-------
1s required, reoccupled. Records that should be kept are 1) positioning method
and equipment used, 2) names of responsible persons and their duties, 3) loca-
tion of equipment on board the sampling vessel, 4) modifications In Methods
or equjpment used from those described In the sample plan, and 5) data on the
calibration procedures and frequency of calibration. In addition, the occupied
stations should be recorded In the log book, and plotted and numbered on the
most accurate and up-to-date map of the area. Such maps can be obtained from
the U.S. Geological Survey and National Ocean Survey.
3.1.2.2 Operation of Sampler and Criteria far Sample Acceptance
To minimize twisting of the sampler and to ensure proper contact of the
sampler with the bottom, the sampling device should be attached to the cable
using a ball bearing swivel. The sampler should be lowered through the water
column at a controlled rate of approximately 1 ft/s and never allowed to free
fall. Free falling could result 1n premature triggering of the device,
excessive bow wake, or Improper contact with the sediment surface. Ideally,
the sampler should only gently contact the sediment with minimal disturbance
and be forced Into the sediment only by the weight of the sampler. After the
sample 1s collected, 1t should be raised slowly from the bottom at a controlled
speed of approximately 1 ft/s. To minimize swinging of the sampler when it
breaks the surface, the sample vessel should head Into the waves. When the
sample 1s brought to the surface, the outside of the sampler should be care-
fully rinsed with clean water 1) to remove any material on the outside that
could contaminate the sample during removal and 2) to permit the sampler to be
visually Inspected to determine the sample acceptability. The sampler should
be secured.Immediately after 1t Is brought on board to avoid sample tipping,
spilling, or disturbance. Excessive swinging of the sampler, striking the
vessel, or sampler tipping could result In unacceptable sample disturbance.
If the sample falls any of the following acceptance criteria 1t should
be rejected and another sample taken:
» •
• Sampler 1s not'leaking.
• Desired penetration depth Is achieved.
15
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• Sediment surface 1s flat and does not show signs of disturbance or
washout. •
• Sample surface Is not pressed against the top of the sampler.
• • e Overlying water 1s present-
• Overlying water 1s not turbid.
• Anthropogenic material 1s not evident (I.e., bottles, cans, etc.).
I
3.1.2.3 Sample Collection
After the sample 1s determined to be acceptable, Information on the
!
sample should be recorded 1n the field log book (Section 3.1.2.7).
i
Before sediment samples are taken for analysis or compositing and
mixing, the overlying water must1be removed from the sampler by slowly siphoning
1t off near one side of the sampler. Care should be taken to ensure that the
sediments are not disturbed, and:that fine-grained surfldal sediment and
organic matter are not lost while reaovlng the overlying water. Once the
overlying water 1s removed, the sediment can be sampled. To prevent contami-
nation during sample collection, samples should be taken only from the center
p-jftlc-a of the saiplsf tc- avoid ^ct^otlal €ont.as1?!£t1on frow- contact with the
" _ • s
sampler. In addition, the sawples should only come 1n contact with the cleaned
sampling equipment and should not be touched with hands that are not gloved.
The samples are placed efther 1ni the appropriate sample container or in a
stainless steel bowl for compositing and mixing.
3.1.2.4 Mixing of Samples
I
When removing subsaaples for different chemical analyses of the same
sediment sample or when combining samples from several sediment grabs to
provide sufficient material for [analysis, the sample should be thoroughly
mixed. The samples can be composited and mixed by transferring them to a
dryi solvent-rinsed stainless s^eel bowl and stirring with a clean stainless
steel spoon or spatula until achieving homogeneous color and texture, the
bowl and all utensils used for islxlng should be changed after each sample or
at least solvent rinsed with methylene chloride between uses and covered with
foil to prevent airborne or other contamination. The compositing, mixing,
16
-------
and subsampUng should be completed as soon as the samples are collected.
However, 1f a clean room or clean area 1s not available, then compositing and
mixing should not be attempted due to the potential for contamination, but
should be completed as soon as possible at the laboratory or other clean
facility.
3.1.2.5 Total Organic Carbon and Dry Weight Sample
A minimum of 25 g of sediment sample should be collected 1n either a
glass or plastic container that has been properly cleaned.
3.1.2.6 Contaminant Sample
The sediment sample to be used for analyzing semi volatile compounds,
Including the nonpolar organic contaminants, should be collected 1n 240-mL or
larger, wide-mouth glass Jars with Teflon-lined screw Hds. The sample jar
must be properly cleaned to prevent contamination of the sample. The sediment
sample should be at least 200 g (wet weight). Filling the wide-mouthed Jar
approximately three-quarters full will ensure obtaining at least this amount
of material.
3.1.2.7 Sample Custody Information
After the sample has been collected and stored In the appropriate con-
tainer, all relevant data pertaining to Its collection should be documented
1n the field log book. Information to be documented should Include the
following:
• unique sample number
• station location
• date of collection
• depth of water
• gross characteristics, Including texture, color, presence of
organisms, presence of debris, presence of oily sheen, and odor
• gross characteristics of the vertical profile of the sediment.
Including changes 1n characteristics and presence and depth of a
redox potential discontinuity layer
17
-------
• penetration depth of sampler
• depth of sample
• comments on sample quality.
«•
Cha1n-of-custody procedures and forms should be prepared that allow for
the documentation of the samples! and their status at every stage In the process
from collection through final analysis. An example of a chaln-of-custody and
analysis request fora was given jln Figure 1. In addition, waterproof labels
containing the sample number, preservation techniques, date and time of collec-
tion, location of sample, and signature of the collector should be affixed to
each bottle. All writing on these labels should be done with Indelible Ink.
3.1.3 Sample Preservation and Shipping
- Immediately after collection, the samples should be refrigerated (4°C)
or placed 1n shipping containers; with dry 1ce and stored 1n the dark. Samples
should be analyzed within 7 days>* however, 1f the analysis cannot be performed
within 7 days, the samples should be frozen to -20*C or to -80eC, 1f possible.
Freezing Is required to reduce the potential for m1crob1a1 activity. Care
must be taken with frozen samples to prevent container breakage by leaving
headspaee fsr tha satsr ts osp4B&. Usually, this Is accomplished by freezing
the containers at an angle rather than In'an upright position. Appropriate
handling times have not been established for frozen sediment, although 6 to
12 months are generally conslderisd to be acceptable. Although freezing may
alter the sediment matrix, TetraiTech (1986) presents data that suggest that
the effects are minimal.
If possible, all samples should be delivered to the analytical
laboratory as soon as sampling 1s completed to ensure that the samples are
analyzed within 7 days. If sample delivery to the laboratory 1s delayed,
then storage procedures described previously must be followed aind documented.
If the samples contain hazardous)materials, guidance for shipping can be found
In U.S. Department of Transportation (1984). Procedures that should be followed
1n preparing the samples for shipping or transportation are as follows:
• Containers should be durable and be able to withstand rough
treatment during shipping.
t "|
i 18
-------
• Samples should be tightly packed 1n shipping containers with
dividers and, depending on the shipping time, the space between
the bottles filled with packing material or dry 1ce.
• 'The original cha1n-of-custody and analysis request form should be
enclosed 1n protective packaging and placed within the shipping
container. Copies of the form should be retained.
• After the samples and forms have been placed 1n the shipping
container, a custody seal, and •FRAGILE' and -THIS END UP' labels
should be placed on the outside of the shipping container.
• If the samples must be shipped to a distant city, carriers that
provide tracking of shipments and delivery receipts should be
used to confirm that the shipment was delivered as required and
to serve as additional cha1n-of-custody Information. All shipping
charges should be prepaid by the shipper to prevent delay 1n
shipping.
• Notice of receipt of the samples should be received from the
analytical laboratory.
3.2 ANALYTICAL PROCEDURES
Before analyzing the sediment sample, the analytical laboratories must
be evaluated to determine that they can perform the desired analyses within
established guidelines. The review should Include an evaluation of the labora-
tory's Instrument capability and the level of staff experience. In addition,
the program manager will need to establish minimum QA/QC requirements with
the laboratory before the work 1s done. Evaluation procedures should Include
analysis of standard reference materials and analysis of replicate samples to
establish the quality of a laboratory's analytical capability (e.g., accuracy
and precision of their data). Based on the review of data from the available
laboratories, the most appropriate analytical laboratory can be selected.
The minimum QA/QC requirements should be consistent with thos.e established by
the EPA's Contract Laboratory Program.
Figure 2 depicts the general approach that 1s recommended for the deter-
mination of the water and carbon content of the sediments, and for the
19
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Analysis of
Moisture
Content
(Appendix A)
H
Sediment
Preparation
(Text)
Sediment Extraction
! and Concentration
i (Appendix O
Extract Cleanup
(NOAA)
Preliminary
Samnta
Screening (Text)
Extract Analysis
(NOAArEPA)
Organic Carbon
Analyses
(Appendix B>
I
EPA - U.S. Environmental Prediction Agency
NOAA » National Oceanic and Atmoepneric Administration
FIGURE 2. General Approach to Sediment Preparation, Extraction,
and Analysis i
20
-------
Isolation, characterization, and quantification of nonpolar organic compounds
1n the sediments. For nonpo'lar organic compounds, our recommendation 1s based
on a selection process that emphasized the following criteria:
• . state-of-art level of the methods and analytical techniques
• ability of the methods to address the analysis of all compounds of
Interest (Table 3) ' .
• extent of method(s) validation and documentation
• availability of the Information on the Methods to the user.
Based on a review of the available methods and application of these criteria,
we recommend that the standard analytical procedures published by the National
Oceanic and Atmospheric Administration's National Analytical facility (MacLeod
et al. 1985), referred to as the NOAA method, be used as the main resource
for guidance on Isolation and analysis of nonpolar organic contaminants from
sediments. The NOAA (MacLeod et al. 1985) nethod 1s chosen because 1t satisfac-
torily meets all of the criteria. The EPA has published several methods for
analyzing environmental samples that contain the coapounds of Interest (see
Table 3); however, these methods do not meet one or more of the above criteria.
