ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF ENFORCEMENT
EPA-330/9-81-001
NEIC
PESTICIDE SAMPLING GUIDE
March 1981
John T. Ellison
NATIONAL ENFORCEMENT INVESTIGATIONS CENTER
Denver, Colorado
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Mention of trade names or commercial sources in this manual are for
identification only and do not constitute endorsement by this Agency.
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CONTENTS
I INTRODUCTION
II GENERAL SAMPLING CONSIDERATIONS
III AIR
IV DUST, SOIL, AND SEDIMENT
V WATER
VI FISH
VII BENTHOS
VIII PLANTS
IX WILDLIFE AND LIVESTOCK
X HUMAN TISSUE
XI SAMPLE CONTROL PROCEDURES
XII BIBLIOGRAPHY
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ACKNOWLEDGMENTS
In 1975, a draft pesticide sampling manual was written by Henry Bell,
National Enforcement Investigations Center (NEIC). Parts of the 1975 draft
were incorporated into the EPA Pesticide Inspector's Handbook, prepared by
EPA Pesticide and Toxic Substances Enforcement Division. This 1981 update
of the 1975 manual incorporates material gathered by Bruce Binkley and Rob-
ert Schneider of NEIC. John Ellison is the principal author.
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I. INTRODUCTION
Pesticide workshops and training courses for Federal, state, and local
regulatory personnel have been and continue to be conducted by the National
Enforcement Investigations Center (NEIC). This pesticide sampling guide
provides these regulatory personnel and their colleagues with general in-
formation about pesticide sampling, and an overview of NEIC pesticide sam-
pling methods.
Applied pesticides can move through the air, water, or soil, affecting
both target and non-target flora and fauna. Thus, the sections of this
guide describe sampling for pesticides in air, soil, water, fish, plants,
wildlife and livestock, and human tissues. To assist the manual user in
comparing alternative collecting techniques, sampling methods are presented
in a standard format listing apparatus then procedures.
Because study objectives and environmental variables will influence
the choice of sampling methods, this guide cannot cover each set of circum-
stances that may be encountered. The user is advised to exercise judgment,
ingenuity, and common sense in designing a sampling regime. Advice on sam-
pling techniques or site-specific situations can be obtained from:
Compliance Investigations Branch
EPA-National Enforcement Investigations Center
Denver Federal Center, Box 25227
Denver, Colorado 80225
(303) 234-2336
The NEIC has been involved in conducting pesticide use observations since
1976 at the request of the EPA Pesticide and Toxic Substances Enforcement
Division. NEIC has conducted studies to document pesticide use and user
compliance with pesticide label instructions and permits in numerous
states including Arizona, California, Colorado, Delaware, Florida, Georgia,
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Indiana, Maine, Minnesota, Mississippi, Montana, Oregon, Tennessee, Texas,
*
Utah, and Washington . Techniques evaluated during these studies are pre-
sented in this guide.
Reports detailing pesticide studies conducted by NEIC are available upon
request.
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II. GENERAL SAMPLING CONSIDERATIONS
Pesticide sampling should be goal-oriented: a pesticide application
is going to or has occurred, and sampling is desired to measure real or po-
tential threats to human health and the environment. Each case must be
considered individually but the following paragraphs present a general or-
der of events.
When you receive a request for pesticide sampling, the first step is
to collect background information. This might require an onsite reconnais-
sance, and one should ask these questions. What pesticide is involved? On
what and where is it being applied? How is the application to be made?
Who are the local contacts? Are there rules and regulations to be followed
other than label restrictions? What is the goal of the sampling? The goal
of the sampling should dictate the kinds of samples to be collected.
The goal of pesticide application sampling is usually to measure
drift. For drift studies, air sampling equipment like high-volume samplers
and drift cards are appropriate. Water sampling is necessary if the pesti-
cide might have drifted into water. To measure possible harm to fish, fish
exposures and acetylcholinesterase inhibition (AChE) testing could be done.
Soil and vegetation sampling is used to check suspected pesticide damage to
foliage. Each type of sampling is selected as necessary to provide infor-
mation about pesticide movement and its effects.
After the background information is assembled, develop a study plan to
define study objectives, sampling to be conducted, analyses required, per-
sonnel requirements, and equipment needs. At this time, and possibly when
you are gathering background information, consult the chemists to determine
analytical requirements and quality control considerations, especially
those which specifically impact sampling methodology and other field con-
siderations. It is necessary to ensure that the pesticide of interest can
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be collected by the method attempted. Test sampling procedures as thor-
oughly as possible before the survey.
To prepare the field equipment, calibrate it as necessary, or learn
the procedures for field calibration. Clean the sampling equipment by
rinsing or wiping with an appropriate solvent. Wash glassware with hot,
soapy water, rinse well and, prior to use, rinse all equipment and glass-
ware with the appropriate solvent.
The field work involves as much preparation as actual sampling. Care-
fully observe spray sites and note any sensitive areas near the spray site
like schools, subdivisions, bodies of water, or major roads. Contact pro-
perty owners, the applicator, and local regulators. Make a map or sketch
of the area and select sampling sites and parameters. Pesticide applica-
tions are often made very early in the morning or late in the evening.
Rarely is any set schedule kept and weather delays are common. Cooperation
with the applicator is almost a necessity if sampling and observations are
to coincide with spraying. Pesticide applications are often made on short
notice, so equipment and personnel have to be ready for quick response.
This requires that everyone be familiar enough with the operation to set up
the equipment in the dark, which is often necessary.
The actual application is often anti-climactic; the real work is in
the preparation. Still, notes, observations, and photos taken during the
actual application can enhance data interpretation and any report presen-
tation. Samples need to be carefully collected and properly preserved and
shipped for desired analyses.
Laboratory analyses should be requested to provide data that will ful-
fill the objectives of the study. Data interpretation will be easier if
proper planning has resulted in suitable sampling and appropriate analyses
to address the study objectives.
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III. AIR
This section describes the supplies, equipment, and techniques re-
quired to collect air and drift samples for pesticide residue analysis.
Spray droplet cards, solid adsorbent samplers, and high-volume samplers
have proven most useful for NEIC.
Method development work before the field work is conducted is especi-
ally important in air sampling. Sampling methods must be tested to make
sure the pesticide of interest can be extracted from the environment into
or onto an artificial media. This is a mechanical or chemical process that
is not applicable for all pesticides in all environmental conditions.
