume with methylene chloride. Prepare dilutions in methylene chloride to obtain solutions containing 2, 10, and 20 ng/gL.

Acenaphthene-------------------------------- 34205

Acenaphthylene ------------------------------34200 3. Interferences

Anthracene---------------------------------- 34220

Benzidine -----------------------------------39120 Any compound having chemical and physical

Benzo(a)anthracene ---------------------------34526 properties similar to an analyte of interest may inBenzo(b)fluoranthene-------------------------- 34230 terfere.

Benzo(k)fluoranthene-------------------------- 34242

Benzo(g,hJ)perylene --------------------------- 34521 4. Apparatus

Benzo(a)pyrene------------------------------- 34247

4-Bromophenylphenyl ether--------------------- 34636 4.1 Boiling chips, micro, carbon chips: Rinse

Butyl benzyl phthalate -------------------------34292 with hexane, air dry, and heat at 300 ° C overnight.

bis(2-Chloroethoxy)rnethane--------------------- 34278 4.2 Concentrator, Kuderna-Danish (K-D),

bis(2-Chlororethyl) ether -----------------------34273 500 mL, all glass, with ground-glass joints, a 10.0- bis(2-Chloroisopropyl) ether ---------------------34283 mL receiver, and a one-ball Snyder column . 2-Chloronaphthalene --------------------------34581

4-Chlorophenylphenyl ether--------------------- 34641 4 .3 Evaporative concentrator, Organomation

Chrysene------------------------------------ 34320 N-Evap, or equivalent: Water bath must be mainDibenz(a,h)anthracene -------------------------34556 tained at 50' to 55 ° C.

1,2-Dichlorobenzene--------------------------- 34536 4.4 Gas chromatograph/mass spectrome1,3-Dichlorobenzene ---------------------------34566 ter/data system, (GC/MS) Hewlett-Packard 5985 B

1,4-Dichlorobenzene--------------------------- 34571

3,3'-Dichlorobenzidine -------------------------34631 GUMS, or equivalent: The gas chromatograph is

Diethyl phthalate -----------------------------34336 used with one of the following options:

Dimethyl phthalate ----------------------------34341 4.4 .1 Columns, fused silica capillary colDi-n-butyl phthalate ---------------------------39110 umn, 25 m x 0.20 mm id (inside diameter), SE-54 2,4-Dinitrotoluene----------------------------- 34611 bonded column, 0.33-gm film thickness .

2,6-Dinitrotoluene----------------------------- 34626

Di-n-octyl phthalate ---------------------------34596 Detector, mass spectrometer (MS).

bis(2-Ethylhexyl) phthalate ---------------------39100 Injection temperature, 260'C.

Fluoranthene --------------------------------34376 Carrier gas, 1mL/min, helium.

Fluorene ------------------------------------34381 Transfer line temperature, 285°C.

Hexachlorobenzene---------------------------- 39700 Mode, splitless injection.

Hexachlorobutadiene --------------------------39702

Hexachlorocyclopentadiene---------------------- 34386 Program rate, 45' to 300°C, 2.5-min iniHexachloroethane----------------------------- 34396 tial hold, 6 ° C/min, 15-min final hold.

Indeno(1,2,3-cd)pyrene------------------------- 34403 4.4 .2 Columns, two fused silica capillary

Isophorone ----------------------------------34408 columns, 25 mx0.20 mm id, SE-54 bonded column, Naphthalene---------------------------------34696 0.33-gm film thickness .

Nitrobenzene --------------------------------34447

N-Nitrosodimethylamine------------------------ 34438 Detectors, mass spectrometer, FID 285'C.

N-Nitrosodiphenylamine ------------------------ 34483 Injection temperature, 260° C.

N-Nitrosodi-n-propylamine---------------------- 34428 Mode, splitless injection .

Phenanthrene-------------------------------- 34461 Carrier gas, 1 mUmin, helium.

Pyrene -------------------------------------34469 Transfer line, 285'C.