The EPA 600- and 1600-serfes methods (EPA 1984) are not recommended, because
they are specifically directed toward analysis of the compounds 1n water only,
and extraction procedures for sediments are not adequately addressed. The
EPA solid waste methods (EPA 1986) are not recoanended, because they are not
presented 1n an easily followed, single-analysis scheme from sediment prepara-
tion through analysis? they contain too many options to permit the final results
to be compared; and they recomend the use of reagents or approaches that are
not currently considered appropriate or state of the art. Methods that apply
to each of the steps 1n the general scheme are described In detail 1n Appendixes
A through 0 and 1n MacLeod et al. (1985). Recommended departure from the
methods 1n the NOAA report (MacLeod et al. 1985) for sediment dewaterlng and
extraction of large and wet sediment samples are described 1n Appendix C.
Because the NOAA report does not provide a procedure for quantification by
21
-------
TABLE 3. Nonpolar Organic Priority Pollutants^
Low-Molecular Weiht
Naphthalene
Acenaphthylene
Acenaphthene
Fl uorene
Phenanthrene
Anthracene
H1h-Moleeu1ar
PAH
Fluoranthene ;
Pyrene |
Senzo (a) anthracene
Chrysene |
Benzof 1 uoranthenes
Benzo(a)pyrene
Indeno(l,2,3-c.d)pyrene
01 benzo (a , M anthracene
Benzo (g , h , 1 ) peVy 1 ene
PCBs i
DOT, 000, DOE
Aldrln j
Chlordane (technical mixtures and metabolites)
D1eldr1n >
Heptachlor and metabolites
Undane !
Aldrln and netabolltes
(a) Fro» Table 1, ?ect1on 307, Clean Water Act of 1977
i
gas chromatography/mass spectro»;etry (GC/MS), EPA method 8270 (EPA 1986) or
EPA Method 1624 (EPA 1984) may be used until a validated approach 1s developed.
Implementation of the analysis approach (see Figure 2) begins with general
sample preparation. During this! step, factors that could adversely affect
later steps In the analysis schejne are addressed and minimized. As stated
previously, all analyses should be performed within 7 days of collection of
22
-------
the samples. If the analyses cannot be completed within 7 days, 'then the
sample should be frozen (see Section 3.1.3). All sediment preparation proce-
dures and handling should be similar for the samples used for the dry weight,
total organic carbon, and contaminant analyses. Following general sample
preparation, subsamples of sediment may be analyzed for dry weight (Sec-
tion 3.2.1 and Appendix A) and total organic carbon content (Section 3.2.2 and
Appendix B). Another subsample of the sediment sample 1s subjected to organic
solvent extraction (Section 3.2.3).
Removal of organic constituents from the sediment 1s achieved by sample
dewaterlng followed by Soxhlet extraction (Appendix C) or by centrlfugatlon
and extraction (MacLeod et al. 1985). A key Issue that must be addressed
during the extraction step Is extraction efficiency, which 1s affected by the
ability of the organic solvent to come Into Intimate contact with sediment
particles. Because extraction efficiency 1s directly related to the presence
of water 1n the sample, aspects of the methods describe approaches for removal
(dewaterlng with methanol) or containment (addition of sodium sulfate) to
enhance extraction efficiency. Internal standards are addressed and recom-
mendations are made on compounds to be used for quantification of selected
compound classes. Calibration and spiked blanks art also discussed relative
to method Integrity. Finally, an approach 1s recommended for concentrating
sample extracts that minimizes contaminant losses.
Concentrated sample extracts are then subjected to the very Important
step of sample cleanup (MacLeod et al. 1985). The cleanup process Involves^
subjecting each extract to a series of treatments that selectively remove
materials that would Interfere with the analysis of the compounds of Interest.
Interfering materials that art removed during extract cleanup Include elemen-
tal sulfur, polar compounds (e.g., adds and bases), parafflnlc hydrocarbons,
Uplds, and other blogenlc compounds.
After Interfering materials have been removed, It-Is recommended that
samples be subjected to a preliminary screening by gas chromatography to obtain
a qualitative measure of sample complexity and the range 1n contaminant concen-
tration. Information derived from the screening analysis will assist the
researcher 1n determining sample dilution or concentration requirements, In
23
-------
selecting the appropriate detector, and In determining the extent to which
mass spectrometry may be required for chemical characterization and
quantification. ;
3.2.1 Dry Weight Determl nation
Dry weight of the sediment is determined on an unfrozen sample of the
sediment by heating a sample of known weight (5 to 10 g) overnight (or at
least 16 hours) at 70CC. The analysis procedure Is shown In Figure 3. A drying
temperature of 7Q°C, rather than 105«C, Is chosen to provide consistency between
the dry weight basis used to report the results of the organic carbon and
contaminant analyses, because drying to 105°C could result 1n loss of some of
the sem1volat11e organic compounds that are targeted for analysis. Before
drying, all bulk objects (e.g., sticks, leaves, and rocks) are removed and
the sample homogenized. Preparation procedures should be similar to those
used In the total organic carhop and contaminant analysis. After the dried
sample 1s allowed to cool In a desiccator, the sample Is weighed. A desiccator
1s used to prevent relncorporatjon of moisture Into the sample during cooling.
The dry weight 1s determined us>ng the following formula: .
X dry weight - 100 x g of r* sa"°le
g or sample
The details of the procedure ar« described 1n Appendix A.
The QA/QC procedures and clean laboratory practices should be followed
to ensure accuracy of the analysts. The analytical balance should be Inspected
and calibrated on a preasslgned schedule. Desiccators should be checked fre-
quently for proper sealing and for replacement of deslccant to prevent moisture
from accumulating during cooling. At least 10X of the samples should be
analyzed 1n duplicate.
The dry weight should be reported as percent to at least 1 decimal place
(I.e.; 0.1X). |
3.2.2 Total Organic
.
Total organic carbon 1s a measure of the total amount of nonvolatile,
partially volatile, volatile, and paniculate organic material 1n a sample,
i
24
-------
Prepare Sediment
Sample*
i
Place 5 to 10 g
in Tared Crucible
i
i
Weigh to Nearest
0.1 g
i
r
Dry Overnight
«70°C
i
r
Cool to Room
Temperature in
Oeeiccator
i
Weigh to Nearest
0.1 g
•Portion of same sample • uaod for total
organic carbon analyse*.
FIGURE 3. Flow Chart for Determining the Dry Height of a Sediment Sample
25
-------
Many methods for determining the total organic carbon content of soils and
sediments have been reported (Black et al. 1965). All the commonly used methods
Involve either wet or dry combustion of samples and quantitative determination
of the organic carbon content of the sediment from the C02 that evolves.
Quantification methods Include vol;imetr1c, t1tr1metr1c, gravimetric, or con-
-------
Prepare Sediment
Sample*
Dry it 70°C;
Cool in
Desiccator
Place 0.2 to
0.3 g in
Combustion Boat
Weigh to Neareat
0.1 g
±
Add HO to
Remove Carbonate
•Portion of same sample is uaed for
' percent moisture determination.
Dry ft 70°C;
Cool in
Desiccator
Add Cupric Oxide
or Equivalent
Material
±
Combust Sample at
980°CWhae
Measuring Amount
of CO Evolved
FIGURE 4. Flow Chart for Detenrining the Total Organic Carbon
Content of a Sediment Sample
27
-------
calibrated on a preasslgned schedule. If.a carbon analyzer 1s used, 1t should
be calibrated dally, and a standard reference material should be analyzed at
least once with each set of sediment samples. Desiccators should be checked
i
frequently for proper sealing and for replacement of deslccant to prevent
moisture from accumulating during drying. In the conduct of sediment total
organic carbon analysis, you should 1) use thoroughly homogenized samples,
2) cool all equipment and samples In a desiccator and 3) analyzt replicate
samples. At least 10X of the saijaples should be analyzed 1n duplicate.
The amount of carbon 1n the sediment sample should be reported as a percent
based on the dry weight of the sediment to the nearest 0.1S.
3.2.3 Analysis' of Sediments for Semi volatile Priority Pollutants
Procedures for analyzing sediments to determine contaminant concentrations
will be limited to those emphasizing sealvolatile priority pollutants (see
Table 3). These procedures have been developed to detect these pollutants at
trace levels 1n sediments [1 to 50 ng/g dry weight for neutral compounds (e.g.,
PAHs) and 0.1 to 15 ng/g dry weight of pesticides and PCBs'J. As mentioned
previously, the recommended procedures are primarily documented In MacLeod et
al. (1985). although EPA (1984) is recoranendad for GC/MS analyst of PAHs.
3.2.3.1 Sample Preparationiand Extraction
Figure 5 shows the steps Involved 1n the preparation and extraction of
samples. Specific procedures, reagents, materials, and apparatus to perform
this step are described 1n MacLeod et al. (1985) and Appendix C. A sample
size of approximately 50 to 100 g (wet weight) of sediment for extraction and
a concentrated volume of 0.5 «L are considered adequate to attain the low-
level detection limits required for setilvolatlle organic compounds. However,
concentration to as little as 20 fiL may be required for characterization of com-
pounds by GC/MS. Smaller sample!sizes can adversely affect detection limits,
and smaller final voluaes can result In excess loss of target compounds because
of volatilization. During sample preparation, excess water Is decanted from
the sample and bulk objects (e.g,>, sticks, leaves, and rocks) are removed.
28
-------
Prepare
Sampto
1
Add Watar
Partition and Dry
Organic Fraction
antrata
Organic Fraction
I
Procaadte
Sampia Oaanup
D«watar
Sampla
with Mathanoi
*
AddlntamaJ
Standarda
*
SoxhJat Extract
Scdimant
*
Combined SoxMat
andMathanot
Extract!
•»
Mathanol/Wataf
Extract
•;;:-
FIGURE 5. Sample Preparation and Extraction
29
-------
Failure to perform these manipulations could result 1n poor extraction perfor-
mance and Increase variability ib sediment data. It Is also Important to use
a homogenized sediment sample to ensure representativeness of the results.
Prior to Soxhlet extraction, water remaining 1n the sediment after
decanting 1s removed by contacting the sediment with methanol (Appendix C).
