&EPA
United States
Environmental Protection
Agency
Environmental Monitoring
Systems Laboratory
P.O. Box 93478
Las Vegas NV 89193-3478
EPA/600/4-89/039
November 1989
Research and Development
Single- Laboratory
Evaluation of
Method 8060-
Phthalate Esters
Project Summary/
Project Report
-------
September 1989
SINGLE-LABORATORY EVALUATION
OF METHOD 8060-PHTHALATE ESTERS
Project Summary
By
Viorica Lopez-Avila, Franklin Constantine,
June Mi lanes, and Robert Gale
Acurex Corporation
Environmental Systems Division
485 Clyde Avenue
Mountain View, California 94039
and
Werner F. Beckert
Quality Assurance Division
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89193-3478
EPA Contract Nos. 68-03-3226 and 68-03-3511
• U.S. EPA Library
Las Vegas, NVf m 19
Environmental Monitoring Systems Laboratory—Las Vegas
Office of Research and Development
U.S. Environmental Protection Agency
Las Vegas, Nevada 89193-3478
-------
PROJECT SUMMARY
SINGLE-LABORATORY EVALUATION OF METHOD
8060 "PHTHALATE ESTERS"
by
Viorica Lopez-Avila, Franklin Constantine,
June Milanes, and Robert Gale
Acurex Corporation
Mountain View, California 94039
and
Werner F. Beckert
Quality Assurance Division
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89193-3478
ABSTRACT
SW-846 Method 8060 for the determination of phthalate esters in aqueous
and solid matrices was modified and evaluated in a single laboratory. The
range of compounds of interest was expanded to 16 phthalate esters. A study
to determine the sources of phthalate esters contamination in the laboratory,
its extent, and ways to minimize background contamination was conducted as
part of the evaluation. The packed columns specified for gas chromatographic
analysis were replaced with two fused-silica open tubular columns of
dissimilar stationary phases. The two fused-silica open tubular columns are
connected to an inlet splitter and two electron capture detectors; this setup
allows the primary and confirmatory analyses to be conducted simultaneously.
Extract cleanup was performed on alumina or on Florisil, however, three of the
target compounds were not recovered from the 10-g Florisil column
(Method 3620). The use of commercially available Florisil cartridges was
evaluated. Our results indicate that this approach is feasible for all 16
compounds. The interferences represented by organochlorine pesticides were
evaluated, and possible internal standards and surrogate compounds were
identified. The revised method was tested with an estuarine water, a
leachate, a groundwater, an estuarine sediment, a municipal sludge, and a
sandy loam soil. The results obtained indicated acceptable accuracy and
precision for most of the target analytes.
-------
INTRODUCTION
Regulations for hazardous waste activities under the Resource
Conservation and Recovery Act (RCRA) of 1976 and its elements require
analytical methodologies that provide reliable data. The document "Test
Methods for Evaluating Solid Waste, Physical/Chemical Methods," Office of
Solid Waste Manual SW-846 (1), provides a compilation of methods for
evaluating RCRA solid wastes for environmental and human health hazards. One
of the methods in this document, Method 8060, addresses the determination of
phthalate esters. This method provides conditions for sample extraction
(Methods 3510, 3520, 3540, 3550), sample extract cleanup (Methods 3610, 3620,
3640) and gas chromatographic (GC) determination of six phthalates in
environmental matrices including groundwater, liquids, and solids. Analyses
are performed by gas chromatography (GC) using two packed columns at various
temperatures, and the compounds are determined with a flame-ionization (FID)
or an electron-capture detector (ECD).
Problems with the current Method 8060 include:
• The primary column specified, a 1.8-m x 4-mm ID glass column packed
with 1.5 percent SP-2250/1.95 percent SP-2401 on Supelcoport
(100/120 mesh), needs to be operated at two temperatures (180°C and
220°C) in order to chromatograph the six compounds.
• The confirmatory column specified, a 1.8-m x 4-mm ID glass column
packed with 3 percent OV-1 on Supelcoport (100/120 mesh), also
needs to be operated at two temperatures (200°C and 220°C) in order
to chromatograph the six compounds.
• Only six phthalate esters are currently listed, but other
phthalates have been found in environmental samples.
• Surrogate compounds are required to be spiked into the sample
matrix prior to extraction, yet no compounds are specified or
recommended for this purpose. Likewise, internal standards are
required whenever internal standard calibration is used for
quantification purposes, yet no internal standards are specified
or recommended.
• Extract cleanup is performed according to Method 3610 or 3620, yet
no data are included on the recovery of the six compounds from the
extract cleanup step.
• Many phthalate esters are present as contaminants in or on
laboratory equipment and in solvents and reagents (2). Procedures
on how to clean glassware and how to remove phthalate esters from
solvents and materials should be tested and incorporated in the
protocol. Also, examples of typical background contamination of
some common laboratory items should be given to make the analyst
aware of such problems.
Acurex, under contract to the Environmental Monitoring Systems
Laboratory in Las Vegas (EMSL-LV), conducted an evaluation and improvement
1
-------
study of Method 8060. Since the current protocol was inadequate in certain
areas (e.g., in addressing background contamination) and was lacking
information in other areas such as the sample cleanup and the GC analysis, the
method evaluation and improvement study was approached in two phases.
Phase I, the developmental phase, addressed the following:
• Literature review to gather the relevant information
• Assessment of background contamination of solvents, materials used
in sample cleanup, and apparatus used for sample extraction
• Selection and evaluation of capillary columns for use in the
analysis of 16 phthalate esters (Table 1)
• Evaluation of sample extraction procedures
• Evaluation of GC/ECD and GC/FID for the analysis of samples
containing the test compounds
• Evaluation of alumina (Method 3610) and Florisil chromatography
(Method 3620)
• Selection of surrogate and internal standards for use in
Method 8060
• Sample preservation studies.
TABLE 1. PHTHALATE ESTERS INCLUDED IN THE EVALUATION
Compound CAS No.
Dimethyl phthalate (DMP) 131-11-3
Diethyl phthalate (DEP) 84-66-2
Diisobutyl phthalate (DIBP) 84-69-5
Di-n-butyl phthalate (DBP) 84-74-2
Bis(4-methyl-2-pentyl) phthalate (BMPP) 146-50-9
Bis(2-methoxyethyl) phthalate (BMEP) 117-82-8
Diamyl phthalate (DAP) 131-18-0
Bis(2-ethoxyethyl) phthalate (BEEP) 605-54-9
Hexyl 2-ethylhexyl phthalate (HEHP) 75673-16-4
Dihexyl phthalate (DHP) 84-75-3
Benzyl butyl phthalate (BBP) 85-68-7
Bis(2-n-butoxyethyl) phthalate (BBEP) 117-83-9
Bis(2-ethylhexyl) phthalate (DEHP) 117-81-7
Dicyclohexyl phthalate (DCP) 84-61-7
Di-n-octyl phthalate (OOP) 117-84-0
Dinonyl phthalate (DNP) 84-76-4
-------
Upon completion of the experimental work in Phase I, the protocol was
tested in Phase II on three aqueous matrices and three solid matrices.
Performance data generated during the evaluation of the revised Method 8060
include:
• Measures of precision and accuracy
• Evidence of analyte identification
• Evidence of resolution of analyte from interfering substances
• Ruggedness study
• Method detection limits.
EXPERIMENTAL
Apparatus
(a) Glassware—Essentially as specified in Methods 3510, 3520, 3540,
3550, 3610, and 3620.
(b) Mixxor—Lidex Technologies, Inc.
(c) Sonicator—Heat Systems Ultrasonics, Inc., Model W-375.
(d) Gas Chromatographs—Varian 6000 with constant-current/pulsed-
frequency ECD, interfaced with a Varian Vista 402 data system;
Varian 6500 with FID, interfaced with either a Spectra Physics 4290
integrator or a Varian Vista 402 data system. For the simultaneous
injection on the DB-5 and DB-1701 columns, the Varian 6000 was
equipped with a J&W Scientific press-fit, Y-shape, glass-splitter,
and with dual ECDs.
(e) Autosampler—Varian Model 8000.
(f) GC Columns—(1) DB-5, (2) Supelcowax-10, (3) DB-210, (4) DB-608, (5)
DB-1701, (6) RT -5, 30 m x 0.25 mm ID or 30 m x 0.53 mm ID and
different film thickness.
Materials
(a) Solvents and other reagents—As specified in Methods 3510, 3520,
3540, 3550, 3610, and 3620.
(b) Florisil—J. T. Baker, Lot No. 442707, 60/80 mesh, activated at
400"C for 16 hours, then deactivated with water (3 percent by
weight).
(c) Alumina—Alumina WoelmN Super I, activated/deactivated as described
for Florisil. ,
(d) C18 membrane disks—Analytichem International
3
-------
(e) Florisil disposable cartridges—Supelclean SPE tubes consisting of
serological-grade 6-mL polypropylene tubes, packed each with 1 g
LC-Florisil (40-/xm particles, 60-A pores) held between polyethylene
frits.
(f) Standards—DEP was obtained from Scientific Polymer Products, all
other phthalates, as well as benzyl benzoate and diphenyl
terephthalate, were obtained from Chem Service (distributed by
Bryant Laboratories, Inc.). Purities were stated to be greater than
98 percent. Stock solutions of each compound at 1 mg/mL were
prepared in isooctane (Baker Resi-Analyzed, J. T. Baker); working
calibration standards were prepared initially in isooctane and later
in hexane by serial dilutions of a composite stock solution prepared
from the individual stock solutions.
(g) Materials used in contamination evaluation (solvents and other
materials used in sample preparation)—Various grades purchased from
a variety of suppliers.
(h) Environmental materials—
• Sandy loam soil, obtained from Soils Incorporated, Puyallup,
Washington, with the following characteristics: pH 5.9 to 6.0;
89 percent sand, 7 percent silt, 4 percent clay; cation exchange
capacity 7 meq/100 g; total organic carbon content
1,290 ± 185 mg/kg.
• A sediment sample of unknown origin. Analysis of the extract
by GC/MS indicated the presence of petroleum hydrocarbons.
• NBS SRM-1572, Citrus leaves.
. NBS SRM-1632a, Coal.
. NBS SRM-1633a, Coal flyash.
• Estuarine water and sediment collected form San Francisco Bay,
South San Francisco, California.
• Leachate prepared by Method 1310 from a soil contanimated with
lead.
• Ground water collected at a semiconductor plant in Sunnyvale,
California.
• Municipal sludge collected from Santa Clara Valley Water
District, San Jose, California.
• Sandy loam soil obtained by mixing 20 percent organic soil with
80 percent sand.
-------
Contamination Study
Solvent samples (acetone 150 mL, hexane 150 mL, diethyl ether 30 ml,
methylene chloride 180 mL) were individually concentrated by K-D evaporation to
10 mi and further reduced to 1 ml with high-purity nitrogen; only isooctane was
not concentrated. At least two replicate samples of each solvent were prepared
and analyzed.
Samples of Florisil (20 g), silica gel (20 g), anhydrous sodium sulfate
(50 g) and glass wool (5 g) were immersed overnight in solvent which was then
separated and concentrated to 1 ml for GC analysis. Two washings were performed
in each case and the concentrates analyzed separately. The effect of baking at
400°C for 4 hours was evaluated for anhydrous sodium sulfate and glass wool.
Samples of filter paper (10 g), paper thimbles (10 g) and aluminum foil
(5 g) were cut into 0.5-in x 0.5-in pieces and immersed overnight in solvent
which was then separated and concentrated to 1 ml for GC analysis. Two washings
were performed in each case and the concentrates analyzed separately.
Gas Chromatography
Operating conditions: DB-5—120'C to 160°C (hold 16 min) at 15°C/min,
injector temp. 275°C, detector temp. 320"C; Supelcowax-10—150°C (hold 2 min)
to 220°C at 15°C/min, then 260'C (hold 16 min) at 4°C/min, injector temp. 270'C,
detector temp. 270°C; DB-210—1258C (hold 1 min) to 240'C (hold 16 min) at
5'C/min, injector temp. 250°C, detector temp. 250°C; DB-5/DB-608; DB-608/DB-
1701; DB-608/RT -5; DB-5/DB-1701—150°C (hold 0.5 min) to 220'C at 3eC/min, then
to 2756C (hold 15 min) at 5'C/min, injector temp. 250°C, detector temp. 320°C.
Sample Extraction
The extraction efficiencies of Methods 3510 (separatory funnel) and 3520
(continuous liquid-liquid extraction) for the target compounds were determined
with reagent water. Microextraction of 50 ml samples using a Mixxor device and
hexane (10 mL) was also tested. Preconcentration of phthalate esters onto C1B-
membrane disks (Analytichem International) followed by elution with acetonitrife
resulted in quantitative recoveries for all 16 phthalate esters.
Solid samples were extracted either in a Soxhlet extractor with
hexane/acetone (1:1) (Method 3540) or by sonication with methylene
chloride/acetone (1:1) (Method 3550).
Extract Cleanup
Florisil and alumina Chromatography: glass columns were packed each with
10 g deactivated Florisil or alumina and topped with 1 cm of precleaned anhydrous
sodium sulfate. The charged columns were first eluted with 40 ml hexane which
was discarded; the phthalate esters were eluted with 4:1 hexane/diethyl ether
(100 ml for the Florisil column, 140 ml for the alumina column).
Florisil disposable cartridges: the cartridges were washed with 4 mL
pesticide-grade hexane prior to use. The eluting solvents used were hexane,
mixtures of hexane and diethyl ether, and mixtures of hexane and acetone.
-------
Removal of organochlorine pesticides in the presence of phthalates was attempted
with mixtures of 5-percent, 20-percent, 25-percent, and 30-percent methylene
chloride and hexane.
Surrogate Compound and Internal Standard Evaluation
Ten compounds were evaluated as possible internal standards and
surrogates for Method 8060.
RESULTS AND DISCUSSION
Phthalate Ester Contamination Study
Only a brief summary of the results is presented here. Detailed results
of the study will be published elsewhere (3).
Solvents
Five organic solvents from up to six different commercial suppliers were
analyzed for 11 phthalate esters. As can be seen from the summary results listed
in Table 2, six phthalate esters were detected in some or all of these solvents.
The only phthalate ester detected in any of the methylene chloride samples above
6 ng/mL was DOP at 8.8 ng/mL in one sample.
Since typical volumes of hexane and acetone used in sample preparation
are 200 to 300 mL, the amounts of phthalate esters that can be introduced as
contaminants with solvents could be considerable.
Materials
The phthalate contamination summary values (averaged across brands) for
the materials listed in Table 3 represent averages of second washings. Florisil,
alumina and silica gel showed significant levels of phthalates even in the second
washing. Florisil disposable cartridges (not listed in Table 3) in the first
washing showed levels from 10 to 460 /*g per cartridge for 8 of the 11 phthalate
esters listed in Table 3. However, washing of the cartridges just prior to use
with 4 ml hexane resulted in acceptable method blanks. Washing alone is not
sufficient for sodium sulfate and glass wool, but baking these materials at 400°C
for 4 hours followed by solvent washing gave acceptable blanks. High levels
were found in filter paper, paper thimbles, and aluminum foil.