MAGNESIUM DIOXIDE SLIDE
Magnesium dioxide slides are used to detect droplet deposits from
overspray or drift from pesticide applications. Drawbacks for field use
include fragility of glass slides and sensitivity of the Mg02 coating to
dust, abrasion, and moisture. Cost is approximately $l/slide. A stand
holding four slides and slide box is pictured below.
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Apparatus
1. Glass microscope slides, 1" x 3", with sintered ends for labeling
2. Magnesium foil strip for Mg02 coating
3. Wooden stand with clips to hold slides
4. Slide box for transporting slides
5. Data sheets and identification tags (see NEIC Policies and Pro-
cedures Manual for examples)
Procedure
1. Coat slides with Mg02 by burning magnesium ribbon beneath each
slide. Protect the worker from the noxious fumes and extremely
bright light by performing the procedure with adequate ventila-
tion and using dark protective glasses.
2. Handle coated slides carefully and protect them in a slide box
until ready to use.
3. Using clips, for example spring-type clothespins, attach slides
to a stake at approximately vegetation height, with one coated
slide facing each direction (N, S, E, W), thus using 4 slides
per station.
4. Inspect exposed slides for visible craters in the Mg02 coating,
at time of collection. Check sintered glass label portion of
each slide for proper identification. Place exposed slides in
slide box for transport to the laboratory for analysis.
5. Count and measure craters microscopically and note in which
direction droplet deposits were moving on impact. Measure
craters to the nearest micron with lOOx magnification.
Comments
A pesticide droplet will splatter or spread when it hits a sampling
device. For a given sampling device the amount of droplet spread, known as
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the spread factor, will vary for different pesticides and carriers. By us-
ing published spread factors, the volume median diameter or VMD of measured
droplets can be determined. The VMD can then be used to predict how far
pesticide droplets would theoretically travel. By sampling at numerous
locations, empirical data will show distances pesticide droplets actually
travelled. However, empirical data is limited by the sensitivity of the
sampling device. Nineteen references on pesticide drift are listed in Sec-
tion XII. Several of the references deal with spread factors and VMD cal-
culations.
AGAR PLATES
Agar plates are inexpensive devices for detecting overspray or drift
of biological insecticides like Bacillus thurinqiensis* used for integrated
pest management. The cost is approximately $l/plate.
Apparatus
1. Agar plate or petri dish
2. Trypticase soy agar
3. Wooden upright platforms high enough to be placed at or slightly
above surrounding vegetation
4. Data sheets and identification tags
5. Laboratory incubation facilities
Procedure
1. Prepare trypticase soy agar plates.
2. Place prepared plates atop wooden platforms at approximately
the height of surrounding vegetation.
* Stager, William, Bt Methodology, unpublished paper, 1980, 4 pp.
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3. Remove petri dish lid to expose media to ambient air during
spray operation. Keep exposure time to 2 hr or less because
ambient bacteria may also be collected and interfere with analy-
sis.
4. After exposure, replace lids and transport labeled plates to
the laboratory for analysis.
5. To confirm Bacillus thurinqiensis, incubate plates 24 hr at
37°C and record bacterial growth; then develop colonies and ex-
amine microscopically.
Spray droplet cards are used to detect droplet deposits from overspray
or drift from pesticide applications. The sensitivity to different chemi-
cal compounds varies with the card type. Several card types are exposed -
simultaneously at each sampling site. A sample card is pictured below.
1. Kromecote cards, 4" x 5", finely-textured, glossy photographic
paper. Sensitive to parathion and dyes. Card sources are listed
in Section XII, Equipment References.
Apparatus
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2. Linagraph 480 cards, cut size 4" x 6", coarse-textured, non-
glossy photographic paper. Very sensitive to water but also
heat, light, and oil sensitive. Sunlight causes paper to dis-
color but does not interfere with its sensitivity to water.
3. Multi-spectrum 209 copy type 640 paper cut to 4" x 5", medium-
textured copy paper. Senstivie to oils and dyes.
4. Posterboard base, 12" x 12"
5. Upright wooden platform, approximate height of surrounding
vegetation
6. Tape, stapler, tacks, rubber bands
7. Plastic bags or aluminum foil
8. Data sheets and identification tags
Procedure
1. Assemble drift card by stapling or taping one of each type card
onto 12" x 12" posterboard.
2. Tack or rubber band assembled drift card to wooden platform.
3. Expose card during and after pesticide application. Pick up
the card after pesticide mist settles; do not leave it overnight.
4. After exposure, collect drift card and place in plastic bag or
aluminum foil with label for transport to the laboratory.
5. Analyze by measuring droplet diameters to the nearest micron and
counting droplets/cm2 at 30x magnification. See Mg02 Slide com-
ments for discussion of spread factors and VMD. For chemical
verification of pesticide presence, arrange for chemical analysis
in advance.
AERIAL SCREENS
*
Aerial screens can be used for collecting dust-transported pesticides
* Tessari and Spencer, 1971, "Air Sampling in the Human Environment,"
Jour, A.O.A.C., 54 (6):1376-1382.
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and detecting particle and vapor drift at a cost of approximately $5/cloth
screen.
Apparatus
1. 0.5 m2 nylon chiffon cloth precleaned with N-hexane
2. Wooden frame mounted on poles
3. Solvent appropriate for desired pesticide. Check with chemists.
A mixture of 10% ethylene glycol and 90% acetone is often used.
4. Glass jars
5. Data sheets and identification tags
Procedure
1. Install frames in desired locations.
2. Suspend pretreated nylon chiffon cloths securely in wooden
frame.
3. Expose screens for entire spray period and remainder of day.
4. Transport collected screens in clean and labeled jars to the
laboratory for pesticide analysis.
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SEQUENTIAL PESTICIDE SAMPLER
The sequential pesticide sampler is used to detect vapor drift (< 0.1pm).
Its use is limited because the instrument is bulky, fragile, electric-powered,
and heavy. The cost is approximately $2,500/sampler as pictured below.
Apparatus
1. Sequential pesticide samplers with extra glassware and vacuum
pump (MISCO model 88)
2. 100 mSL solvent appropriate for desired pesticide. Check with
chemists; reagent-grade ethylene glycol is often used.
3. Aluminum foil
4. 125-m£ glass, screw-top bottles with teflon or foil-lined lids,
pre-rinsed with acetone
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5. Electrical power source and cord
6. Data sheets and identification tags
7. Shipping container with crushed ice
Procedure
1. Pour 100 m£ of appropriate solvent into each impinger. Assemble
sampling train and insert glass wool to prevent moisture being
drawn into vacuum pump.