2,3,7,8-Tetrachlorodibenzo-p-dioxin ---------------34675 Program rate, 45° to 300°C, 2.5-min ini- 1,2,4-Trichlorobenzene -------------------------34551

tial hold, 6°C/min, 15-min final hold.
DETERMINATION OF ORGANIC SUBSTANCES IN WATERANDFLUVIAL SEDIMENTS

5.2 DFTPP (Decafluorotriphenylphosphine)

solution, 50 ng/gL: Dilute 20 gL DFTPP (Supelco,

or equivalent, 25-mg/mL solution) to 10 mL in a

volumetric flask with methylene chloride .

5.3 Internal standards, perdeuteronaphthalene (naphthalene-ds), Aldrich Chemical Co., or

equivalent; perdeuterophenanthrene (phenanthrene

dio), Kor Isotopes, Division of Kor, Inc., or equivalent; and perdeuterochrysene (chrysene-d 12 ), Kor

Isotopes, Division of Kor, Inc., or equivalent. Weigh

about 20 mg of perdeuteronaphthalene, perdeuterophenanthrene, and perdeuterochrysene to three

significant figures, quantitatively transfer to a 100-

mL volumetric flask, and dilute to volume with

methylene chloride.

5.4 Sodium chloride, reagent grade: Heat at

300'C overnight and store in a closed glass container.

5.5 Sodium hydroxide, 37 percent (weight/

volume) : Dissolve 185 g sodium hydroxide pellets in

500 mL organic-free water and reflux 8 hr. Cool and

store at VCC.

5.6 Sodium sulfate, granular, anhydrous, reagent grade: Heat at 300°C overnight and store in a

glass-stoppered Erlenmeyer flask at 130'C.

5.7 Solvents, hexane, isopropanol, and methylene chloride, glass distilled, pesticide analysis

quality, Burdick and Jackson, or equivalent.

5.8 Surrogate standards, 1-fluoronaphthalene, Aldrich Chemical Co., or equivalent; 2,2'-

difluorobiphenyl, Pfaltz and Bauer Co., or equiva

lent; and p-dibromobenzene, FDA, or equivalent.

Weigh 4 mg of each of the three compounds to three

significant figures, quantitatively transfer to a 100-

mL volumetric flask, and dilute to volume with isopropanol .

5 .9 Water, organic-free .

6. Procedure

All glassware must be washed in a warm detergent solution, rinsed with organic-free water, and

heated at 300°C overnight. Immediately before use,

it must be rinsed with methylene chloride . Stopcock

grease should not be used on ground-glass joints .

6.1 Immediately upon receipt of the sample,

store at 4'C. Extraction must begin within 48 h

following receipt of the sample.

6.2 A blank must accompany each group of

samples. For each sample and blank, rinse a 2-L

separatory funnel and a 500-mL Erlenmeyer flask

with methylene chloride .

77

6.3 Weigh the capped sample bottle to three

significant figures and record the weight for subsequent calculations .

6.4 Adjust the sample to pH 11, as indicated

by pH paper, by the addition of sodium hydroxide

solution.

6.5 Pour the sample into a separatory funnel

containing 100 g sodium chloride. Allow the sample

bottle to drain into the separatory funnel for several

minutes . Stopper and shake until the salt is dissolved .

6 .6 Weigh the empty, capped sample bottle to

three significant figures, calculate and record the

net sample weight.

6 .7 Add 1 mL of the surrogate standard (step

5.8) to the sample in the separatory funnel .

6 .8 Add 100 mL methylene chloride to the

empty sample bottle and swirl to wash the sides of

the container with the solvent. The Teflon liner is

not rinsed because of the potential of contamination

from solvent that has contacted the cap threads and

the surface beneath the liner. Pour the contents of

the bottle into the separatory funnel .

6 .9 Stopper the separatory funnel and shake

for at least 1 min, venting often to relieve pressure.

6 .10 Drain the organic layer into a 500-mL

Erlenmeyer flask containing approximately 30 g sodium sulfate .

6 .11 Extract the sample two more times with

50 mL methylene chloride by repeating steps 6.8

through 6.10, collecting the two organic extracts in

the Erlenmeyer flask containing the sodium sulfate.

NOTE: The aqueous phase may be retained

and extracted for the acidic extractable organic

compounds after adjusting the pH to 2 (method

0-3117) .

6.12 Cover the Erlenmeyer flask with aluminum foil and allow to stand at room temperature for

approximately 4 h.

