Work Plan for Determining the Occurrence of 

Glyphosate, Its Transformation Product AMPA, 

Other Herbicide Compounds, and Antibiotics 

in Midwestern United States Streams, 2002

By W.A. Battaglin, E.M. Thurman, D.W. Kolpin, E.A. Scribner, 

M.W. Sandstrom, and K.M. Kuivila

Denver, Colorado

2003

U.S. GEOLOGICAL SURVEY

Open-File Report 03-69

Prepared in cooperation with the

U.S. ENVIRONMENTAL PROTECTION AGENCY

OFFICE OF PESTICIDE PROGRAMS
District Chief

U.S. Geological Survey

Box 25046, Mail Stop 415

Denver Federal Center

Denver, CO 80225–0046

For additional information write to:

District Chief

U.S. Geological Survey

Box 25046, Mail Stop 415

Denver Federal Center

Denver, CO 80225–0046

U.S. DEPARTMENT OF THE INTERIOR

GALE A. NORTON, Secretary

U.S. GEOLOGICAL SURVEY

Charles G. Groat, Director

The use of firm, trade, and brand names in this report is for identification purposes only and does 

not constitute endorsement by the U.S. Geological Survey.

Copies of this report can be purchased 

from:

U.S. Geological Survey

Information Services

Box 25286

Denver Federal Center

Denver, CO 80225
CONTENTS III

CONTENTS

Abstract.................................................................................................................................................................................. 1

Introduction............................................................................................................................................................................ 1

Glyphosate Toxicity..................................................................................................................................................... 2

Changes in Herbicide Use ........................................................................................................................................... 3

Herbicide Transformation Products............................................................................................................................. 4

Antibiotics in Streams.................................................................................................................................................. 4

Work Plan .............................................................................................................................................................................. 4

Objectives and Hypotheses.......................................................................................................................................... 4

Sampling Sites ............................................................................................................................................................. 9

Sampling Schedule ...................................................................................................................................................... 9

Sampling Procedures ................................................................................................................................................... 11

Analytical Procedures.................................................................................................................................................. 11

Quality Assurance........................................................................................................................................................ 11

Data Analysis Methods and Reports...................................................................................................................................... 12

References Cited.................................................................................................................................................................... 12

Appendix................................................................................................................................................................................ 15

FIGURES

1. Graph showing herbicide use estimates in millions of pounds of active ingredient applied .................................... 3

2. Map showing locations of sampling sites and associated contributing drainage areas............................................. 10

TABLES

1. Toxicity of technical-grade glyphosate to aquatic organisms ................................................................................. 2

2. Midwestern streams to be sampled in 2002, and streamflow conditions for samples collected in 1998................ 5

3. Physical properties and chemical compounds to be determined on samples collected in 2002 ............................. 6

4. Schedule for quality-assurance samples.................................................................................................................. 12

CONVERSION FACTORS AND ABBREVIATIONS

Multiply By To obtain

cubic foot per second (ft3/s) 0.02832 cubic meter per second

mile (mi) 1.609 kilometer

pound (lb) 453.6 grams

pound per acre (lb/acre) 1.121 kilogram per hectare

square mile (mi2) 2.590 square kilometer

gallon (G) 3.785 liter (L)

milligrams per liter (mg/L) 1,000 micrograms per liter (μg/L)

CONTENTS
IV CONTENTS

Abbreviated Water-Quality Units

cubic meters per second (m3/s)

micrograms per liter (μg/L)

milligrams per liter (mg/L)

milliliter (mL)

millimeter (mm)

microsiemens per centimeter at 25 degrees Celsius (μS/cm)

Miscellaneous Abbreviations and Acronyms

AMPA – amino methyl phosphonic acid

DEA – deethylatrazine

DDA – didealkylatrazine

DIA – deisopropylatrazine

GCS – GCMS analysis method for herbicides and herbicide transformation products

LCGY – LCMS analysis method for glyphosate, glufosinate, and AMPA

LC50 – contaminant concentration at which 50 percent of test organisms die 

LCAA – LCMS analysis method for herbicide transformation products

LCAN – LCMS analysis method for antibiotics

LCEA – LCMS analysis method for herbicides and herbicide transformation products

LCMS – Liquid chromatography mass spectrometry

LOEC – lowest observed effect concentration

NASQAN – National Stream Quality Accounting Network

NAWQA – National Water-Quality Assessment

NOEC – no observed effect concentration
Abstract 1

Work Plan for Determining the Occurrence of 

Glyphosate, Its Transformation Product AMPA, 

Other Herbicide Compounds, and Antibiotics in 

Midwestern United States Streams, 2002

By W.A. Battaglin, E.M. Thurman, D.W. Kolpin, E.A. Scribner, M.W. Sandstrom, and

K.M. Kuivila

Abstract

Changes in herbicide use in the Midwestern 

United States have been substantial over the last 

5 years. Most significant is a tripling in the use of 

glyphosate (N-[phosphonomethyl]glycin). Over 

this same time period (1997–2001), atrazine use 

increased by 20 percent and acetochlor use 

increased by 10 percent, while cyanazine use 

decreased by 99 percent, alachlor use decreased 

by 70 percent, and metolachlor use decreased by 

55 percent. Previous studies have documented 

that herbicide flushes occur in midwestern 

streams during runoff events for several weeks to 

several months following application, and that 

herbicide concentrations in midwestern streams 

during flushing events are related to rates of 

herbicide use. 

The objective of this study is to determine 

the distribution of glyphosate and its primary 

transformation product aminomethylphosphonic 

acid (AMPA) in midwestern streams during postapplication and harvest-season runoff events. 

Water samples will be collected in 2002 during 

two post-herbicide-application runoff events and 

one harvest-season runoff event from 53 sites on 

streams in the Midwestern United States. All 

samples will be analyzed at the U.S. Geological 

Survey Organic Geochemistry Research Laboratory in Lawrence, Kansas, for glyphosate and 

20 other herbicides. Samples will also be 

analyzed for a glyphosate transformation product 

(AMPA) and 26 other herbicide transformation 

products, using GC/MS or HPLC/MS. Selected 

samples will be analyzed for 36 antibiotics or 

antibiotic transformational products. Results from 

this study will represent the first broad-scale 

investigation of glyphosate and AMPA in U.S. 

water resources.

INTRODUCTION

This report describes a plan of study to determine the distribution of glyphosate and its primary 

transformation product aminomethylphosphonic acid 

(AMPA) during runoff events in midwestern streams. 

The study will also determine if recent changes in 

herbicide use are affecting herbicide concentrations, 

and the distribution of didealkylatrazine (DDA), other 

herbicide transformation products, and selected antibiotics in midwestern streams. 

Since 1989, the U.S. Geological Survey (USGS) 

Toxic Substances Hydrology Program has conducted 

reconnaissance studies of streams in the Midwestern 

United States (U.S.) to determine the geographic and 

seasonal distribution of herbicide compounds. Studies 

in 1989 and 1990 documented that large amounts of 

atrazine, cyanazine, alachlor, and metolachlor were 

flushed into streams during post-herbicide-application 

runoff events (Thurman and others, 1991, 1992; 

Goolsby and Battaglin, 1993). Additional studies in 

1990-1992 using automatic samplers (Scribner and 

others, 1994) showed that the peak herbicide concentrations tend to occur during the first runoff event after 

herbicide application. Smaller herbicide concentration 

peaks continue to occur during runoff events for 
2 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern United States Streams, 2002

several weeks to several months following application. 

