PB85-169597
PAH (Polycyclic Aromatic Hydrocarbons)
Uptake by Plants: Methodology and
Initial Investigations
Clemson Univ., SC
Prepared for
Environmental Research Lab., Athens, GA
Feb 85
I
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EPA/600/D-85/036
February 1985
PAH UPTAKE BY PLANTS
Methodology and Initial Investigations
by
John Coatesl, Alan W. Elzermanl, and A. Wayne tidirlson?
Environmental Systems Engineering
Clemson University
Clemson, SC 29631
^Environmental Research Laboratory
U.S. Environmental Protection Agency
Athens, GA 30613
ENVIRONMENTAL RtSfcARCH LABORATORY
OFFICt OK RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
ATHENS, (iA 30613
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NOTICE
This document has been reviewed in accordance with
U.S. Environmental Protection Agency policy and
approved for publication. Mention of trade naaes
or coaaercial products does not constitute endorse-
ment or recoMendation for use.
ii
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PAH UPTAKE BY PUNTS; METHODOLOGY AND INITIAL INVESTIGATIONS
JOHN COATESi, ALAN W. ELZERMANl, A. WAYNE GARRISON2
(1 Environmental Systems Engineering, Clemson University,
Clemson, SC 29631; (2)Environmental Research Laboratory,
U.S. EPA, Athens, OA 30613.
. INTRODUCTION
Polycycl1c aromatic hydrocarbons (PAKs) are formed during
the pyrolysis of naturally occurring organic materials.
Pyrolytic reactions may be induced anthropogenlcally or by
natural events (e.g., forest and grassland fires or volcanic
activity). Background levels and the ublgultous occurrence
of PAHs in the environment may result from synthesis 1n
terrestrial vegetation, microbial systhesls and volcanic
activity (Andelman and Suess, 1970; Suess, 1976; Shabad,
1980). The greatest amounts of PAHs released Into the environ-
ment, however, come from anthropogenic sources (Andelman and
Suess, 1970). Fossil fuel burning for power production
appears to be the most significant source, with motor vehicle
fuel combustion and anthropogenic forest and agricultural
fires contributing an additional IX and 81 of the total,
respectively (Suess, 1976). Recent reviews by Howard and
Fazio (1980) and Edwards (1983) provide summaries addressing
the sources, occurrence, plant uptake, degradation and analyti-
cal methodologies for PAHs 1n the environment.
Although a large data base has accumulated on the presence
of PAHs in food products and plants, routine measurement of
the uptake and accumulation of PAHs 1n vegetation has been
hampered by analytical techniques. Sensitivity Is often a
problem with these techniques and the majority of the proce-
dures cited in the literature are tedious, length/ and expen-
sive to perform and fheiefore are not amenable to routine
analyses. For example, Winkler et al_. (1977) described a
method for the determination of TOHs with three to seven
rings In wet and dry maize. Samples were extracted using a
Soxhlet apparatus; extracts were rotary evaporated, saponified,
filtered over silica gel, evaporated for a second tipe and
partitioned with Sephadex LH 20. Howard et al_. (1968) described
a method of similar complexity for root vegetables using
saponification, multiple solvent partitioning, solvent replace-
ment, and adsorption chromatography on FloMsil. Recently,
Coates et aK (1984) have reported a simpler method suitable
for routine analysis of PAHs 1n plant tissues with sufficient
sensitivity for screening PAH uptake by plants growing under
conditions of low to moderate exposure to PAH contamination,
1
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PAHs IN PLANTS
Such as might occur next to a roadway, upon Irrigation with
treated wastewater, or during overland Mow treatment of
wastewater. As reviewed brlow, the sonlflcatlon extraction
procedure coupled to a silicic acid clean-up scheme was found
to be reliable and convenient.
The new sonlflcatlon method of Coates et al. (1984) was
applied In a field study of plants 1 rrlgateT~wTTh PAH-
spiked domestic wastewater or grown on contaminated soil. The
method was found appropriate to measure concentrations In
hybrid grain sorghum and fescue down to the 25 ugAg level.
Apparent uptake of some PAHs from contaminated soil and/or
Irrigation water was demonstrated using this technique.