6 .13 Quantitatively transfer the dried extract

into a K-D apparatus, add a boiling chip, and attach

a Snyder column.

6.14 Concentrate the extract to about 5 mL

by heating the apparatus on a 80'C water bath in a

fume hood.

6.15 Allow the K-D apparatus to cool. Dry the

apparatus with a towel, especially around the

ground-glass joint of the receiver.

6.16 Separate the Snyder column from the KD flask and rinse the walls of the K-D flask with

approximately 2 mL methylene chloride. Separate

the receiver from the K-D flask and rinse the
78

ground-glass joint of the K-I) flask into the receiver

with methylene chloride .

6 .17 Reduce the volume of the methylene

chloride in the receiver to less than 0.9 mL on the

evaporative concentrator. During the concentration,

rinse the receiver walls two or three times with

small portions of methylene chloride.

6.18 Stopper the receiver with a ground-glass

stopper and store the extract at 4'C until analysis

can proceed .

NOTE: The extract can be combined with the

extract from the acidic extractable organic compounds (method 0-3117) immediately before injection into the GC/MS.

6.19 Immediately before analysis, add 0.05

mL of internal standard . Adjust the final volume of

the sample extract in the receiver to 1 .0 mL.

6.20 Mass spectrometer tuning:

6.20.1 Use perfluorotributylamine to tune

the mass spectrometer in a manner that results in

satisfactory calibration of mass assignments as well

as agreement with the criteria listed in step 6.20.2 .

6.20 .2 Set the MS to scan the mass range

40-450 amu (atomic mass units) . Temperature program the GC from 45' to 275'C at 20'C/min with

an initial hold of 1 .5 min . Inject 50 ng (1 gL of

solution, step 5 .2) . Obtain a background corrected

mass spectrum of DFTPP and verify that all of the

following criteria are met:

Mass Ionabundance criteria

51 ------------------------- 30-60 percent of mass 198

68 ------------------------------<2 percent of mass 69

70 ------------------------------<2 percent of mass 69

127 ------------------------- 40-60 percent of mass 198

197 -----------------------------<1 percent of mass 198

198 ------------- Base peak, 100 percent relative abundance

199 --------------------------- 5-9 percent of mass 198

275 ------------------------- 10-30 percent of mass 198

365---------------------------->1 percent of mass 198

441 ------------------------------------- <mass 443

442 --------------------------- >40 percent of mass 198

443 ------------------------- 17-23 percent of mass 442

6 .21 Analyze the extract by injection of an

aliquot into the GC/MS system optimized as follows :

6.21 .1 For systems configured with option

A, analyze the mass range 40 to 450 amu at a scan

rate sufficient to obtain a minimum of 5 scans per

chromatographic peak. Record the total ion current

chromatogram and the mass spectrum of each peak.

6.21 .2 For systems configured with option

B, analyze the mass range 40 to 450 amu at a scan

TECHNIQUES OF WATER - RESOURCES INVESTIGATIONS

rate sufficient to obtain a minimum of 5 scans per

chromatographic peak. Record the total ion current

chromatogram and the mass spectrum of each peak

as well as the retention time and integrated area of

each peak from the FID.

6.22 Process the data to determine the identity of the extractable base/neutral compounds including the priority pollutants in the following manner :

6.22.1 Identification of the target compounds is accomplished by a computerized reverse

search procedure employing a 25-scan retention

time window.

6 .22 .2 Identification of the extractable

base/neutral compounds that are not target compounds is accomplished by a computerized library

search versus the National Bureau of Standards library reference spectra on each peak.

6.23 Determine the largest characteristic ion

and quantitate the area on this ion for any identified

peak, including the internal standard peak and the

surrogate standards peaks. Alternatively, if the system is configured with option B, the quantitation can

be carried out on the FID response rather than on

the mass spectrometer response . The integrated

area of an identified peak from the FID is recorded

for subsequent calculations . The better chro

matogram (FID or MS) is used for quantitation .

6.24 Confirm the surrogate compounds found

in the sample by injecting an aliquot of the corresponding surrogate standards (step 5 .8) into the gas

chromatograph and analyze according to steps 6.21

through 6.23. Record the integrated area obtained.