By late summer, herbicide concentrations are generally low (less than 0.05 μg/L) and tend to remain low 

until the process repeats during the following planting 

season.

Recent studies have focused on collecting water 

samples during these post-application runoff events. 

The same 53 stream sites have been sampled in 1989, 

1990, 1994, 1995, and 1998. Results from these 

studies demonstrate that the concentrations of several 

herbicides in midwestern streams have decreased since 

1989 (Battaglin and Goolsby, 1999; Scribner and 

others, 2000). In some cases, these decreases correspond to declines in herbicide application amounts, 

whereas in other cases, the reason for the decrease 

more likely is changes in application practices. These 

studies also found a widespread occurrence of herbicide transformation products in midwestern streams. 

In the 1998 stream reconnaissance, the median 

concentrations of acetochlor, alachlor, and metolachlor 

were all less than the median concentration of one or 

more of their transformation products. This concentration pattern would be expected later in the summer 

during low-flow conditions but was not expected 

during higher flows in spring and early summer 

(Battaglin and others, 2001). 

Glyphosate (N-[phosphonomethyl]glycin) is a 

broad-spectrum nonselective systemic herbicide that 

was first used in the early 1970’s. Glyphosate is 

among the most widely used herbicides in the world. 

Glyphosate has not been extensively monitored in the 

United States even though it is commonly used for 

agricultural, silvicultural, and nonagricultural weed 

control. In Germany, both glyphosate and its primary 

transformation product (AMPA) have been detected at 

low concentrations in rivers. Glyphosate, however, is 

both highly polar and water-soluble, making analysis 

for it in water samples difficult (Skark and others, 

1998). In 1997 (the last year with available estimates), 

more than 50 million pounds of glyphosate were sold 

for agricultural and nonagricultural weed control 

(Aspelin and Grube, 1999).

Glyphosate Toxicity

Glyphosate is a nonselective herbicide that 

inhibits aromatic amino acid biosynthesis. It is 

considered to be only slightly toxic to birds, fish, and 

aquatic invertebrates and does not bioaccumulate. The 

toxicity of chemicals to nontarget organisms can be 

compared using various endpoints including LC50

values (concentration that kills 50 percent of test 

organisms), NOEC (no observed effect concentration), 

and LOEC (lowest observed effect concentration). The 

48-hour LC50 values for freshwater fish and aquatic 

invertebrates range from 55 to 780 mg/L (table 1). The 

LOEC’s for freshwater green algal growth were less at 

4.00 and 1.55 mg/L, respectively. Several studies 

examined the effect of glyphosate on soil bacteria and 

found no adverse effects on soil microbial biomass or 

on nitrification; but in fact, microbial activity

Table 1. Toxicity of technical-grade glyphosate to aquatic organisms

[LC50, 50 percent lethal concentration; LOEC, lowest observed effect concentration; FETAX, Frog Embryo Teratogenesis Assay; 

NOEC, no observed effect concentration]

Organism Endpoint Value

(mg/L)

Daphnia magna a 48-hour LC50 780

Midge (Chironomus plumosus)

a 48-hour LC50 55

Bluegill sunfish a, b 48-hour LC50 24–150

Rainbow trout a, b 48-hour LC50 85–140

Fathead minnow b 48-hour LC50 97

Freshwater green algae c LOEC on growth 1.55, 4.00

Amphibian d FETAX 5,407 (acid equivalent)

Coho salmon smolt e NOEC osmoregulation and growth 2.78

a

 U.S. Environmental Protection Agency, 1993.

b Folmer and others, 1979.

c

 Saenz and others, 1997.

d

 Perkins and others, 2000.

e

 Mitchell and others, 1987.
INTRODUCTION 3

increased with glyphosate as a carbon source 

(Stratton, 1990; Stratton and Stewart, 1991; Busse and 

others, 2001; Haney and others, 2002). The effect of 

glyphosate on amphibians was evaluated using the 

Frog Embryo Teratogenesis Assay (FETAX), and the 

resulting LC50 was 5,407 mg acid equivalent/L 

(Perkins and others, 2000). Mitchell and others (1987) 

found the NOEC for sublethal effects of glyphosate on 

coho salmon smolt osmoregulation and growth was 

2.78 mg/L. Some of the surfactants in glyphosate 

formulations, particularly the ethylated amines and 

contaminants associated with them, are thought to be 

more toxic than glyphosate.

Changes in Herbicide Use

Several substantial changes in herbicide use 

have occurred since the last stream reconnaissance 

study in 1998. The most significant is the rapid 

increase in the use of glyphosate that corresponded 

with the introduction of “Roundup Ready” crops. 

Between 1997 and 2001 (the newest data available) 

agricultural glyphosate use in nine Midwestern States 

(Illinois, Indiana, Iowa, Kansas, Minnesota, Missouri, 

Nebraska, Ohio, and Wisconsin) more than tripled 

from 10.3 to 36.1 million pounds applied (fig. 1). Over 

this same time period, atrazine use increased from 

43.7 to 52.2 million pounds, acetochlor use increased 

from 26.1 to 28.7 million pounds, and simazine use 

increased from 0.8 to 1.3 million pounds; while cyanazine use decreased from 15.7 to 0.05 million pounds, 

alachlor use decreased from 7.8 to 2.3 million pounds, 

metolachlor use decreased from 46.1 to 20.5 million 

pounds, and metribuzin use decreased from 1.2 to 

0.2 million pounds (U.S. Department of Agriculture, 

2002). 

60

50

40

30

20

10

0

1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001

metolachlor

alachlor

acetochlor

glyphosate

atrazine

metribuzin simazine

cyanazine

YEAR

HERBICIDE USE, IN MILLIONS OF POUNDS

Figure 1. Herbicide use estimates in millions of pounds of active ingredient applied.
4 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern United States Streams, 2002

Assuming that these trends in herbicide use 

continued through 2002, we suspect glyphosate and its 

primary transformation product AMPA will be 

detected in a majority of midwestern streams during 

2002 post-application runoff events. We also suspect 

that when compared with the distribution of concentrations measured in the 53 midwestern streams in 

1998, those measured in 2002 will be about the same 

for atrazine and acetochlor and significantly lower for 

alachlor, cyanazine, and metolachlor.

Herbicide Transformation Products

The half-life of glyphosate in soils typically 

found in the Midwestern United States is approximately 25–40 days. Glyphosate can be transformed 

(degraded) in soil, water, and plants to AMPA. 

Glyphosate loses an acetic acid moiety to form the 

AMPA transformation product. AMPA retains the 

phosphate group and many of the physical properties 

of glyphosate. AMPA is soluble and easily dissolved 

but will precipitate in the presence of iron and calcium 

after formation of insoluble phosphate salts (Royal 

Society of Chemistry, 1998). AMPA is most likely 

more persistent than glyphosate and also should be 

monitored as part of any study of glyphosate.