In the NOAA method (MacLeod et al. 1985), excessive water Is reaoved by
centrlfugatlon, and residual sediment moisture 1s contained through the addition
of anhydrous sodium sulfate to the sediment sample prior to sediment tumbling.
The efficacy of the NOAA method with large sediment samples (50 to 100 g)
needs to be evaluated.
After sediment dewaterlng, Internal standards are added to the sediments
to adjust for analyte losses during sample workup. The standards contain
mixtures of deuterated aromatic hydrocarbons or halogenated compounds (Table 4).
The Internal standard solution fiar PAHs 1s prepared In hexane to a concentra-
tion of approximately 50 ng/^L for each compound; the Internal standard for PCBs
and pesticides Is prepared In hexane to a concentration of approximately
1 ng//tl.. Along with the sediment samples, calibration and spiked blank samples
are carried through the extraction procedure to Identify and account for poten-
tial contamination and losses. Compounds that ar« r*cn?>mended for use 1n the -
blank samples are given 1n Tables 5 through 8. the calibration solutions are
TABLE A. List of Compounds used as Internal Standards
Internal Standards for PAH^
I
j
Naphthalehe-d8
Acenaphthene-dlO
Perylene-dl2
Internal Standard for PCBs and Pesticides
4.4'-01bromooctaf1uorob1pheny1
(a)
(a) Adapted from MacLeod et al. (1985)
30
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IABLL-5. List of Compounds used in the PAH Calibration Solution^
Hexamethylbenzene
Naphthalene
2-Methyl naphthalene
1-Methylnaphthalene
Blphenyl
2,6-01methylnaphthalene
Acenaphthene
F1uorene
Phenanthrene
Pyrene
Benzo(a)anthracene
Chrysene
Benzo(e)pyrene
Benzo(a)pyrene
?erylene
01benzo(a,h)anthracene
Naphthalene-d8
Acenaphthene-dlO
Pery1ene-dl2
(a) Adapted from MacLeod et al.
(1985)
prepared 1n hexane to a concentration of approximately 5 ng/jtL for each
compound; the spiked solutions are prepared 1n hexane to a concentration of
approximately 50 ng/0L.
The second step 1n the analysis 1s removal of the organic compounds from
the sediment samples by Soxhlet extraction. Host Soxhlet extraction procedures
reported 1n the literature use a mixture of solvents that range 1n polarity.
Our recommended Soxhlet extraction solution 1s benzene/methanol (3:2). However,
some laboratories may have safety regulations limiting laboratory worker expo-
sure to benzene, thereby making use of this extraction solution Impossible.
In this case, we recommend using either methylene chlorlde/methanol (2:1),
methylene chlorlde/methanol (9:1), or the NOAA method to extraction of the
sediment (Figure 6). The EPA method 3540 (EPA 1986) recommends that either
toluene/methanol, acetone/hexane, or acetone/methylene chloride mixtures be
used. Hunchak and Suffet (1987) showed that acetone and mixtures of acetone
31
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TABLE 6. List of Compounds usto\1n the PCS and Pesticide
Calibration Solution^*'
Tetrachloro-m-xylene
Hexachlorobenzcine
Llndane (i-BHC)
Heptachlor
Heptachlor-epoxlde
Aldrln
e-Chlordane
Trans-nonachlor
01eldr1n
M1rex
o,p'-DDE i
P.P'-ODE [
o.p'-DDO !
p.p'-DOD
o.p'-DDT ;
p.p'-DDT
2,4I-D1chlorob1pheny1
2,5,4'-Tr1chlorob1phenyl
2,4,2' ,4'-Tetrfflchlorob1phenyl
2,4,5,2' ,5'-Pentachlorob1phenyl
2,4,5,2',4',5l4Hexachlorob1phenyl
2,3,4,5,6,2l,5<-Heptachlorob1phenyl
2,3,4,5,2' ,3' ,r ,5'-Octachlorob1pheny1
luufobipnenyi
(a) Adapted frxDis MacLeod et al. (1985)
i
and hexane contain numerous artifacts that can Interfere with full scan analysis
of environmental sanples; therefore, we do not recommend use of any of these
extraction solutions. .
Following extraction, the »ethanol/water solution fron the sediment
dewaterlng step 1s combined with 'the Soxhlet extract and partitioned to obtain
the final organic extract. The resulting extract Is then dried by elutlon
through a sodlua sulfa-te column and concentrated either by Snyder column or
rotary evaporation. We recouaend the Snyder column procedure, because 1t Is
most effective In removing solvent while «1n1a1z1ng the loss of analyte,
although the rotary evaporation Method may be faster and less costly. Before
32
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TABLE 7. List of Compounds used 1n the PAH Spike Solution^4!)
Naphthalene
2»Methylnaphtha!ene
1-Methylnaphtha!ene
Blphenyl
2,6-01methyl naphtha!ene
Acenaphthene
Fluorene
Phenanthrene
Anthracene
1-Methylphenanthrene
Fluoranthene
Pyrene
Benzo(a)anthracene
Chrysene
Benzo(e)pyrene .
Benzo(a)pyrene
Perylene
D1benzo(a,h)anthracene
(a) Adapted from MacLeod et a!.
(1985)
the rotary evaporation method 1s used routinely 1n a suite of chemical analyses-,.
the recoveries of target compounds Bust be reported and should be within the
acceptable limits as defined by the scientific community for the Snyder column
method.
If methylene chloride Is used as one of the extraction solvents, then.
the final step 1n the extract preparation 1s solvent exchange with hexane
during the final sample concentration step. Removal of methylene chloride 1s
necessary to minimize chromatographlc effects during the sample cleanup phase.
Final extract volume Is targeted at 1 to 2 «L.
3.2.3.2 Sample Cleanup
In addition to the nonpolar organic contaminants of Interest, the sediment
extracts contain a variety of different materials, Including polar compounds
(e.g., adds and bases), I1p1ds, parafflnlc hydrocarbons, Inorganic consti-
tuents, and other blogenlc naterlals. The objective of the cleanup phase Is
33
-------
L1st of Compounds used 1n the PCS and Pesticide Spike Solut1on.(a)
Hexachlorobenzene
Undane (T-SHC)
Heptachlor
Heptach 1 or-tpox1 de
Aldrln
c-Chlordane
Trans-nonachlor
D1eldr1n ;
M1rex
o,p'-DOE
p,p'-DOE .
o,p'-DOD
p.p'-DDD • |
o.p'-DDT
p.p'-DDT
2,4'TDIchloroblphenyl
2,5,4'-Tr1chlbrob1phenyl
2,4f2l,4'-Tetrachlorob1phenyl
2,4,5,2',5I-Pentachlorob1phenyl
2,4,5,2',4a,5'-Hexachlorob1phenyl
2,3,4,5,6,2l,£l-Heptachlorob1phenyl
2,3,4,5,2l,3',4l,5'-0ctachlorob1phenyl
2,3,4,5,6,2l,3l,4',5'-Nonach1orob1phenyl
(a) Adapted from MacLeod et ai - (1985)
to selectively remove some of these extraneous »ater1als and to reduce/elimi-
nate Interferences caused by these materials prior to the chemical analysis
phase. Figure 7 depicts, 1n general terms, the sequence of steps required to
clean solvent extracts In preparation for chealcal analysis. In the first
cleanup step, polar coapoiinds and parafflnlc hydrocarbons are removed by
subjecting the sample to chromatography on a nlxture of silica gel and alumina.
The operating mechanise for removal of unwanted compounds In this cleanup
step 1s adsorption. Parafflnlc!hydrocarbons precede the elutlon of PAHs,
PCBs, and pesticides from the column, and polar compounds are retained by the
column. Next, the recovered fraction from the adsorption chroraatography step
1s subjected to chromatography on Sephadex where I1p1ds and other blogenlc
materials are removed. The prlwary mechanism for removal of these materials
34
-------
Prepare
Sampto
Add
Standards
Add Sodium
Sulfata and
Turhbto
Sampia
Tumbla
Sampta
Tumbta
Sampta
Dacant
Extract
Dacant
Extract
Occam
Extract
Combined
Extracts
i
Concamrata
Extract
Procwdto
Sampia Claanup
(NOAA) •
NOAA » National Ooaanic and Atmoapharic AdmWatration
FIGURE 6. Saaple Preparation and Extraction (NOAA Method)
35
-------
Concentrated •
Sample Extract* *
Acids. BUM, Neutral*,
U'pkJs, and Btogenic Materials
|
Sulfur
I
Chromatograph on Potar
Adaorbant SBien Gel/
..Alumina*. .. .....
1
1
Paraffinic Hydrocarbons,
Acids, and Bmea
Sulfur
I
Aromatic and ChtorinateeJ
Hydrocarbons, Liptds, and
Btogenie Materials ;
Chroma tographic
on Sepnadex
lipidsand
Blogeroc Materials
Aromatic and \
Chionnatad Hydroearbonc
Preliminary Screen by Sao
Chromatograpny/ Flame lor*-
ration, /Bectron Capture!
and /Mass Spectrometry!
Detector (NOAA-EPA)
*Bam*ntal sulfur can ba rsmowd from tha sample at tnaac stagm by
sevvfai dlffaram tachniquas (Appartdix 0 and MacLeod et al. 1966). If a
samplers) is suspected of having large quantities of sulfur, treatment (to
remove the sulfur in advance of adsorption chromatography • racommenijed.
I
EPA » U.S. Environ mental Protection Agency
NOAA - Nationai Oceanic »nd Atmospheric Administration
FIGURE 7. ' General Scheme for Sample Cleanup
-------
1n this step 1n the extract cleanup 1s size exclusion. High-molecular weight
I1p1d and blogenlc materials precede the elutlon of nonpolar compounds from
the column.
Elemental sulfur can also be a major Interference 1n the analysis of
nonpolar organic compounds by GC. Specifically, sulfur Interferes with the -
analysis of 'Individual peaks using gas chromatography/electron-capture detec-
tion (GC/ECD). Sulfur can be removed from the sample prior to chroaatography
through reaction with mercury, tetrabutylammonium sulfate, or activated copper.