2. Adjust flow to 1.0 ft3/min.
3. Set timer to control switching air flow from one impinger as-
sembly to another; 6-hr and 24-hr clocks are available. Run
one impinger prior to application, the second impinger during
application, and the third and fourth impingers post-application.
If spraying is delayed, the sampler may have to be shut off after
the pre-application sample is collected and restarted just prior
to application.
4. After sampling, remove solvent from each impinger and place in
separate labeled 125-mi glass bottles.
5. Transport on ice in a chest to laboratory for pesticide analysis.
Avoid sunlight striking samples as much as possible.
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HIGH-VOLUME SAMPLER
The high-volume air sampler is used to detect particle or vapor drift
by capturing material on dry fiberglass filters. Highly volatile pesti-
cides may evaporate from filter. The standard unit costs about $450 while
the smaller, portable, battery-operated model costs about $300 without bat-
tery. The standard unit is pictured below.
Apparatus
1. High-volume air sampler
2. Fiberglass filter 20 x 25.4 cm
3. Envelope for filter
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4. Electric power or battery
5. Data sheets and identification tags
6. Shipping container for samples
Procedure
1. Place fiberglass filter in high-volume sampler.
2. Start instrument. If quantitative sample is desired, instrument
should be calibrated according to manufacturer's directions.
3. Operate pre-, during, and post-application until pesticide has
settled. High-volume samplers collect dust as well as pesticide.
During long operating times, dust might clog filter pores and in-
terfere with chemical analysis.
4. Remove exposed filter from sampler, fold exposed, "dirty" side
on itself, and place in labeled envelope or folder. Keep exposed
filter in the dark.
5. Transport to laboratory in container that keeps filters in the
dark.
SOLID ADSORBENT COLLECTORS
Solid adsorbent samplers are used to detect particle and vapor drift.
The collectors use various packing materials including C-18, XAD-2, carbon,
and silica. The samplers are inexpensive, costing up to a few dollars
each. A vacuum pump to pull air through the sampler costs about $200. A
sample train and examples of different solid adsorbent samples are pictured
below.
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Apparatus
1. Solid adsorbent column
2. Flexible tubing and "Y" fittings
3. Vacuum pump capable of producing an air flow of 10£/min through
solid adsorbent trains
4. Plastic bag for samples
5. Data sheets and identification tags
Procedures
1. Set up solid adsorbent columns in parallel and attach to vacuum
pump with flexible tubing. Note direction of air flow so chem-
ists can extract in same direction if desired.
2. Operate for desired period. Operation for 24 hr or longer is
possible.
3. After sampling, disconnect solid adsorbent columns and place
in plastic bag or foil with identification. Store in sample
chest for transport to laboratory.
4. Include one or more unused solid adsorbent columns as a blank
sample.
5. In the laboratory, extract pesticides with appropriate solvent
and analyze for the desired pesticide.
Comments
Related Information, such as temperature, wind speed and direction,
and precipitation, along with information about the sampling locality and
pesticide use is helpful in the interpretation of ambient air sampling re-
sults. Need for or value of such information must be predetermined and
arrangements made to obtain this information at the time of sampling.
Numbering and Labeling samples in the field should be given careful
attention to prevent mistaken identification, either by the collector or
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the analyst. Record each field number in the sampler's notes by date and
time collected, method of collection, location of sample point, method
of preservation, and witnesses present.
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IV. DUST, SOIL, AND SEDIMENT
This section describes the supplies, equipment and techniques re-
quired to collect, preserve and ship dust, soil and sediment samples for
analysis of pesticide residues.
DUST SAMPLING
Dust sampling for pesticide residues is essential during investiga-
tions of areas that are used for storage of foodstuffs or in areas fre-
quently occupied by humans or animals. Cost of a suitable vacuum is less
than $50.
Apparatus
1. Small vacuum cleaner that uses disposable bags (e.g. Hoover
"Handi Vac")
2. Clean, spare bags for vacuum cleaner
3. Acetone-washed aluminum foil
4. Data sheets, identification tags
5. Shipping container and crushed ice
Procedure
1. Obtain dust sample by thoroughly vacuuming the site including
the floor, walls, ceiling, and other areas in which dust may be
found.
2. After vacuuming, empty the contents of disposable bag onto a
sheet of foil. Dispose of the used bag and clean the vacuum be-
tween uses. Obtain, if possible, one cupful or about 100 to
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200 g of dust. Securely wrap the sample, attach the identifi-
cation tag and pack in shiping container for shipment. Preser-
vative and holding times depend on the pesticide and chemical
analysis.
SOIL SAMPLING
Soil sampling has two uses. Surface samples can detect pesticide
overspray or drift into non-target areas. Deeper samples can detect past
pesticide practice. A coring device as pictured below costs about $100 to
make. Ready-made corers are also available commercially.
Apparatus
1. Metallic coring device capable of collecting a soil sample 2 in.
in diameter by 3 in. deep. A coring device can be constructed by
welding a 36-in T-handle to a 3-in piece of steel pipe (2-in. ID).
One end of the pipe should be beveled to form a cutting edge.
2. Acetone-washed aluminum foil
3. Plastic bags
4. Small spatula, cake slicer, or 4-in. blade knife
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5. Data sheets and identification tags
6. Shipping container with crushed ice
Procedure
1. Place cutting edge of coring device on the ground and force into
soil to a 3-in. depth using twisting or rocking motions to free
corer and withdraw sample.
2. Carefully push core out of sampler onto a pre-cleaned sheet of
aluminum foil. Core may be sectioned if desired. Rinse corer
with solvent after each sample.
3. Wrap each sample in foil and place with sample identification
into whirl-pac type plastic bag being careful to seal each sam-
ple.
4. Keep labeled samples in shipping container with crushed ice for trans-
port to laboratory.
SEDIMENT SAMPLING
Sediment sampling is used to detect pesticide-contaminated material
that settles out of water and accumulates on stream beds or lake bottoms.
Sampling water overlying these deposits might not reveal significant quan-
tities of pesticides, yet pesticides may be found in the sediment which
acts as a sink. Numerous samplers are suitable for sediment sampling, in-
cluding the Petersen dredge ($250), Ekman dredge ($150), tube corer ($300),
mud snapper ($100), and the Emery bacteriological sampler ($275). In many
instances, a simple core tube may be used costing less than $3. Biological
or scientific supply companies like Wildco sell sediment sampling gear.