The 1998 reconnaissance (Battaglin and others, 

2001) and other recent investigations (Thurman and 

others, 1994; Boyd, 2000; Kalkhoff and others, 1998; 

Phillips and others, 1999; Porter and others, 2001) 

have documented the widespread occurrence of herbicide transformation products in water. Still, scientists 

have not analyzed natural water samples for many of 

the herbicide transformation products that are likely to 

occur in the environment. Thurman and others (1994) 

showed that both the deethylatrazine (DEA) and 

deisopropylatrazine (DIA) transformation products of 

atrazine are commonly found in soils, ground water, 

and surface water of the Midwestern United States. 

Both DEA and DIA form by losing either an ethyl or 

isopropyl group—which is a common degradation 

pathway in soil. It is hypothesized that the continued 

transformation of DEA and DIA with the loss of either 

the remaining ethyl or isopropyl group will result in 

didealkylatrazine (DDA). Its occurrence was 

suspected in the past, but methods were not available 

until now to isolate and identify DDA. DDA retains 

the chlorine atom and probably retains some of the 

herbicidal activity and toxicity of the parent atrazine. 

No published data exist for the occurrence of DDA in 

surface or ground water. The authors hypothesize that 

both AMPA and DDA will be detected in most 

midwestern streams.

Antibiotics in Streams

Recent studies have shown antibiotics to be 

present in U.S. streams (Kolpin and others, 2002). A 

number of the antibiotics found are those used in livestock production, but the transport mechanisms of 

antibiotics to streams are not yet understood. Samples 

collected during runoff events in midwestern streams 

will help to identify the dominant source of antibiotics 

in streams. If point sources such as sewage-treatment 

discharges or confined animal feeding operation 

(CAFO) discharges are the dominant source, then 

concentrations should be much lower during runoff 

events than during lower flow conditions. If nonpoint 

sources (such as manure applications to fields) are 

dominant, then concentrations should be about the 

same or higher during runoff events. Because manure 

applications also take place during the post-harvest 

period, fall and early winter storms may be “flushing” 

events for antibiotics.

WORK PLAN

Approximately 180 samples will be collected 

during the 2002 reconnaissance study. Samples will be 

collected from 53 sites (table 2) on small to mediumsized midwestern streams. All 2002 reconnaissance 

samples will be sent to the USGS Organic Geochemistry Research Laboratory (OGRL) in Lawrence, 

Kansas, and analyzed for 21 herbicides, 27 herbicide 

transformation products, and 36 antibiotics (table 3). 

The analytical reporting limits for all compounds are 

at or below known biologically significant levels 

(Battaglin and Fairchild, 2002).

Objectives and Hypotheses

The principal objective of this study is to determine the distribution of glyphosate and its primary 

transformation product AMPA in midwestern streams 

during runoff events. The proposed study also 

provides a plan to determine (1) if recent changes in
WORK PLAN 5

Table 2. Midwestern streams to be sampled in 2002, and streamflow conditions for samples collected in 1998

[mi2, square miles; ft3/s, cubic feet per second]

Map 

no. State ID Station name

Drainage 

area

(mi2)

Sampling dates and flow, in ft3/s

Pre-emergence Post-emergence

1 IA 05411850 Turkey River near Eldorado 640 not sampled in 1998

 2 IA 05421000 Wapsipinicon River at Independence 1,048 5/27 595 6/20 1,300

 3 IA 05455100 Old Mans Creek near Iowa City 201 6/10 209 6/30 840

 4 IA 05472500 N. Skunk River near Sigourney 730 5/21 1,140 6/10 1,810

 5 IA 05474000 Skunk River at Augusta 4,303 5/26 15,600 6/18 14,400

 6 IA 05480500 Des Moines River at Fort Dodge 4,190 5/16 7,880 6/12 5,190

 7 IA 05484500 Raccoon River at Van Meter 3,441 5/17 6,330 6/10 9,160

 8 IA 06606600 Little Sioux River at Correctionville 2,500 5/27 1,400 6/18 2,760

 9 IA 06607200 Maple River at Mapleton 669 5/29 891 6/9 889

10 IA 06609500 Boyer River at Logan 871 5/22 1,070 6/9 2,890

11 IL 03378000 Bonpas Creek at Browns 228 5/23 834 7/7 499

12 IL 03381495 Little Wabash River at Carmi 3,088 5/23 9,340 7/9 4,020

13 IL 05439500 S. Branch Kishwaukee R. at Fairdale 387 5/20 1,620 7/29 61

14 IL 05526000 Iroquois River near Chebanse 2,091 6/10 1,340 7/8 8,490

15 IL 05540500 Dupage River near Shorewood 324 6/9 284 7/29 166

16 IL 05569500 Spoon River at London Mills 1,072 5/20 1,270 7/1 1,780

17 IL 05576500 Sangamon River at Riverton 2,618 5/26 6,800 7/9 4,070

18 IL 05587000 Macoupin Creek near Kane 868 not sampled in 1998

19 IL 05592100 Kaskaskia River near Cowden 1,330 5/20 1,400 7/8 2,700

20 IL 05594000 Shoal Creek near Breese 735 5/22 768 7/8 761

21 IN 03275000 Whitewater River near Alpine 522 5/26 727 7/8 652

22 IN 03302800 Blue River at Fredericksburg 283 5/27 447 7/8 490

23 IN 03328500 Eel River near Logansport 789 5/21 701 6/30 688

24 IN 03333450 Wildcat Creek near Jerome 146 5/21 126 6/30 563

25 IN 03335000 Wildcat Creek near Lafayette 794 5/20 883 7/1 1,130

26 IN 03351000 White River near Nora 1,219 5/28 1,260 7/1 1,780

27 IN 03362500 Sugar Creek near Edinburgh 474 5/26 1,610 7/20 1,490

28 IN 03371500 E. Fork White River near Bedford 3,861 5/28 14,300 7/6 6,050

29 KS 06885500 Black Vermillion River near Frankfort 410 5/5 80 6/9 162

30 KS 06890100 Delaware River near Muscotah 431 5/5 111 6/10 100

31 MN 05317000 Cottonwood River near New Ulm 1,280 5/26 525 7/21 673

32 MN 05476000 Des Moines River at Jackson 1,220 5/26 339 8/24 267

33 MN 05320270 Little Cobb River near Beauford 130 5/18 166 8/17 2.8

34 MO 06817700 Nodaway River near Graham 1,320 6/18 5,030 8/19 282

35 NE 06803000 Salt Creek at Roca 167 5/15 470 6/10 148

36 NE 06804000 Wahoo Creek at Ithaca 271 5/15 118 6/8 495

37 NE 06880800 W. Fork Big Blue River near Dorchester 1,206 5/23 247 6/10 231

38 NE 06815000 Big Nemaha River at Falls City 1,340 5/26 333 6/8 438
6 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern United States Streams, 2002