MATERIALS ANO METHOOS
Chemicals
(a) Acetonltri1e and pentane (pesticide grade)
b) Sodium sulfate (anhydrous granular, ACS quality)
c) Sodium chloride (granular, ACS quality)
(d) Mercuric chloride (granular, ACa quality)
(e) Silicic acid (Unlsi 1R activated silicic acid, 100-
200 mesh, CI arkson Chemical Co., Inc.)
(f) PAHs (Aldrlch Chemical Co.)
Acenaphthylene, 95X
Flouranthene, 98%
Pvrene, 99+%
Bemo[a]Pyrer.e, 98%
Benzo[g,h,1]Perylene, M.P. 277-279°C
(g) PAH standards (National Bureau of Standards SRM
1647, 16 PAHs In acetonltrlle)
(h) Laboratory blank water (twice distilled In glass)
Apparatus
(a) Sample preparation (HobartR food processor Model
8181 D)
(b) Sonlflcatlon extraction (Brlnkman PolytronR with
PT-20 ST probe generator)
(c) Centrifuge (Sorvall Superspeed RC-2B automatic,
refrigerated, with GSA heat)
(d) Sample concentration (Kuderna-Oanlsh concentrator,
500 ml)
(e) Sample clean-up columns (Pasteur plpets)
(f) Gas-liquid chromatography (Hewlett-Packard Model
5880 Capillary GC equipped with an FID detector and
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PAHs IN PLANTS
a 30M x 0.2Iran 10 SE-54 capillary col win having a
0.25-i«i film thickness). Instrument conditions:
linear velocity, )3 cm/sec; Injector temperature,
250°C; detector temperature, 350°C; temperature
program, 70°C for 2 minutes then 20*C/m1n to 125°C
then 4°C/m1n to 290#C, held for 15 minutes.
Plant Extraction Cleanup Technique
As reported by Coates et aK (1984), acetonltrlle (75 mL)
was added to the centrifuge tube containing the plant mulch.
The sonlflcation probe generator was Immersed Into the suspen-
sion tube and the mixture sonicated for 2 minutes at maxlmun
speed. Contents wsre then centrlfuged at 5500 g for 20
minutes and the centrlfugate quantitatively transferred to a
500 mL separator? funnel. The solid was washed with 25 mL of
acetonltrlle and centrlfuged; t*e centrlfugate was combined
with the first extract. Pent' (50 mL) was added to the
separator? funnel containing t oetonltrlle and shaken
vigorously for 2 minutes. Water & iturated with sodium chloride
(10 mL) was then added to the separator? funnel, followed by
350 mL of laboratory blank water, and again shaken for 30-40
seconds. The 1mr?s1sc1ble phases were separated, and the
pentane layer was dried by passing through a sodium sulfate
column and drained Into a 5C0-mL K-D apparatus.
The pentane extract «s concentrated to a volume of 3 to
4 mL 1n a water bath at 65°C, removed, and allowed to cool
for 10 minutes. The K-D flask was rinsed with 3 mL of dry
pentane and the 10-mL concentrator tube was removed from the
flask. All ground glass joints were rinsed with 2 mL of
pe.itane, which also was collected 1n the concentrator tube.
The extract was then blown down to 0.5 mL with dry nitrogen
at room temperature and quantitatively transferred to a
prewashed micro clean-up column containing 1.5 gm of Un1s11
silicic acid; the column was then eluted with 5 mL of dry
pentane to remove primarily aliphatic hydrocarbons *-orn the
extract. The second and third 5-mL fractions were ch
eluted with a 85:15 solvent mixture of pentane and methylene
chloride (DCM). All fractions were again concentrated to
0.5hhL with dry nitrogen and analyzed by GC-FIO. All PAHs
appeared 1n the second 5-mL fraction collected. No PAH* were
ever found in the first (pentane) fraction nor the third
(pentane:DCM) fraction. The clean-up proceduih was sufficient
for extracting up to 15 gm of air dried plant material;
additional silicic acid was required to remove Interfering
polar constituents when extracting more than 15 gms of plant
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PAHs IN PLANTS
material. The method Mas scaled up when necessary on the
basis of silicic acid pore volume to plant mass ratios.