In the case of activated copper, the copper can be added to the top of the
adsorption column and sulfur can be removed as part of the sorptlon chromato-
graphy step. For samples that are suspected of containing large quantities
of sulfur, 1t 1s recommended that the sample(s) be treated prior to adsorption
chromatography to remove sulfur using one of the methods given 1n Appendix 0.
Although mercury and copper are commonly used to remove sulfur (methods are
described 1n Appendix D), these methods have limitations, Including degradation
of endrln by mercury and loss of heptachlor 1n the copper column that preclude
their use on certain environmental samples. Following completion of cleanup,
the sample 1s ready for analysis. Each of the cleanup procedures will now be
discussed 1n more detail.
.fJ' s'
3.2.3.2.1 Cleanup Via Adsorption Chromatography. This section describes
1n more detail the specific method 1n which two adsorbents (silica and alumina)
are combined to perform the first cleanup step. In this procedure, the
recovered extract contains both the aromatic and chlorinated organic compounds
of Interest. Specific details describing apparatus, reagents, and procedures
for this method are described 1n Section 5 of MacLeod et al. (1985).
The steps depicting the approach are shown 1n Figure 8. In this approach,
slurries of alumina and silica gel are sequentially packed Into a glass column.
The mixed sorbent 1s then topped with a layer of acid-washed sand. If sulfur
removal 1s required and sulfur concentrations are suspected to be relatively
low (I.e., trace quantities), then a layer of copper powder placed on top,of
the sand 1s used to remove sulfur during this chromatographlc step. If sulfur
concentrations In the extracts are suspected to be relatively high (e.g.,
sulfur 1s observed precipitating out of solution), then sulfur cleanup using
37
-------
Concaiiuaiad Sampla
Exnet Exehangrt
into Ktearw
Slurry-Pack
Cotumn with
Alumina
Add Slurry of
Sifea Gat Atop
Alumina
I AddLayarof
'Sand Atop SSca
Add Uayarof Coppar
Powder Atop Sand
Column with
end
ilmreduea Sarnpia
to Too of Column
Bute Csiumn witri
Parmrwand CaMact
in Raak
±
Butn Column with
CHjC^/Pwnan
ondCoKaesin
±
Buta
20% MaOH in CHjCJj
andCotectin
Conuntv
ol 400 Ott
Polv Compound!
PwffiiMc HydroGafftoons
Raalt Comamit
PAH, PCS. end
±
Extract
Procaad to Sizift
Exduaion.
Chrematograpny Stap
(NOAA)
NOAA « National Qeaante and Atntoapnarto AdmMamnion
PAH • Porynudaar Aromatic Hyt'iroiavton
PCS - Poryentoonatad Biphany41
FIGURE 8. Cleanup of PAH, PCB, and Pesticide-Containing Sauples via
Silica Gel/Alumina Chroraatography (Adapted from MacLeod
et al. 1985)
38
-------
one of the methods In Appendix 0 should be completed before this cleanup step
1s Initiated. The sample concentrate 1n hexane 1s Introduced to the top of
the column and the column sequentially eluted with pentane (fraction contains
parafflnlc hydrocarbons) followed by a 50/50 mixture of methylene chloride/
hexane.' The fraction elutlng with the latter solvent (designated SA2 in MacLeod
et al. 1985). contains the compound classes of Interest (I.e., PAHs, PCBs, and
pesticides). This fraction Is concentrated and then subjected to size exclusion
chromatography.
3.2.3.2.2 Sample Cleanup Via Sire-Exclusion Chromatography. The second
step In the cleanup Is size-exclusion chromatography of sediment extracts
that have been subjected to adsorption chromatography treatment. The method
1s designed to remove I1p1ds and other blogenlc materials from the sample,
further reducing the potential for Interference during the screening and
chemical analysis steps.
Figure 9 describes the steps that are Important to the conduct of the
method. Specific details describing apparatus, reagents, and procedures are
described In Section 6 of MacLeod et al. (1985). In the first step, the column
1s packed with a slurry of Sephadex that has been allowed to swell overnight
by placing 1n solvent [cyc"ohe~ane:methanol:methyl*ne chloride, 6:4:3 (V:V:V)].
The packed column Is then allowed to settle overnight to.provide additional
time for the solvent to Bake Intimate contact with all surfaces of the porous
media and to eliminate air bubbles. If air bubbles persist, pass warm solvent
through the column. If this action does not remove the bubbles, then the 5
column will have to be repacked. Prior to column calibration, the height of
the Sephadex 1s adjusted to 26.5 cm. To simulate b1ogen1c/!1p1d material, a
tissue extract that had been subjected to silica gel/alumina chromatography
1s used to accurately establish Its elutlon profile. The method also makes
use of an aromatic hydrocarbon standard (azulene/perylene) 1n the column's
calibration to bracket the elutlon time of the compounds of Interest.
The NOAA report (MacLeod et al. 1985) emphasizes the need to maintain
sample solubility. This solubility Is achieved by maintaining the sample 1n
a mixed solvent [cyclohexane:methanol:methy1ene chloride, 6:4:3 (V:V:V)].
Following sample Introduction, two lead fractions are eluted from the column
39
-------
Cancan tratad
Sampia in>
Oia
R
Contanti
Swal Packing
MatariaJ (Saphaoaxl
Pack
Mataritf into Column
Co*jn*i
CsUxmi Column
H
Introduca Sampia
to Top of Column
@uw Column with
23 mL of SoKwie
Suta Column wMi
CoMaetin
Suw Column
[Coaaetin
Suw Column **rth
90 mL of Sonwic
CotectinRaak
Oiaeard <
of Gnouaad
CyHnoar
R«ak Contanti SA2-L1
Upiea and Stoganie
Matarioto
Raak Contanti SA2-U
Arematie«nd
Chterinntud
Hydrwarbooi
±
Sampto
FIGURE 9. Sample Cleanup via Gel Permeation Chnxaatography
(Adapted frws MacLeod et al. 1985)
40
-------
and discarded. These fractions contain I1p1d and other blogenle materials.
A subsequent fraction (Fraction SA2-L2) contains the compounds of Interest
and 1s concentrated and subjected to analytical screening and analysis.
3.2.3.3 Analytical Screening and Analysis of Sample Extracts
* *™**^"^^^h*«»^fc™i«^™^"""^"~—"——"^"^^•™^~^^^^^^^^^™"™^^™""^^ ' j^^^—
The decisions facing the analytical chemist once the extracts have been
processed through the cleanup steps are governed by experimental needs and by
limitations that may be Imposed by the make-up of the samples. To address
the first point, the researcher will need to have characterization or quanti-
tative data on several classes of compounds 1n the extract to make decisions
on which type of Instrument to use In the quantification. Also, concentration
of compounds of Interest 1n the extract might be quite low; thereby, taxing
to the limit the analyst's skills to obtain the necessary data. Figure 10
depicts the logic for selecting the appropriate analytical tools and for
preparing the samples for characterization and quantification of Individual
constituents by gas chromatography/flame 1on1zat1on detector (GC/FID), GC/ECD,
or GC/MS. In the first step, all samples are screened by GC to determine the
relative range 1n concentration present 1n all of the samples. Sample volumes
at this stage are usually 1 ml. If the researcher 1s Interested 1n the analysis
of PAHs, PCBs, or pesticides, then the screening should Include both GC/FID
and GC/ECO techniques. Quantitative examination of each chromatogram will
allow the analyst to determine whether adjustments 1n the volume of the sample
are required to Increase or decrease sensitivity. If Increased sensitivity
1s required, additional sample concentration can be performed. If additional
sample concentration (approximately 100 j»L) falls to result 1n detectable
peaks (above background), then detection Units will need to be reported for
the sample and no additional analysis Is required. Alternatively, samples
determined to be too concentrated may be diluted by an appropriate amount to
ensure accurate quantification. During the qualitative examination of the
chromatograms, the researcher may detect familiar patterns characteristic of
certain classes of organic compounds. The patterns may Include those of PCBs
(In the form of specific aroclors) or PAHs (In the form of petroleum). Such
Information may assist the researcher In analytical standard selection.
41
-------
Scr*MT) Extracts
by GC/.RD
!
Ara Quantifiabla
Paaks Praeant?
No
Yaa
Screan Extracts
by GC/ECD
Furthar
Sampia
No
Ara QuamWaM
Panics Praaant?
latfviitB foe
IsSampia too
Concarintad?
OHuta
Cnafvctanatic Profw
Praaant? Anaiybeal
Stondantt Adaquata to
Quamrfy Conpounda
of Intaraat?
Parform 6C/MS
Y*»«
Uthizalaorapa
Dilution Tachntqua
n
OC/MS s* S«npiaa
Raqutrad/Oaairad?
Uaa GC/MS
Charactanzxoon to
Labai Paaks in GC
ChrematOQrama
S^aet10%of
Skicnptoe for GC/MS
±
Ouwitify Conatituan
by GC/RD
and/or GC/ECO
Sandaniii
Evakutaand
Report RaauitB
GC/ECD • Gaa Chrematography/aaetron Captura Dataetor
GC/RD - Gaa Chrernatography/Rama lonaatkm Dataetor
GC/MS - Gaa Chromatograpny/Maat Spacromatry Dataetor
FIGURE 10. Analytical Screening and Analysis of Samples
42
-------
Once the researcher has a qualitative picture of the chemistry of each
sample, sound decisions on the best quantitative tool can be made. The
researcher may have found that the Initial screening by GC/FID and/or GC/ECD
on some samples provided chromatograns of sufficient quality that the samples
»
will not have to be rerun. If quantification of samples by GC/MS Is required
or desired, -additional adjustments 1n sample concentration may be required to
take Into account the lesser sensitivity of the mass selective detector.