6 .25 Confirm any identified extractable

base/neutral compounds found in the sample by injecting an aliquot of the corresponding base/neutral

standard into the gas chromatograph and analyze

according to steps 6.21 through 6 .23 . Record the

integrated area obtained . If a base/neutral standard

is not available, quantitate relative to the internal

standard (see step '7 .6) .

7. Calculations

7 .1 Calculations of response factors and relative response factors:

7.1 .1 Calculate the response factor for each

compound in the base/neutral standard or surrogate

standard (step 6.23) using the following equation :

Asi

RF, =

Csi
DETERMINATION OF ORGANIC SUBSTANCES IN WATER ANDFLUVIAL SEDIMENTS 79

where AxCl2x Vax 1,000

RFi = response factor of compound i in standard, Concentration (pg/L) =RRFixAI2x Wx V4

in area/ng,

Csi = amount of compound i injected, in ng/gL, where

and A = area of identified peak in sample extract,

Asi = area of compound i peak in the standard.

CI2 = amount of internal standard injected, in

7.1 .2 Calculate the response factor of the ng,

internal standard using the following equation: RRFi = relative response factor of compound i,

AI2 = area of internal standard peak in sample

RFI =

AI, extract,

Cl, W= weight of sample extracted, expressed in

mL (1 .000 g = 1 .000 mL),

where Vs = final volume of extract, in mL, and

RF1= response factor of internal standard, in V4 = volume injected, in gL.

area/ng, 7..4 Calculations of concentrations of other

CI, = amount of internal standard in base/neu- compounds: The concentrations of all other identitral standard, in ng injected, and fied extractable base/neutral compounds in the origAI, = area of internal standard peak.

inal water sample for which there are no standards,

7.1 .3 Calculate a relative response factor are calculated relative to the concentration of the

by the following equation:

internal standard, and are semiquantitative for the

RFi purposes of a general organic scan. The response

RRFi = RFI factor of the compound is assumed to be exactly

equal to that of the internal standard. Calculate the

where concentration using the following equation:

RRFi = relative response factor, compound i,

RFi = response factor of compound i , and

RFI = response factor of internal standard.

7 .2 Calculations of recoveries of surrogates:

Calculate the percent recovery of each surrogate

standard recovered from the original Water- sus

pended-sediment mixture using the following equation :

Percent recovery =

A xCI2 x Vs x 100

RRFi xAI2 x CS x V4

where

A = area of identified surrogate peak in sample extract,

CI2 = amount of internal standard injected, in

ng,

RRF; = relative response factor of compound i,

AI2 = area of internal standard peak in sample

extract,

CS = concentration of standard, in ng/~tL,

Vs = final volume of extract, in mL, and

V4 = volume injected, in gL.

7.3 Calculation of concentrations of analytes :

Calculate the concentration of each identified extractable base/neutral priority pollutant in the origi

nal water-suspended-sediment mixture using the

following equation :

8. Report

Concentration (lag/L) =

A x Va x 1,000

RFIxWxV4

where

RFI= response factor of internal standard, in

area/ng,

A = area of identified peak in sample,

W=weight of sample extracted, expressed in

mL (1 .000 g = 1 .000 mL),

Va = final volume of extract, in mL, and

V4 = volume injected, in gL.

8.1 Report concentrations of extractable

base/neutral organic compounds (except chrysene,

benzo(a) anthracene, dioctyl phthalate, benzo(b)

fluoranthene, benzo(k) fluoranthene, benzo(a) pyrene, indeno(1,2,3-cd) pyrene, dibenz(a,h) anthracene, and benzo(g,h,i) perylene) in water and

water-suspended-sediment mixtures as follows: less

than 5.0 ~tg/L, as "less than 5.0 pg/L" ; 5.0 gg/L

and above, two significant figures.

8.2 Report concentrations of chrysene,

benzo(a) anthracene, dioctyl phthalate, benzo(b)

fluoranthene, benzo(k) fluoranthene, benzo(a) py

rene, indeno(1,2,3-cd) pyrene, dibenz(a,h) anthracene, and benzo(g,h,i) perylene as follows: less than
80 TECHNIQUES OF WATER- RESOURCES INVESTIGATIONS

10 gg/L, as "less than 10 wg/L"; l0gg/L and above,

two significant figures.