Sam)le Collection and Spiking for Recovery Studies
Immature hybrid grain sorghum Mas harvested from the
Clemson University Agricultural Experiment Station - Simpson
Farm diving the early fall of 1982 and stored at -I0°C until
recovery studies Mere Initiated. All plants Mere removed
from storage and the aerial plant parts homogenized 1n a food
processor prior to spiking for recovery studies.
A weighed portion (2*10 gm) of plant mule*". was placed In
a 150-fliL glass centrifuge tube with I ml of a PAH mixed
standard (NBS SRM 1647). Laboratory blank water (150 mL) and
1 rr.L of 185 inmolar mercuric chloride as a bactericide were
added to the tti>e. The tube was capped and placed on a slide
shaker 1n a dark room for 24 hours. After shaking, the
contents were centrlfuged at 5500 g for 20 minutes. The
aqueous phase was transferred to a separatory funnel and
extracted 3 times w4th 30-mL portions of pentane. Extracts
were combined and concentrated using a K-0 apparatus to
approximately 3 to 4 mL, then by dry nitrogen to 0.5 mL. The
centrifuge tube wall was extracted 3 times with 10-mL portions
of pentane and concentrated to 0.5 m*. above. The plant
residue was extracted by sonlflcatlo..
Experimental Design for Field Study.
A field study was originally Initiated (Coates et al..
1964) to demonstrate that the sonlflcatlon technique was
suitable to extract plant tissue that had accumulated PAHs
during growth as well as with spiked samples. The additional
data reported here were obtained to make an Initial assessment
or uptake of selected PAHs and to Investigate whether uptake '
could occur from contaminated soil as well as from Irrigation
water. In the Irrigation water study, hybrid grain sorghum
and fescue were grown from seed In 20-L containers and Irrigated
dally with PAH-spiked municipal sewage effluent for an 8-weyk
period during the simmer of 1983. Treated activated sludge
effluent wastewater was obtained from the Coneross Wasto
Treatment Plant, Oconee County, South Carolina, and stored
with aeration at 4°C during the study period. The ? xpsr Intents 1
design was a completely -ando.n design with triplicate controls
and triplicate PAH treatment representing two levels of PAH
contamination for both sorghum and fescue. Polycycllc aromatic
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PAHs IN PLANTS
hydrocarbons used In this 'nvestlgatlon Mere acenaphthylene,
fluoranthene, pyrene, benzo[a]pyrene, and benzo[g,h,1]perylene.
The concentration of the five PAHs In the spiked wastewater
and the total mass of PAHs Introduced to the growing plants
during the study period are listed in Table 1.
TABLE i
CONCENTRATIONS AND MASSES OF PAHs USED IN THE IRRIGATION STUDY
PAH
Concentration
1n Spiked Wastewater
Total Mass Applled
In Spiked Waste-
water to Each
Container
Treatment
1 2
ug/L
Treatment
1 2
u9
Acenaphthylene
2.0
1000
640
320,000
F1 uoranthene
2.0
200
640
64,000
Pyrene
1.0
100
320
32,000
Benzo[a]pyrene
0.05
1.0
16
320
Benzo[g,h,1jperyl ene
0.10
0.5
32
160
plants were irrigated using hand-pump pressurized spray
cans. Containers were moved frequently to eliminate position
In the growing area as a variable. The aerial part of each
plant was harvested at the end of the growing period and
frozen until extracted. The seeds were planted 1n July,
Irrigation with spiked wastewater was begun 1n September, and
the plants were harvested In November.
To obtain some Indication of the -«o*»nt1al for root
uptake of PAHs, the soil In two 20-L to liners was contami-
nated with three PAHs (acenaphthylene, f.joranthene and
pyrene). One gram of each PAH was added to the top two Inches
of soil and thoroughly mixed. The soil density was estimated
to be 1.25 gm/cm3, which translated Into a calculated 217 ppm
PAH concentration In soil (not verified by extraction).
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PAHs IN PLANTS
Contaminated soil was incubated for 1 month prior to seeding.