Although GC/MS has the advantage of more absolute compound Identification and
Increased capability over GC alone to Minimize Interference/quantification
problems, this method 1s more time consuming and more expensive than GC/ECD
or GC/FID methods. Also for most analytes, GC/MS 1s less sensitive than GC/ECD
or gas chromatography/halogen-spedflc detector (GC/HSD) methods for pesticides
and PCBs and GC/FID for PAHs. For example, a sample may have sufficient
concentration of PCB to be quantified by GC/ECD, but not by GC/MS. For these
reasons, full quantification of all samples by GC/MS may not be required or
desired. In this case, the researcher should select 10Z of the samples (samples
that typify the diversity 1n complexity of all sauries to be analyzed) for
GC/MS analysis to conflra the presence of specific compounds, ind quantify
all samples using either GC/FID or GC/ECD. Samples analyzed by ~C/ECD that
show the presence of PCBs my also show other peaks from those corresponding
to the PCB standards. Representative samples need to be analyzed by GC/MS to
Identify these peaks and to verify the presence of these other analytes that
were Indicated by retention time comparisons. In addition, multlcomponent
peaks are often present (e.g., overlapping PCB and PCB/pest1c1de combinations)
requiring resolution by single 1on monitoring. During the quantitative analysis
process, the researcher should give serious consideration to using reference
standards supplied by the EPA or National Bureau of Standards.
The recommended method for the quantitative analysis of extracts for
PAHs, PCBs, and pesticides using GC/FID and GC/ECD 1s described In detail 1n
Section 12 of MacLeod et al. (1985). Using a capillary column Instead of the
packed column for gas chronatography 1s recommended to ensure the resolution
of anthracene and phenanthrene, chrysene and benzo(a)anthracene,
benzo(b)fluoranthene and b«nzo(k)fluoranthene, and d1benzo(a,h)anthracene and
1ndeno(l,2,3-cd)pyrene pairs, and to maximize the resolution of PCB Isomers.
43
-------
Capillary column capability 1s a'lso required to resolve the deuterated forms
of naphthalene, acenaphthene, anp perylene that are used as Internal standards
for PAHs from the nondeuterated compounds In the sediment extracts. To calcu-
late the concentration of chlorinated analytes (I.e., PCBs and pesticides),
dlbromcroctafluoroblphenyl 1s used as the Internal standard for the GC/ECD
analysis. T.he ECO 1s very sensitive to halogenated compounds; however, the
detector requires careful calibration and appropriate use of Internal and
i
calibration standards to ensure quality data.
At' this time, validated methods for the quantification of nonpolar organic
compounds of Interest by GC/MS employing Isotope dilution techniques are limited
to PAH (EPA Method 1624, Rev. B Of EPA 1984). If quantification by GC/MS of
this class of compounds 1s required, we recommend EPA Method 1624. For PCBs
and pesticides, we recommend the same method with quantification performed
employing selective recovery staWard(s) (e.g., dlbromooctafluoroblphenyl).
i
3.2.4 Quality Assurance/Quality Control Procedures
To produce analytical results of high quality and reproduceb1!1ty, QA/QC
procedures must b« followed to ensure accurate, nrteise, and reliable measure-
ments. Any laboratory performing these analyses must keep"records of equipment
calibration, and follow clean laboratory practices. Specific QA/QC proce-
dures Include proper preparation of the analytical containers to ensure that
no sample contamination will occur from these sources, analysis of all sol-
vents to determine their purity, and calibration and regular maintenance of all
equipment. The analytical procedures also Include the addition of Internal
standards to allow for quantification of the analytes of concern and to account
for any losses that occur during sample preparation. The compounds used 1n
the Internal standards were chosen to ensure representativeness of the compounds
of Interest and also to ensure that these Internal standards will not occur
1n a GC peak within 0.1 m1n of the analyte peak. Also Included In the analysis
are calibration and spiked blank samples to Identify and allow the analyst to
account for potential contaminant and losses. If the recovery of any Internal
standard 1s less than SOX, then the sample must be reanalyzed. A portion of
the calibration samples are analyzed In tandea with the extract samples and
are used as reference for determining the concentration of the analyte
44
-------
(Section 3.2.5). 'Finally, the container and procedural blanks (see Section
3.1.1.1) should be analyzed periodically to assess the level of contamination
that occurred during sample collection.
3.2.5 flata Reporting
The final data are reported as the ng of analyte/g of the sediment sample
on dry weight basis using the following equation (Section 12 of Macleod et
al. 1985):
no of analyte l x ng I-Std added 100
no o anaye -
g of sediment! dry weight * R^ A sample weignt I dry wt
R analyte peak area In extract
wnere KJ « i-5td
pea* area in extract
analvte concentration 1n reference vial (nq/uL)
R2 " I-Std concentration in reference vial (n$/itL)
m analyte peak area In reference vial
R3 * i-sld peak area 1n reference vial
I-Std - Internal standard
XDry wt « percent dry weight determined using the '-nhod 1n Appendix A,
45
-------
4.Q DATA CAI qf|
After determining the dr^ weight, organic carbon content, and contaml-
nant concentration, the organic carbon-normalized concentration of -the contami-
nant 1rf the sediment can be calculated and compared with the numerical sediment
quality criteria. Because organic carbon Is the primary sortent phase on the
sediment and the quantity of t!he organic carbon affects the toxldty and
accumulation of the associated contaminants, a numerical set of criteria that
can be used In a cross section of sediment types will be expressed as normalized
to sediment organic carbon content. Use the following formula to calculate
the organic carbon-normalized Concentration of the contaminant:
(contamlbant concentration) x
li organic carbon
100
When the contaminant concentration 1s In units of ng/g and the total organic
carbon content 1s reported as percent, then the units for the organic carbon
normalized concentration will be W7 g of contam1nant/g of organic carbon.
46
-------
5.0 CONCLUSION
Methods have been recommended for collecting sediment samples and for
analyzing dry weight, total organic carbon content, and concentration of
nonpola'r organic contaminants in these samples. The primary concern In applying
any of the recommended methods or potential alternatives 1s that proper QA/QC
procedures be followed, and that the performance of the sampling and analysis
methods be monitored. If variations 1n sample matrices require some
modifications to the proposed methods, all modifications must be thoroughly
documented and their performance must be compared to referenced methods. It
1s also critical that the procedures be performed only by experienced personnel
who follow clean laboratory practices. If these guidelines are followed, the
sampling and analysis will meet these criteria and will be useful 1n applying
sediment quality criteria for nonpolar organic contaminants.
47
-------
6.0 REFERENCES
Adams, W. J.r R. A. Klramerle and R. 6. Mosher. 1986. "Aquatic Safety
Assessment of Chemicals Sorted to Sediments." In Aouatie Tn^minny and
Hazard Assessment; Sevftnth Svimwiinn. ASTM STP 854, eds., R. D. cardwen, R.
£!r?yi ?n^R* £* Bahner- PP- 429-453. American Society of Testing Materials,
Philadelphia, Pennsylvania. \
C*uA:: 5' Di !V??SI J; L; Wh1tc' L- E- 2ns«1nger, "and F. E. Clark.
5. Methods Of Soil Analyst. American Society of Agronomy, Madison,
Wisconsin.
Bolton, H. S.f R. J. Bretler, B. W. V1gon, J. A. Scanlon, and S. L. Clark.
1985. National Perspective on. Sediment nuaTi^y- Criteria and Standards
Division, U.S. Environmental Protection Agency, Washington, D.C.
DIToro, D. M., J. S. Jerls, and b. C1arc1a. 1985. "Diffusion and Partitioning
of Hexachloroblphenyl 1n Sed1»?nts." Environ . sei. Tachnol. 19:1169-1176.
Hunchak, K., and I. H. Suffet. J987. "Analysis of Acetone-Hexane Artifacts
Produced 1n the Soxhlet Extraction of Solid Env1 ronn»ental Samples.' J^
Chromatography 392:185-198.
Karlckhoff, S. W. 1981. "Seal -Empirical Estlaatlon of Sorptlon of Hydrophoblc
Pollutants on Natural Sediments and Soils." Chgnosphgr* 10:833-846.
roiiusint Sorptiofl In Aquatic Systems "
J. Hydraulic FngT 110(6) :707-735.
Karlckhoff, S. W., D. S. Brown aiid T. A. Scott. 1979. "Sorptlon of Hydropho-
blc Pollutants on Natural Sedlwent." Water Res. 13:241-284.
•
MacLeod, W. D., D. W. Brown, A. JJ. Friedman, D. G. Burrows, 0. Maynes, R. W.
Pearce, C. A. Wlgren, and R. Gi. Bogar. 1985. Fjttractabie TQ*^ Organic
Compgunds, Second Edition. Standard Analytical Procedures of the NOAA
National Analytical Facility 1985-1986. NOAA Technical Memorandum NMFS
F/NWC-92, Seattle, Washington.
Schwarzenbach, R. P. and J. Westall. 1981. "Transport of Non-Polar Organic
Compounds from Surface to Groundwatef." Environ, sei. Tgehnoi,, 15:1360-1367.
Tejra I«ch. 1986. Recommended Protocols for Measuring Organic rnmpotmH^ in
Puoet Sound Sediment and Ti*s.»» ^mpix, Prepared for Office of Puget Sound,
Region 10, U.S. Environmental Protection Agency, Seattle, Washington.
U.S. Department of Transportation. 1984. "Hazardous Materials Regulations."
49 Fed. Rgq. Chapter 1, Subchapter C, pp. 52-792.
48
-------
U.S. Environmental Protection Agency (EPA). 1986. Test MethoH« fnr Evaluating
SoUd Waste. SW-846. Office of Solid Waste and Emergency Response,
Washington, O.C.
U.S. Environmental Protection Agency (EPA). 1984. Guidelines fnr
T»
-------
-------
APPENDIX A
METHOD FOR DETERMINING THE DRY WEIGHT OF A SEDIMENT SAMPLE
-------
-------
APPENDIX A
METHOD FOR DETERMINING THE DRY WEIGHT OF A SEDIMENT SAMPLE
A.I SUMMARY OF METHOD
The sediment sample 1s homogenized and a portion removed for dry weight
determination. The sample is dried overnight at 70°C. After cooling in a
desiccator, the weight loss is determined and used to calculate the dry weight
of the sample. A flow chart depicting the analytical scheme is shown in
Figure A.I.
A.2 EQUIPMENT
• drying oven capable of sustaining a 70°C temperature
• desiccator
• porcelain crucibles
• apparatus for grinding samples, such as Fisher Mortar Model 155
Grinder, Fisher Scientific Company, Catalogue Number 8-323, or an-
equivalent brand and model
V.