Precleaning of these materials is a must when phthalate esters at low
nanogram levels are to be quantified.
GC Column Evaluation
Of the six fused-silica capillary columns evaluated, the DB-210 column
was found to be the least desirable because of a significant baseline drift
during column programming and was therefore eliminated from further
consideration.
The retention times of the 16 phthalates of interest on the DB-5 fused-
silica capillary column and the Supelcowax-10 fused-silica open tubular column
-------
TABLE 2. PHTHALATE ESTER CONTAMINATION RANGES IN COMMON SOLVENTS* (ng/mL)
Phthalate
Dimethyl
Di ethyl
Diisobutyl
Di-n-butyl
Diamyl
Dihexyl
Benzyl butyl
Bis(2-ethylhexyl)
Dicyclohexyl
Di-n-octyl
Dinonyl
No. of diff. brands
Below det. limit
Concentr. factor
Acetone
<0.10
<0.10 - 0.40
<0.10 - 0.35
<0.10 - 0.50
<0.10
<0.10 - 0.45
<0.10 - 0.46
<0.10 - 0.45
<0.10
<0.10
<0.10
8
1
150
Hexane
<0.10
<0.10
<0.10 - 0.35
<0.10
<0.10
<0.10 - 0.87
<0.10
<0.10 - 0.40
<0.10
<0.10
<0.10
8
2
150
Di ethyl
ether Isooctane
<0.20 - 3.45 <10
<0.20 <10
<0.20 <10
<0.20 - 2.9 <10 - 103
<0.20 <10
<0.20 - 0.75 <10 - 42
<0.20 <10
<0.20 - 2.2 <10 - 69
<0.20 <10
<0.20 <10
<0.20 <10
8 8
0 0
30 1
Methyl ene"
chloride
<6
<6
<6
<6
<6
<6
<6
<6
<6
8.8
<6
8
7
180
"Averages of two to four determinations.
"Analyzed by GC/FID which resulted in high detection limits.
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TABLE 3. REPRESENTATIVE PHTHALATE ESTER CONTAMINATION VALUES OF LABORATORY MATERIALS (ng/g)
oo
Phthalate FlorisiT
Dimethyl
Diethyl
Diisobutyl
Di-n-butyl
Di amyl
Dihexyl
Benzyl butyl
Bis(2-ethylhexyl)
Dicyclohexyl
Di-n-octyl
Dinonyl
No. of diff. brands
6.2
5.3
0.9
<0.5
<0.5
0.8
<0.5
2.3
<0.5
1.5
2.3
2
Alumina6
129
28
29
14
5.8
1.2
0.6
4.8
<0.5
<0.5
<0.5
2
Silical
gele
i
15
0.6
0.8
<0.5
3.8
18
1.3
<0.5
0.7
<0.5
2
Sodium
sulfate"
0.7
1.8
0.8
3.5
0.5
3.3
4.7
1.0
<0.5
<0.5
<0.5
2
Glass
wool*
27.4
2.2
3
<2.0
<2.0
13
4
4.4
<2.0
5
<2.0
2
Filter
paper1
67.5
<1.0
11.5
6.5
<1.0
15.5
3.3
11
<1.0
<1.0
<1.0
1
Paper
thimbles9
35
2.3
2.3
<2.0
<2.0
17.0
3.0
2.8
<2.0
<2.0
<2.0
1
Aluminum
foil"
24
0.6
0.7
0.5
<0.5
3.0
2.5
2.5
<0.5
1.1
1.0
2
"20 g Florisil, second immersion with 200 ml hexane/diethyl ether (4:1).
"20 g alumina, second immersion with 300 ml hexane/diethyl ether (4:1).
C20 g silica gel, second immersion with 300 ml acetone.
"40 g anhydrous sodium sulfate, second immersion with 300 ml hexane/acetone (1:1).
"5 g glass wool, second immersion with 300 ml hexane/acetone (1:1).
'10 g filter paper, second immersion with 100 ml hexane/acetone (1:1).
810 g paper thimbles, second immersion with 100 ml hexane/acetone.
"5 g aluminum foil, second imersion with 200 ml hexane/acetone (1:1).
'Not able to quantify because of interference.
-------
and the DB-5 and DB-1701 fused-silica open, tubular columns are presented in
Table 4. The GC conditions were chosen such that all compounds are resolved
and the total analysis time is approximately 35 min. All phthalate esters
including surrogates were resolved on the DB-5 and DB-1701 columns; these columns
were proposed for incorporation in the revised method 8060 since they can be used
in the dual-column/dual-detector approach for the determination of Method 8060
phthalate esters.
Sample Extraction
The extraction of reagent water spiked with each of the 16 phthalate
esters at 50 ng/L per component for separatory funnel and continuous liquid-
liquid extraction and 1 mg/L for the Mixxor extraction gave the following
results:
• The continuous liquid-liquid extraction technique had unacceptable
reproducibilities for all compounds; for five of the phthalate
esters the average recoveries were only 20 to 45 percent.
• Extraction with hexane in the Mixxor device gave unacceptable
recoveries and reproducibilities.
• The separatory funnel extraction produced recoveries >70 percent
for most compounds, and reproducibilities were better than
10 percent for two-thirds of the compounds.
Further evaluation of the separatory funnel extraction technique at
lower spiking levels (25, 10, and 1 /Kj/L) confirmed its usefulness. At 25 fig/L,
the recoveries ranged from 90 to 130 percent, with 11 recoveries between 90 and
110 percent, and at 10 /jg/L, the range was 73 to 117 percent, with 10 recoveries
between 90 and 110 percent. At 1 /tg/L, the recoveries ranged from 53 to
152 percent; only four values were between 90 and 110 percent.
Method Performance
Method performance, as used here, includes method accuracy (percent
recovery), method precision (percent relative standard deviation), and method
detection limits. In the case of aqueous samples, the method accuracy given as
percent recovery of the 16 phthalate esters spiked into an estuarine water, a
leachate, and a groundwater at 20 /ig/L and 60 /ig/L ranged from 59.5 to
117 percent (Figure 1). In the case of solid samples (an estuarine sediment,
a municipal sludge, and a sandy loam soil) the recoveries were distributed over
a much wider range (Figure 2) indicating that method accuracy is a function of
matrix and concentration. Method precision for aqueous samples (Figure 3) was
better than 27.5 percent. Method precision for solid samples (Figure 4) varied
from matrix to matrix (Figure 4).
The method detection limits (MDL) were determined for HPLC-grade water
from the standard deviations (SD) of seven replicate measurements. They
represent the minimum concentrations that can be measured and reported with
99 percent confidence. They ranged from 22 to 640 ng/L for water samples
subjected to Florisil cleanup and 26 to 320 ng/L for water samples not subjected
to Florisil cleanup; in both cases a DB-5 capillary column (single-column
approach) was used.
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TABLE 4. GC RETENTION TIMES FOR THE PHTHALATES8
Phthalate
Retention time (min)
Supelco-
DB-5b wax-10b
30 m x 30 m x
0.25 mm ID 0.53 mm ID
DB-5C DB-17010
30 m x 30 m x
0.53 mm ID 0.53 mm ID
Dimethyl
Diethyl
Diisobutyl
Di-n-butyl
Bis(4-methyl-2-pentyl)
Bis(2-methoxyethyl)
Diamyl
Bis(2-ethoxyethyl)
Hexyl 2-ethylhexyl
Dihexyl
Benzyl butyl
Bis(2-n-butoxyethyl)
Bis(2-ethylhexyl)
Dicyclohexyl
Di-n-octyl
Dinonyl
Benzyl benzoate (IS)
3.
3.
6.
7.
7.
7.
.42
.45
.48
14
.96
.40
8.41
8.17
8.63
9.62
9.69
10.53
11.13
10.98
13.03
16.00
5.77
5.62
6.11
7.26
8.43
8.14
12.05
10.15
12.41
11.13
12.21
16.36
16.94
13.31
16.66
17.25
20.73
7.87
7.06
9.30
14.44
16.26
18.77
17.02
20.25
19.43
21.07
24.57
24.86
27.56
29.23
28.88
33.33
38.80
12.71
6.37
8.45
12.91
14.66
16.27
16.41
18.08
18.21
18.97
21.85
23.08
25.24
25.67
26.35
29.83
33.84
11.07
Diphenyl phthalate (SU)
Diphenyl isophthalate (SU)
Dibenzyl phthalate (SU)
d
d
d
d
d
d
29.46
32.99
34.40
28.32
31.37
32.65
aThe GC conditions have been specified under "Gas Chromatography."
"Single-column approach.
°Dual-column/dual-detector approach. The two columns were connected to a
J&W Scientific press-fit, Y-shape, glass-splitter.
dNot available.
10
-------
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o
o
u
UJ
o
a:
rm
140
130
120
110
100
90
80
70
60
40 -
30 -
20 -
10
DMP DEP DIBP DBP BMPP BMEP DAP BEEP HEHP DHP BBP BBEP DEHP DCP OOP DNP
M1C2
M2C1
M2C2
M3C1
M3C2
Figure 1.
Method accuracy for aqueous matrices (M1—estuarine water;
M2—leachate; M3—groundwater; C.—concentration at 20 ng/L per
component; C2—concentration at 60 /ig/L per component).
-------
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fTTI M1C1
M1C2
M2C1
M2C2
M3C1
M3C2
Figure 2. Method accuracy for
M2 — municipal sludge; 3
1 /ig/g per component; C2
solid matrices (M1 — estuarine sediment;
M — sandy loam soil; Ct — concentration at
oncentration at 3 /jg/g per component).
-------
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M1C1 | \\| M1C2
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M2C2
M3C1
M3C2
Figure 3.
Method precision for aqueous matrices (M,—estuarine water;
M2—leachate; M3—groundwater; C.—concentration at 20 /zg/L per
component; C2—concentration at 60 ng/L per component).
-------
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M1C1 | \\| M1C2
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M3C1
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Figure 4. Method precision for solid matrices (Mt—estuarine sediment;
M2—municipal sludge; M3—sandy loam soil; Ct—concentration at
per component; C2—concentration at 3 jzg/g per component).
-------
Phthalate recoveries from soil samples, spiked at 1 ppm with the
16 phthalates, using Method 3540 (Soxhlet extraction), ranged from 54 percent
for BEEP to 135 percent for DHP with 11 recoveries >70 percent. When sonication
was used, the recoveries ranged from 32 percent for BMPP to 112 percent for DMP,
with 13 recoveries >70 percent.
Extract Cleanup
Alumina and Florisil chromatography were performed with standards in
hexane according to Methods 3610 and 3620, respectively (Table 5). For the
Florisil cartridge cleanup, various solvents and solvent combinations were tried
on standards in hexane and on standards in the presence of organochlorine
pesticides. It was found that the organochlorine pesticides can be removed
efficiently from the cartridges with hexane/methylene chloride (4:1); under these
conditions, the phthalate esters are retained on the Florisil cartridge and can
be recovered with hexane/acetone (9:1). The recoveries are presented in Tables
5 and 6. Additional details on the Florisil cartridge cleanup method can be
found in Reference 4.
Surrogate Compound and Internal Standard
Of ten compounds evaluated, benzyl benzoate was selected as internal standard
and diphenyl phthalate, diphenyl isophthalate, and dibenzyl phthalate were
considered as surrogate compounds. The selection was based primarily on the
observation that both compounds are resolved from the other phthalate esters
under the conditions of the GC analysis.
CONCLUSIONS
Contamination from solvents, reagent materials, and glassware used in the
analysis limit the detection of phthalate esters at trace levels (ppt-ppb range).
Consequently, their determinations in environmental samples at ppt-ppb range
require pesticide-grade solvents, thorough cleaning of the glassware followed
by heat-treatment (for those items that can withstand 400eC temperatures), and
the minimum number of steps in sample workup.
Extraction of water samples in a separatory funnel was desired over the
continuous liquid-liquid extraction since it gave good recoveries and reproduc-
ibilities for most target analytes, greatly reduced the extraction time, and also
minimizes contamination. Preconcentration of aqueous samples on C<8-membrane
disks followed by extraction of the phthalate esters with acetonitrile gave
quantitative recoveries and good reproducibilities and was therefore incorporated
in the revised Method 8060.
Extract cleanup using Florisil disposable cartridges and elution with
hexane/acetone (9:1) gave quantitative recoveries for all 16 phthalate esters
proposed for incorporation in Method 8060. Organochlorine pesticides overlap
with the phthalate esters when the GC is performed on a DB-5 fused-silica
capillary column. Use of Florisil disposable cartridges and elution with
20 percent methylene chloride in hexane helps to remove the organochlorine
pesticides. Phthalate esters are then recovered from the Florisil cartridge
with hexane/acetone (9:1). The use of Florisil disposable cartridges was
included as an option since it results in quantitative recoveries, reduces
contamination, saves chemicals, and reduces laboratory waste.
15
-------
TABLE 5. EXTRACT CLEANUP RECOVERIES (IN PERCENT)
Florisil Cartridges'
Phthalate
Dimethyl
Diethyl
Diisobutyl
Di-n-butyl
Bis(4-methyl-2-pentyl)
Bis(2-methoxyethyl)
Diamyl
Bis(2-ethoxyethyl)
Hexyl 2-ethylhexyl
Dihexyl
Benzyl butyl
Bis(2-n-butoxyethyl)
Bis(2-ethylhexyl)
Dicyclohexyl
Di-n-octyl
Dinonyl
Florisil*
43
57
80
85
85
0
82
0
105
74
90
0
82
84
115
73
Alumina*
65
62
77
77
89
70
75
67
91
72
87
63
91
84
108
71
Fraction 1
0
0
0
12
0
0
3.3
0
0
0
0
0
0
0
0
0
Fraction 2
130
88
118
121
123
32
94
82
94
126
62
98
110
106
123
102
(52)
(2.8)
(16)
(13)
(5.7)
(31)
(8.3)
(19)
(8.3)
(6.4)
(15)
(6.5)
(2.7)
(3.3)
(7.0)
(8.7)
aAverage of two determinations.
"Averages of three determinations; RSDs given in parentheses.
Fraction 1 was eluted with 5 ml hexane/methylene chloride (4:1) and
Fraction 2 with 5 mL hexane/acetone (9:1).