Fescue Mas planted 1n one container and hybrid grain sorghum
was planted 1n the second container. Two controls wr.re
planted at the same time and all four containers wert frr1gat°d
wit* tapwater during the 8-week growing period. During this
tirce the soil surf* ; was not allowed to dry, thus reducing
the effect of "dust.ng," which could contaminate aerial plant
parts.
Evaluation of Analytical Technique
As reported by Coates et ai_. (1964), the elutlon ¦ .n .
and corresponding relative retention times for the 16 • s
present In the N6S SRM 1647 standard are lasted in Table 2.
The chromatographic conditions used 1n this Investigation are
listed 1n the Materials and Methods section. Retention
orders for PAHs having different molecular weights were
verified using gas chromatography on an SE-54 capillary
column and selected 1on monitoring mass spectrometry (GC-MS).
The retention order for the remaining PAH molecular weight
pairs (phenanthrene-anthracene, fluoranthene-pyrene,
chrysene-B[a]A, and B[b]F-B[k]F) were determined by spiking
the NBS standard with an EPA quality control sample containing
one of the PAHs of the paired combinations and analyzing by
the GC-MS technique.
RETENTION TIMES FOR 16 PAHs PRESENT IN THE NBS SRM 1647
STANDARD*
RESULTS ANO DISCUSSION
TABLE 2
PAH
Relative Retention
Mol. Wt. (referenced to
Pyrene)
1. Naphthalene
2. Acenaphthylene
3. Acenaphthene
4. Fluorene
5. Phenanthrene
6. Anthracene
128.2
152.2
154.2
166.2
178.2
178.2
0.272
0.457
0.485
0.566
0.733
0.741
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PAHs IN PLANTS
TABLE 2 (Continued)
PAH
Relative Retention
Mol. Wt. (referenced to
Pyrene)
7. Fluoranthene
8. Pyrene
9. Benzo[a]anthracene
10. Chrysene
11. Benzo[b]fluoranthene
12. Benzofluoranthene
13. Benzopyrene
14. Indeno[l,2,3-cd]pyrene
15. D[a,h]anthracene
16. BenzoCg.hJ ]perylene
202.3
202.3
228.3
228.3
252.3
252.3
252.0
276.3
278.4
276.3
0.959
1.000
1.238
1.246
1,439
1.443
1.494
1.739
1.749
1.805
~After Coates et al., 1984
Plant extracts when concentrated typically contain high
percentages of polar organic compound* that co-elute and
significantly Interfere with many chromatographic analyses.
Many of the available clean-up schemes are complex and tedious
to Implement for routine analyses. The single sorbent clean-up
scheme used by Coates et aj_. (1984) coupled with solvent
partitioning was sufficient to allow quantification of all
PAHs listed in Table 2 at the 25 yg/kg level. PAHs extracted
by acwtonltrile were partitioned Into pentane, dried on
sodium sulfate, concentrated to 0.5 mL and cleaned up with a
micro-silicic acid column, as noted In the Materials and Methods
Section. Two 5huL fractions were eluted with a pentane:DCM
(85:15) solvent mixture. Optimization of the solvent ratio
was necessary to maintain the usefulness of the single adsorbent
clean-up step (Coates et al_., 1984).
Figure la Is a chromatogram of the PAH standard and
figure lb 1s a chromatogran of the first 5-ml eluant fraction
of a plant extract using pentane:DCM (85:15) as eluant.
Coates et al. (1984) showed quantitative recovery for most of
the PAHs"from hybrid grain sorghum extracts for the concentra-
tion and cleanup steps with the exception of 1ndeno[l,2,3-cd]
pyrene and benzo[g,h,1]perylene, which showed losses of 12
and 17 percent, respectively.
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P/ ..IN PUNTS
(a)
J L
U
it u
is
I j.*. i
(b)
„]JL-L
FIGURE 1. Chromatograra of (a) NES 1647 standard (numbered
peaks appear 1n order as In Table II) and (b) sample eluted
with 85:15 pentanerDCM.