Apparatus
1. Commercial or homemade sampler
2. Widemouth glass quart jars with teflon or foil lid liners
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3. Wash tub or bucket
4. Data sheets and identification tags
5. Shipping container with crushed ice
Procedure
1- Lower the sampling device through the water and into the sedi-
ment. Retrieve collected sediment sample.
2. Empty contents of sampler directly into precleaned glass jars
or into bucket.
3. As needed, remove leaves, sticks, and rocks from sediment before
transfer to quart widemouth jar. Store on ice for shipment to
laboratory.
Comments
A bottom-material sampler must be capable of collecting and retaining
the "fines" which sometimes contain the highest concentration of pesticide
residues. In large lakes or estuaries, bottom sediment samples are usually
collected with various types of dredges, including the Ekman and Petersen
dredges. The Ekman dredge is widely and sucessfully used for soft bottoms.
The Petersen dredge is less capable of retaining fine sediments but is more
suited for collecting samples of hard bottoms such as sand, grave, marl and
similar materials. The dredge-type weighted sampler is acceptable for use
in lakes and slow-moving streams having little suspended sediment. A cor-
ing tube is excellent for most shallow sediment sampling as it provides a
relatively undisturbed sample and can be sectioned if desired. Scuba div-
ers can collect relatively undisturbed samples using coring tubes in deeper
waters.
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V. WATER
This section describes the supplies, equipment and techniques required
to collect, preserve, and ship fresh water samples for analysis of pesti-
cide residues.
GRAB WATER SAMPLING
Grab water sampling is an effective method to determine runoff prob-
lems or overspray into streams by detecting pesticide contamination in wa-
ter bodies. Time and location of sampling may be critical and needs care-
ful consideration, but collection techniques are simple. Cost varies with
equipment from < $1 for a jar to $200 for a Kemmerer, Van Dorn, or APHA
sampler. These samplers are available from scientific equipment supply
companies like Kahl Scientific.
Apparatus
1. Water sampling device, which in the case of a widemouth glass
quart jar with teflon-lined lid, also becomes the sample container.
Do not use plastic bottles, and avoid any sample contact with
plastics.
2. For sampling where bottle cannot be dipped directly, a commercial
water sampler, including weighted bacteriological water sampler,
Kemmerer, Van Dorn, or APHA sampler.
Procedure
1. Fill quart jar with water by dipping at the water surface. Many
pesticides remain on the surface of water, especially oil-based
types. In some instances, begin sampling downstream and collect
series of samples progressively closer to the source.
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2. If commercial sampler is used, collect sample at location and
depth desired and transfer to widemouth glass quart jar with
teflon-lined lid. Clean sampler after each use.
3. In general, keep samples cool, preferably on ice and in the
dark. Holding time will depend on the pesticide and chemical
analysis required.
COMPOSITE WATER SAMPLING
Composite water sampling is an effective method to collect a series of
water samples to detect pesticide contamination in surface waters. For
manual sampling the only cost is for the sampling container, but with an
automatic sampler such as a Manning pictured below, the cost is much high-
er ($3,200).
1. Same as for grab sampling if done manually
2. Automatic sampling requires sampler such as Manning with glass-
ware
3. Data sheets and identification tags
4. Shipping container with crushed ice
Apparatus
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V-3
Procedure
1. Manually dip each sample and add to composite sample container.
2. With automatic sampler, assemble glass jars and set timing mechan-
i sm.
3. Composite collected samples based on application events and/or
time-of-travel studies.
4. Seal jar, label, and place on ice for transport to the laboratory.
TIME-OF-TRAVEL STUDIES
Time-of-travel studies aid in establishing water sampling locations
and time of sampling by determining water flow rate over desired distance.
Cost for surface floats and a stopwatch can be less than $50, while a flu-
orometer and dye can cost more than $2,000.
Apparatus
1. Surface floats: chips, oranges, styrofoam or similar objects,
stopwatch
2. Tracer dye: fluorescein or rhodamine dye, fluorometer
Procedure
1. Surface Floats: Average the time of three trips by surface
floats placed mid-stream over entire distance or a representative
portion.
2. Tracer dye: Inject dye at mid-stream and time travel to desired
point until initial, highest, and/or last reading is obtained.
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VI. FISH
This section describes the supplies, equipment, and techniques re-
quired to perform acetylcholinesterase inhibition tests on fish and to col-
lect, preserve, and ship fish samples for analysis of pesticide residues.
ACETYLCHOLINESTERASE INHIBITION TESTS (AChE)
The AChE test can demonstrate sublethal effects of cholinesterase in-
hibiting substances such as carbamate and organo-phosphate pesticides on
fish, sometimes at or below laboratory residue analysis detection limits.
Cost can be less than $10 for fish and cages but laboratory equipment (pH
stat) is expensive. A basic instrument suitable for AChE tests costs about
$3,000. Glassware and accessories will cost another $1,000 to $1,500.
Apparatus
1. Ten fish per sample station (4- to 6-inch)
2. Fish cages capable of holding ten fish
3. Live box with aerator
4. Field laboratory equipped with pH-stat, chemicals, glassware,
balance, etc.
Procedure
1. Expose 10 fish per cage in stream for a minimum of 96 hr including
the day of spraying and the 3 following days.
2. After exposure, remove fish from cages and transport them live
to a field laboratory.
3. For laboratory analyses, remove fish brains, weigh them, and
measure AChE activity.
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4. Compare results to control fish and published values in the liter-
ature.
Comments
Acetylcholine (ACh) is a key compound in nerve function. It is most
probably connected with the transmission of impulses along nerves and
across synoptic spaces which are small clefts between one neuron and the
next. The widith of these clefts is approximately 150-500 Angstroms. When
a nerve is stimulated, ACh is released from an inactive bound form and is
then quickly converted to its non-active components, choline and acetic
acid, by the hydrolyzing enzyme acetylcholinesterase (AChE). Specific in-
hibitors of the enzyme AChE, such as organophosphate and carbamate pesti-
cides, permit toxic amounts of free acetylcholine to accumulate. This
leads to a continual stimulation of major portions of the nervous system
resulting in convulsion, coma, and finally death.
The normal level of activity for AChE (expressed as the rate at which
the enzyme can hydrolyze acetylcholine) in the nervous tissue, particularly
the brain, is fairly constant for a particular species of animal. Any ap-
preciable decline in this level is a strong indication of the presence of a
specific inhibitor.