16
-------
TABLE 6. PERCENT RECOVERIES OF PHTHALATE ESTERS FROM VARIOUS MATRICES BY
FLORISIL CARTRIDGE CLEANUP WITH HEXANE/METHYLENE CHLORIDE (4:1) AND
HEXANE/ACETONE (9:1) AS ELUANTS"
Phthalate
Dimethyl
Diethyl
Diisobutyl
Di butyl
Bis(4-methyl-2-pentyl)
Diamyl
Bis(2-ethoxyethyl)
Hexyl 2-ethylhexyl
Dihexyl
Benzyl butyl
Bis(butoxyethyl)
Bis(2-ethylhexyl)
Dicyclohexyl
Dioctyl
Dinonyl
Sandy
Loam
Soil
78
79
79
74
77
82
37
80
78
82
86
74
91
80
84
Sediment
75
79
82
78
84
86
24
88
88
99
94
85
96
92
96
NBS
SRM-1572
80
89
90
84
102
100
62
95
86
114
98
108
106
104
106
NBS
SRM-1632a
76
79
108
83
91
76
32
93
92
102
106
88
98
95
111
NBS
SRM-1633a
82
84
86
83
86
89
33
81
80
98
98
112
95
88
92
aSpiking level is 50 ng/mL for each compound. Data shown are for Fraction 2
which was eluted with 5 mL hexane/acetone (9:1).
17
-------
Preservation of aqueous samples at neutral and acidic pH and 4°C is
adequate for 21 days. Preservation of water samples at pH 9 and 4°C should be
avoided since most compounds show significant decrease in concentration after
14 days of storage. Storage of spiked soil samples at -10'C is preferred over
refrigeration at 4°C, since it minimizes loss of the lower-molecular-weight
esters.
The dual-column/dual-detector approach for the analysis of phthalate
esters increases sample throughput by allowing the primary and confirmatory
column analyses to be performed simultaneously. Excellent reproducibilities of
the retention times and detector responses were achieved with two 30 m.x 0.53-
mm ID fused-silica open tubular columns of dissimilar stationary phases connected
to an injection tee and two ECDs.
REFERENCES
1. Test Methods for Evaluating Solid Waste (1986), 3rd Ed., SW-846, U.S.
Environmental Protection Agency, Washington, DC.
2. Giam, C. S., H. S. Chan, and G. S. Neff. Anal. Chem. 47, 225-229 (1975).
3. Lopez-Avila, V., J. Milanes, and W. F. Beckert. "Phthalate Esters as
Contaminants in Gas Chromatography." In preparation.
4. Lopez-Avila, V., J. Milanes, N. S. Dodhiwala, and W. F. Beckert.
"Cleanup of Environmental Sample Extracts Using Florisil Solid-Phase
Extraction Cartridges." J. Chromatogr. Sci. 27(5), 209-215 (1989).
Werner F. Beckert is the EPA Project Officer (see below).
The complete report, entitled "Single-Laboratory Evaluation of
Method 8060 Phthalate Esters" will be available only from:
National Technical Information Service
5285 Port Royal Road
Springfield, VA 22161
Telephone: (703) 487-4650
The EPA Project Officer can be contacted at:
Environmental Monitoring Systems Laboratory
U.S. Environmental Protection Agency
P.O. Box 93478
Las Vegas, NV 89193-3478
18
-------
SINGLE-LABORATORY EVALUATION
OF METHOD 8060 -- PHTHALATE ESTERS
By
Viorica Lopez-Avila, Franklin Constantine,
June Mi lanes, and Robert Gale
Acurex Corporation
Environmental Systems Division
Mountain View, California 94039
Contract Numbers 68-03-3226 and 68-03-3511
Project Officer
Werner F. Beckert
Quality Assurance Division
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89193-3478
ENVIRONMENTAL MONITORING SYSTEMS LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
LAS VEGAS, NEVADA 89193-3478
-------
NOTICE
The information in this document has been funded wholly or in part by
the United States Environmental Protection Agency under Contracts
No. 68-03-3226 and 68-03-3511 to Acurex Corporation. It has been subject to
the Agency's peer and administrative review, and it has been approved for
publication as an EPA document. Mention of Trade names or commercial products
does not constitute endorsement or recommendation for use.
ii
-------
PREFACE
This is the final report for Work Assignments 2-14 and 3-16, EPA Contract
No. 68-03-3226, and Work Assignment 1-16, EPA Contract No. 68-03-3511,
"Single-Laboratory Evaluation of Method -- Phthalate Esters," conducted at
Acurex Corporation, Project Nos. 8007, 8009, and 8113. These projects were
directed by Dr. Viorica Lopez-Avila.
This report was written by Dr. Viorica Lopez-Avila. Technical support
for both projects was provided by Mr. Franklin Constantine, Mr. June Milanes,
and Dr. Robert Gale.
iii
-------
ABSTRACT
SW-846 Method 8060 for the determination of phthalate esters in aqueous
and solid matrices was modified and evaluated in a single laboratory. The
range of compounds of interest was expanded to 16 phthalate esters. A study
to determine the sources of phthalate esters contamination in the laboratory,
its extent, and ways to minimize background contamination was conducted as
part of the evaluation. The packed columns specified for gas chromatographic
analysis were replaced with two fused-silica open tubular columns of
dissimilar stationary phases. The two fused-silica open tubular columns are
connected to an inlet splitter and two electron capture detectors; this setup
allows the primary and confirmatory analyses to be conducted simultaneously.
Extract cleanup was performed on alumina or on Florisil, however, three of the
target compounds were not recovered from the 10-g Florisil column
(Method 3620). The use of commercially available Florisil cartridges was
evaluated. Our results indicate that this approach is feasible for all
16 compounds. The interferences represented by organochlorine pesticides were
evaluated, and possible internal standards and surrogate compounds were
identified. The revised method was tested with an'estuarine water, a
leachate, a groundwater, an estuarine sediment, a municipal sludge, and a
sandy loam soil. The results obtained indicated acceptable accuracy and
precision for most of the target analytes.
Included in this report as an appendix is an extensive literature review
covering analytical methods for the determination of phthalate esters in
water, soil, and sediment samples.
-------
TABLE OF CONTENTS
Section Page
Notice ii
Preface iii
Abstract iv
Figures vi
Tables x
1. INTRODUCTION 1
2. CONCLUSIONS 6
3. RECOMMENDATIONS 8
4. LITERATURE REVIEW 9
5. EXPERIMENTAL PROCEDURES 10
5.1 SAMPLE ACQUISITION 10
5.2 BACKGROUND 17
5.3 EVALUATION OF GAS CHROMATOGRAPHY 17
5.4 EXTRACTION TECHNIQUES 19
5.4.1 Sample Extraction 19
5.4.2 Soil Spiking 19
5.5 EXTRACT CLEANUP 21
5.5.1 Florisil Chromatography 21
5.5.2 Alumina Chromatography 21
5.6 SAMPLE PRESERVATION 21
5.7 GC/MS METHODOLOGY 22
5.8 GLASSWARE CLEANUP 22
6. RESULTS AND DISCUSSION 23
6.1 BACKGROUND CONTAMINATION STUDY 23
6.1.1 Organic Solvents 23
6.1.2 Reagent Water 29
6.1.3 Materials 29
6.1.4 Glassware 45
6.1.5 Sample Vials 45
6.2 EVALUATION OF GAS CHROMATOGRAPHY 45
6.2.1 Single-Column Approach 50
6.2.2 Dual-Column/Dual-Detector Approach 68
6.3 SAMPLE EXTRACTION 68
6.4 EXTRACT CLEANUP 97
6.5 PRESERVATION STUDY 104
6.6 REVISED METHOD 8060 112
6.6.1 Reproducibility of the GC Technique 112
6.6.2 Instrument Calibration 126
6.6.3 Method Accuracy 142
6.6.4 Method Detection Limits 142
6.6.5 Ruggedness Testing 159
6.6.6 Confirmation by GC/MS 173
-------
TABLE OF CONTENTS (continued)
Section Page
REFERENCES 179
APPENDIX A -- SINGLE-LABORATORY EVALUATION OF METHOD 8060 -- PHTHALATE
ESTERS LITERATURE REVIEW A-l
APPENDIX B -- METHOD 8061 PHTHALATE ESTERS B-l
APPENDIX C -- GC/MS CHROMATOGRAMS AND MASS SPECTRA (PLOT AND LIST) FOR
METHOD 8060 COMPOUNDS C-l
APPENDIX D -- RESULTS OF INORGANIC ANALYSIS FOR THE ESTUARINE WATER,
LEACHATE, AND GROUNDWATER SAMPLES D-l
vi
-------
FIGURES
Number Page
1 GC/ECD chromatograms of solvent washings obtained from two
different brands of Florisil 40
2 GC/ECD chromatograms of solvent washings obtained from two
different brands of alumina 41
3 GC/ECD chromatograms of solvent washings of filter paper
(Whatman No. 41) before precleaning and after precleaning . . 42
4 GC/ECD chromatograms of solvent washings obtained from paper
thimbles before precleaning and after precleaning 43
5 GC/ECD chromatograms of solvent washings from All tech pest
grade glass wool before baking at 400"C and after baking ... 44
6 GC/ECD chromatogram of composite phthalate esters standard
analyzed on a 30 m x 0.25 mm ID DB-5 fused-silica capillary
column 53
7 GC/ECD chromatograms of diisohexyl phthalate, diisooctyl
phthalate, and diisononyl phthalate analyzed on a 30 m x 0.25
mm ID DB-5 fused-silica capillary column 54
8 GC/ECD chromatograms of diisodecyl phthalate and hexyl decyl
phthalate analyzed on a 30 m x 0.25 mm ID DB-5 fused-silica
capillary column 55
9 GC/ECD chromatogram of butyl decyl phthalate analyzed on a
30 m x 0.25 mm ID DB-5 fused-silica capillary column 56
10 GC/FID chromatogram of composite phthalate esters standard
analyzed on a 30 m x 0.53 mm ID Supelcowax-10 fused-silica
open tubular column 59
11 GC/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular column . 60
12 GC/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular column . 61
13 GC/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular
column 62
vn
-------
FIGURES (continued)
Number Page
14 GC/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular
column 63
15 GC/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular
column 64
16 GC/ECD chromatogram of composite phthalate esters standard
analyzed on a DB-210 fused-silica open tubular
column 66
17 GC/ECD chromatograms of the phthalate esters standard
analyzed on the DB-608/DB-5 column pair 73
18 GC/ECD chromatograms of the phthalate esters standard
analyzed on the DB-608/DB-1701 column pair 74
19 GC/ECD chromatograms of the phthalate esters standard
analyzed on the DB-608/RTx-5 column pair 75
20 GC/ECD chromatograms of the phthalate esters standard
analyzed on the DB-5/DB-1701 column pair 76
21 Average recovery of Method 8060 compounds from HPLC-grade
water as a function of time at pH 7 119
22 Average recovery of Method 8060 compounds from HPLC-grade
water as a function of time at pH 9 120
23 Average recovery of Method 8060 compounds from HPLC-grade
water as a function of time at pH 121
24 Recovery of Method 8060 compounds from sandy loam soil as a
function of time at -10*C 123
25 Recovery of Method 8060 compounds from sandy loam soil as a
function of time at -10'C and +4'C 124
26 Method accuracy for aqueous matrices 145
27 Method precision for aqueous matrices 146
viii
-------
FIGURES (concluded)
Number Page
28 Method accuracy for solid matrices 147
29 Method precision for solid matrices 148
30 GC/ECD chromatogram of WP-482 Sample 2 155
31 GC/ECD chromatogram of WP-482 Sample 3 156
32 GC/ECD chromatogram of WP-485 157
33 GC/ECD chromatogram of WP-281 Sample 2 158
34 GC/ECD chromatogram of a hexane blank analyzed on a DB-5
fused-silica capillary column 161
35 GC/ECD chromatogram of composite phthalate esters standard
at 0.5 pgM 162
36 GC/ECD chromatogram of composite phthalate esters standard
at 1 pgM 163
37 GC/ECD chromatogram of composite phthalate esters standard at
5 pgM 164
38 GC/ECD chromatogram of composite phthalate esters standard at
10 pgM 165
39 GC/ECD chromatogram of composite phthalate esters standard
at 25 pgM 166
40 GC/ECD chromatogram of composite phthalate esters standard
at 50 pgM 167
41 GC/MS chromatogram of composite phthalate esters standard . . 178
ix
-------
TABLES
Number Page
1 Phthalate Esters Investigated in this Study 2
2 Identification of the Solvent Samples Investigated 11
3 Materials Investigated for Possible Contamination with
Phthalate Esters 15
4 NIST Standard Reference Materials and Other Solid Matrices
Used in the Method Evaluation 16
5 GC Operating Conditions for the Single-Column Approach .... 18
6 GC Operating Conditions for the Dual-Column/Dual-Detector
Approach 20
7 Background Levels of Phthalate Esters in Various Brands of
Acetone 24
8 Background Levels of Phthalate Esters in Various Brands of
Hexane 25
9 Background Levels of Phthalate Esters in Various Brands of
Isooctane 26
10 Background Levels of Phthalate Esters in Various Brands of
Diethyl Ether 27
11 Background Levels of Phthalate Esters in Various Brands of
Methylene Chloride 28
12 Background Levels of Phthalate Esters in Various Brands of
Reagent Waters 30
13 Typical Levels of Phthalate Esters Removed from Florisil,
Alumina, and Silica Gel by 20 Percent Diethyl Ether in
Hexane and Acetone 32
14 Phthalate Esters Detected in Procedural Blanks from
Florisil Cartridges 34
15 Typical Levels of Phthalate Esters Removed from Filter
Paper by Hexane/Acetone (1:1) 35
-------
TABLES (continued)
Number Page
16 Typical Levels of Phthalate Esters Removed from Paper
Thimbles by Hexane/Acetone (1:1) 36
17 Typical Levels of Phthalate Esters Removed from Aluminum
Foil by Hexane/Acetone (1:1) 37
18 Typical Levels of Phthalate Esters Removed from Anhydrous
Sodium Sulfate by Hexane/Acetone (1:1) 38
19 Typical Levels of Phthalate Esters Removed from Glass Wool
by Hexane/Acetone (1:1) 39
20 Typical Levels of Phthalate Esters Contamination Remaining
on Laboratory Glassware that was Subjected to Common
Laboratory Cleanup Procedures Before and After Baking at
400°C Temperatures for 4 Hrs 46
21 Typical Levels of Phthalate Esters Contamination on
Laboratory Glassware Subjected to Common Laboratory Cleanup
Procedures, Before and After Storage in an Analytical
Laboratory Environment 47
22 Typical Levels of Phthalate Esters Contamination in Hexane
Stored in Sample Vials for up to 61 Days at 4°C 48
23 Typical Levels of Phthalate Esters Contamination in
Isooctane Stored in Sample Vials for up to 61 Days of 4"C . . 49
24 Retention Times of Method 8060 Compounds Analyzed on a DB-5
Fused-Silica Capillary Column 51
25 Retention Times of the Additional Phthalate Ester Standards
Analyzed on a DB-5 Fused-Silica Capillary Column 52
26 Retention Times of the Method 8060 Compounds Analyzed on a
Supelcowax-10 Fused-Silica Open Tubular Column 57
27 Retention Times of 16 Phthalate Ester Standards Analyzed on
a Supelcowax-10 Fused-Silica Open Tubular Column 58
28 Retention Times of the Phthalate Ester Standards Analyzed
on a DB-210 Fused-Silica Open Tubular Column 65
XI
-------
TABLES (continued)
Number Page
29 Ratio of EC Detector Absolute Response at 350°C and 320°C . . 67