Overall method calibration and recovery also was
evaluated (Coates et al., 1984). Initial calibration studies
for the 16 PAHs use? Tn this Investigation Indicated detector
!1near1ty over the analytical working range so a single point
calibration curve was used to quantify PAH concentration and
calculate recovery efficiencies. Standards (NBS SRM 1647)
were injected in the gas chromatograph every 4 to 5 samples
to monitor system drift. Results of overall method recovery
studies are listed in Table 3. These data were corrected for
residual naphthalene, acenaphthylene and acenaphthene concen-
trations remaining 1n the aqueous phase after spiking and
Incubation with the plant mulch. Aqueous phase concentrations
of the remaining PAHs ware below detectable limits (C.05-1
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PAHs IN PLANTS
ng/ml). See Materials and Methods for spiking details.
Concentrations of PAHs spiked Into the plant tissue
homogenates were between 600 and 4500 ug/kg.
TABLE 3
OVERALL METHOD RECOVERY EFFICIENCIES (PAH PLANT CONCENTRATION
MO-WOO *A«1
PAH
Mass 1n
Spike
Percent
Recovered*
%
1.
Naphthalene
22.5
2.
Acenaphthylene
13.1
3.
Acenaphthene
21.0
4.
Fluorene
4.92
5.
Phenanthrene
5.06
6.
Anthracene
3.29
7.
Fluoranthene
10.1
8.
Pyrene
9.84
9.
B[a]A
5.03
10.
Chrysene
4.68
11.
B[b]r
5.11
12.
B[k]F
5.02
13.
B[a]P
5.30
14.
I[l,2,3-cd]P
4.06
15.
D[a,h]A
3.68
16.
B[g,h,1]Perylene
4.01
47*5 n
81*(8)
80+(8)
8Z+(8)
90+(9
82?(8
83+12
837(13
72+(6
717(9
62+16
617(15
57+(5)
49+(17)
45+(17
617(14
•Average of four replicates with standard deviations in
parentheses.
Data 1n Table 3 suggest that the son1f1cat1on procedure
was generally efficient 1n removing most of the PAHs from the
plant matrix. Recovery efficient as, however, for the more
volatile naphthalene and for a few of the more hydrophobic
PAHs were lower (45-62%). The practical detection limit was
approximately C.025 u9/9*
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PAHs IN PLANTS
Field Application Studies
Irrigation Study. At the end of the 8-week growing
period, the hybrid grain sorghum and fescue that were Irrigated
with PAH-sp1ked domestic wastewater Were harvested and stored
frozen until extracted. All plants were extracted using the
son1f1cat1on technique with the clean-up scheme outlined
above, and extracts were analyzed by GC-FID.
Results are listed 1n Tables 4 and 5 for hybrid grain
sorghum and fescue, respectively. Concentrations for each
plant type are 1n units of microgram PAH per gram of air dried
plant extracted. Error estimates for the data presented 1n
Table 4 were not available because replicates for both
control and treatments 1 and 2 had to be combined to have
enough plant material to extract. Error estimates for the
control and the two treatment levels for the fescue uptake
study were calculated and were used 1n significance analysis.
TABLE 4
PAH CONCENTRATIONS IN HYBRID GRAIN SORGHUM IRRIGATED WITH
PAH-SHkEB DOMESTIC WASTEWATER
PAH
Control
vg/g
Treatment
1
m9/9
Treatment
2
ug/g
Acenaphthylene
ND*
ND
ND
Fluoranthene
0.05
0.82
1.32
Pyrene
0.02
0.49
0.63
Benzo(a)pyrene
ND
ND
ND
Benzo(g,h,1)perylene
ND
ND
ND
*ND means not detected (less than approximately 0.02 yg/g).
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PAHs IN PLANTS
TABLE 5
PAH CONCENTRATIONS IN FESCUE IRRIGATED WITH PAH-SPIKED
PflHESTlC WASTEWKTEE
PAH
Control Treatment Treatment
1 2
(avg. _+ standard deviation)
Acenaphthylene ND* ND 0.58+0.16
Fluoranthene 0.12+0.03 0.20+0.07 50+13
Pyrene 0.07+0.03 0.10+0.04 24+6
Benzo(a)pyrene KD" TlD TiD
Benzo(g,h,1)perylene ND ND 0.13+0.03
*ND means not detected (less than approximately 0.025 wg/g).