Acetylcholinesterase levels of activity are assayed using the pH stat
method.* This method measures the acetic acid formed by the hydrolysis of
the substrate acetylcholine by the enzyme AChE. The brains of five fish
are pooled, weighed wet, and homogenized in distilled water. The brains
are then further diluted with distilled water to a tissue concentration of
5 mg/m£. Then 4 m£ of the diluted homogenate are mixed with 4 m£ of 0.03 M
acetylcholine iodide which provides a specific substrate for the enzyme
AChE. A recording pH stat is used to titrate the liberated acetic acid
with 0.01 N NaOH. The reaction is carried out at the predetermined
* Coppage's Procedure - U.S. Environmental Protection Agency. Gulf Breeze
Environmental Research Laboratory - Gulf Breeze, FL
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optimum pH and temperature with a nitrogen purge over the surface of the
reaction vessel to prevent adsorption of atmospheric carbon dioxide. The
micromoles of acetylcholine hydrolyzed are calculated; from the micromoles
of NaOH required to neutralize the free acetic acid. AChE activity is ex-
pressed as micromoles of acetylcholine hydrolyzed per hour per mg of brain
ti ssue.
All samples are analyzed in duplicate, and mean values are calculated.
FISH COLLECTION
The first step in an investigation involving the suspected contamina-
tion of fish by pesticides is to immediately collect samples of the fish
and water. Next, the stream or lake should be inspected to delineate the
affected area and to determine the possible source(s) of the pesticide con-
tamination. Sources to be considered are sewage outfalls, irrigation re-
turn water, spray and mixing operations, industrial water, industrial ef-
fluents, and mining wastes. Tributaries should be examined if the source
of pollution cannot be found on the main stream.
In the event of a fish kill, the investigator must determine: (1)
cause of kill which is often determined only after analytical results are
available, (2) areal extent of kill, (3) numbers and kinds of fish in-
volved, (4) time and circumstances of kill, (5) other water quality parame-
ters extant at the time of kill and during the investigation, and (6) other
immediate hazards to downstream users including consumers of the water.
Several parameters are useful in describing a fish kill. Important
ones that should be measured are temperature, dissolved oxygen, pH, alka-
linity, hardness, conductivity, and turbidity.
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Methods of sample collection vary with the type of analyses to be per-
formed. Ideally, samples of the suspected source should be collected along
with water both upstream from the suspected discharge and downstream of
this point after the discharged material has become thoroughly mixed in the
stream.
Collecting fish can help determine the cause of a fish kill or bioac-
cumulation in fish living in waters chronically exposed to pesticides.
Cost can vary from less than $20 for a dip net to $1,000 for an electro-
shocking unit.
Apparatus
1. Long-handled dip net (Turtox #105 A28) or 35 ft. tied bag seine
(Nichols Net and Twine Co., East Saint Louis, Illinois) or Elec-
troshocking outfit (Coffelt Electronics, Englewood, Colorado) or
fishing tackle
2. Acetone-washed aluminum foil
3. Data sheets and identification tags
4. Shipping container with dry or crushed ice
Procedure
1. Collect fish by most appropriate method
2. Collect about 1 lb of each type of sample/species from similar
size fish within each species. Wrap muscle, viscera, or whole
fish samples in aluminum foil.
3. Transport on ice or freeze the samples if shipment or analysis
must be delayed
-------
VI-5
Comments
Useful information in tish-kill or pesticide-contaminated fish inves-
tigations can be obtained by examining fish collected from the area. Fish
samples taken upstream of the suspected toxicant source allow the analyst
to establish normal pesticide background levels in critical tissues, and
these sample fish should be labeled and tagged with the same care as other
samples.
Often it is useful to collect blood for analytical purposes from fish
obtained both upstream of, and in the immediate pesticide-polluted area.
Blood analyses of fish from these sources is often helpful in documenting
that pesticides are a real or potential problem. For suspected chronic
pesticide contamination, collect viscera and fat from fish for analytical
purposes.
Blood may be collected in clean vials obtained at a drug store.
Ideally, a minimum of 5 mSL should be collected from each fish; otherwise
composite the blood from two or more fish until the 5 m£ volume is reached.
Blood is easily collected from live fish by drying them with paper
towels, severing the tail 5 to 10 mm in front of its base, and allowing the
blood to drip from the caudal artery into the vial. Fish blood clots ra-
pidly, and should a clot appear before the fish is thoroughly bled, it can
be removed by recutting or wiping the severed surface with a paper towel.
Vials should be capped, labeled, tagged, packed in ice (but not frozen),
and transported to the laboratory within 24 hr for analysis or refrigerated
storage.
The number of distressed or killed fish is also important to deter-
mine. Ascertain the number of miles of stream affected by the pesticide
pollution and record the information on a topographical or other suitable
map. Count all dead or distressed fish in the area, if possible or practi-
cal. Any partial count must be unbiased. The following is an appropriate
procedure:
-------
Vl-6
Make 100-yard counts at half-mile intervals for the entire
stream distance involved, beginning with the first distressed
or dead fish observed. In the case of a very large kill cover-
ing many miles of a river, it may be necessary to make counts
at one-mile intervals. Pictures and aerial photographs may be
useful. Regardless of interval, proceed as follows:
1. Identify and count all fish in each 100-yd segment of
stream
2. Calculate by species the total number of fish killed and
the total number distressed in each count area
3. Multiply each total of each species by 8.8 or 17.6 (the
number of 100-yd segments/half-mile or mile)
4. Numbers obtained in item 3 represent an estimation of
the total kill and total distressed fish for each species.
This count method has many drawbacks, problems, or weaknesses Dead
fish tend to accumulate in stream areas where flow is low, such as backwa-
ters or eddys. Distressed fish are almost impossible to count without net-
ting. The same fish may be recounted as it resurfaces. Extrapolating the
count to a per-mile number may lead to large errors.
-------
VII-1
VII. BENTHOS
This section describes the supplies, equipment, and techniques re-
quired to perform aquatic invertebrate sampling. Benthos are excellent for
evaluating environmental impact of pesticides in water but require signifi-
cant background sampling and available taxonomic expertise. Benthic sam-
pling devices include the Surber sampler ($125), Ekman dredge ($150), Pe-
tersen dredge ($250), and Ponar dredge ($220).
Apparatus
1. Benthic sampling device (dip net, Surber sampler, Petersen dredge,
etc.)
2. #30 mesh seive
3. Enamel pan, 6" x 12" x 2"
4. Forceps
5. Small glass jars or vials
6. Alcohol or dry ice
7. Data sheets and identification tags
8. Shipping container
Procedure
1. Collect sample with device chosen.
2. Sieve out sediment, pick out rocks, sticks.
3. Use enamel pan to field pick and separate sample into basic
taxonomic groups.