30 Retention Times and Relative Retention Times of Phthalate
Esters on the DB-5/DB-608 Column Pair 69
31 Retention Times and Relative Retention Times of Phthalate
Esters on the DB-608/DB-1701 Column Pair 70
32 Retention Times and Relative Retention Times of Phthalate
Esters on the RTx-5/DB-608 Column Pair 71
33 Retention Times and Relative Retention Times of Phthalate
Esters on the DB-5/DB-1701 Column Pair 72
34 Reproducibility of the Detector Response Using the
DB-5/DB-608 Column Setup (J&W Scientific Inlet Splitter) ... 77
35 Reproducibility of the Detector Response Using the
DB-608/DB-1701 Column Setup (8-in Injection Tee from Supelco) 78
36 Reproducibility of the Detector Response Using the
DB-5/DB-1701 Column Setup (J&W Scientific Inlet Splitter) . . 79
37 Method Linearity Using the DB-5/DB-1701 Setup 80
38 GC Method Evaluation—Internal Standard Summary 81
39 Average Recoveries of Method 8060 Compounds Using Separatory
Funnel Extraction (Method 3510) 82
40 Average Recoveries of Method 8060 Compounds Using Continuous
Liquid-Liquid Extraction (Method 3520) 83
41 Average Recoveries of Method 8060 Compounds Using Mixxor
Extractor and Hexane 84
42 Results of Method 3510 and 8060 Performance
(Concentration 1) 86
43 Results of Methods 3510 and 8060 Performance
(Concentration 2) 87
44 Results of Methods 3510 and 8060 Performance
(Concentration 3) 88
xii
-------
TABLES (continued)
Number Page
45 Recovery of Surrogate Compounds Spiked into Reagent Water
Samples (Method 3510) 89
46 Evaluation of C8-Membrane Disks for Preconcentration of
Phthalate Esters 90
47 Evaluation of C8-Membrane Disks for Preconcentration of
Phthalate Esters (Multiple Use of Disks) 91
48 Evaluation of C18-Membrane Disks for Preconcentration of
Phthalate Esters (Multiple Use of Disks) 92
49 C18-Membrane Disks Method Evaluation 93
50 C18-Membrane Disks Method Evaluation—Surrogate Recoveries . . 94
51 Results of Methods 3550 and 8060 Performance 95
52 Results of Methods 3540 and 8060 Performance 96
53 Recovery of Surrogate Compounds Spiked into Soil Samples
(Method 3550) 98
54 Percent Recoveries of Method 8060 Compounds Using Alumina
(Method 3610) and Florisil (Method 3620) Column
Chromatography 99
55 Elution Patterns and Percent Recoveries of the Method 8060
Compounds from the Florisil Cartridges Using 10-Percent
Acetone in Hexane 100
56 Elution Patterns and Percent Recoveries of the Method 8060
Compounds from the Alumina Cartridges Using 10-Percent
Acetone in Hexane and 20-Percent Acetone in Hexane 101
57 Elution Patterns and Percent Recoveries of the Method 8060
Compounds from Alumina Cartridges of Various Sizes by Elution
with 20-Percent Acetone in Hexane 102
58 Percent Recoveries of the Method 8060 Compounds from Florisil
and Alumina Cartridges when Interferents are Present 104
xiii
-------
TABLES (continued)
Number Page
59 Elution Patterns and Percent Recoveries of Method 8060
Compounds from Florisil Cartridges with Hexane/Diethyl Ether
(1:1) 105
60 Results of the Florisil Cartridge Cleanup Evaluation Study . . 106
61 Experimental Design for Cartridge Florisil Cleanup Method
Development 107
62 Recovery of Phthalate Esters from the 1-g Florisil Cartridge . 110
63 Recovery of Phthalate Esters from the 1-g Florisil Cartridge
in the Presence of the OCPs Ill
64 Interferences in the Determination of Phthalate Esters Caused
by OCPs 112
65 Results of Method Blank Analyses for the Florisil
Cartridge 113
66 Percent Recoveries of Phthalate Esters from Various Matrices
by Florisil Cartridge Cleanup with Hexane/Methylene Chloride
(4:1) and Hexane/Acetone (9:1) as Eluants 114
67 Concentration of the 16 Phthalate Esters as a Function of
Time at pH 68 to 6.9 . . '. 115
68 Concentration of the 16 Phthalate Esters as a Function of
time at pH 9 116
69 Concentration of the 16 Phthalate Esters as a Function of
Time at pH 2 117
70 Recovery of the 16 Phthalate Esters as a Function of Time
at -10'C (soil matrix) 122
71 Reproducibility of the Absolute Area and Retention Time of
Benzyl Benzoate 125
72 Absolute Areas of the Internal Standard 127
73 Reproducibility of the GC/ECD Response Factors Used in
Quantitation 128
xiv
-------
TABLES (continued)
Number Page
74 Comparison of the Multilevel Calibrations Performed on
August 18, 1987, and August 28, 1987 129
75 Comparison of the Multilevel Calibrations Performed on
November 17, 1987, and November 25, 1987 130
76 Multilevel Calibration Performed on December 8, 1987 ..... 131
77 Daily Response Factor at 500 pg//iL 132
78 Daily Response Factor at 1,000 pg//iL 133
79 Daily Response Factor at 1,500 pg//iL 134
80 Daily Response Factor at 500 pg//iL 135
81 Daily Response Factor at 1,000 pg/ML 136
82 Daily Response Factor at 1,500 pg//iL 137
83 Daily Response Factor at 500 pg//*L 138
84 Daily Response Factor at 1,000 pg//iL 139
85 Daily Response Factor at 1,500 pg//*L 140
86 Comparison of the Multilevel Calibrations Performed on
November 25, 1987, and December 8, 1987 141
87 Accuracy and Precision Data for Method 3510 and
Method 8061 143
88 Accuracy and Precision Data for Method 3550 and
Method 8061 144
89 Results of Method 8060 Analysis for EPA WP-482 Sample 4 ... 149
90 Results of Method 8060 Analysis for EPA WP-482 Sample 1 ... 150
91 Results of Method 8060 Analysis for EPA WP-482 Sample 2 ... 151
92 Results of Method 8060 Analysis for EPA WP-482 Sample 3 ... 152
xv
-------
TABLES (concluded)
Number Page
93 Results of Method 8060 Analysis for EPA WP-485 153
94 Results of Method 8060 Analysis for EPA WP-481 Sample 1 ... 154
95 Estimation of the Instrument Detection Limit 160
96 Results of the Method Blank Analyses for the MDL Study for
Water Samples 168
97 Method Detection Limit Study -- Florisil Disposable
Cartridges Method Blanks 169
98 Method Detection Limits for Water Samples not Subjected to
Florisil Cartridge Cleanup 170
99 Method Detection Limits for Water Samples Subjected to
Florisil Cartridge Cleanup 171
100 Conditions Varied and Assigned Values for Gas Chromatographic
Analysis (Method 8060) for Ruggedness Test 172
101 Design for Ruggedness Test of Experimental Conditions .... 174
102 Ruggedness Test for Method 8060 -- Concentrations of Test
Compounds Found for Each Experiment 175
103 Ruggedness Test for Method 8060 -- Group Differences for the
Test Compounds 176
104 Retention Times (Scan Numbers) and Three Most Intense Peaks
in the Mass Spectra of Method 8060 Compounds 177
xv i
-------
SECTION 1
INTRODUCTION
Regulation of hazardous waste activities under the Resource Conservation
and Recovery Act (RCRA) of 1976 and its elements requires use of analytical
methodologies that provide reliable data. The document "Test Methods for
Evaluating Solid Waste," Office of Solid Waste Manual SW-846, revised recently
(1), provides a compilation of methods for evaluating RCRA solid wastes for
environmental and human health hazards. One of the methods in this document,
Method 8060, addresses the determination of phthalate esters. This method
provides sample extract cleanup and gas chromatographic conditions for the
determination of six compounds (Table 1) in a variety of environmental samples
including groundwater, liquids, and solids. Aqueous samples are extracted
with methylene chloride in a separatory funnel (Method 3510) or in a
continuous liquid-liquid extractor (Method 3520); solid samples are extracted
with hexane/acetone (1:1) or methylene chloride/acetone (1:1) using a Soxhlet
extractor (Method 3540) or a sonicator (Method 3550). The extracts are
cleaned using alumina or Florisil chromatography according to Methods 3610 or
3620, respectively. Organic liquids may be analyzed directly or diluted
according to Method 3580. The analysis is performed by gas chromatography
(GC) with electron capture detection (ECD) or flame ionization detection
(FID). The sensitivity of Method 8060 depends on the level of interferences
rather than on instrumental limitations. The method detection limits in
reagent water, in the absence of interferences, for the six compounds listed
in Method 8060 range from 0.29 ng/l to 3 /Kj/L. The detection limits for the
same compounds in liquid and solid wastes are highly matrix dependent.
The purpose of this study was to evaluate, in a single laboratory, and
improve Method 8060 to the extent possible and test the revised protocol on
a variety of matrices. A review of the most recent revision of Method 8060
was performed prior to the initiation of this study to identify the areas
that required further developmental work. The following comments were made:
• Only six phthalate esters are currently listed in Method 8060, and
no CAS registry numbers are given for any of these compounds.
• It is not clear whether the compounds listed in Method 8060 are the
only phthalates to be determined by this method. Since the presence
in the environment of the phthalate esters probably parallels their
production, other phthalate esters such as diisodecyl phthalate and
n-hexyl n-decyl phthalate should be considered. Furthermore, several
other phthalate esters were reported in water and air samples
collected over the United States (2). These include: diisobutyl
phthalate, dicyclohexyl phthalate, didecyl phthalate, and dimethyl
1
-------
TABLE 1. PHTHALATE ESTERS INVESTIGATED IN THIS STUDY
Compound name
Compound
abbreviation
CAS No.
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Dihexyl phthalate
Hexyl 2-ethylhexyl phthalate
Butyl benzyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis(2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Diisohexyl phthalate
Diisooctyl phthalate
Diisononyl phthalate
Diisodecyl phthalate
Hexyl decyl phthalate
Bis(2-ethylhexyl)hexahydro phthalate
Dimethyl cyclohexyl phthalate
Benzyl 2-ethylhexyl phthalate
Bi s[2- (2-ethoxyethoxy)ethyl ] phthal ate
Butyl cyclohexyl phthalate
Butyl decyl phthalate
Butyl isodecyl phthalate
Butyl octyl phthalate
Dicapryl phthalate
Diundecyl phthalate
2-Ethylhexyl isodecyl phthalate
Isodecyl tridecyl phthalate
Octyl decyl phthalate
Octyl isodecyl phthalate
Isohexyl benzyl phthalate
Dimethyl isophthalate
Di octyl isophthalate
DMPa'b
DEpa,b
DIBPa>b
DBpa,b
BMPPa>b
BMEPa'b
DAP8|b
BEEpa,b
DHpa,b
HEHPa'b
BBpa,b
BBEPa'b
DEHPa'b
DCpa,b
D0pa,b
DNPb
DIHP
DIOP
DINP
DIDP
HDP
BEHHP
DMCP
BEHP
BEEEP
BCP
BDP
BIDP
BOP
DCAP
DUP
EHIDP
IOTP
OOP
OIDP
IHBP
DMIP
DOIP
131-11-3
84-66-2
84-69-5
84-74-2
146-50-9
117-82-8
131-18-0
605-54-9
84-75-3
75673-16-4
85-68-7
117-83-9
117-81-7
84-61-7
117-84-0
84-76-4
c
27554-26-3
28553-12-0
26761-40-0
25724-58-7
c
c
c
c
c
89-19-0
42343-36-2
84-78-6
c
c
c
c
119-07-03
c
c
c
4654-18-6
aCompound is currently listed in Method
bCompound considered for incorporation
CCAS number not available.
8060 (SW-846,
in the revised
3rd Revision)
Method 8060.
-------
terephthalate. Thus, diisobutyl phthalate, dicyclohexyl phthalate,
and didecyl phthalate should also be considered for incorporation in
Method 8060.
• The use of ECDs and FIDs is stipulated in the protocol. However, no
explanation or suggestion is given when to use one detector versus
the other. Such an explanation should be made in relation to the
method detection limits.
• Compound confirmation is currently done using a different packed
column, and the provisions made in Method 8060 for second column
confirmation may not be sufficient, especially when the analysis is
performed by GC/FID; GC/MS confirmation should be included in the
method as an option.
• Interferences should be addressed in detail. Because of the
ubiquitous nature of phthalate esters, it is difficult to perform
accurate analyses free of artifacts. Procedures on how to clean
glassware and how to purify the solvents and the materials in order
to have control over the background contamination should be
incorporated. Furthermore, examples of typical background
contamination of some common laboratory items should be given in
order to make the analysts aware of such problems.
• The ECD temperature should be specified. The variation of sensitivity
of the electron capture with detector temperature is well documented.
D. Russell and B. McDuffie (3) showed that phthalate ester responses
on ECDs decrease with an increase in temperature between 250°C and
340°C. The dibutyl phthalate response increased slightly between
255°C and 260°C, but then decreased from 260°C to 310°C, above which
temperature no response was observed. Other phthalate esters
(dimethyl, diethyl, dihexyl, diphenyl, dioctyl) also exhibited this
phenomenon at temperatures above 300°C to 310'C.
• GC analysis is performed on packed columns. Column 1 (1.5%
SP-2250/1.95% SP-2401) requires two isothermal analyses (at 180°C
and 220 C), and Column 2 (3% OV-1) also requires two isothermal
analyses (at 200°C and 220°C). Isothermal temperature analyses with
packed columns are not practical when dealing with complex matrices
because material injected onto the column will tend to accumulate at
the head of the column, and the higher-boiling materials will emerge
from the column at a later time, contributing to cross-contamination
of samples. Furthermore, the resolution when using the packed columns
is far below that achieved with capillary columns and, consequently,
substitution of the packed columns by capillary columns should be
investigated.
• No internal standards and surrogate compounds are specified for use
in Method 8060.
• The acceptance criteria for accuracy (compound recovery) are quite
broad. Four of the six phthalate esters have recovery ranges from
-------
detected (D) to as high as 159 percent. Narrower ranges are desirable
at least for clean water samples.
• The sample cleanup procedures (Methods 3610 and 3620) specify the
use of large volumes of solvents that may increase the contamination
of sample extracts by impurities in the solvents (e.g., 140 ml of
20 percent diethyl ether in hexane for Method 3610 and 100 ml of
20 percent diethyl ether in hexane for Method 3620) and methods
requiring less adsorbent and less solvents are therefore desirable.