Fluoranthene and pyrene were found 1n the sorghum at
concentrations significantly greater than the control. Four
of the five PAHs were present 1n trie fescue Irrigated with
spiked wastewater at concentrations greater than 1n the
controls. Concentration differences between treatment 1 and
treatment 2 and the controls were significant at the <0.01
level as determined by the Student's T test.
Table 1 gives the total mass of each PAH applies to the
plants for each treatment through the spiked Irrigation
water. No attempt was made to account for losses of PAHs due
to sorption, blodegradatlon, photolysis, volatilization from
soil surface or transpiration, but only to determine whether
measureable quantities of PAHs could be found associated with
growing plants Irrigated with PAH contaminated wastewater.
Similarly, although the experiment Indicated uptake by the
plants, 1t does not distinguish between foliar or root uptake
or a combination of the two. As expected, given different
physical and chemical characteristics, some PAHs accumulated
more than others. Further work 1s required to elucidate mecha-
nisms of uptake and accumulation, however.
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PAHs IN PLANTS
Contaminated Soil Study
Results are listed In Tables 6_and 7 for hybrid grain
sorghum and fescue and Indicate the magnitude of plant uptake
for the PAHs used 1n this study. It 1s Important to note that
treatments were not replicated and the nunbers should only be
viewed as Indications of potential accumulation. No attempt
was made to correlate uptake or accumulation as a function of
plant growth. The data strongly suggest, however, that some
PAHs may be taken up by plant roots and translocated to the
aerial plant. Two of the three PAHs studied, acenaphthylene
and fluoranthene, exhibit greater aqueous solubilities and
lower hydrophobic character than pyrene and were found at
higher levels 1n the plant tissue than pyrene 1n the contami-
nated soil study. In addition, acenaphthylene was the most
volatile of the PAHs studied and may have been lost from the
system. If the results presented are order of magnitude
indicators of plant accumulation tendencies, they are consis-
tent with the above characteristics of PAHs and Indicate
lower uptake of PAHs with lower aqueous phase concentrations.
TABLE 6
PAH CONCENTRATIONS IN SORGHUM GROWN ON CONTAMINATED SOIL
PAH -
Control
Contaminated
Soil
wg/y
wg/g
Acenaphthylene
ND*
NO
Fluoranthene
5.23
73.49
Pyrene
NO
2.38
*ND means not detected (approximately less than 0.02 ug/g)
12
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PAHs IN PLANTS
TABLE 7
PAH CONCENTRATION IN FESCUE GROWN ON CONTAMINATED SOIL
PAH
Control
Contaminated
Soil
vg/g
v9/9
Acenaphthylene
ND*
3.09
F1uoranthene
0.13
10.32
Pyrene
0.06
3.29
*NU means not detected (approximately less than 0.02 yg/g)
Further work 1s required to elucidate and verify uptake
mechanisms and to determine the Impact of application rates
and methods, volatilization, photodegradatlon, blodegradatlon,
plant metabolism and translocation as possible explanations
for the ranges of PAH concentrations observed 1n aerial plant
parts.
ACKNOWLEDGEMENTS
This work was supported primarily by a cooperative
agreement to Environmental Systems Engineering, Clemson
University from the U.S. Environmental Protection Agency
through the Athens Environmental Research Laboratory, for
which appreciation 1s expressed. Partial support through NSF
grant No. ISP-8011451, "Environmental Engineering Chemistry" 1s
also gratefully acknowledged. Finally, our thanks to
Elaine McGarlty and Sherry Jarrard for their excellent work on
the manuscript.
NOTE: Mention of trade names or commercial products
does not constitute endorsement or recommendation for use by
the United States Environmental Protection Agency.
REFERENCES
1. Andelman, J.B., and Suess, M.J. (1970): Polynuclear
aromatic hydrocarbons 1n the water environment, Bui 1. WHO,
43: 479-508.
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PAHs IN PLANTS
2. Coates, J. T., Elzerman, A. W., and Garrison, A. W.
(1984). Analysis of selected PAHs 1n plant tissues.
Final Report, Grant No. CR810496, Analytical Techniques
for Selected Organlcs In Plant Tissue, Athens Environ-
mental Research Lab, US-EPA. ("Manuscript 1n press).
3. Edwards, N.T. (1983): Polycycllc aromatic hydrocarbons
(PAHs) In the terrestrial environment - A Review, J.