4. Place sample in jars or vials, label, and either freeze for
residue analysis or preserve in alcohol for population analysis.
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VIII-1
VIII. PLANTS
This section describes the supplies, equipment, and techniques re-
quired to collect, preserve, and ship plant samples to be analyzed for pes-
ticide residues. Plant samples are useful for determining overspray or
pesticide drift into sensitive areas by detecting pesticides on or in
plants. Cost should be less than $10 in miscellaneous supplies.
Apparatus
1. Shovel, spade, cultivating fork, and clippers
2. Acetone-washed aluminum foil
3. Plastic bags
4. Data sheets and identification tags
5. Shipping container with crushed ice
Procedure
1. Remove plant part or uproot entire plant.
2. Wrap in foil and place in plastic bag with identification.
3. Ship to laboratory in container with crushed ice.
Comment
Pesticide may occasionally be visibly apparent on leaf surfaces.
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IX-1
IX. WILDLIFE AND LIVESTOCK
This section describes the supplies, equipment, and techniques required
to collect, preserve, and ship wildlife and livestock samples for analysis
of pesticide residues.
Apparatus
Tissues
1. Widemouth glass bottles, 2V16" high x l1/4" diameter, (Owens-
Illinois mold number AN-6764), with screw caps having teflon or
foil liners
2. Data sheets, identification tags
3. Shipping container with crushed ice
Blood
1. Glass vials, 60 x 17 mm, 10 m£ (Arthur H. Thomas Co. catalog num-
ber 9802-G, Caps are catalog number 2849-A, size 15). Use caps
with teflon or foil liner.
2. Ten-mi Vacutainer tubes (Scientific Products number B2985-4A),
tube holder (Scientific Products number B-3023-110) and needles
(Scientific Products number B-3029)
3. Data sheets, identification tags
4. Shipping container with crushed ice
Urine
1. Glass urine specimen bottle (Scientific Products number B-7925 with
B-7931 caps)
-------
IX-2
2. Data sheets, identification tags
3. Shipping container with crushed ice
Stomach Contents
1. Widemouth glass quart jars (Mason type) with teflon or foil-lined
lids
2. Data sheets, identification tags
3. Shipping container with crushed ice
Procedure
The Agency representative must make arrangements to have a local vet-
erinarian collect blood and tissue samples from live animals. This step is
necessary to protect the Agency against possible legal charges of trespass
which might be brought by the farmer or his agents. Obtain permission from
the livestock owner. In the case of wildlife samples, notify the State
Fish and Game Department of the purpose of the sample collection, and ob-
tain appropriate collection permits.
For the following classes of compounds, sampling requirements are as
listed:
Organophosphate Compounds
1. Two 10-m.fc Vacutainer tubes of blood from live animals
2. 25 m£ of urine from live animals
3. 25 m£ of blood from dead animals (probably clotted)
4. 25 m£ of urine from dead animals, if possible
5. 1 to 2 oz of adipose tissue from dead animals
6. 1 to 2 oz of brain tissue from dead animals, if possible
7. 1 lb of stomach content material (from the rumen)
-------
IX-3
Orqanochlorine Compounds
Sampling requirements are identical to those for organophos
phates, except that brain tissue is not needed.
Carbamate Compounds
25 m£ of urine from live animals
Other Compounds
Other compounds such as herbicides, rodenticides, fungi-
cides, etc., are highly variable and no general guidelines are
suitable. Under these conditions, alternate instructions should
be solicited from the National Enforcement Investigations Center
staff (see page 1-1 of this document).
With the samples collected and properly labeled, place in shipping
container with crushed ice and transport to laboratory within 24 hr.
-------
x-l
X. HUMAN TISSUES
This section deals with methods for obtaining blood and urine samples
from human subjects, types of equipment used, and sample preservation and
shipping.
Apparatus
Blood
1. 10-m£ Vacutainer® tubes (Scientific Products Number B2985-4A),
tube holder (S.P. number B-3023-110), and needles (S.P. number
B-3029)
2. Data sheets, identification tags
3. Shipping container with crushed ice
Urine
1. Glass urine specimen bottle (S.P. number B-7925 with B-7931
caps)
2. Data sheets, identification tags
3. Shipping container with crushed ice
Procedure
Where pesticide applications are suspected of causing a health hazard
or actually poisoning humans, it may be necessary to collect samples of
blood, urine, or tissues from the persons affected.
The following general guidelines constitute only suggested information
and procedures. In all cases, before obtaining samples from human subjects,
-------
X-2
obtain legal counsel and medical assistance to protect all parties in-
volved.
The legal aspects are very important in obtaining blood and urine sam-
ples from human subjects. In any medical research involving human sub-
jects, the legal considerations focus on informed consent of the individ-
ual. Consent alone is not sufficient to protect the researcher. A subject
must give "informed consent". Informed consent is clearly defined in the
HEW "Guidelines on Protection of Human Subjects" (Appendix A), which have
been adopted by the Environmental Protection Agency as follows:
"Informed consent means the knowing consent of an individual or his
legally authorized representative, so situated as to be able to exer-
cise free power of choice without undue inducement or any element of
force, fraud, deceit, duress, or other form of constraint or coercion.
The basic elements of information necessary to such consent include:
1) A fair explanation of the procedures to be followed, and their
purposes, including identification of any procedures which are
experimental;
2) A description of any attendant discomforts and risks reasonably
to be expected;
3) A description of any benefits reasonably to be expected;
4) A disclosure of any appropriate alternative procedures that might
be advantageous for the subject;
5) An offer to answer any inquiries concerning the procedures; and
6) An instruction that the person is free to withdraw his consent
and to discontinue participation in the project or activity at
any time without prejudice to the subject. (HEW "Guidelines",
546.3(c))
Adherence to the following procedures is recommended; however, as sta-
ted earlier, legal counsel should be solicited case-by-case:
In a suspected pesticide poisoning case, request the patient's
doctor to sample blood and urine. A written consent from the
-------
X-3
patient should be obtained by the doctor. Such consent should
be specific about reasons for the sampling and should meet the
requirements of the HEW definition above. In cases where a pa-
tient is unconscious, an informed consent must be obtained by
someone legally authorized to provide such consent.
In general, consent forms are given to a patient by his/her physician
or by hospital personnel. It is safest, legally, to follow standard proce-
dures. Speak with the patient's doctor about sample needs and obtain his
approval of the Agency's plans. The doctor will explain the request to the
patient and ask his/her permission.