Thuren (4) reported a procedure using 3 g of 3-percent
water-deactivated Florisil; the extract was loaded to the Florisil
column and eluted with 10 mi petroleum ether followed by 25 ml of
20 percent diethyl ether in petroleum ether. The first fraction (0
to 15 ml) contained polychlorinated biphenyls (PCBs) and DDT, and the
second fraction (16 to 35 ml) contained the phthalates esters.
• The protocol lacks any information regarding the recoveries of the
six phthalate esters from the cleanup procedures given in Methods
3610 and 3620. Advantages and disadvantages of each method should be
stated.
Since the current protocol was inadequate in certain areas (e.g., in
addressing background contamination) and was lacking information in other
areas such as sample cleanup and GC analysis, the method evaluation and
improvement study was approached in two phases. Phase I, the developmental
phase, addressed the following:
• Literature review to gather relevant information
• Assessment of background contamination of solvents, materials used
in sample cleanup, and apparatus used for sample extraction
• Selection and evaluation of capillary columns for use in the analysis
of phthalate esters
• Evaluation of sample extraction procedures
• Evaluation of GC/ECD and GC/FID for the analysis of samples containing
the test compounds
• Evaluation of alumina (Method 3610) and Florisil chromatography
(Method 3620)
• Selection of surrogate and internal standards for use in Method 8060
• Sample preservation studies.
Upon completion of the experimental work in Phase I, the protocol was
revised and then tested in Phase II on a variety of samples. Performance data
generated during the evaluation of the revised Method 8060 include:
• Measures of precision and accuracy
-------
• Evidence of analyte identification
• Evidence for resolution of analyte from interfering substances
• Ruggedness study
• Method detection limits.
Subsequent sections of this report present the conclusions and
recommendations of this study, a brief literature review, details of the
experimental procedures, and the results and discussion. Appendix A contains
the literature review. The revised protocol is included as Appendix B.
Appendix C presents the mass spectra of 16 phthalate esters and 6 additional
compounds proposed as surrogates and internal standards for Method 8060.
Appendix D contains the results of the inorganic analyses of the various
aqueous matrices used in the method evaluation.
-------
SECTION 2
CONCLUSIONS
Based on the results presented in this report, several important
conclusions can be drawn concerning the determination of phthalate esters in
the environmental samples.
• Revision of Method 8060 for the determination of the phthalate esters
in environmental samples was necessary because: the packed columns
did not have enough resolving power to handle complex environmental
samples; the number of compounds listed in the method was limited to
six, whereas several other phthalate esters were reported in air and
water samples collected throughout the United States; confirmation of
compounds by GC/MS was not mentioned; interferences were not addressed
in detail; the acceptance criteria for method accuracy were too broad;
and sample cleanup procedures did not give any recovery information
for the six phthalate esters, and large columns and large volumes of
eluting solvents are used which may potentially contaminate samples
upon extract concentration.
• Contamination from solvents and reagent materials used in the analysis
and glassware limits the detection of phthalate esters at trace levels
(ppt-ppb range). Consequently, their determination in environmental
samples at ppt-ppb range requires pesticide-grade solvents, thorough
cleaning of the glassware followed by heat-treatment (for those that
can withstand 400*C temperatures), and a minimum number of steps in
sample workup.
• Extraction of water samples in a separatory funnel was desired over
the continuous liquid-liquid extraction since it gave good recoveries
and reproducibilities for most target analytes, greatly reduces the
extraction time, and also minimizes contamination. Preconcentration
of aqueous samples on C18-membrane disks followed by extraction of
phthalate esters with acetonitrile gave quantitative recoveries and
good reproducibilities and was therefore incorporated in the revised
Method 8060.
• Extract cleanup using Florisil disposable cartridges and elution with
hexane/acetone (9:1) gave quantitative recoveries for all 16 phthalate
esters proposed for incorporation in Method 8060. Organochlorine
pesticides overlap with the phthalate esters when the GC is performed
on a DB-5 fused-silica capillary column. Use of Florisil disposable
cartridges and elution with 20 percent methylene chloride in hexane
helps to remove the organochlorine pesticides. Phthalate esters are
-------
then recovered from the Florisil cartridges with hexane/acetone (9:1).
The use of Florisil disposable cartridges was included as an option
since it results in quantitative recoveries, reduces contamination,
saves chemicals, and reduces laboratory waste.
Preservation of aqueous samples at neutral and acidic pH and 4°C is
adequate for 21 days. Preservation of water samples at pH 9 and 4°C
should be avoided since most compounds show significant decrease in
concentration after 14 days of storage. Storage of spiked soil
samples at -10'C is preferred over refrigeration at 4"C, since it
minimizes the loss of the lower-molecular-weight esters.
A dual-column/dual-detector approach for the analysis of phthalate
esters increases sample throughput by allowing the primary and
confirmatory column analyses to be performed simultaneously.
Excellent reproducibilities of the retention time and detector
response were achieved with two 30 m x 0.53 mm ID fused-silica open
tubular columns of dissimilar stationary phases connected to an
injection tee and electron capture detectors.
-------
SECTION 3
RECOMMENDATIONS
• The revised Method 8060 presented in this report has been evaluated
in a single laboratory with a few relevant aqueous and solid samples.
However, the revised method should be evaluated by other laboratories
and with additional samples. This would help identify the
applicability range of the method and would define the interlaboratory
method performance.
• When applying the revised Method 8060 to samples which have not been
previously characterized, identification and quantification of the
16 phthalate esters using the dual-column/dual-detector approach is
very reliable. However, determination of the phthalate esters by
6C/MS in the selected ion monitoring mode should be considered for
future investigation since it would certainly improve the method
detection limits, and extract cleanup may not be required.
• Other internal standards and surrogate spiking compounds need to be
investigated, and criteria for the selection of internal standards and
surrogates need to be developed.
• Standard reference materials certified for phthalate esters need to
be developed so interlaboratory method evaluation studies can be
conducted.
-------
SECTION 4
LITERATURE REVIEW
In the initial phase of this study, a literature review concerning
analytical methods for the determination of phthalate esters in water, soil,
sediment, and sludge samples was conducted. This literature review was
performed by using the Computerized Chemical Abstracts search as well as
several review articles dealing specifically with the analysis of phthalate
esters. Furthermore, recent issues of Analytical Chemistry, the Journal of
ChromatographyT the Journal of Agricultural and Food Chemistry, the
Association of Official Analytical Chemists Journal, and Environmental Science
and Technology were searched for information that had not yet been entered in
the computer data bases.
The computer searches were done using DIALOG. Chemical Abstracts Files
were searched back to 1977 for all references containing "phthalate esters,"
"gas chromatography," "extraction," and "cleanup." Eighty articles judged to
be scientifically relevant to the objectives of this study'were retrieved from
the literature.
The literature review summary is included as Appendix A and presents the
material gathered in the following order:
• Sample preservation techniques
• Extraction techniques for water, soil, and sediment samples
• Sample extract cleanup techniques
• GC analysis (columns, retention time information, chromatographic
problems, etc.)
• Compound confirmation
• Background contamination by phthalate esters.
From the review presented in this report, it is evident that there is
sufficient information in the literature on sample extraction, cleanup, and
analysis of phthalate esters to form a data base upon which to design future
experiments. However, none of the information retrieved discusses
specifically how to determine all of the phthalate esters listed in
Method 8060 or those listed in Table 1 in hazardous liquid and solid samples.
-------
SECTION 5
EXPERIMENTAL PROCEDURES
The method development tasks which are presented in this report include
evaluation of the extent of background contamination by phthalate esters,
sample analysis by GC/ECD and GC/FID using the single column and the dual
column approach the sample extraction, and extract cleanup.
5.1 SAMPLE ACQUISITION
All phthalate esters listed in Table 1 were obtained from Chem Service
(distributed by Bryant Laboratories, Inc.), except for diethyl phthalate and
diisooctyl phthalate which were obtained from Scientific Polymer Products.
The purity of the compounds was stated in the catalog to be greater than
98 percent. Stock solutions of each test compound were prepared initially in
isooctane (Baker Resi-Analyzed, J. T. Baker) at concentrations of 1 mg/mL.
Working calibration standards were prepared initially in isooctane and later
in hexane by serial dilution of a composite stock solution prepared from the
individual stock solutions.
The organic solvents and the reagent water samples listed in Table 2 were
investigated for possible contamination with phthalate esters. Other
materials that were investigated included Florisil, alumina, silica gel,
filter paper, paper thimbles, aluminum foil, sodium sulfate, and glass wool
purchased from various suppliers (Table 3).
The aqueous samples used in the method evaluation included an estuarine
water sample taken from the San Francisco Bay, a groundwater sample, and a
leachate sample. The leachate sample was prepared from a soil sample, highly
contaminated in lead, as follows: 100 g (net weight) of soil was mixed with
1,600 mL deionized water, adjusted to pH 5.2 with 0.5 N acetic acid, and
shaken for 24 hrs on a mechanical shaker. Details of procedure are given in
Method 1310 of SW-846 Methods Manual. The leachate was filtered through a
0.45-pm Millipore filter (Fisher Scientific, Pittsburgh) prior to extraction
with methylene chloride. The results of the inorganic analyses of the three
aqueous matrices are included as Appendix D of this report.
The solid matrices used in the method evaluation included three NIST
Standard Reference Materials, two sandy loam samples, a sediment of unknown
origin contaminated with petroleum hydrocarbons, an estuarine sediment, and
a municipal sludge (Table 4).
10
-------
TABLE 2. IDENTIFICATION OF THE SOLVENT SAMPLES INVESTIGATED
Solvent
Acetone
Acetone
Acetone
Acetone
Acetone
Acetone
Acetone
Acetone
Hexane
Hexane
Hexane
Hexane
Purchased from
VWR Scientific
VWR Scientific
American Scientific
American Scientific
Fisher Scientific
Sargent Welch
Fisher Scientific
Caledon Laboratories
VWR Scientific
VWR Scientific
American Scientific
American Scientific
Identification
Brand
Omnisolve, glass-distilled,
Lot No. 6352, EM Science
Baker Resi -Analyzed,
pesticide residue,
Lot No. 643165, J. T. Baker
Chemical Co.
Nanograde, Lot No. 0018-KAXN,
Mallinckrodt
American Burdick & Jackson,
Product 010 Lot No. AQ529
Pesticide grade, UN 1090,
Lot No. 870359,
Fisher Scientific
Mallinckrodt Nanograde
Lot No. 0018 KBDY
Optima
Lot No. 870881
Distilled in glass
Lot No. 706037
Omnisolve, glass-distilled,
Lot No. 6349, EM Science
Baker Resi -Analyzed,
Lot No. 604100,
J. T. Baker Chemical Co.
Nanograde, Lot No. 4159KBEZ,
Mallinckrodt
American Burdick & Jackson,
Product 217, Lot No. AP783
Acurex ID
8707-032-1
8707-032-2
8707-032-3
8707-032-4
8707-032-5
8707-032-6
8707-032-7
8707-032-8
8707-032-9
8707-032-10
8707-032-51
8707-032-52
8707-032-63
8707-032-64
8707-032-71
8707-032-72
8707-032-11
8707-032-12
8707-032-13
8707-032-14
8707-032-15
8707-032-16
8707-032-17
8707-032-18
(continued)
11
-------
TABLE 2. (continued)
Solvent
Hexane
Hexane
Hexane
Hexane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Purchased from
Fisher Scientific
Sargent Welch
Fisher Scientific
Caledon Laboratories
VWR Scientific
VWR Scientific
American Scientific
American Scientific
Fisher Scientific
Sargent Welch
Fisher Scientific
Caledon Laboratories
Identification
Brand
Pesticide grade, UN 1208,
Lot No. 761495,
Fisher Scientific
Mallinckrodt Nanograde
Lot No. 4159 KAXD
Optima
Lot No. 873245
Distilled in glass
Lot No. 706302
Omnisolve, glass-distilled,
Lot No. 6323, EM Science
Baker Resi -Analyzed,
Lot No. 627718,
J. T. Baker Chemical Co.
Nanograde, Lot No. 6051KBBR
Mallinckrodt
American Burdick & Jackson,
Product 362, Lot No. AP915
Pesticide grade, 0-297,
Lot No. 864429,
Fisher Scientific
Mallinckrodt Nanograde
Lot No. 6051 KAPM
Optima
Lot No. 872894
Distilled in glass
Lot No. 707031
Acurex ID
8707-032-19
8707-032-20
8707-032-53
8707-032-54
8707-032-65
8707-032-66
8707-032-73
8707-032-74
8707-032-31
8707-032-32
8707-032-33
8707-032-34
8707-032-35
8707-032-36
8707-032-37
8707-032-38
8707-032-39
8707-032-40
8707-032-57
8707-032-58
8707-032-67
8707-032-68
8707-032-77
8707-032-78
(continued)
12
-------
TABLE 2. (continued)
Solvent
Purchased from
Identification
Brand
Acurex ID
Diethyl Ether VWR Scientific
Diethyl Ether American Scientific
Diethyl Ether American Scientific
Diethyl Ether Fisher Scientific
Diethyl Ether Fisher Scientific
Diethyl Ether Sargent Welch
Diethyl Ether Atomergic
Diethyl Ether Caledon Laboratories
Methylene
chloride
Methylene
chloride
Methylene
chloride
Methylene
chloride
VWR Scientific
VWR Scientific
American Scientific
American Scientific
Baker Analyzed
Lot No. A23099
J. T. Baker Chemical Co.
Nanograde, Lot No. 3434KBBA
Mallinckrodt
American Burdick & Jackson
Product 106, Lot No. AQ476
HPLC grade, E198-4
Lot No. 871047
Spectraanalyzed, E193
Lot No. 861047
Nanograde, Lot No. 3434KBBA
Mallinckrodt
Ultrar
Lot No. 378950
Distilled in glass
Lot No. 706145
Omni solve, glass-distilled,
Lot No. 7057, EM Science
Baker Resi-Analyzed,
Lot No. A03137,
J. T. Baker Chemical Co.
Nanograde, Lot No. 3023KBED-A
Mallinckrodt
American Burdick & Jackson,
Product 300, Lot No. A0174
8707-032-21
8707-032-22
8707-032-23
8707-032-24
8707-032-25
8707-032-26
8707-032-27
8707-032-28
8707-032-29
8707-032-30
8707-032-55
8707-032-56
8707-032-61
8707-032-62
8707-032-75
8707-032-76
8707-032-41
8707-032-42
8707-032-43
8707-032-44
8707-032-45
8707-032-46
8707-032-47
8707-032-48
(continued)
13
-------
TABLE 2. (concluded)
Solvent
Methyl ene
chloride
Methyl ene
chloride
Methyl ene
chloride
Methyl ene
chloride
Water
Water
Water
Water
Water
Purchased from
Fisher Scientific
Sargent Welch
Fisher Scientific
Caledon Laboratories
American Scientific
American Scientific
Fisher Scientific
VWR Scientific
VWR Scientific
Identification
Brand
Pesticide grade
D142-4, Lot No. 870226
Nanograde, Lot No. 3023
KAGR-A, Mallinckrodt
Optima
Lot No. 870483
Distilled in glass
Lot No. 707192
Chrom AR Mallinckrodt
HPLC grade, Lot No. 6795
American Burdick & Jackson
Product 365, Lot No. AP643
HPLC grade
Lot No. 872758
Baker analyzed HPLC water
Lot No. A19097
Omni solve EM Science
Lot No. 7020
Acurex ID
8707-032-49
8707-032-50
8707-032-59
8707-032-60
8707-032-69
8707-032-70
8707-032-79
8707-032-80
8707-034-1
8707-034-2
8707-034-3
8707-034-4
8707-034-5
8707-034-6
8707-034-7
8707-034-8
8707-034-9
8707-034-10
14
-------
TABLE 3. MATERIALS INVESTIGATED FOR POSSIBLE CONTAMINATION WITH
PHTHALATE ESTERS
Material
Purchased from
Identification
Florisil
Florisil PR
Alumina
Alumina
Silica gel
Silica gel
VWR Scientific
VWR Scientific
VWR Scientific
Universal Scientific, Inc.