9. Precision

9.1 Surrogate recoveries must be from 40

percent to 130 percent unless a matrix effect can be

demonstrated . Water and water-suspended-sedi

ment samples were spiked with surrogate standards

and recoveries were determined by two operators in

a single laboratory over an 8-mo period . Results are

as follows:

9.2 Deionized water samples were spiked

with base/neutral extractable compounds and recoveries were determined by two operators in a single

laboratory over a 1-yr period. Results are as follows:

Relative

stanIon used Concen- Number dard

for trattion ofsam- Average device

quanti- range ples an- recovery tion

Compound tation (ng/pL) alyzed (percent) (percent)

Selected references

Glaser, J.A., Foerst, D.L ., McKee, G.D ., Quave, S.A ., and Budde,

W.L., 1981 Trace analysis for wastewaters: Environmental

Science and Technology, v. 15, p. 1426-1435.

Sauter, A.D ., Betowski, L.D ., Smith, T.R ., Strickler, V.A .,

Beimer, R.G ., Colby, B.M ., and Wilkison, J.E ., 1981, Fuse

column GC/MS for the analysis of priority

rnal of High Resolution Chromatography and

munications, v. 4, p. 366-384.

Protection Agency, 1979, Base/neutral acids,

-Method 625: Federal Register, v. 44, no .

~U .S . G .P .O . 1987 - 181-407 :40093

Compound

Ion used

for

quantitation

Concentrattion

range

(ng/gL)

Number

ofsamples analyzedAverage

recovery

(percent)

Relative

standard

device

tion

(percent)

4-Chlorophenyl phenyl ether 204 50 1 111 ---

Chrysene--------------- 228 50-113 6 42 46

Dibenz(a,h)anthracene ---- 278 69 1 71 ---

Di-n-butyl phthalate------ 149 100 5 53 19

1,2-Dichlorobenzene------ 146 50-100 5 56 43

1,3-Dichlorobenzene------ 146 100-201 6 97 30

1,4-Dicholorobenzene----- 146 102 1 51 ---

Diethylphthalate --------- 149 50-100 14 69 37

Dimethyl phthalate - - - - -- - 163 199 1 19 ---

2,4-Dinitrotoluene ------- 165 50-106 6 63 19

Dimethyl phthalate ------ 149 100 15 69 42

bis(2-ethylhexyl) phthalate - 149 100 2 42 ---

Fluorene -------------- 166 50-197 5 99 11

Fluoranthene----------- 202 224 1 98 ---

Hexachlorobenzene ------- 284 50-76 3 91 ---

Hexachlorobutadine------ 225 50-74 3 94 ---

Indeno(1,2,3-cd)pyrene --- 276 50 1 104 ---

Naphthalene ----------- 128 50-130 6 81 17

Nitrobenzene----------- 77 219 1 50 ---

N-Nitrosodimethylamine -- 74 50 1 68 ---

N-Nitrosodiphenylamine--- 169 50 1 48 ---

Phenanthrene ---------- 178 218 1 94 ---

Pyrene---------------- 202 50-334 16 94 16

1,2,4-Trichlorobenzene---- 180 100 5 78 24

Acenaphthylene--------- 152 190 2 105 --- silica capillary Anthracene ------------ 178 100-177 5 66 45 pollutants : Jo Benzo(a)anthracene------ 228 69 5 79 23

Benzo(k)fluoranthene----- 252 50-71 2 98 ---

Chemical Co

Benzo(a)pyrene --------- 252 50--100 5 115 22 U.S . Environmental

bis(2-chloroethoxy)methane 93 50--100 3 64 --- and pesticide

4-Bromophenyl phenyl ether 248 100 5 43 6 233, p. 69,540 .

Relative

stanIon used Concen- Number dard

for tration of sam- Average deviaquanti- range ples am recovery tion

Compound tation (ng/pL) alyzed (percent) (percent)

1-Fluoronaphthalene ----- 146 48--104 45 65 29

p-Dibromobenzene------- 236 51--107 46 67 26

2,2'-Difluorobiphenyl ----- 190 43-79 46 68 20