While it is clear that Agency personnel cannot directly collect a sam-
ple, the dictation of types of samples would still be an Agency preroga-
tive. For the following classes of compounds sample needs are:
Qrqanophosphate and Organochlorine Compounds
®
1. Two 10-m£ Vacutainer tubes of blood
2. 25 m£ of urine
Carbamate Compounds
1. 25 m£ of urine
Other Compounds
1. Two 10-m£ Vacutainer tubes of blood
2. 25 m£ of urine
-------
XI-1
XI. SAMPLE CONTROL PROCEDURES
Standard sample collection procedures, as outlined in this guide, will
ensure sample integrity during collection, transportation, storage, and
analysis. Sampling procedures, as well as documented chain-of-custody pro-
cedures, protect against misidentification, loss, or error of data relating
to collection theft, loss, damage, or alteration of the sample.
The chain-of-custody procedure may vary depending on the type of sam-
ple collected. The goal is to provide a traceable record of what happened
to the sample from time of collection to introduction as evidence in legal
proceedings. Sampling procedures and sample possession both need to be
documented. The NEIC Policies and Procedures Manual details NEIC chain-of-
custody procedures (See Appendix).
-------
XII-1
XII. BIBLIOBRAPHY
PESTICIDE DRIFT REFERENCES
1. Akesson, N.B. and W.E. Yates. 1964. Problems relating to application
of agricultural chemicals and resulting drift residues. Ann. Review
Entomol. 9:285-318.
2. Akesson, N.B., S.E. Wilce, and W.E. Yates. 1971. Continuing aerial
applications to treated fields - a realistic goal. Agrichemical Age,
December 1971. pp. 11-14.
3. Akesson, N.B., W.E. Yates, and S.E. Wilce. 1972. Needed: Better
drift control. Agrichemical Age, December 1972.
4. Cheng, L. 1977. Stain method for measurement of drop size. Environ.
Sci. Technol., 11(2):192-194.
5. Christensen, P., W.E. Yates, and N.B. Akesson. 1971. Meteorology and
drift. Proc. 4th Int. Agric. Aviat. Congr. (Kingston, 1969). pp. 337-
345.
6. Courshee, R.O. 1960. Some aspects of the application of insecticides.
Ann. Review of Entomol. 5:327-352.
7. Coutts, H.H. and W.E. Yates. 1968. Analysis of spray droplet distri-
bution from agricultural aircraft. Trans. Am. Soc. Agric. Engin. 11(1):
25-27.
8. Davis, J.M. 1953. A rapid method for estimating aerial spray depos-
its. J. Econ. Entomol. 46(4);696-698.
9. Maybank, J. and K. Yoshida. 1969. Delineation of herbicide-drift haz-
ards on the Canadian praries. Trans. Am. Soc. Agricul. Engin. 12:759-
752.
10. Maybank, J. et al. 1977. Spray drift and swath deposit pattern from
from agricultural pesticide applications; report of the 1976 field tri-
al program. Physics Division, Saskatchewan Research Council, Saskatoon.
60 pp.
11. Maybank, J., K. Yoshida, and R. Grover. 1978. Spray drift from agri-
cultural pesticide applications. 0. Air Pollut. Conf. Assn. 28(10):
1009-1014.
12. Rathburn, C.B. 1970. Methods of assessing droplet size of insectici-
dal sprays and fogs. Mosquito News, 30(4):501-513.
-------
XI1-2
13. Scriven, R.A. and B.E.A. Fisher. 1975. The long-range transport of
airborn material and its removal by deposition and washout - I. Gen-
eral considerations. Atmospheric Environ. 9:49-58.
14. Scriven, R.A. and B.E.A. Fisher. 1975. The long-range transport of
airborn material and its removal by deposition and washout - II. The
effect of turbulent difussion. Atmospheric Environ. 9:59-68.
15. Wallace, K. and K. Yoshida. 1978. Determination of dynamic spread
factor of water droplets impacting on water-sensitive paper surfaces.
J. Colloid Interface Sci. 63(1):164-165.
16. Yates, W.E. and N.B. Akesson, 1963. Fluorescent tracers for quantita-
tive microresidue analysis. Trans. Am. Soc. Agric. Engin. 6:104-114.
17. Yates, W. E. and N.B. Akesson. 1966. Selection of spray equipment and
operating techniques for agricultural aircraft. Proc. Third Interna-
tional Agric. Aviaition Congress. March 15-18. Arhem, Netherlands,
pp. 251-277.
18. Yates, W.E., N.B. Akesson, and R.F. Cowden. 1974. Criteria for mini-
mizing drift residues on crops downwind from aerial applications.
Trans. Am. Soc. Agric. Engin. 17:627-632.
19. Yoshida, K. and J. Maybank. 1969. Determination of herbicide spread
factors. Can. Agricut. Engin. 11(2):66"70.
EQUIPMENT REFERENCES
1. Kromecote cards, 4" x 5", approximately $10/100 sheets from Industrial
Chemical Corporation, 4711 W. 58th Avenue, Arvada, CO 80002; phone
(303) 427-2727.
2. Linagraph paper, 6" x 200'-roll, approximately $30/200 ft.-roll from
Eastman Kodak; 2800 Forest Lake, Dallas, IX 75234; phone (214) 241-1611.
3. Multi-spectrum 209 copy type 640 paper, 8%" x 11", approximately $25/700
sheets from 3M office supply distributors.
4. Information on other equipment available upon request.
-------
APPENDIX
SAMPLE CONTROL PROCEDURES
(Excerpts from NEIC Policies and Procedures Manual)
-------
A-l
11-11
(10/79)
SAMPLE CONTROL
A sample* is physical evidence collected from a facility or from
the environment. An essential part of all NEIC enforcement investi-
gations is that the evidence gathered be controlled. To accomplish
this, the following sample identification and Chain-of-Custody pro-
cedures have been established.
SAMPLE IDENTIFICATION
The method of identification of a sample depends on the type of
measurement or analyses performed. When in-situ measurements are
made, the data are recorded directly in logbooks or Field Data Records
(FORs), with identifying information (project code, station numbers,
station location, date, time, samplers), field observations and
remarks. Examples of in-situ measurements are pH, temperature,
conductivity, flow measurement, continuous air monitoring, and stack
gas analysis.
Samples, other than in-situ measurements, are identified by a
sample tag (page 11-12) or other appropriate identification (herein-
after referred to as a sample tag).