A.3 EQUIPMENT PREPARATION
• Crucible should be cleaned by heating to 950°C, cooled in a desiccator,
and weighed to the nearest 0.1 g.
A.4 SAMPLE PREPARATION
• If possible, the dry weight and other chemical analyses .should be
performed on fresh sediment samples. However, if the samples must
be frozen, then the dry weight should be determined on a frozen sample.
Before analysis, the frozen sample should be thawed slowly at room
temperature. All bulk materials (e.g., sticks, leaves, and rocks)
should be removed from the sample and the sample homogenized to uniform
A.I
-------
Prepare Sadimant
Sampia*
Placa 5 to 10 g
to Tarad Crucitta
Weigh to Naaratt
0.1 g
Dry Ovamight
«70"C
1
r
Cooi to Room
Daaiccator
}
r
Wwgh to N««r««t
0.1 g
•Portion of sama sampte ia uaad for total
organic carbon anaiyaaa.
FIGURE A.I. Flow Ch^rt for Determining the Dry Weight of
a Sediment Sample
A.2
-------
texture and color. If fresh samples are used, then overlying water
should be discarded before homogenization. If frozen samples are
used, water that results from freezing should be incorporated into
the sample during hooogenization. If the sample will not pass through
a 1-nro. sieve, it should be processed with the sediment grinder or
equivalent equipment to homogeneous texture and color before analysis.
The sample preparation procedures should be the same as those used
in preparing samples for total organic carbon and contaminant analysis.
The sample taken for dry weight determination is one subsample from
that collected for total organic carbon analysis.
A. 5 ANALYTICAL PROCEDURE
• Place 5 to 10 g of homogenized sample in tared crucible.
• Determine weight to nearest 0.1 g.
• Place crucible with sample in drying oven at 70"C.
• Dry overnight or at least 16 h.
• Cool sample to room temperature in desiccator.
• Determine weight of dried sample to nearest- D.I g.
A. 6 CALCULATION
* dry M1|tt - 100 x 1
A. 7 QUALITY CONTROL
• Crucibles should not be touched with bare hands after cleaning.
• All analytical balances should be inspected and calibrated on a
preassigned schedule.
• Desiccators should be checked frequently for proper sealing and for
replacement of desiccunt.
• 10% of samples should be analyzed in duplicate.
A.3
-------
-------
APPENDIX B
METHOD FOR DETERMINING THE TOTAL ORGANIC CARBON
CONTENT OF A SEDIMENT SAMPLE
-------
-------
APPENDIX B
METHOD FOR DETERMINING THE TOTAL ORGANIC CARBON
CONTENT OF A SEDIMENT SAMPLE
B.I SUMMARY OF METHOD
The sediment sample is homogenized and a portion removed for total organic
carbon concentration. The sample is first dried at 70°C, then a portion of
the sample (0.2 to 0.5 g) is treated with HC1 to remove carbonates. The
carbonate-free sample is heated to 950°C, and the amount of COa evolved is
measured. The analytical scheme is shown in Figure B.I. This method is adapted
from Tetra Tech (1986).
B.2 EQUIPMENT
• induction furnace, such as Leco HR-12, Dohrmann DC-50, Coleman CH
Analyzer, Perkin Elmer 240 elemental analyzer, Carlo-Erba 1106
• analytical balance with 0.1 mg accuracy
• desiccator
• combustion boats *
• 10% hydrochloric acid
• cupric oxide fines or equivalent reference material
• benzole acid or other carbon source as a standard, if necessary
B.3 EQUIPMENT PREPARATION
• Clean the combustion boats by placing them in the induction furnace
at 950°C. After cleaning, the combustion boats should not be touched
with bare hands.
• Cool boats to room temperature in a desiccator.
B.I
-------
Prepare Sediment
Sample*
1
Dry «t 70°C;
Cool in
Desiccator
±
Place 0.2 to
0.5 gin
Combustion Boot
Wtigh to NMreet
0.1 g
I
Add HO to
J.
•Portion of Mm* sample is used for
percent moisture determination.
Dry at 70°C;
Coo- in
Deekicatof
Add Cupric Oxide
or Equiveient
Materiel
Combust Sample at!
9eO°CWhae'
Meesurkig Amount;
of CO Evolved
FIGURE B.I.
Flow Chart for beteralning th« Total Organic Carbon
Content of a Sediment Sample
3.2
-------
• Weigh each boat to the nearest 0.1 mg. .
• If an ascarite tube is used to capture the O>2, weigh the tube.
• Set up and calibrate the inductive furnace and associated carbon
dioxide analytical equipment according to manufacturer's instructions.
B.4 SAMPLE PREPARATION
A minimum of 25-g samples of the sediment should be collected in a glass
or plastic container and stored on ice, if storage times are less than 7 days,
or frozen, if storage times are longer. If the samples are frozen, they should
be thawed slowly to room temperature before analysis. Homogenize each sample
to uniform texture and color.
B.5 ANALYTICAL PROCEDURE
• Transfer 5 to 10 g of sample to a clean container.
• Dry sediment sample to constant weight at 70°C * 2°C. The drying
temperature is kept low to minimize the loss of senivolatile and
volatile compounds. S"
• Cool dried samples to room temperature in a desiccator.
• Grind sample using a mortar and pestle to break'up aggregates.
• Transfer 0.2 to 0.5 g of the sample to a cleaned, preweighed combustibn
boat. ,
• Determine the sample weight to the nearest 0.1 g.
• Add several drops of 10* HC1 to the dried sample to remove the
carbonates. Wait until effervescing is completed before adding more
acid. Continue adding acid until effervescing no longer occurs when
acid is added. The add should be added slowly and in small quantities
to prevent loss of sample due to effervescing.
• Dry the HC1-treated sample to constant weight at 70°C * 2'C.
• Cool the sample to rooii temperature in a desiccator.
B.3
-------
• Add previously ashed cupric oxide or equivalent reference material to
the combustion boat. I
• Combust the sample in the inductive furnace at a minimum temperature
of 50°C * 10°C, collecting or measuring the C0£ evolved.
i •
i
B.6 CALCULATIONS i
• If an ascari to-filled tube is used to capture the C02> the carbon
content of the sample can be calculated as follows:
percent carbon
where A » weight in grams of C02 determined by weighing .the
ascarite tube before and after combustion of the sample.
B » dry weight in grams of the unacidified sample in the
combustion boat
0.2729 » ratio of molecular weight of carbon to the molecular
weight of carbon dioxide.
A silica trap should be placed in the line before the ascarite tube
to prevent moisture from entering the tube. An additional silica
trap may also be placed at the exit end of the ascarite tube to trap
any water formed by reaction of the COa with the NaOH in the tube.
If an elemental analyzer is used, the amount of C02 will be measured
directly. To ensure accuracy, the instrument should be calibrated
daily, using the empty boat as a blank for the zero value and at least
two standards covering the expected range of the samples. The carbon
content is calculated as follows:
percent carbon - £m99l
B.4
-------
where C » amount of carbon that is evolved in grams as determined
by the carbon analyzer
B * dry weight in grams of the unacidified sample in the^
combustion boat.
B.7 QUALITY CONTROL
• Crucibles should not be touched with bare hands after cleaning.
• All analytical balances should be inspected and calibrated on a
preassigned schedule.
• Desiccators should be checked frequently for proper sealing and for
replacement of desiccant.
« If carbon analyzer is used, it should be calibrated daily with standard
reference material.
• 10% of samples should be analyzed in duplicate.
B.8 REFERENCE
Tetra Tech. 1986. Recommended Protocols for Measuring Organic Compounds in
Puget Sound Sediment and Tissue Samples.Prepared for Office of Puget Sound,
Region 10, U.S. Environmental Protection Agency, Seattle, Washington.
B.5
-------
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APPENDIX C
SEDIMENT DEWATERING AND EXTRACTION
-------
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APPENDIX C
SEDIMENT DEWATERING AND EXTRACTION
C.I SUMMARY OF METHOD
The method combines attributes of procedures described in Tetra Tech
(1986a, 1986b). The sediment; sample is dried with methanol, placed in an
extraction thimble or between two plugs of glass wool, and extracted using an
appropriate solvent in a Soxhlet extractor. The combined extract (methanol
and Soxhlet) is subjected to liquid-liquid partitioning. The organic phase
is then dried, concentrated, and, as necessary, exchanged into a solvent
compatible with the cleanup procedures. The analytical scheme is shown in
Figure C.I.
C.2 APPARATUS AND MATERIALS
• Soxhlet extractor • 40-on I.D., with 500 ml round-bottom flask.
• drying column - 20-mra I.D. Pyrex chromatographic column with Pyrex
glass wool at bottom and a Teflon stopcock.
NOTE: Fritted glass disks are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without frits
may be purchased. Use a small pad of Pyrex glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone followed
by 50 ml of elution solvent prior to packing the column with adsorbent.
• Kuderna-Danish (K-D) apparatus
- Concentrator tube « 10 «L, graduated (Kontes K-570050-1025 or
equivalent); ground-glass stopper is used to prevent evaporation
of extracts ,
- Evaporation flask •• 500 «L (Kontes K-570001-500 or equivalent);
attach to concentrator tube with springs
C.I
-------
Prapert
Sample
Dawatar
Sampto
with Mathanol
Addlntamal
Standards
Soxhfet Extract
S^
Combined
•nd
Exuwiv
Add Wttar
Partition and Dry
Organic Fraction
Concantratto
Organic
I
to
Sampia CJo«iu®
Mathanol/Watar
Extract
FIGURE C.I. Sample Preparation and Extraction
C.2
-------
- snyder column • three-ball macro (Kontes K-503000-0121 or
equivalent)
.- snyder column - two-ball micro (Kontes K-569001-0219 or
equivalent)
• boiling chips - solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent)
• water bath - heated, with concentric ring cover, capable of temperature
control (*5°C); the bath should be used in a hood
• vials - glass, 2-mL capacity, with Teflon-lined screw cap
• glass or paper thimble or glass wool - contaminant free
• heating mantle - rheostat controlled
• syringe - 5 mL ,
• apparatus for grinding - If the sample will not pass through a 1-mm
standard sieve or cannot be extruded through a 1-nm opening, it should
be processed into a homogeneous sample that meets these requirements.