Environ. Qua!., 12: 427-441.
4. Geqer, W. and Schnaffner, C. (1978): Determination of
polycyclic aromatic hydrocarbons 1n the environment by
glass capillary gas chromatography, Anal. Chem., 50: 243-
249. . .
5. Grimmer, G. and Hlldebrandt, A. (1972): Concentration
and estimation of 14 polycycllc aromatic hydrocarbons at
low levels In high-protein foods, oils, and fats, J.
Assoc. Offlc. Anal. Chem., 55: 631-635.
6. (toward, J.W. and Fazio, T. (1980): Review of polycycllc
aromatic hydrocarbons 1n foods, J. Assoc. Offlc. Anal.
Chem., 63: 1077-1104.
7. Howard, J.W., Fazio, T., White, R.H., and Kllmeck, B.A.
(1968): Extraction and estimation of polycycllc aromatic
hydrocarbons In total diet composites. J. Assoc. Offlc.
Anal. Chem., 51: 122-129.
8. Kolarovic, L. and Traltler, H. (1982): Determination of
polycycllc aromatic hydrocarbons In vegetable oils by
caffeine complexatlon and glass capillary gas chromatography,
J. Chromatog., 237: 263-272.
9. Schamp, N. and vanWassenhove, F. (1972): Determination
of benzo(a)pyrene 1n bitumen and plants, J. Chromatog.,
69: 421-425.
10. Shabad, L.M. (1980): Circulation of carcinogenic
polycycllc aromatic hydrocarbons 1n the human environment
and cancer prevention, J. Natl. Cancer Inst., 64: 405-410.
11. Suess, M.J. (1976): The environmental load and cycle of
polycycllc aromatic hydrocarbons, Sc1. Total Environ., 6:
239-250.
12. Winkler, E., Buchele, E., and Mueller, 0. (1977):
Method for the determination of polycycllc aromatic
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chromatography, J. Chromatog., 138: 151-lb4.
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TECHNICAL REPORT DATA
/Phase rmJ liuiructiont on the rtrrne beforr completingi
1 REPORT NO 7
EPA/600/D-85/036
4. title and Subtitle
PAH UPTAKE BY PLANTS: Methodology and Initial
Investigations
ft. REPORT DATE
February 1985
6. PERFORMING ORGANIZATION CODE
»
7 AUTHORlS)
John T. Coates, Alan W. Elzerman and A. Wayne Garrison1
• PERFORMING ORGANIZATION REPORT NO
B PERFORMING OROANIZA HON NAME AND AOORESS
Environmental Systems Engineering, Clemson University,
CI emson SC 29631
~Environmental Research Laboratory, U.S. Environmental
Protection Agency, Athens GA "'OSIS
10 PROGRAM ELEMENT NO
CCUL1A
68-01-2281
12. SPONSORING AGENCY NAME AND ADDRESS
U.S. Environmental Protection Agency—Athens GA
Office of Research and Development
Environmental Research Laborato",
*.thens GA 30613
13. TYPE OF REPORT AND PERIOO COVEREO
Published paper
14 SPONSORING AGENCY CODE
EPA/600/01
19 SUPPLEMENTARY NOTE-
Contact: A.W. Garrison, H-2b0-Jl8j
IS. ABSTRACT
An analytical protocol was developed that allows qiantlf1cat1on of 16 PAHs 1n
grain sorghum and fescue grass. Compounds are extracted from the plant stem and
foliage by homo&enatlon/sol1cat1on using acetonltrlle as the primary solvent. The
extract 1s cleaned up by solvent partitioning Into pentane followed by adsorption
chromatography on silicic acid, then analyzed by GC-FID. This method can be used to
neasure PAH concentrations at the 25 ug/kg level 1n the plant.
17 Kt* ftORDS ANO DOCUMENT ANALYSIS
i DESCRIPTORS
h IDENTIFIERS/OPEN ENOEO TERMS
l COSATI 1 kill liloup
1* DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
It SECURITY CLASS (rhilRtport)
UNCLASSIFIED
J1 NO OF PAGES
18
MW[AWf!6r""w'
JJ PRICE
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