These samples are removed from the sample location and trans-
ported to a laboratory or other location for analysis. Before re-
moval, however, a sample is often separated into portions depending
upon the analyses to be performed. Each portion is preserved in
* For purposes of this manual, the term 'sample' includes remote
sensing imagery.
-------
A-2
Pioieci Code
Station No
Monih/Day/Year
Time
Designate
Com© Crao
z
1
O £
to I
4^
00
r~
a
3
o
o
2
o
Station Location
Samplers (Signatures)
Remarks
00
tu
o
n
o_
o
c
»<
Mutagenicity
"0
o
l/l
o
Q.
CO
00
Volatile Organics
Priority Pollutants
o
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O)
o
(/>
O
O
N.
2
CO
Oil and Grease
(Cyanide
Metals
5
CO
o
c:
»<
| Phenolics
COD. TOC Nutrients
00 CD
O o
a o
(A
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§
—
ANALYSES
Preservative:
Yes ~ No ~
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
NATIONAL ENFORCEMENT INVESTIGATIONS CENTER
Building 53, Box 25227, Denver Federal Center
Denver, Colorado 80225
®EPA
SAMPLE TAG
Updated from 10/79 Manual
-------
A-3
11-13
(10/79)
accordance with applicable procedures and the sample container is
identified by a sample tag. The information recorded on the sample
tag includes:
Project Code
Station Number
Date
Time
Station Location
Samplers
Tag Number
Remarks
- A three-digit number assigned by NEIC
- A two-digit number assigned by the Project
Coordinator and listed in the project plan
- A six-digit number indicating the year,
month, and day of collection.
- A four-digit number indicating the military
time of collection - for example: 0954
The sampling station description as specified
in the project plan
Each sampler is identified.
A unique serial number is stamped on each tag
The samplers record pertinent observations.
The tag used for water samples (also soil, sediment and biotic
samples) contains an appropriate place for designating the sample as
a grab or a composite, and identifying the type of sample collected
for analyses. The tag used for air samples requires the sampler to
designate the sequence number and identify the sample type. The
Project Coordinator will detail procedures for completing tags used
for soil, sediment and biotic samples. The sample tags are attached
to or folded around each sample.
After col lection, separation, identification, and preservation,
the sample is maintained under Chain-of-Custody procedures discussed
below. If the composite or grab sample is to be split, it is aliquo-
ted into similar sample containers. Identical sample tags are com-
pleted and attached to each split and marked " Split". The
tag identifies the split sample for the appropriate government agency,
-------
11-14
(10/79)
facility, laboratory, or company. In a similar fashion, all tags on
blank or duplicate samples will be marked "Blank" or "Duplicate" re-
spectively.
CHAIN-OF-CUSTODY PROCEDURES (March 29, 1978)
Due to the evidentiary nature of samples collected during en-
forcement investigations, possession must be traceable from the time
the samples are collected until they are introduced as evidence in
legal proceedings. To maintain and document sample possession,
chain-of-custody procedures are followed.
Sample Custody
A sample is under custody if:
1. It is in your possession, or
2. It is in your view, after being in your possession, or
3. It was in your possession and then you locked it up to
prevent tampering, or
4. It is in a designated secure area.
Field Custody Procedures
1. In collecting samples for evidence, collect only that
number which provides a good representation of the media
being sampled. To the extent possible, the quantity and
types of samples and sample locations are determined prior
to the actual field work. As few people as possible should
handle samples.
-------
A-5
11-15
(10/79)
2. The field sampler is personally responsible for the care
and custody of the samples collected until they are trans-
ferred or dispatched properly.
3. Sample tags shall be completed for each sample, using
waterproof ink unless prohibited by weather conditions. For
example a logbook notation would explain that a pencil was
used to fill out the sample tag because a ballpoint pen
would not function in freezing weather.
4. The Project Coordinator determines whether proper custody
procedures were followed, during the field work and decides
if additional samples are required.
Transfer of Custody and Shipment
1. Samples are accompanied by a Chain-of-Custody Record (see
pages 11-16 and 11-17). When transferring the possession of
samples, the individuals relinquishing and receiving will
sign, date, and note the time on the Record. This Record
documents sample custody transfer from the sampleroften
through another person, to the analyst in a mobile labora-
tory, or at the NEIC laboratory in Denver.
2. Samples will be packaged* properly for shipment and dis-
patched to the appropriate NEIC laboratory** for analysis,
with a separate custody record accompanying each shipment
(e.g., one for each field laboratory, one for samples
* See Appendix B
** See Appendix C for Safety Precautions When Accepting Samples
From Outside Sources.
-------
ENVIRONMENTAL PROTECTION AGENCV NATIONAL ENFOflCEMflPJT INVESTIGATIONS CENTER
OHtce of Enforcement Bwild'n9 53 Bo* 20227 Denver Federal Comer
CHAIN OF CUSTODY RECORD Denver Colorado E02"25
Relinquished by fS'gnjturtt
Oaie
t Time
Received Dy fSisnituref
Relinquished by (S'fnjturtf
Dale
Time
Received by (Signnurei
Relinquished by tS>9»*tuteS
Date,
Time
Received by IS'enjtunl
Relmqunhed by tS'sntrvrtf
03ie / Time
Received by fSignstur*}
flelinquishccl by {S
-------
A-7
11-18
(10/79)
driven to Denver). Shipping containers will be padlocked
for shipment to the Denver laboratory. The method of ship-
ment, courier name(s) and other pertinent information is
entered in the "Remarks" box.
3. Whenever samples are split with a source or government
agency, a separate Chain-of-Custody Record is prepared for
those samples and marked to indicate with whom the samples
are being split. The person relinquishing the samples to
the facility or agency should request the signature of a
representative of the appropriate party, acknowledging
receipt of the samples. If a representative is unavailable
or refuses to sign, this is noted in the "received by" space.
When appropriate, as in the case where the representative
is unavailable, the custody record should contain a statement
that the samples were delivered to the designated location
at the designated time.
4. All shipments will be accompanied by the Chain-of-Custody
Record identifying its contents. The original Record will
accompany the shipment, and a copy will be retained by the
Project Coordinator.
5. If sent by mail, the package will be registered with return
receipt requested. If sent by common carrier, petty cash
will be used for expenditures of less than $100, otherwise
a Government Bill of Lading will be used. Air freight
shipments are sent collect. Freight bills, post office
receipts and Bills of Lading will be retained as part of
the permanent documentation.
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