39 NE 06882000 Big Blue River at Barneston 4,447 5/15 1,110 6/9 1,170

40 NE 06884000 Little Blue River near Fairbury 2,350 5/12 261 6/8 239

41 OH 03157000 Clear Creek near Rockbridge 89 6/10 36 6/30 84

42 OH 03219500 Scioto River near Prospect 567 6/2 128 6/29 1,040

43 OH 03223000 Olentangy River at Claridon 157 6/9 13 6/29 3,770

44 OH 03230500 Big Darby Creek at Darbyville 534 6/3 208 6/29 11,400

45 OH 03234500 Scioto River at Higby 5,131 6/4 2,620 7/8 5,030

46 OH 03240000 Little Miami River near Oldtown 129 6/10 97 6/30 191

47 OH 03267900 Mad River at Eagle City 310 6/10 228 6/30 454

48 OH 04185000 Tiffin River at Stryker 410 6/1 98 7/7 40

49 OH 04186500 Auglaize River near Fort Jennings 332 6/17 670 7/8 751

50 WI 04087240 Root River at Racine 190 6/1 98 8/3 5

51 WI 05340500 St. Croix River at St. Croix Falls 6,240 6/3 4,220 8/5 1,690

52 WI 05407000 Wisconsin River at Muscoda 10,400 6/16 12,900 8/7 8,890

53 WI 05430500 Rock River at Afton 3,340 6/17 2,140 7/21 1,830

Table 2. Midwestern streams to be sampled in 2002, and streamflow conditions for samples collected in 1998—Continued

[mi2, square miles; ft3/s, cubic feet per second]

Map 

no. State ID Station name

Drainage 

area

(mi2)

Sampling dates and flow, in ft3/s

Pre-emergence Post-emergence

Table 3. Physical properties and chemical compounds to be determined on samples 

collected in 2002

[ft3/s, cubic feet per second; μS/cm, microsiemens per centimeter at 25 degrees Celsius; 

°C, degrees Celsius; mg/L, milligrams per liter; μg/L, micrograms per liter]

Property or compound Method Reporting 

limit

Physical properties

streamflow meter or rating 0.01 ft3/s

specific conductance meter 1.0 μS/cm

pH electrometric 0.1 unit

water temperature thermometer or thermister 0.1 °C

Herbicides

acetochlor GCS 0.05 μg/L

alachlor GCS 0.05 μg/L

ametryn GCS 0.05 μg/L

atrazine GCS, LCEA 0.05 μg/L

cyanazine GCS, LCEA 0.05 μg/L

dimethenamid GCS 0.05 μg/L

diuron LCEA 0.05 μg/L

flufenacet GCS 0.05 μg/L

fluometuron LCEA 0.05 μg/L

glyphosate LCGY 0.10 μg/L

glufosinate LCGY 0.10 μg/L

linuron LCEA 0.05 μg/L

metolachlor GCS 0.05 μg/L

metribuzin GCS 0.05 μg/L
WORK PLAN 7

Herbicides—Continued

pendimethalin GCS 0.05 μg/L

prometon GCS 0.05 μg/L

prometryn GCS 0.05 μg/L

propachlor GCS 0.05 μg/L

propazine GCS, LCEA 0.05 μg/L

simazine GCS, LCEA 0.05 μg/L

terbutryn GCS 0.05 μg/L

Herbicide transformation products

acetochlor ESA LCAA 0.05 μg/L

acetochlor OXA LCAA 0.05 μg/L

acetochlor SAA LCAA 0.05 μg/L

alachlor ESA LCAA 0.05 μg/L

alachlor OXA LCAA 0.05 μg/L

alachlor SAA LCAA 0.05 μg/L

AMPA LCGY 0.10 μg/L

cyanazine acid LCEA 0.05 μg/L

cyanazine amide GCS, LCEA 0.05 μg/L

deethylcyanazine LCEA 0.05 μg/L

deethylcyanazine acid LCEA 0.05 μg/L

deethylcyanazine amide LCEA 0.05 μg/L

deethylatrazine GCS, LCEA 0.05 μg/L

deethylhydroxyatrazine LCEA 0.05 μg/L

deisopropylatrazine GCS, LCEA 0.05 μg/L

demethylfluometuron LCEA 0.05 μg/L

desiopropylhydroxyatrazine LCEA 0.05 μg/L

didealkylatrazine LCEA 0.05 μg/L

dimethenamid ESA LCAA 0.05 μg/L

dimethenamid OXA LCAA 0.05 μg/L

flufenacet ESA LCAA 0.05 μg/L

flufenacet OXA LCAA 0.05 μg/L

hydroxyatrazine LCEA 0.05 μg/L

metolachlor ESA LCAA 0.05 μg/L

metolachlor OXA LCAA 0.05 μg/L

propachlor ESA LCAA 0.05 μg/L

propachlor OXA LCAA 0.05 μg/L

Antibiotics

Beta lactams

amoxicillin LCAN 0.20 μg/L

ampicillin LCAN 0.10 μg/L

cefotaxime LCAN 0.10 μg/L

cloxacillin LCAN 0.10 μg/L

oxacillin LCAN 0.10 μg/L

penicillin G LCAN 0.10 μg/L

penicillin V LCAN 0.10 μg/L

Table 3. Physical properties and chemical compounds to be determined on samples 

collected in 2002—Continued

[ft3

/s, cubic feet per second; μS/cm, microsiemens per centimeter at 25 degrees Celsius; 

°C, degrees Celsius; mg/L, milligrams per liter; μg/L, micrograms per liter]

Property or compound Method Reporting 

limit
8 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern United States Streams, 2002

Antibiotics–Continued

Macrolides

erythromycin LCAN 0.10 μg/L

lincomycin LCAN 0.05 μg/L

roxithromycin LCAN 0.10 μg/L

trimethoprim LCAN 0.05 μg/L

tylosin LCAN 0.10 μg/L

virginiamycin LCAN 0.10 μg/L

Quinolines

ciprofloxacin LCAN 0.05 μg/L

clinafloxacin LCAN 0.05 μg/L

flumequine LCAN 0.05 μg/L

lomefloxacin LCAN 0.05 μg/L

norfloxacin LCAN 0.05 μg/L

ofloxacin LCAN 0.05 μg/L 

oxolinic acid LCAN 0.05 μg/L

sarafloxacin LCAN 0.05 μg/L

Sulfonamides

sulfachlorpyridazine LCAN 0.05 μg/L

sulfadiazine LCAN 0.05 μg/L

sulfadimethoxine LCAN 0.05 μg/L

sulfamerazine LCAN 0.05 μg/L

sulfamethazine LCAN 0.05 μg/L

sulfamethoxazole LCAN 0.05 μg/L

sulfathiazole LCAN 0.05 μg/L

Tetracyclines

chlorotetracycline LCAN 0.10 μg/L

doxycycline LCAN 0.10 μg/L

minocycline LCAN 0.20 μg/L

oxytetracycline LCAN 0.10 μg/L

tetracycline LCAN 0.10 μg/L

Tetracycline transformation 

products

anhydrochlortetracycline LCAN 0.10 μg/L

anhydrotetracycline LCAN 0.20 μg/L

demeclocycline LCAN 0.10 μg/L

Table 3. Physical properties and chemical compounds to be determined on samples 

collected in 2002—Continued

[ft3

/s, cubic feet per second; μS/cm, microsiemens per centimeter at 25 degrees Celsius; 

°C, degrees Celsius; mg/L, milligrams per liter; μg/L, micrograms per liter]

Property or compound Method Reporting 

limit
WORK PLAN 9

herbicide use are affecting herbicide concentrations in midwestern streams, (2) the distribution 

of DDA and other herbicide transformation products with limited environmental data, and (3) the 

distribution of selected antibiotics. 