VWR Scientific
VWR Scientific
Filter paper VWR Scientific
Filter paper VWR Scientific
Paper thimbles Cal Glass
Paper thimbles VWR Scientific
Aluminum foil Alpha Beta Supermarket
Aluminum foil Alpha Beta Supermarket
Sodium sulfate Fisher Scientific
Sodium sulfate VWR Scientific
Glasswool All tech Associates, Inc.
Glasswool
All tech Associates, Inc.
J. T. Baker, 60/100 mesh,
Lot No. 606721
Supelco, Inc., 60/100 mesh
Lot No. 575
J. T. Baker, aluminum oxide,
neutral, Brockman activity,
grade I, Lot No. 634733
Alumina Woelm N-Super I,
activity grade Super I, Woelm
Pharma
J. T. Baker
Baker-analyzed, 60/200 mesh,
Lot No. 610728
EM Science, 100/200 mesh,
grade 923, Stock No. 7091
VWR product, grade 54, 18.5 cm
Whatman No. 41, ashless,
18.5 cm
Schleicher & Schuell,
43 x 123 mm, Ref. No. 350267
Whatman 43 mm x 123 mm
Reynolds Heavy, 75 ft2
Skaggs Alpha Beta Heavy Duty,
37.5 ft2
Fisher Scientific,
Lot No. 870988
EM Science, Stock No. 6183
Pest grade, Lot No. 7753,
Stock No. 4034
DMCS treated, Lot No. 7651
Stock No. 4037 AA8
15
-------
TABLE 4. NIST STANDARD REFERENCE MATERIALS AND OTHER SOLID MATRICES
USED IN THE METHOD EVALUATION
Material
Description
SRM-1572
SRM-1632a
SRM-1633a
Sandy loam
soil
Sediment
sample
Estuarine
sediment
Municipal
sludge
Citrus leaves from Lake Alfred area of central Florida. The
material was air-dried, ground to pass through a 425-/im screen,
dried at 85'C, mixed in a feed blender, and sterilized with
Cobalt-60 radiation.
Coal obtained from the Humphrey No. 7 mine and coal preparation
plant of the Consolidation Coal Co., Osage, West Virginia.
Contains approximately 1.8 to 1.9 percent sulfur and was ground to
pass through a 60-mesh sieve.
Coal flyash, obtained from a coal-fired power plant that uses
Pennsylvania and West Virginia coals. The material was sieved to
pass through a 90-/jm screen.
Mixture of 20 percent organic soil and 80 percent sand.
Unknown origin, contaminated with petroleum hydrocarbons.
Collected from the San Francisco Bay Area off Seaport Blvd., South
San Francisco, California (highly contaminated with elemental
sulfur).
Obtained from the Santa Clara Valley Water District, San Jose,
California.
16
-------
The revised method 8060 was also evaluated with a series of EPA
performance evaluation samples identified as WP 482 Sample 1, WP 482 Sample 2
WP 482 Sample 3, WP 482 Sample 4, WP 485, and WP 281 Sample 2. These samples
consisted of concentrated acetone solutions containing some of the target
compounds and other organics (e.g., phenols, polynuclear aromatic
hydrocarbons, chlorinated benzenes, nitrotoluenes, etc.) at known
concentrations. These solutions were diluted with hexane and analyzed by
GC/ECD to determine if other organics present in the sample interfere with the
target compounds.
5.2 BACKGROUND CONTAMINATION STUDY
A portion of each solvent (150 mL for acetone and hexane, 30 mi for
diethyl ether, and 180 ml for methylene chloride) was condensed by
Kuderna-Danish evaporation to approximately 10 ml, and then to 1 ml using a
gentle stream of high-purity nitrogen (99.999 percent), which was directed
across the mouth of the Kuderna-Danish receiver. No concentration was
performed for isooctane. At least two replicate samples of each solvent were
prepared and analyzed by GC/ECD, with the exception of methylene chloride
concentrates which were analyzed by GC/FID. The volumes of solvents used are
those specified in Method 8060 and in the 3500 series methods. For example,
Method 3510 specifies extraction of aqueous sample with three 60-mL portions
of methylene chloride and Method 3620 specifies elution of a Florisil column
with 140 ml of 20 percent diethyl ether in hexane.
Materials such as Florisil, alumina, silica gel, anhydrous sodium
sulfate, and glass wool were immersed overnight in 200 or 300 mL of solvent.
The exact volume and the solvent used are specified in the Results and
Discussion section. The solvent was then decanted and concentrated to 1 ml
for GC analysis. The amounts of Florisil, alumina, and anhydrous sodium
sulfate are twice as large as those recommended in Method 3620, 3610, and
3510, respectively. For glass wool we used arbitrarily 5 g.
Other materials such as filter paper, paper thimbles, and aluminum foil
were cut into 0.5 in. x 0.5 in. pieces and immersed into 100 or 200 ml of
hexane/acetone (1:1). The solvent was then separated and concentrated to 1 mL
for GC analysis.
5.3 EVALUATION OF GAS CHROMATOGRAPHY
GC (with ECD and FID was evaluated only with fused-silica capillary
columns.' The gas chromatographs used throughout this study were a Varian 6000
equipped with a constant current pulsed-frequency ECD and interfaced with a
Varian Vista 402 data system, a Varian 6500 gas chromatograph equipped with
a FID and interfaced with either a Spectra Physics 4290 integrator or a Varian
Vista 402 data system, and a Varian 3400 gas chromatograph equipped with a
constant current pulsed-frequency ECD and interfaced to a Varian Vista 402
data system. All injections were performed with Varian Model 8000
autosamplers.
Three fused-silica capillary columns were investigated for the single
column approach. The GC operating conditions for each column are given in
Table 5.
17
-------
TABLE 5. GC OPERATING CONDITIONS FOR THE SINGLE-COLUMN APPROACH
00
Instrument
Column dimensions
Liquid phase
Film thickness (tan)
Carrier gas
Carrier flowrate (cm/sec)
Makeup gas
Makeup flowrate (mL/min)
Temperature program:
Injector temperature (°C)
Detector temperature (*C)
Injection volume (pL)
Type of injection
Splitless time (sec)
Split flow (mL/min)
Column 1
Varian 6000 (ECD)
30 m x 0.25 mm ID
DB-5
(J&W Scientific)
0.25
Hel ium
5' (33 psi)
Nitrogen
65
(1) 150*C to 275°C at
15'C/min
(2) 120*C to 260'C at
15'C/min, then 260'C
to 300'C at 2'C/min
(3) 120'C to 260°C at
15'C/min, then 260°C
to 280'C at 2'C/min
275
285 and 310
1.0 (isooctane)
Splitless
45
60
Column 2
Varian 6500 (FID)
30 m x 0.53 mm ID
Supelcowax-10
(Supelco Inc.)
1.0
Helium
(40 psi)
Helium
30
100'C (2-min hold) to
240'C (14-min hold) at
10'C/min
250
250
1.0 (isooctane)
Splitless
45
100
Column 3
Varian 3400 (ECD)
30 m x 0.53 mm ID
DB-210
(J&W Scientific)
1.0
Hel ium
11.5 mL/min
Nitrogen
28
125'C (1-min hold) to
240'C (16-min hold) at
5'C/min
250
250
1.4 (hexane)
On-column
--
--
•Measured at 120'C.
-------
Four 30 m x 0.53 mm ID fused-silica open tubular columns were
investigated for the dual-column/dual-detector approach. The four columns
were paired as shown in Table 6, using two different injection tees.
5.4 EXTRACTION TECHNIQUES
5.4.1 Sample Extraction
The extraction efficiencies for the Method 8060 compounds were determined
for HPLC-grade water at pH 2, 7, and 9, using separatory funnel extraction
with methylene chloride (Method 3510) and continuous liquid-liquid extraction
(Method 3520). The compounds were spiked into each water sample at 5 /ig/L;
quantification was performed by GC/ECD using a DB-5 fused-silica capillary
column and external standard calibration.
Microextraction of 50-mL spiked HPLC-grade water samples using a Mixxor
device (Lidex Technologies, Inc.) and hexane (10 ml) as the extraction solvent
was also tested. The solvent and the aqueous sample were added to the upper
reservoir, the cap was tightened, and the piston was moved up and down in the
mixing chamber five or six times. The hexane layer was decanted into a 40-mL
vial and dried with precleaned anhydrous sodium sulfate. The hexane layer was
then transferred to another vial and concentrated to 1 ml under a gentle
stream of high-purity nitrogen. Quantification was performed by GC/ECD using
a DB-5 fused-silica capillary column and external standard calibration.
Evaluation of Cfi- and C18-membrane disks (Analytichem International) was
performed with HPLC-grade water and groundwater spiked at 25 ng/L and
100 ^g/L. The aqueous matrix (500 mL) was filtered through the membrane disk
installed in a Millipore filtration apparatus at a flowrate of approximately
25 mL/min. The Cg,- or the C^-membrane disk was preconditioned, immediately
prior to use, with 10 mL metnanol followed by 10 mL HPLC-grade water. To
improve the extraction efficiency of the membrane disk, 2.5 mL methanol were
added to each aqueous sample prior to filtration. The target compounds were
extracted from the membrane disk with 10 mL acetonitrile. The acetonitrile
extract was concentrated to a final volume of 1 mL using a gentle stream of
high-purity nitrogen (99.999 percent) and analyzed by GC/ECD using the dual-
column/dual -detector approach.
Soil or sediment samples were extracted either with hexane/acetone (1:1)
in a Soxhlet extractor (Method 3540) or with methylene chloride/acetone (1:1)
using a sonic probe (Heat Systems Ultrasonics, Inc., Model W-375) following
the procedures specified in Method 3550.
5.4.2 Soil Spiking
The following procedure was used for spiking soil samples that were used
in comparing Methods 3540 and 3550. One hundred grams potting soil and 400 g
sand were mixed with 200 mL deionized water and were blended at full speed
for 2 min in a Waring laboratory blender, Waring Products Division, Dynamics
Corporation of America, New Hartford, Connecticut. Twenty milliliters of an
isooctane solution containing the Method 8060 compounds at concentrations of
35 ng/ml were added, and blending was continued for 10 min with intermittent
cooling to yield a smooth slurry. Immediately after blending, the slurry was
19
-------
TABLE 6. GC OPERATING CONDITIONS FOR THE DUAL-COLUMN/DUAL-DETECTOR
APPROACH
ro
O
Instrument
Column dimensions
Liquid phase (File) thickness)
Carrier gas and flowrate
Makeup gas
Makeup flowrate (at/Bin)
Temperature Program:
Injector temperature (*C)
Detector temperature (*C)
Injection volume (
-------
separated into 35-g portions. The slurry was stirred 10 sec after each
portion removal from the blender.
The portions were serially labeled in the order in which the removal was
done. Those portions that carried even numbers were analyzed immediately.
The portions with odd numbers were either frozen for up to 1-1/2 months at
-10°C or stored at 4°C in a walk-in refrigerator.
5.5 EXTRACT CLEANUP
5.5.1 Florisil Chromatography
Florisil from J. T. Baker (60/80 mesh size; Lot No. 442707) was activated
for 16 hrs at 400°C, allowed to cool in a precleaned amber glass jar, and then
deactivated with distilled water (3 percent by weight). The glass jar was
rolled for 10 min and then allowed to sit for at least 2 hrs. Ten grams of
the deactivated Florisil were loaded to a chromatographic column (20 mm ID x
500 mm length) and topped with 1 cm (bed height) of precleaned anhydrous
sodium sulfate. The columns were first eluted with 40 mL hexane which was
discarded; hexane standards or sample extracts were loaded to the columns, and
the phthalate esters were eluted with 100 mL of 20 percent diethyl ether in
petroleum ether. The fractions were concentrated to 10 mL by Kuderna-Danish
evaporation.
Florisil disposable cartridges (Supelco, Inc.) containing LC-Florisil
40 urn particles (60 A pores) were each prewashed with 4 mL pesticide grade
hexane prior to use. Florisil cartridges were eluted in sets of 12 on a
Supelclean™ vacuum manifold (Supelco, Inc.) that provided increased sample
throughput while the volume of the eluting solvent was kept to a minimum. The
eluting solvents evaluated with the Florisil disposable cartridges are given
in Section 6.4.
5.5.2 Alumina Chromatography
Alumina Woelm N Super I, W200 series was activated for 16 hours at 400°C,
allowed to cool in a precleaned amber glass jar, and then deactivated with
distilled water (3 percent by weight). The glass jar was rolled for 10 min
and then allowed to sit for at least 2 hrs. Ten grams of the deactivated
alumina were loaded to a chromatographic column (20 mm ID x 500 mm length) and
topped with 1 cm (bed height) of precleaned anhydrous sodium sulfate. The
columns were first eluted with 35 mL hexane which was discarded; hexane
standards or sample extracts were loaded to the columns and the phthalate
esters were eluted with 140 mL of 20 percent diethyl ether in hexane. The
fractions were concentrated to 10 mL by Kuderna-Danish evaporation.
5.6 SAMPLE PRESERVATION
Preservation studies were carried out for both water and soil samples.
Fourteen 1-L reagent water samples were spiked with the test compounds at one
concentration and stored in the dark at 4*C for up to 21 days. Two samples
were extracted immediately; other samples were removed at day 1, 3, 7, 10, 14,
and 21 and analyzed for the 16 test compounds. Duplicate measurements were
performed at each time event. In addition, six 1-L samples were spiked with
21
-------
the test compounds at one concentration and were adjusted to pH 2 with 6N
H?S04. Two samples were extracted immediately; the other samples were stored
at 4°C for 7 days and 14 days. This experiment was repeated at pH 9.