Fisher Mortar Model 185 Grinder, Fisher Scientific Co., Catalogue
Number 8-323, or an equivalent brand and model, is recommended for
sample processing. This grinder should handle most solid samples,
except gummy, fibrous, or oily materials.
• analytical balance
C.3 REAGENTS
• reagent water - reagent water is defined as water in which an
contamination is not observed at the method detection limit of the
compounds of interest.
• methanol - pesticide quality or equivalent
• extraction solvents - sediment samples shall be extracted using either
of the following solvent systems:
- methylene chloride/methanol 2:1 (V:V), pesticide quality or
equivalent
C.3
-------
- methylene chloride/raethanol 9:1 (V:V), pesticide quality or
equivalent
.- Benzene/methanol 3:2 (V;V), pesticide quality or equivalent.
• exchange solvent • hexane that is pesticide quality or equivalent.
C.4 SAMPLE PREPARATION , • '
If a fresh sediment sample is used, decant and discard any water layer on
a sediment sample. If a frozen sample is used, thaw slowly at room temperature
and incorporate overlying water.; Mix sample thoroughly, especially composited
samples. Discard any foreign objects (e.g., sticks, leaves, and rocks).
Homogenize to uniform texture and color.
C.5 ANALYTICAL METHODS
* Sediment Dewaterinq. Weigh 50- to 100-g sample of sediment to nearest
0.1 g. Mix sample of wet sediment with 50 wL of methanol in a glass
centrifuge bottle. The mixture is centrifuged at 163 X gravity
(1000 rpm) for 10 ain. The supernatant is removed and saved. The
methane! wash is repeated and the two supernatants combined. The
supernatant solution is filtered through glass wool and the glass
wool is rinsed with .an additional 10 ml of methanol. The filtered ::
solution is saved. Alternatively the sediment and methanol solution
may be placed in an extraction thimble and the methanol removed by
draining. A glass wool plug above and below the sample in the Soxhlet
extractor is an acceptable alternative for the thimble. The glass
wool plug will filter the methanol solution. Save the filtered
methanol sample for combining with the Soxhlet extract.
• Addition of Internal Standard Spike. Place dewatered sediment sample
in Soxhlet thiable. Add llo »L. of the internal standard spiking
solution(s) Into the sample. Cover the sample with a thin layer of
solvent-cleaned glass wool; An internal standard (i.e., i chemically
inert compound not expected to-occur in an environmental sample)
should be added to each sample and the blank sample just prior to
C.4
-------
extraction or processing. The recovery of the internal standard is
used to monitor for unusual matrix effects, gross sample processing
errors, etc. Standard recovery 1s evaluated for acceptance by
determining whether the measured concentration falls within the
acceptance limits. Recommended standards for different analyte groups
follow; however, these compounds or others that better correspond to
the analyte group Bay be used if past experience warrants.
• Polynuclear aromatic hydrocarbons (PAH) internal spiking
solutions: The recommended Internal standards are the deuterated
compounds of naphtha!ene-d8, acenaphthalene-dlO, and perylene-
d!2. Prepare the internal standard spiking solution in hexane
that contains the compounds at a concentration of approximately
50 ng/pL. The final concentration that is used will depend on
the nature of the sanple.
- Organochlorine pesticide and polychlorlnated biphenyl (PCS)
Internal spiking solution: The recommended internal standard for
organochlorine pesticides and PCBs 1s p,p' dibromooctafluorobi-
phenyl. l-repare the Internal standard spiking solution at a
concentration of 1 ng/pL in hexane. The final concentration -.
that is used will depend on the nature of the sample.
Extraction. Place 200 til of the extraction solvent and one or two •.
clean boiling chips Into the Soxhlet tillable. Attach the flask to the"
extractor and extract the sample for 16 to 24 h. Stir the sample in
the thimble at least twice (after the second cycle and after
approximately 12 h) to prevent solvent channeling. (The glass wool
should be removed during stirring and then replaced.) The Soxhlet
apparatus should be wrapped up to the condenser with aluminum foil
to ensure even heating during cycling. Allow the extract to cool
after extraction Is completed.
Liquid-Liquid Extraction. Transfer the cooled extract to i 500-mL
separator/ funnel. Rinse the Soxhlet flask twice with clean extraction
solvent and add this rinse to the extract in the separatory funnel.
Add the filtered methanol solution from the dewatering step. Wash
C.5
-------
the solvent extract with approximately 100 mi of 50% Na2S04 saturated
organic-free water. Collect and store the organic layer. Re-extract
the aqueous phase twice with 60 ml of clean hexane and add both
extracts to the initial organic fraction. If benzene/aethanol was
used in. the extraction process, the organic layer will be the top
phase in the funnel* If one of the methylene chloride/methanol
mixtures was used, the organic layer will be the bottom phase.
- Formation of emulsions or precipitates during liquid-liquid
extraction should be noted and considered when reviewing the
results. The addition of Na2$04 nay reduce emulsions; however,
if the emulsion interface between layers is more than one-third
the volume of the solvent layer, the analyst must employ
mechanical techniques to complete the phase separation. The
optimal mechanical technique depends on the sample and may include
stirring, filtration of the emulsion through precleaned glass
wool, or centrifugatiort.
Dry and Concentrate the Extract. Assemble t K-3 concentrator by
attaching a 10-mL concentrator tube to a 500-mL evaporation flask.'
- Dry the organic layer by pouring it through an anhydrous NaaS04
drying column (approximately 30 cm by 2 cm). Use approximately
30 ml of hexane to rinse the drying column and combine this
solution with the dried extract. Collect the dried extract in
a K-D concentrator.
j
- Add one or two clean boiling chips to the flask and attach a
three-ball macro Snyder column. Prewet the Snyder column by
adding approximately IjoL of methylene chloride to the top of
the column. Place the K-D apparatus in a hot water bath (15°C
to 20°C above the boiling point of the solvent), so that the
concentrator tube Is partial1y immersed in the hot water and
the entire lower rouncUrd surface of the flask is bath®d wi.th
hot vapor. Adjust the vertical position of the apparatus and
the water temperature, as required/to complete the concentration
in 10 to 20 Bin. At the proper rate of distillation, the balls
C.6
-------
of the column will actively chatter, but the chambers will not
flood. When the apparent volume of liquid reaches 1 ml, remove
the K-D apparatus from the water bath and allow it to drain and
cool for at least 10 min.
• Solvent Exchange. The solvent exchange step is required before the
cleanup of the samples if methylene chloride was used in the extraction
solution. To perform the solvent exchange, remove the Snyder column,
add 50 ml of hexane, and reattach the Snyder column. Concentrate the
extract as described in the previous paragraph.
• Calibration and Spiked Standard Solutions. The calibration solutions
for PAHs, PCBs, and pesticides are prepared in hexane to a
concentration of approximately 50 ng/^L for each of the compounds
given in Tables C.I and C.2. The spiked standard solutions for PAHs,
PCBs, and pesticides are also prepared in hexane to a concentration
of approximately 50 ng/fiL for each of the compounds given in Tables C.3
and C.4. These solutions are subjected to the sane analytical
procedures as the actual samples starting at the Soxhlet extraction
step.
- The extracts obtained may now be cleaned up to remove interfer-
ences and analyzed using the methods in MacLeod et al. (1985;
starting at Section 5) or if sulfur cleanup is required by us.ing
one of the methods in Appendix 0. If cleanup and analysis of
the extract will not be performed immediately, stopper the
concentrator tube and refrigerate. If the extract will be stored
longer than 2 days, it should be transferred to a Teflon-sealed
screw-cap vial and labeled appropriately.
C.6 QUALITY CONTROL
• All calibration and spiked standard solutions should be subjected to
exactly the same analytical procedures as those used on actual samples.
• Solvents, reagent, glassware, and other sample processing hardware may
yield artifacts and/or interferences to sample analysis. All these
C.7
-------
I
TABLE C.I. List of Compounds used in the PAH Calibration Solution
Hexamethylbenzene
Naphthalene
2-Methytnaphthalene
1-Methylnaphthalene
Biphenyl
2,6-Oi methylnaphthalene
Acenapht'hene
Fluorene
Phenanth'rene
Pyrene
Benzo(a)anthracene
Chrysene!
Benzo(e)pyrene
Benzo(a)pyrene
Perylene
Oi benzo(a,h)anthracene
Naphthalene-d8
Acenaphthene-dlO
Perylene-dl2
^
(a) Adspts-j frCsa Hacleod et al.
(1985)
materials must be demonstrated to be free fron interferences under the
conditions of the analysis by analyzing method blanks. Specific
selection of reagents and purification of solvents by distillation .
in all-glass systems nay be required.