Specific hypotheses:

1. Glyphosate will be detected at a majority of the 

stream sites in post-emergence runoff samples, 

but not in the majority of harvest-season runoff 

samples.

2. AMPA will occur with a frequency and in concentrations similar to that of acetochlor ESA. The 

frequency of AMPA detection will be larger in 

harvest-season runoff samples than in post-application runoff samples.

3. The ratio of AMPA to glyphosate will be greater 

than one in pre-emergence and harvest-season 

samples but less than one in post-emergence 

samples.

4. The AMPA in midwestern streams is from transformation of glyphosate and not transformation 

of detergents in sewage-treatment plants. 

5. When compared to concentrations measured in 

1998 samples, those measured in post-application runoff in 2002 will be about the same for 

atrazine and acetochlor and significantly lower 

for alachlor, cyanazine, and metolachlor. 

6. Didealkylatrazine (DDA) will be detected in most 

midwestern streams in concentrations similar to 

that of de-ethylatrazine.

7. The herbicide transformation product to parent 

ratios for alachlor, cyanazine, and metolachlor 

will be larger in 2002 than in 1998 because of 

declining use of these products.

8. Selected antibiotics will be detected in about onehalf of the 53 midwestern streams.

9. Because of the timing of spring manure applications, antibiotic concentrations will be higher in 

the pre-emergence runoff samples than in the 

post-emergence runoff samples or harvestseason runoff samples.

Sampling Sites

The sampling sites are representative of unregulated streams that drain small, agriculturally dominated watersheds in the Midwestern United 

States. All but 2 of the 53 stream sites sampled in 

this study have been sampled in the prior reconnaissance studies (Goolsby and others, 1994; 

Scribner and others, 1998). The Turkey River near 

Eldorado site (05411850) is about 15 miles downstream from the previously sampled site at 

Spillville (05411600). The Little Cobb River near 

Beauford site (05320270) was sampled in 1998 

but not in the previous reconnaissance studies, 

which had sampled the Rock River at Luverne 

(06483000). The locations of the sites to be sampled and their drainage basins are shown in figure 

2. Sampling stations, drainage areas, 1998 sampling dates, and streamflow are given in table 2.

Sampling Schedule

Three samples will be collected at each site. The 

first set of samples will be collected after pre-emergence herbicides have been applied (usually after 

planting but before crop plants emerge in May or June) 

and following a precipitation event that produces a 

significant increase in streamflow. Ideally, streamflow 

should represent runoff conditions with flow at or 

above the 50th percentile (50 percent exceeds streamflow for the period of record, published in annual 

USGS Water Resources Data reports). These samples 

will be referred to as “pre-emergence runoff samples.” 

The second set of samples will be collected after postemergence herbicides like glyphosate have been 

applied (usually to the foliage of weeds in June or 

July), again following a precipitation event that 

produces runoff conditions and streamflows at or 

above the 50th percentile. These samples will be 

referred to as “post-emergence runoff samples.” The 

third set of samples will be collected during crop 

harvest (usually September to November), and 

following a precipitation event that produces runoff 

conditions and rising streamflows. These samples will 

be referred to as “harvest-season runoff samples.” 
10 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern United States Streams, 2002

MINNESOTA NORTH

DAKOTA

SOUTH

DAKOTA

NEBRASKA

KANSAS

ILLINOIS

INDIANA

MICHIGAN

WISCONSIN

KENTUCKY

OHIO

IOWA

0 100 200 MILES

0 100 200 KILOMETERS

45°

95° 85°

40°

EXPLANATION

Missouri River Ohio River

Mississippi River

53 50

13

15

14

16

17

18

20

19

12

22

46

47

42

43 49

48

44

45

41

28

21

27

26

25

24

23

11

5

6

7

2

52

51

1

33

31

32

36

37 35

40 39

29

30

38

34

39

8

9

10 4

3

1 - Turkey River near Eldorado, IA

2 - Wapsipinicon River at Independence, IA

3 - Old Mans Creek near Iowa City, IA

4 - N. Skunk River near Sigourney, IA

5 - Skunk River at Augusta, IA

6 - Des Moines River at Fort Dodge, IA

7 - Raccoon River at Van Meter, IA

8 - Little Sioux River at Correctionville, IA

9 - Maple River at Mapleton, IA

10 - Boyer River at Logan, IA

11 - Bonpas Creek at Browns, IL

12 - Little Wabash River at Carmi, IL

13 - S. Branch Kishwaukee River at Fairdale, IL

14 - Iroquois River near Chebanse, IL

15 - Dupage River near Shorewood, IL

16 - Spoon River at London Mills, IL

17 - Sangamon River at Riverton, IL

18 - Macoupin Creek near Kane, IL

19 - Kaskaskia River near Cowden, IL

20 - Shoal Creek near Breese, IL

21 - Whitewater River near Alpine, IN

22 - Blue River at Fredericksburg, IN

23 - Eel River near Logansport, IN

24 - Wildcat Creek near Jerome, IN

25 - Wildcat Creek near Lafayette, IN

26 - White River near Nora, IN

27 - Sugar Creek near Edinburgh, IN

28 - E. Fork White River near Bedford, IN

29 - Black Vermillion River near Frankfort, KS

30 - Delaware River near Muscotah, KS

31 - Cottonwood River near New Ulm, MN

32 - Des Moines River at Jackson, MN

33 - Little Cobb River near Beauford, MN

34 - Nodaway River near Graham, MO

35 - Salt Creek at Roca, NE

37 - W. Fork Big Blue River near Dorchester, NE

38 - Big Nemaha River at Falls City, NE

39 - Big Blue River at Barneston, NE

40 - Little Blue River near Fairbury, NE

41 - Clear Creek near Rockbridge, OH

42 - Scioto River near Prospect, OH

43 - Olentangy River at Claridon, OH

44 - Big Darby Creek at Darbyville, OH

45 - Scioto River at Higby, OH

46 - L. Miami River near Oldtown, OH

47 - Mad River at Eagle City, OH

48 - Tiffin River at Stryker, OH

49 - Auglaize River near Fort Jennings, OH

50 - Root River at Racine, WI

51 - St. Croix River St. Croix Falls, WI

52 - Wisconsin River at Muscoda, WI

53 - Rock River at Afton, WI

Stream sampling site 36 - Wahoo Creek at Ithaca, NE

MISSOURI

Figure 2. Locations of sampling sites and associated contributing drainage areas.
WORK PLAN 11

Sampling Procedures

Samples will be collected using methods 

described in Shelton (1994). The equal-width-increment sampling method (Edwards and Glysson, 1988) 

will be used for all samples. The step-by-step 

sampling procedure is given in the Appendix. All 

pesticide samples will be filtered through 0.7-mm 

pore-size baked glass-fiber filters, using an aluminum 

plate filter holder and a ceramic piston fluid-metering 

pump with all Teflon tubing, into precleaned 125-mL 

amber glass bottles. Eight 125-mL amber glass bottles 

from each site will be filled and sent to the ORGL. 