Thirty-gram portions of sandy loam soil spiked with the 16 phthalate
esters at 1 /ig/g per component were stored frozen at -10°C for 1 month. At
day 3, 7, 14, and 28 they were removed and analyzed for the target compounds.
5.7 GC/MS METHODOLOGY
A Finnigan 4510B GC/MS system interfaced to a Finnigan Nova 4X data
system was used in this study. The GC was equipped with a split/splitless
injector. The column was a 30 m x 0.25 mm ID DB-5 fused-silica capillary
column (0.25 urn film thickness) supplied by J&W Scientific, Inc. The GC
instrumental conditions were as follows:
• Temperature program -- 40'C to 300°C at 8'C/min
• Injector temperature -- 250'C
• Transfer line temperature -- 260*C
• Injection volume -- 1 /tl_
• Solvent -- methylene chloride
• Carrier gas -- helium at 10 psi at 40°C
The MS conditions were as follows:
• Ion source tuning -- as per EPA DFTPP requirement
• Ion source temperature -- 190eC
• Scanning mass range -- 45 to 450 amu
• Scan rate -- 1 sec/cycle
• Electron energy -- 70 eV
• Multiplier voltage -- 1,400 eV
5.8 GLASSWARE CLEANUP
All glassware used in this study, except the continuous liquid-liquid
extractors, Soxhlet extractors, and the volumetric glassware, were washed with
Alconox detergent and hot water, were rinsed with tap water and with deionized
water, and were finally baked at 400"C for 2 hrs unless otherwise indicated.
The continuous liquid-liquid extractors, the Soxhlet extractors, and the
volumetric glassware were washed as described above and were rinsed with
American Burdick & Jackson acetone immediately after washing and again prior
to use.
22
-------
SECTION 6
RESULTS AND DISCUSSION
6.1 BACKGROUND CONTAMINATION STUDY
This study was undertaken in conjunction with the evaluation of EPA
Method 8060 for the determination of phthalate esters in environmental
samples. Since contamination of samples could not be avoided even with the
highest degree of precaution, we wanted to know the sources of contamination
and the conditions under which we might be able to reduce the background
contamination. Various solvents recommended for use in Method 8060 were
obtained from six commercial suppliers and were concentrated up to 180-fold
to determine background levels of 11 phthalate esters. Reagent water samples
from three commercial suppliers, totaling five different brands, were analyzed
for 16 phthalate esters. Materials used in sample workup that were
investigated in this study include: Florisil, alumina, silica gel, filter
paper, paper thimbles, aluminum foil, anhydrous sodium sulfate, and glass
wool. In each case, a minimum of two brands of each material was obtained
from commercial suppliers. Solvent washing, as well as high temperature
(400eC) treatment followed by solvent washing, was employed to remove traces
of phthalate esters. The solvent washings were analyzed for 11 phthalate
esters. Finally, the likelihood of contamination of laboratory glassware
subjected to common laboratory cleanup procedures, and the likelihood of
contamination of hexane and isooctane stored in sample vials of different
sizes and capped with metal foil or Teflon-lined crimp-top caps were
addressed.
6.1.1 Organic Solvents
Five organic solvents from 6 different commercial suppliers were analyzed
for 11 of the most common phthalate esters. The results are summarized in
Tables 7 through 11. Five phthalate esters that were detected in these
solvents include: DIBP, DBF, DHP, BBP, and DEHP. Their levels range from 0.1
to 0.87 ng/mL for hexane and acetone, 0.2 to 3.6 ng/mL for diethyl ether, and
10 to 115 ng/mL for isooctane. No phthalate esters were detected in any of
the methylene chloride solvents, except for OOP in one sample; however, the
detection limits were much higher than for the other solvent, since the
condensates were analyzed by 6C/FID.
Since typical volumes of hexane and acetone used in sample workup are 200
to 300 mL, then the amounts of the phthalate esters which will be introduced
as contaminants with the organic solvent may be as high as 260 ng per
compound. This amount will result in concentrations of 26 ng/g of sample for
a 10 g sample extracted with hexane/acetone.
23
-------
TABLE 7. BACKGROUND LEVELS OF PHTHALATE ESTERS IN VARIOUS BRANDS OF
ACETONE
ro
Concentration (ng/aL)*
Suppl ier
VUR Scientific
VUR Scientific
American Scientific
American Scientific
Fisher Scientific
Fisher Scientific
Caledon Laboratories
Sargent Welch
Brand
Owl solve, glass distilled,
EN Science
Baker Resi -Analyzed, pesticide
residue, J. T. Baker Cheelcal Co.
Nallinkrodt, Nanograde
American Burdick & Jackson,
Product 010
Fisher Scientific, pesticide grade,
UN 1090
OptiM
Distilled in glass
Nallinkrodt, nanograde
Lot No.
6352
643135
0018 KAXN
AQ529
870359
870881
706037
0018 KBDY
DNP
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
b
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
OEP
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
b
b
0.40
<0.10
<0.10
<0.10
<0.10
<0.10
DIBP
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
0.33
0.37
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
DBP
0.20
0.24
0.47
0.40
0.44
0.40
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
0.50
0.50
-------
TABLE 8. BACKGROUND LEVELS OF PHTHALATE ESTERS IN VARIOUS BRANDS OF HEXANE
Concentration
Suppl 1«r
VHR Scientific
VWR Scientific
American Scientific
American Scientific
Fisher Scientific
Fisher Scientific
ro
en Caledon Laboratories
Sargent Welch
Brand
Omni solve, glass distilled,
EN Science
Baker Resl-Analyzed, J. T. Baker
Chemical Co.
Halltnkrodt, Nanograde
American Burdick » Jackson,
Product 217
Fisher Scientific, pesticide grade,
UN 1208
Optima
Distilled in glass
Nallinkrodt, nanograde
Lot No.
6349
604100
4159KBEZ
AP783
761495
873245
706302
4159 KBXD
ONP
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
DEP
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
<0.10
DIBP
-------
TABLE 9. BACKGROUND LEVELS OF PHTHALATE ESTERS IN VARIOUS BRANDS OF
ISOOCTANE
Supplier
VUR Scientific
VUR Scientific
American Scientific
American Scientific
Fisher Scientific
Fisher Scientific
ro
°* Caledon Laboratories
Sargent Welch
Brand Lot Ho.
Omnisolve, glass distilled, 6323
EN Science
Baker Analyzed, 627718
J. T. Baker Chemical Co.
Nallinkrodt, Nanograde 6051KBBR
American Burdlck & Jackson, AP915
Product 362
Fisher Scientific, pesticide grade, 864429
0-297
Optima 872894
Distilled In glass 707031
Nallinkrodt, nanograde 6051 KAPN
DHP OEP DIBP DBP
<10 <10 <10 93
<10 <10 <10 81
<10 <10 <10 91
<10 <10 <10 79
<10 <10 <10 110
<10
-------
TABLE 10. BACKGROUND LEVELS OF PHTHALATE ESTERS IN VARIOUS BRANDS OF
DIETHYL ETHER
ro
Concentration
Supplier
VUR Scientific
American Scientific
American Scientific
Fisher Scientific
Fisher Scientific
Atoaerglc
Caledon Laboratories
Sargent Helen
Brand
Baker Analyzed,
J. T. Baker Cheat cal Co.
Mallinkrodt, Nanograde
American Burdick I Jackson,
Product 106
HPLC grade E198-4
Spectra-Analyzed E193
Atowrgic
Distilled in glass
Mallinkrodt, nanograde
Lot No.
A23099
3434KBBA
AQ476
871047
861047
378950
706145
3434 KBBA
OHP
<0.80
<0.80
3.6
3.3
<0.80
<0.80
<0.80
<0.80
1.1
1.5
<0.10
<0.10
<0.80
<0.80
2.0
2.2
OEP
<0.80
<0.80
<0.80
<0.80
<0.80
<0.80
<0.80
<0.80
<0.80
<0.80
<0.10
<0.10
<0.80
<0.80
<0.80
<0.80
DIBP
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.10
<0.10
<0.20
<0.20
<0.20
<0.20
DBP
1.8
2.8
2.9
2.9
2.5
1.8
2.5
1.7
2.7
3.1
<0.10
<0.10
<0.20
<0.20
<0.20
<0.20
DAP
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.20
<0.10
<0.10
<0.20
<0.20
<0.20
<0.20
OHP
0.70
0.60
0.83
0.67
0.59
<0.20
<0.20
<0.20
0.55
0.60
<0.10
<0.10
1.6
1.4
0.20
0.20
(ng/«L)'
BBP
0.18
0.14
0.16
0.13
0.14
<0.05
<0.05
-------
TABLE 11. BACKGROUND LEVELS OF PHTHALATE ESTERS IN VARIOUS BRANDS OF METHYLENE CHLORIDE8
Concentration
Supplier
VWR Scientific
VWR Scientific
American Scientific
American Scientific
Fisher Scientific
Fisher Scientific
Caledon Laboratories
ro
00
Sargent Welch
Brand
Oranl solve, glass distilled,
EH Science
Baker Rest -Analyzed,
J. T. Baker Chemical Co.
Halllnkrodt, Nanograde
American Burdlck & Jackson,
Product 300
Pesticide grade,
D142-4
Optima
Distilled In glass
Halltnkrodt, nanograde
Lot No.
7057
A03137
3023KBED-A
A0174
870226
870483
707192
3023KAGR-A
ONP
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
OEP
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
DIBP
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
DBP
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
<3.0
DAP
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
DHP
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
(ng/roL)
BBP
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
*
DEHP
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
DCP
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
OOP
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
8.8
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
DNP
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
<6.0
•Concentration factor Is 180. Analyses were performed by GC/FID using a 30 m x 0.53 on ID Supelcowax-10 fused-silica open tubular column. Two replicate samples
Mere analyzed from each brand of Mthylene chloride.
-------
6.1.2 Reagent Water
No phthalate esters were detected in any of the five different brands of
reagent water at levels above 0.5 or 1.0 /jg/L. The results are presented in
Table 12. Analyses were performed by GC/FID.
The HPLC-grade water samples from Fisher Scientific (Lot No. 874716) were
selected to be analyzed by GC/ECD following extraction with methylene chloride
(American Burdick & Jackson, lot AT 348) and solvent-exchange with hexane
(American Burdick & Jackson, log AQ 406). Four phthalate esters, DBP, DHP,
BBP, and DHP, were detected in seven samples at levels ranging from 0.1 to
0.2 /jg/L. None of the other 12 phthalate esters were detected. The method
detection limits, estimated from the instrument detection limits, were
approximately 0.02 /ig/L.
6.1.3 Materials
Tables 13 through 19 present data on the background levels of the
phthalate esters in various materials such as Florisil, alumina, silica gel,
filter paper, paper thimbles, aluminum foil, anhydrous sodium sulfate, and
glass wool. GC/ECD chromatograms of solvent washings obtained from these
materials are shown in Figures 1 through 5.
Florisil, alumina, and silica gel (Table 13) showed significant levels,
and, therefore, use of these materials in sample cleanup should be employed
cautiously. Washing of these materials, prior to use, with the solvent(s)
used for elution during the extract cleanup step was helpful.
Use of Florisil disposable cartridges (1 g Florisil per cartridge) has
been investigated to determine if contaminant levels will decrease as the
amount of Florisil and the volume of the eluting solvent are proportionally
decreased. The phthalate esters detected in the method blanks obtained from
Florisil disposable cartridges are shown in Table 14. Their levels ranged
from 10 to 460 ng with 5 phthalate esters in the 105 to 460 ng range. It is
possible that the plastic material used in manufacturing these cartridges
contributed these contaminants.
Complete removal of the phthalate esters does not seem possible, and it
is desirable to keep the steps involved in sample preparation to a minimum.
If cleanup must be used, then method blanks must be obtained prior to sample
workup in order to select the cleanest adsorbent. Furthermore, solvent
volumes should be kept to a minimum, and solvents from various commercial
suppliers should be tested, if information regarding the background levels of
phthalate esters is not available.
The filter paper and paper thimble samples were first precleaned in a
Soxhlet extractor with acetone for 72 hrs, using fresh solvent which was
changed at 24-hr intervals. Following precleaning, samples were cut in small
pieces and were immersed in hexane/acetone (1:1) overnight. Figures 3 and 4
present GC/ECD chromatograms obtained before precleaning and after
precleaning. It is quite obvious from the results presented in Tables 15 and
16 that washing of the paper filter or paper thimble with solvent, prior to
use, is not adequate. Even after Soxhlet extraction for 72 hrs with acetone,
29
-------
TABLE 12. BACKGROUND LEVELS OF PHTHALATE ESTERS IN VARIOUS BRANDS OF REAGENT WATERS
Concentration
Supplier
American Scientific
American Scientific
Fisher Scientific
VWR Scientific
VWR Scientific
Brand Lot No.
Chrom AR 6795
Mallinkrodt
HPLC grade
American AP643
Burdick &
Jackson,
Product 365
HPLC grade 872758"
Baker A19097
Analyzed
HPLC Water
Omni solve 7020
EM Science
DMP
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
DEP
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
DIBP
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
DBP
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
(M9/L)
DNPP
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
•
BNEP
<1.0
<1.0
<1.0
<1.0
<1.0
<1.0
<1.0
<1.0
<1.0
<1.0
DAP BEEP
<0.5 <1.0
<0.5 <1.0
<0.5 <1.0
<0.5 <1.0
<0.5 <1.0
<0.5 <1.0
<0.5 <1.0
<0.5 <1.0
<0.5 <1.0
<0.5 <1.0
(continued)
'One-liter water sample was extracted in a separatory funnel, three times, with 60 mL methylene
chloride (American Burdick & Jackson, Lot No. A0174), the extract dried with anhydrous sodium
sulfate (Fisher Scientific Lot No. 872983, heated at 400°C for 4 hours) and concentrated in a
Kuderna-Danish evaporator to 1 mL. Analyses were performed by GC/FID using a 30 m x 0.53 mm ID
Supelcowax-10 fused-silica open tubular column. Two replicate samples were analyzed from
each brand of reagent water.
"Another lot of HPLC-grade water from Fisher Scientific (lot 874716) was analyzed by GC/ECD.
-------
TABLE 12. (concluded)
Supplier
American Scientific
American Scientific
Fisher Scientific
VWR Scientific
VWR Scientific
Brand Lot No.