Phthalate esters contaminate many types of products commonly found in
the laboratory. Plastics, in particular, must be avoided. Phthalates
are commonly used as plasticizers and are easily extracted from plastic
materials. Serious phthalate contamination may result at
-------
TABLE C.2. List of Compounds used in the PCB and Pesticide
Calibration SolutionU)
Tetrach1oro-ra-xy1ene
Hexachlorobenzene
Lindane (-7-BHC)
Heptachlor
Heptach1or-epoxide
Aldrin
«-Chlordane
Trans-nonachlor
Oieldrin
Mi rex
o.p'-DOE
p,p'-OOE
o.p'-ODO
p.p'-ODD
o(p'-ODT
p.p'-ODT
2,4'-Oich1orobipheny1
2,5,4'-Trichlorobiphenyl
2,4,2',4'»Tetrachlorobipheny1
2,4t5(2',5'-Pentachlorobipheny]
2,4,5,2*,4*,5'-Hexach1orobipheny1
2,3,4,3,6,2*,5*-Heptachlorobiphenyl
2,3,4,5,2*,3*,4*,5«-Octach1orobipheny1
2,3,4,5,6l2',3',4',5l-Nonachlorobipheny1
4,4*-Dibrooooctafluorobiphenyl
(a) Adapted froa MacLeod et al. (1985)
C.9
-------
TABLE C.3. List of Compounds used in the PAH Spike Solution
(a)
Naphthalene
2-Methylnaphthalene
1-Methylnaphthalene
Biphenyl
2,6-Diraethy1naphtha!ene
Acenaphthene
FT uorene
Phenar|ithrene
Anthracene
1-Methy1phenanthrene
Fluoranthene
Pyrene
Benzo(a)anthracene
Chrysene
Benzo|e)pyrene
8enzo(a)pyrene
Perylene
Oibenzo(a,h)anthracene
Adapted from MacLeod et al
(198S)
C.10
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TABLE C.4. List of Compounds used In the PCB and Pesticide Spike Solution
(a)
Hexachlorobenzene
Lindane (-/-BHC)
Heptachlor
Heptachlor-epoxide
Aldriii '
•-Chlordane
Trans-nonachlor
Dieldrin
Hi rex
o.p'-DDE
p.p'-DDE
o.p'-ODD
p.p'-ODD
o.p'-DDT
p.p'-ODT
2,4'-Dichlorobiphenyl
2,5,4'-Trichlorobiphenyl
2 , 4 , 2 ' , 4 ' -Tet rach 1 orob i pheny 1
ZAS.a'.S'-Pentachlorobiphenyl
Z.S.A.S.e.Z'.S'-Heptachlorobiphenyl
2,3,4,5,2' ,3' ,4' .S'-Octachlorobiphenyl
2,3,4,5,6,2',3l,4',5'-Nonachlorobiphenyl
(a) Adapted fron MacLeod et al. (1985)
C.ll
-------
C.7 REFERENCES !
- I
MacLeod, W. D., D. W. Brown, A; J. Friedman, D. G. Burrows, 0. Maynes R. W
Pearce, C. A. Wigren and R. 6. Bogar. 1985. Extractable Toxic Organic
Compounds .. Second Edition. Standard Analytical Procedures of the NdAA
5aSS^A2alytlcal Fac1lity 1985-1986. NOAA Technical Memorandum NMFS
F/NWC-92, Seattle, Washington.
Tech. 1986a. Analytical Methods for U.S. EPA Priority Pollutants and
301 (h) Pesticides in EstuarinT ana Marine Sediments. Prepared for Mar™S
Operations Division, Office of Marine and Sstuanne Protection, U.S.
Environmental Protection Agency, Washington, D.C.
Tetra Tech. 1986b. Recommended Protocols for Measuring Organic Comnounrig in
Puqet Sound Sediment and Tissue Samples. Prepared for 6l?ice of il>un»t
-------
APPENDIX D
METHODS FOR SULFUR CLEANUP OF EXTRACTS
-------
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APPENDIX D
METHODS FOR SULFUR CLEANUP OF EXTRACTS
D.I SUMMARY OF METHOD
Three techniques for the elimination of sulfur are described: 1) the use
of copper powder, 2) the use of mercury, and 3) the use of tetrabutylammonium
(TBA)-sulfite. The TBA-sulfite causes the least amount of degradation of a
broad range of pesticides and organic compounds, while copper and mercury may
degrade organophosphorous and some organocMorine pesticides (Table D.I).
The sample to undergo cleanup is mixed with either copper, mercury, or
TBA-sulfite. The mixture is shaken and the extract is removed from the sulfur
cleanup reagent. The analytical scheme is shown in Figure D.I.
i
•D.2 APPARATUS AND MATERIALS
• mechanical shaker or mixer, such as the Vortex Genie
S
• pipettes, disposable, Pasteur-type
• centrifuge tubes, calibrated, 12 ml
• glass bottles or vials - 10 and 50 ml, with Teflon-lined screwcaps.
D.3 REAGENTS - -"
• reagent water - Reagent water is defined as water in which a
contamination is not observed at the method detection limit of the
compounds of interest.
• nitric acid - dilute.
• acetone, hexane, 2-propanol - pesticide quality or equivalent.
• copper powder - Remove oxides by treating with dilute nitric acid,
rinse with distilled water to remove all traces of acid, rinse with
acetone, and dry under a stream of nitrogen (copper, fine-grained
Mallinckrodt 4649, or equivalent).
D.I
-------
TABLE D.I. Effect of Mercury and Copper on Recovery of Pesticides
Percent Recovery^a' Usings
. Pesticide Mercury Copper
Aroclor 1254 . 97.10 104.26
Lindane 75.73 94.83
Keptachlor 39.84 ; 5.39
Aldrin ; 95.52 93.29
Heptachlor epoxide 69.13 96.55
DDE i 92.07 102.91,
DDT 78.78 . 85.10
BHC , 81.22 98.08
Dieldrin ! 79.11 94.90
Endrin ' 70.83 89.26
Chlorobenzilate 7.14 0.00
Malathion 0.00 0.00
Diazinon i 0.00 0.00
Parathion 0.00 0.00
Ethion i 0.00 0.00
Trithion | 0.00 0.00
(a) Percent recoveries cited are averages based on duplicate
analyses for all compounds other than for aldrin and
BHC* For a]<>i"i foyr and three determinations wens
averaged to obtain the result for mercury and copper,
respectively. Recovery of BHC using copper is based on
one analysis.
mercury - triple distilled. ,
TBA-sulfite reagent • Dissolve 3.39 g TBA hydrogen sulfate in 100 ml
of reagent water. To remove!impurities, extract this solution three
times with 20-raL portions of hexane. Discard the hexane extracts,
and add 25 g sodiuo sulfite to the water solution. Store the resulting
solution, which is saturated;with sodium sulfite, in an amber bottle
with a Teflon-lined screw-cap. This solution can be stored at room
temperature for at least 1 month.
D.2
-------
Cantnfuoa and
Draw OH Samola
Extract
Concamrata
Samola Extract
Concantrata
Samola Extract
Concantrata
SampM Extract
Tranafar Extract
10 Camrifuea
Tuba
Saoaraia Extract
from Coooar
Tranatar Extract via
rlpatta into Concan*
trater Tuba or Vial
AddMarcury:
Agitata
Saearata Sameta
from Marcury
Aperoeriata
Analytical
M«tnod
Tranafar Extract
to Glat» Settta
or Vial
Add TBA-Surfita
and 2-*tooano4;
Shaka
Wmar Sitaka:
\JK Stand: .
Concantrata Extract
TAt « Tameutyiammenum
FIGURE D.I. Analytical Scheme for Removal of Sulfur from Extracts
0.3
-------
D.4 SAMPLE PREPARATION ;
The sample used is the final product from the extraction obtained from
the procedures in Appendix C or from Section 7 of MacLeod et al, (1985).
D.5 ANALYTICAL METHODS ;
• Removal of sulfur using copper
- Concentrate the sample to exactly 1.0 ml in the Kuderna-Danish
(K-D) tube. i
-» If the sulfur concentration is such that crystallization occurs,
centrifuge to settle the crystals, and carefully draw off the
sample extract with a disposable pipette, leaving the excess
sulfur in the K-D tube. Transfer the extract to a calibrated
centrifuge tube.
.» Add approximately 2 g (to the 0.5 mL mark) of cleaned copper
pcvrdsr tc the Centrifuge tube, mx for at least 1 mini on the
mechanical shaker.
•• Separate the extract froa the copper by drawing off the extract
with a disposable pipette and transfer to a clean vial. The
volume remaining still represents 1.0 «L of extract.
NOTE: This separation is necessary to prevent further degradation
of the pesticides. ,
i
• Removal, of sulfur using mercury
NOTE: Mercury is a highly toxic metal and, therefore, must be
used with great care. Prior to using mercury, it is recommended
that the analyst become) acquainted with proper handling and
cleanup techniques associated with this metal.
•• Concentrate the sample extract to exactly 1.0 «l».
« Transfer 1.0 »L of the extract into a clean concentrator tube or
Teflon-sealed vial using a disposable pipette.
Dc4
-------
• Add one to three drops of mercury to the vial and seal. Agitate
the contents of the vial for 15 to 30 sec. Prolonged shaking
(2 h) may be required; if so, use a mechanical shaker.
• Separate the sample from the mercury by drawing off the extract
with a disposable pipette and transfer to a clean vial.
• Removal of sulfur using TBA-sulfite
• Concentrate the sample extract to exactly 1.0 ml.
• Transfer the 1.0 «L to a 50-mL clear glass bottle or vial with
a Teflon-lined screw-cap. Rinse the concentrator tube with
1 ml of hexane, adding the rinsings to the 50-mL bottle.
- Add 1.0 ml TBA-sulfite reagent and 2 ml 2-propanol, cap the
bottle, and shake for at least 1 min. If the sample is colorless
or if the initial color is unchanged, and if clear crystals
(precipitated sodium sulfite) are observed, sufficient sodium
sulfite is present. If the precipitated sodium sulfite
disappears, add pore crystalline sodium sulfite in approximately
100-rag portions until a solid residue remains after repeated
shaking. =;
- Add 5 ml distilled water and shake for at least 1 min. Allow the
sample to stand for 5 to 10 min. Transfer the hexane layer (top)
to a concentrator tube and use the K-0 technique to concentrate
the extract to 1.0 ml.
• Complete the cleanup of the extracts by using the method in Section 5
of MacLeod et al. (1985).
D.6 QUALITY CONTROL
^•^•••••••MaMMMi^^^^^^^^Ha^^^ . ^ ^ |
• All.reagents should be checked prior to use to verify that
interferences do not exist.
• The copper must be very reactive, therefore, all oxides of copper must
be removed so that the copper has a shiny, bright appearance.
0.5
-------
• The sample extract uust be vigorously agitated with the reactive copper
for at least 1 minute.
D.7 REFERENCES
U.S. Environmental Protection Agency (EPA). 1986. Test Methods for Evaluating
Solid Waste. SW-846. Office of Solid Haste and Emergency Response.
Washington, O.C.
MacLeod, W. D., D. W. Brown, A. J. Friedman, D. G. Burrows, 0. Maynes, R. W.
Pearce, C. A. Wigren, and R. G. Bogar. 1985. Extractable Toxic Organic
Compounds. Second Edition. Standard Analytical Procedures of the NOAA—
National Analytical Facility 1985-1986. NOAA Technical Memorandum NMFS
F/NWC-92, Seattle, Washington.
0.6
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