Sample bottles will be clearly labeled with waterproof 

marker or preprinted labels. The minimum information on the label will be the project code, site id, site 

name, date and time of sample collection, sampler 

name(s), and the following phrase "USGS 2002 

RECON." Field measurements for specific conductance, pH, and temperature will be taken for all 

samples and a discharge will be obtained by direct 

measurement, from a rating curve or estimated from a 

nearby USGS gaging station. 

Analytical Procedures

Glyphosate and AMPA. Samples are analyzed for 

glyphosate, glufosinate, and AMPA by using a liquid 

chromatography mass spectrometry (LCMS) method 

as described in Lee and others (2002a). For the LCGY 

analysis, 10-mL water samples are derivatized in the 

laboratory and extracted with solid-phase extraction 

(SPE). The analytes (table 3) are separated by LC and 

identified with an MS in electrospray negative-ion 

(ES–NEG) mode operating in selected-ion mode. The 

concentration of each sample is determined by using 

labeled glyphosate as an internal standard and a standard-addition replicate using the following equation:

C = [ (Ac/Ai)/(Acsa/Aisa – Ac/Ai)] × SAc × DF

where 

C is concentration of the selected compound in 

the sample, 

Ac is area of peak of the ion for the selected 

compound, 

Ai is area of peak for the internal standard, 

Acsa is area of the peak for the selected compound 

in the standard-addition sample, 

Aisa is the area of the peak for the internal standard 

in the standard-addition sample, 

SAc is concentration of the standard addition, and

DF is dilution factor.

Other Herbicides and Transformation Products.

The other herbicide compounds and transformation 

products (table 3) are analyzed using three methods. 

The LCEA analysis (Lee and others, 2002b) uses SPE 

and atmospheric chemical ionization (APCI) positive 

LCMS. The LCAA analysis (Lee and others, 2001) 

uses SPE and ES–NEG–LCMS. The GCS analysis 

(Zimmerman and Thurman, 1999; Kish and others, 

2000) uses SPE and gas chromatography mass spectrometry (GCMS). All three methods use 123-mL 

samples and include surrogate and internal standards. 

Mass spectrometers are operated in selected-ion mode, 

and concentrations of detected analytes are calculated 

from calibration curves.

Antibiotics. A method (LCAN) has been developed 

for the trace analysis (concentrations between 0.05 and 

2.0 μg/L) of five classes of antibiotics consisting of 

seven beta lactams (BLs), six macrolides (MCs), eight 

quinolines (QUs), seven sulfonamides (SAs), and 

eight tetracyclines (TCs) (table 3). These antibiotics 

are commonly are used by humans, livestock, and pets 

or are agricultural feed additives. Some of these antibiotics are suspected to leach into ground and surface 

water. Three slightly different sample preparation 

methods are used for BLs and MCs, QUs and SAs, and 

TCs. All samples are extracted with SPE and determined by LCMS with positive-ion electrospray using 

methods similar to Lindsey and others (2001). Surrogate and internal standards are used in the analysis of 

10-mL samples, and concentrations of detected 

analytes are calculated from calibration curves 

acquired with the samples.

Quality Assurance 

Quality assurance of sample collection and 

handling will consist of 15 concurrent replicate 

samples and 4 field blank samples (table 4). In the 

laboratory, 10 percent or more of the samples will be 

split and analyzed in replicate (including sample preparation) to yield data on the precision of the analysis. 

These same samples are also analyzed as spikes for 
12 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern United States Streams, 2002

determining accuracy. Method blanks are prepared and 

analyzed with each batch of samples. Standard reference materials (SRMs), if available, are routinely used 

as Laboratory Control Standards (LCS). 

DATA ANALYSIS METHODS AND 

REPORTS 

Results from this study will represent the first 

broad-scale investigation of glyphosate and AMPA in 

Midwestern United States streams. Analytical results 

from this study will be published in a USGS open-file 

report. Analysis of the results of this study will be 

published in one or more journal articles to be 

prepared in 2003 and 2004. Some ancillary data in 

addition to streamflow will be collected in association 

with the water-quality data in order to better understand the information provided. The amount of ancillary data collected will be limited by resources but at a 

minimum will include estimates of corn and soybean 

planting date, monthly precipitation and temperature, 

crops planted (will not be available until 2002 Census 

of Agriculture data are available), and pesticide use 

(may only be State level data).

REFERENCES CITED

Aspelin A.L., and Grube, A.H., 1999, Pesticide industry 

sales and usage—1996 and 1997 market estimates: 

U.S. Environmental Protection Agency, Office of 

Prevention, Pesticides, and Toxic Substances, 

Washington, D.C. 733–R–99–001, 39 p.

Battaglin, W.A., and Fairchild, J., 2002, Potential toxicity of 

pesticides measured in midwestern streams to aquatic 

organisms: Water Science and Technology, v. 45, no. 9, 

p. 95–103, available on-line at 

http://co.water.usgs.gov/midconherb/html/

onlinereports.html

Battaglin, W.A., Furlong, E.T., Burkhardt, M.R., and Boyer, 

L.L., 2001, Concentrations of selected sulfonylurea, 

sulfonamide, and imidazolinone herbicides, and other 

pesticides in storm runoff from 71 streams, outflow 

from 5 reservoirs, and ground water from 25 wells in 

Table 4. Schedule for quality-assurance samples

[CR, current replicates; FB, field blanks;--, none]

State ID Station name

Preemergence 

sample

Postemergence 

sample

Harvestseason 

sample

IA 05411600 Turkey River near Eldorado CR -- --

IA 05484500 Raccoon River at Van Meter -- CR --

IA 06607200 Maple River at Mapleton -- -- FB

IL 03378000 Bonpas Creek at Browns CR -- --

IL 03381495 Little Wabash River at Carmi -- CR --

IL 05592100 Kaskaskia River near Cowden -- -- CR

IN 03275000 Whitewater River near Alpine FB -- --

IN 03362500 Sugar Creek near Edinburgh -- CR --

IN 03371500 E. Fork White River near Bedford -- -- CR

KS 06885500 Black Vermillion River near Frankfort CR -- --

MN 05476000 Des Moines River at Jackson -- CR --

MO 06817700 Nodaway River near Graham FB -- --

NE 06880800 W. Fork Big Blue River nr Dorchester -- CR --

NE 06882000 Big Blue River at Barneston -- -- CR

OH 03219500 Scioto River near Prospect CR -- --

OH 03267900 Mad River at Eagle City -- CR --

OH 04185000 Tiffin River at Stryker -- -- CR

WI 04087240 Root River at Racine CR -- --

WI 05430500 Rock River at Afton -- FB --
REFERENCES CITED 13

the Midwestern United States, 1998: U.S. Geological 

Survey Water-Resources Investigations Report 

00–4225, 123 p., available on-line at 

http://co.water.usgs.gov/midconherb/pdf/wrir00-

4225.pdf

Battaglin, W.A., and Goolsby, D.A., 1999, Are shifts in 

herbicide use reflected in concentration changes in 

Midwestern Rivers? Environmental Science and Technology, v. 33, p. 2917–2925. 

Boyd, R.A., 2000, Herbicides and herbicide degradates in 

shallow groundwater and the Cedar River near a 

municipal well field, Cedar Rapids, Iowa: The Science 

of the Total Environment, v. 248, p. 241–253.

Busse, M.D., Ratcliff, A.W., Shestak, C.J., and Powers, 

R.F., 2001, Glyphosate toxicity and the effects of longterm vegetation control on soil microbial communities: 

Soil Biology and Biochemistry, v. 33, p. 1777–1789.

Edwards, T.K., and Glysson, D.G., 1988, Field methods for 

measurement of fluvial sediment: U.S. Geological 

Survey Open-File Report 86–531, 118 p.

Ferrer, I., Thurman, E.M., and Barcelo, D., 1997, Identification of ionic chloroacetanilide-herbicide metabolites in 

surface water and groundwater by HPLC/MS using 

negative ion spray: Analytical Chemistry, v. 69, 

p. 4547–4553.

Folmer, L., Sanders, H., and Julin, A., 1979, Toxicity of the 

herbicide glyphosate and several of its formulations to 

fish and aquatic invertebrates: Archives of Environmental Contaminant Toxicology, v. 8, p. 269–278.

Goolsby, D.A., and Battaglin, W.A., 1993, Occurrence, 

distribution, and transport of agricultural chemicals in 

surface waters of the Midwestern United States, in

Goolsby, D.A., and others, eds., Selected papers on 

agricultural chemicals in water resources of the 

Midcontinental United States: U.S. Geological Survey 

Open-File Report 93–418, p. 1–24.

Goolsby, D.A., Boyer, L.L., and Battaglin, W.A., 1994, Plan 

of study to determine the effects of changes in herbicide use on herbicide concentrations in midwestern 

streams, 1989–94: U.S. Geological Survey Open-File 

Report 94–347, 14 p.

Haney, R.L., Senseman, S.A., and Hons, F.M., 2002, Effect 

of Roundup Ultra on microbial activity and biomass 

from selected soils: Journal of Environmental Quality, 

v. 31, p. 730–735.

Kalkhoff, S.J., Kolpin, D.W., Thurman, E.M., Ferrer, I., and 

Barcelo, D., 1998, Degradation of chloroacetanilide 

herbicides—The prevalence of sulfonic and oxanilic 

acid metabolites in Iowa groundwaters and surface 

waters: Environmental Science and Technology, v. 32, 

p. 1738–1740.

Kish, J.L., Thurman, E.M., Scribner, E.A., and Zimmerman, 

L.R., 2000, Method of analysis by the U.S. Geological 

Survey Organic Geochemistry Research 

Group—Determination of triazine and chloroacetanilide herbicides in water by solid-phase extraction and 

capillary-column gas chromatography/mass spectrometry with selected-ion monitoring: U.S. Geological 

Survey Open-File Report 00–385, 13 p.

Kolpin, D.W., Furlong, E.T., Meyer, M.T., Thurman, E.M., 

Zaugg, S.D., Barber, L.B., and Buxton, H.T., 2002, 

Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999–2000—

A national reconnaissance: Environmental Science and 

Technology, v. 36, p. 1202–1211.

Lee, E.A., Kish, J.L., Zimmerman, L.R., and Thurman, 

E.M., 2001, Methods of analysis by the U.S. Geological Survey Organic Geochemistry Research Group—

Update and additions to the determination of chloroacetanilide herbicide degradation compounds in water 

using high-performance liquid chromatography/mass 

spectrometry: U.S. Geological Survey Open-File 

Report 01–10, 17 p.

Lee, E.A., Strahan, A.P., and Thurman, E.M., 2002a, 

Methods of analysis by the U.S. Geological Survey 

Organic Geochemistry Research Group—Determination of glyphosate, aminomethylphosphonic acid, and 

glufosinate in water using online solid-phase extraction 

and high-performance liquid chromatography/mass 

spectrometry: U.S. Geological Survey Open-File 

Report 01–454, 13 p., available on-line at 

http://ks.water.usgs.gov/Kansas/pubs/abstracts/ofr.01-

454.abs.html

Lee, E.A., Strahan, A.P., and Thurman, E.M., 2002b, 

Methods of analysis by the U.S. Geological Survey 

Organic Geochemistry Research Group—Determination of triazine and phenylurea herbicides and their 

degradation products in water using solid-phase extraction and liquid chromatography/mass spectrometry: 

U.S. Geological Survey Open-File Report 02–436, 

19 p., available on-line at 

http://ks.water.usgs.gov/Kansas/pubs/abstracts/of.02-

436.html

Lindsey, M.E., Meyer, M.T., and Thurman, E.M., 2001, 

Analysis of trace levels of sulfonamide and tetracycline antimicrobials in groundwater and surface 

water using solid-phase extraction and liquid chromatography/mass spectrometry: Analytical Chemistry, 

v. 74, p. 4640–4646.

Mitchell, D., Chapman, P., and Long, T., 1987, Seawater 

challenge testing of Coho salmon smelts following 

exposure to Roundup herbicide: Environmental Toxicology and Chemistry, v. 6, p. 875–878.
14 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern United States Streams, 2002

Perkins, P.J., Boermans, H.J., and Stephenson, G.R., 2000, 

Toxicity of glyphosate and triclopyr using the frog 

embryo teratogenesis assay – Xenopus: Environmental 

Toxicology and Chemistry, v. 19, no. 4, p. 940–945.

Phillips, P.J., Wall, G.R., Thurman, E.M., Eckhardt, D.A., 

and VanHoesen, J., 1999, Metolachlor and its metabolites in tile drain and stream runoff in the Conajoharie 

Creek watershed: Environmental Science and Technology, v. 33, p. 3531–3537.

Porter, S.D., Harris, M.A., and Kalkhoff, S.J., 2001, Influence of national factors on the quality of Midwestern 

streams and rivers: U.S. Geological Survey WaterResources Investigations Report 00–4288, 13 p.

Royal Society of Chemistry, 1998, Metabolic pathways of 

agrochemicals—Part one, herbicides and plant growth 

regulators, T.R. Roberts, ed:, London, Royal Society of 

Chemistry Books, p. 396–400.

Saenz, M.E., DiMarzio, W.D., Alberdi, J.L., and Tortorelli, 

M., 1997, Effects of technical grade and a commercial 

formulation of glyphosate on algal population growth: 

Bulletin of Environmental Contaminant Toxicology, 

v. 59, p. 638–644.

Scribner, E.A., Battaglin, W.A., Goolsby, D.A., and 

Thurman, E.M., 2000, Changes in herbicide concentrations in midwestern streams in relation to changes in 

use, 1989–1998: The Science of the Total Environment, v. 248, no. 2–3, p. 255–263.

Scribner, E.A., Goolsby, D.A., Thurman, E.M., and Battaglin, W.A., 1998, A reconnaissance for selected herbicides, metabolites, and nutrients in streams of nine 

Midwestern States, 1994–95: U.S. Geological Survey 

Open-File Report 98–181, 44 p.

Scribner, E.A., Goolsby, D.A., Thurman, E.M., Meyer, 

M.T., and Pomes, M.L., 1994, Concentrations of 

selected herbicides, two triazine metabolites, and nutrients in storm runoff from nine stream basins in the 

Midwestern United States, 1990–92: U.S. Geological 

Survey Open-File Report 94–396, 144 p.

Shelton, L.R., 1994, Field guide for collection and 

processing stream-water samples for the national 

water-quality assessment program: U.S. Geological 

Survey Open-File Report 94–455, 42 p.

Skark, C., Zullei-Seibert, N., Schottler, U., and Schlett, C., 

1998, The occurrence of glyphosate in surface water: 

International Journal of Environmental Analytical 

Chemistry, v. 70, no. 1–4, p. 93–104.

Stratton, G.W., 1990, Effects of the herbicide glyphosate on 

nitrification in four soils from Atlantic Canada: Water, 

Air, and Soil Pollution, v. 51, no. 373–383.

Stratton, G.W., and Stewart, K.E., 1991, Effects of the 

herbicide glyphosate on nitrogen cycling in an acid 

forest soil: Water, Air, and Soil Pollution, v. 60,

p. 231–247.

Thurman, E.M., Goolsby, D.A., Meyer, M.T., and Kolpin, 

D.W., 1991, Herbicides in surface waters of the 

Midwestern United States—The effect of spring flush: 

Environmental Science and Technology, v. 25, 

p. 1794–1796.

Thurman, E.M., Goolsby, D.A., Meyer, M.T., and Kolpin, 

D.W., 1992, A reconnaissance study of herbicides and 

their metabolites in surface water of the Midwestern 

United States using immunoassay and gas chromatography/mass spectrometry: Environmental Science and 

Technology, v. 26, p. 2440–2447.

Thurman, E.M., Meyer, M.T., Mills, M.S., Zimmerman, 

L.R., Perry, C.A., and Goolsby, D.A., 1994, Formation 

and transport of deethylatrazine and deisopropylatrazine in surface water: Environmental Science and 

Technology, v. 28, p. 2267–2277.

U.S. Department of Agriculture, 2002, National Agricultural Statistics Service (NASS), Agricultural Chemical 

Use Database: Accessed 10/16/2002 at 

http://www.pestmanagement.info/nass/

U.S. Environmental Protection Agency, 1993, Reregistration Eligibility Decision (RED) Glyphosate: Accessed 

10/16/2002 at 

http://www.epa.gov/oppsrrd1/REDs/old_reds/

glyphosate.pdf

Zimmerman, L.R., and Thurman, E.M., 1999, Method of 

analysis by the U.S. Geological Survey Organic 

Geochemistry Research Group—Determination of 

triazine and chloroacetanilide herbicides in water by 

solid-phase extraction and capillary-column gas chromatography/mass spectrometry with selected-ion 

monitoring: U.S. Geological Survey Open-File Report 

98–634, 21 p.
APPENDIX
16 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern U.S. Streams, 2002
APPENDIX 17

Determining where and when to sample

1. Determine which sites are to be sampled in your 

State (table 2).

2. Determine which sites need quality-assurance 

samples collected (table 4).

3. Follow news on progress of planting and weather 

(Battaglin will provide weekly data during the 

growing season, or you can monitor at 

http://usda.mannlib.cornell.edu/reports/nassr/

field/weather/) accessed May, June, and July 

2002. 

4. Collect the first set of surface-water samples (preemergence runoff) after pre-emergence herbicides have been applied (usually May or June), 

when corn is at least 50 percent planted, and 

following a precipitation event that produces a 

significant increase in streamflow. Ideally, the 

sample should represent runoff conditions with 

streamflow at or above the 50th percentile, which 

is equal to the "50 percent exceeds" streamflow 

value for the period of record, published for most 

sites in annual USGS Water Resources Data 

reports.

5. Continue to follow news on local progress of plant 

development and weather. 

6. Collect the second set of surface-water samples 

(post-emergence runoff) after post-emergence 

herbicides, particularly glyphosate, have been 

applied (usually June or July), when soybeans are 

nearing 100 percent emerged, and following a 

precipitation event that produces runoff conditions and streamflow at or above the 50th percentile. 

7. Collect the third set of surface-water samples 

(harvest-season runoff) during or after crop 

harvest (usually September, October, or 

November) following a precipitation event that 

produces runoff conditions and a rise in streamflow at the site. Streamflow does not have to be 

at or above the 50th percentile.

Sample processing

1. Preclean all equipment with a Liquinox/tap-water 

solution, rinse with tap water, deionized water, 

then methanol, and then air dry. 

2. Collect and composite a minimum of ~2 liters of 

water using equal-width-increment sampling into 

a large precleaned glass carboy. 

3. Make field measurements of pH, temperature, and 

specific conductance, in the stream if possible, or 

on the composite sample.

4. Filter 100 mL of sample water through a 0.7-mm 

pore-size baked glass-fiber filter to condition, 

and then filter 1.0 L of sample water into eight 

125-mL baked amber glass bottles. 

5. Label all sample bottles clearly with waterproof 

marker or preprinted labels (a paper label 

completely covered with clear packing tape is 

preferred). The minimum information on the 

label for bottles sent to the ORGL is the project 

code, site ID, site name, date and time of sample 

collection, sampler name(s), and the phrase 

"USGS 2002 RECON” as shown below:

6. Glass bottles will be placed in foam packers to help 

prevent breakage. The eight bottles from each 

site will then be placed in a plastic bag, which 

will help keep them dry in the coolers and keep 

all bottles from the same site together. All 

samples are to be kept chilled on ice and shipped 

within 2 days to the ORGL in Lawrence, Kansas.

7. Notify Bill Battaglin (303-236-4882 extension 256) 

wbattagl@usgs.gov with the date and time of 

sample collection and physical properties as soon 

as possible after samples are shipped.

Quality-assurance samples

Concurrent replicate and field blank samples 

will be collected during all three sampling periods 

(table 4). The intent of these samples is to help determine analytical precision. The concurrent replicate 

Project code DBD

station id 05465500

stream name/locationIowa River at Wapello, IA

date and time 6-18-02 @ 1015

sampler 

name/initials

Joe Sample and Jane 

Stream

study label USGS 2002 RECON

APPENDIX – Detailed sample collection and processing instructions
18 Work Plan for Determining the Occurrence of Glyphosate, Its Transformation Product AMPA, Other Herbicide Compounds, and 

Antibiotics in Midwestern U.S. Streams, 2002

sample will require collecting and compositing a 

larger volume of water from the sites than normally 

needed. The replicates will be processed in the same 

manner as the normal samples using the same filter if 

possible. Record the same site name and ID for both 

samples but a time that is 1 minute later for the replicate sample. Both samples will be sent to the ORGL in 

Lawrence, Kansas, for herbicide analysis. The field 

blank samples will consist of organic-free water that is 

processed through all of the sample collection and 

filtration equipment after it has been cleaned as 

described above. These samples will be labeled as a 

field blank and sent to the ORGL in Lawrence, 

Kansas, for analysis.