Chrom AR 6795
Mallinkrodt
HPLC grade
American AP643
Burdick &
Jackson,
Product 365
HPLC grade 872758"
Baker A19097
Analyzed
HPLC Water
Omni solve 7020
EM Science
HEHP
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
Concentration (/*g/L)"
DHP BBP DEHP BBEP DCP OOP DNP
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
<1.0 <1.0 <1.0 <1.0 <1.0 <1.0 <1.0
'One-liter water sample was extracted in a separatory funnel, three times, with 60 mL methylene
chloride (American Burdick & Jackson, Lot No. A0174), the extract dried with anhydrous sodium
sulfate (Fisher Scientific Lot No. 872983, heated at 400°C for 4 hours) and concentrated in a
Kuderna-Danish evaporator to 1 mL. Analyses were performed by GC/FID using a 30 m x 0.53 mm ID
Supelcowax-10 fused-silica open tubular column. Two replicate samples were analyzed from
each brand of reagent water.
"Another lot of HPLC-grade water from Fisher Scientific (Lot 874716) was analyzed by GC/ECD.
-------
TABLE 13. TYPICAL LEVELS OF PHTHALATE ESTERS REMOVED FROM
FLORISIL, ALUMINA, AND SILICA GEL BY 20 PERCENT
DIETHYL ETHER IN HEXANE AND ACETONE*
Florisil
J. T. Baker
Lot No. 606721
(60/100 mesh)
Florisil PR
Supelco Inc.
Lot No. 575
(60/100 mesh)
Alumina Woe1m N Super I
Woe 1m Pharma
(Universal Scientific,
Inc.)
Compound
DMP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
OOP
DNP
First
washing
110
190
48
140
20
10
24
100
<10
<10
<10
160
110
22
100
<10
12
<10
70
<10
<10
<10
Second
washing
110
160
18
<10
<10
12
<10
62
<10
<10
<10
no
130
12
<10
<10
12
<10
48
<10
<10
<10
First
washing
74
50
22
<10
<10
14
<10
40
<10
58
52
110
120
24
12
<10
12
<10
54
<10
92
96
Second
washing
160
88
20
<10
<10
10
<10
38
<10
34
120
110
48
24
<10
<10
32
<10
38
<10
68
68
First"
washing
420
130
60
14
150
18
<10
68
<10
<10
<10
Second
washing
c
1,300
1,300
880
120
32
18
d
d
d
d
c
420
480
110
140
40
<10
40
<10
<10
<10
(continued)
3 20 percent diethyl ether in hexane was used for Florisil and alumina; 200 and
300 ml, respectively. Acetone (300 mL) was used in the case of silica gel. In
each case 20 g of materials were used. Vextract was 1 mL. Values given are in
ng/mL of extract, and they represent measurements from duplicate experiments.
"Duplicate experiments were performed in this case; however, one of the washings
was lost during solvent concentration.
c Not able to quantify because of interference from solvent.
d Not able to quantify because of high background.
32
-------
TABLE 13. (concluded)
Aluminum oxide,
neutral, J. T. Baker
Brockmann Activity
Grade 1
Lot No. 634733
Silica gel
T. Baker analyzed
Lot No. 610728
(60/100 mesh)
Silica gel
E. M. Science
Grade 923
(100/200 mesh)
Compound
DMP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
OOP
DNP
First
washing
640
52
38
24
160
40
20
600
<10
<10
<10
1,200
66
84
56
110
18
<10
300
<10
<10
<10
Second
washing
1,960
340
220
32
62
14
<10
200
<10
<10
<10
3,200
160
340
88
140
14
<10
46
<10
<10
<10
First
washing
c
<100
2,300
550
650
420
1,300
1,200
340
1,000
1,000
c
<100
2,100
220
990
380
1,800
1,100
400
1,300
600
Second
washing
c
190
11
<10
14
92
780
24
<10
16
<10
c
150
14
<10
<10
130
680
25
<10
18
<10
First
washing
c
<10
<10
31
<10
24
<10
40
<10
25
<10
c
110
33
30
<10
25
<10
349
<10
<10
<10
Second
washing
c
400
<10
36
<10
33
<10
31
<10
<10
<10
c
470
<10
<10
<10
50
<10
22
<10
<10
<10
a20 percent diethyl ether in hexane was used for Florisil and alumina; 200 and
300 ml, respectively. Acetone (300 mL) was used in the case of silica gel. In
each case 20 g of materials were used. Vextract was 1 ml. Values given are in
ng/mL of extract, and they represent measurements from duplicate experiments.
bDuplicate experiments were performed in this case; however, one of the washings
was lost during solvent concentration.
cNot able to quantify because of interference from solvent.
dNot able to quantify because of high background.
33
-------
TABLE 14. PHTHALATE ESTERS DETECTED IN PROCEDURAL BLANKS FROM FLORISIL
CARTRIDGES
Average amount ± standard deviation8
Compound (ng/cartridge)
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
456 ±
159 ±
105 ±
42
66
10
136 ±17
89 ±
262 ±
53 ±
66 ±
24 ±
33 ±
18 ±
41 ±
291 ±
<10
<10
<10
41
72
49
4
17
2
3
12
37
aThe number of determinations was 5. Each Florisil
cartridge was eluted with 5 mL of 20 percent methylene
chloride in hexane (Fraction 1), which was discarded,
followed by 5 mL of hexane/acetone (9:1) which was
concentrated to 1 mL and analyzed by GC/ECD.
34
-------
TABLE 15. TYPICAL LEVELS OF PHTHALATE ESTERS REMOVED FROM
FILTER PAPER BY HEXANE/ACETONE (1:1)*
Concentration (ng/mL extract)
Compound
DMP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
DOP
DNP
First washing
3,100
<10
260
170
44
260
54
160
<10
<10
<10
b
<10
240
210
35
280
72
220
<10
<10
<10
Second washing
710
<10
110
68
<10
140
28
89
<10
<10
<10
640
<10
120
62
<10
170
38
130
<10
<10
<10
Concentration
(ng/g)
380
<1 .0
37
26
4.0
43
9.6
30
<1 .0
<1.0
<1.0
a Whatman #41 (VWR Scientific) filter paper was precleaned
three times in a Soxhlet extractor with American Burdick &
Jackson acetone, each time using fresh solvent which was
changed at 24-hr intervals. Samples were then cut into
0.5 in. x 0.5 in. pieces; 10 g were immersed in 100 ml of
hexane/acetone (1:1) overnight. V ^ was 1 mL. Values
given represent measurements from duplicate experiments.
b Not able to quantify.
35
-------
TABLE 16. TYPICAL LEVELS OF PHTHALATE ESTERS REMOVED FROM
PAPER THIMBLES BY HEXANE/ACETONE (l:l)a
Concentration (ng/mL extract)
Concentration
Compound First washing Second washing (ng/g)
DMP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
OOP
DNP
960
35
94
12
14
470
110
76
<10
<20
<20
1,000
12
89
27
12
800
130
76
<10
<20
<20
330
22
23
<10
<10
140
31
28
<10
22
23
400
24
23
<10
<10
200
29
28
<10
<20
<10
140
4.7
12
2.8
2.2
80
15
10
<1.0
<2.0
<2.0
"Whatman 43 mm x 123 mm paper thimble (VWR Scientific); each
thimble was precleaned three times in a Soxhlet extractor
with American Burdick & Jackson acetone, each time using
fresh solvent which was changed at 24-hr intervals.
Samples were then cut into 0.5 in. x 0.5 in. pieces; 10 g
were immersed in 100 mL of hexane/acetone (1:1) overnight.
v # was 1 mL. Values given represent measurements from
duplicate experiments.
36
-------
TABLE 17. TYPICAL LEVELS OF PHTHALATE ESTERS REMOVED FROM ALUMINUM FOIL
BY HEXANE/ACETONE (l:l)a
Reynolds
heavy-duty aluminum foil
Before baking
at 400°C
After baking at 400'C
Skaggs Alpha Beta
heavy-duty aluminum foil
Before baking
at 400°C
Compound
DMP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
OOP
DNP
Second
290
<10
<10
<10
<10
36
32
36
<10
11
15
washing
260
<10
<10
<10
<10
34
28
13
<10
<10
<10
First
150
33
<10
22
<10
96
84
<10
<10
<10
<10
washing
180
14
<10
<10
<10
26
11
20
<10
<10
<10
Second washing
410
17
14
140
18
140
170
18
<10
105
<10
Second
202
<10
<10
<10
<10
24
19
19
10
<10
13
washing
210
<10
<10
<10
<10
25
21
37
<10
19
<10
aAluminum foil (5 g), before or after baking at 400eC, was cut into 0.5 in.
pieces which were immersed into 200 mL of hexane/acetone (1:1) overnight.
^extract was * mL• Values given are in ng/mL of extract, and they represent
measurements from duplicate experiments.
bThe duplicate extract was lost during solvent concentration.
37
-------
TABLE 18. TYPICAL LEVELS OF PHTHALATE ESTERS REMOVED FROM ANHYDROUS
SODIUM SULFATE BY HEXANE/ACETONE (l:l)a
Compound
Fisher Scientific
Lot No. 870988
Second washing
before baking
at 400°C
First washing
after baking
at 400eC
Second washing
after baking
at 400°C
EM Science
6183
Second washing
before baking
at 400'C
DMP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
DOP
DNP
20
80
11
100
110
22
23
70
10
88
110
22
54
130
120
150
39
100
100
100
11
88
70
39
38
22
10
<23
67
69
62
85
10
55
14
63
16
70
30
aA volume of 300 ml hexane/acetone (1:1) was used for each portion of 50 g
of anhydrous sodium sulfate, Vextract was 1 mL. Values given are in ng/mL of
extract, and they represent measurements from duplicate experiments.
38
-------
TABLE 19. TYPICAL LEVELS OF PHTHALATE ESTERS REMOVED FROM GLASS WOOL BY HEXANE/ACETONE (1:1)'
OJ
vo
Glass wool, All tech Associates
DMCS treated
First
Compound
DHP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
OOP
DNP
washing
86
109
36
14
<10
125
51
63
<10
<10
<10
77
17
30
14
<10
121
43
21
<10
<10
<10
Glass
wool, All tech Associates
Pesticide grade
Average First washing Second washing Average First washing Average
Second concentration before baking before baking concentration after baking concentration
washing
40
10
21
<10
10
89
26
27
<10
27
<10
81
<10
13
<10
11
70
18
21
<10
35
<10
(ng/g) at 400-C
28
13
10
2
<2
41
14
13
<2
6
<2
210
<10
<10
.8 <10
.0 <10
33
<10
51
.0 <10
.2 <10
.0 <10
at 400 «C
290
14
<10
<10
<10
35
17
18
<10
<10
<10
(ng/g)
100
2
<2
<2
<2
6
3
14
<2
<2
<20
.8
.0
.0
.0
.8
.4
.0
.0
at 400'C (ng/g)
85 <24 11
<10 <10 <2
<10 <10 <2
<10 <10 <2
<10 <10 <2
<10 <10 . <2
<10 <10 <2
<10 <10 <2
<10 <10 <2
<10 <10 <2
<10 <10 <2
.0
.0
.0
.0
.0
.0
.0
.0
.0
.0
*5 g of glass wool were weighed out In a 500-mL amber jar, then allowed to soak twice for 16 hours, each time with fresh
hexane/acetone (1:1), 300 ml. The solvent washings were concentrated to 1 mL by Kuderna-Danish evaporation. Values
given are in ng/mL of extract, and they represent measurements from duplicate experiments.
-------
Florisil, VWR Scientific
(Baker Analyzed)
First washing
Florisil, Supelco
First washing
«•>« ft
iss :
Florisil, VWR Scientific
(Baker Analyzed)
Second washing
Florisil, Supelco
Second washing
Figure 1. 6C/ECD chromatograms of solvent washings obtained from two
different brands of FlorisH.
40
-------
Alumina, Woelm Pharma
First washing
Is
Is
r-r
: :
itjft
i S
Alumina, VWR Scientific
(Baker Analyzed)
First washing
Alumina, Woelm Pharma
Second washing
Alumina, VWR Scientific
(Baker Analyzed)
Second washing
Figure 2. GC/ECD chromatograms of solvent washings obtained from two
different brands of alumina.
41
-------
fl
« Before precleaning
Second washing
After precleaning
First washing
After precleaning
Second washing
Figure 3. GC/ECD chromatograms of solvent washings of filter paper (Whatman
No. 41) before precleaning (top) and after precleaning (middle and
bottom).
42
-------
Before precleaning
First washing
After precleaning
First washing
miuiiiiuiiiuiiiiiiiii!
Before precleaning
Second washing
After precleaning
Second washing
Figure 4. GC/ECD chromatograms of solvent washings obtained from paper
thimbles (Whatman 23 x 123) before precleaning and after
precleaning.
43
-------
Before baking
-£
Si-
After baking
i-
:£
Hexane blank
Figure 5. GC/ECD chromatograms of solvent washings from All tech pest grade
glass wool before baking and after baking at 400°C.
44
-------
there were still detectable levels of 6 phthalate esters on the filter paper
and 8 phthalate esters on the paper thimble in the second washing of the
material.
Therefore, paper thimbles and filter paper must be exhaustively washed
with the solvent that is going to be used in the sample extraction. Soxhlet
extraction for 12 hours with fresh solvent should be repeated at least three
times. Method blanks should be obtained before any of the precleaned thimbles
or filter papers are used. Storage of precleaned thimbles and filter paper
in precleaned glass jars covered with aluminum foil is recommended.
Washing of the aluminum foil with hexane/acetone (1:1) seem to be
sufficient in removing any contaminants. The levels of the target compounds
detected in the first washing after baking at 400°C for 4 hrs remained low.
A second washing indicated levels in excess of 100 ng/mL of extract
(Table 17). Possible contamination by the laboratory glassware may be a
reasonable explanation.
Washing alone is not sufficient for sodium sulfate and glass wool, but
baking these materials at 400°C for 4 hrs followed by solvent washing gave
acceptable blanks (Tables 18 and 19).
6.1.4 Glassware
Typical levels (in nanograms) of the 16 test compounds present on
laboratory glassware before and after baking at 400"C for 4 hours are given
in Table 20. The data shown in Table 20 indicate the removal of phthalate
esters by baking the glassware at 400eC for 4 hours is quite adequate.
Storage of glassware in the laboratory introduces contamination even if the
glassware is wrapped in aluminum foil (Table 21). Furthermore, washing of
continuous liquid-liquid extractors and Soxhlet extractors with detergent,
thorough rinsing with hot tap water, followed by deionized water, and acetone,
is not adequate. Even after the Soxhlet extractor was refluxed with acetone
for 3 days, and the solvent changed daily, the levels of DEHP were as high as
500 ng.
6.1.5 Sample Vials
Storage of hexane and isooctane in 1.8 ml- and 15-mL glass vials capped
with metal foil liners or Teflon-lined rubber liners resulted in some
contamination; however, only five contaminants were detected, and their levels
ranged from 20 to 50 ng/mL of uncondensed solvent (Tables 22 and 23).
6.2 EVALUATION OF GAS CHROMATOGRAPHY
The following fused-silica columns were chosen for evaluation: a 30 m
x 0.25 mm ID DB-5 fused-silica capillary column (0.25 /xm film thickness), a
30 m x 0.53 mm ID Supelcowax-10 fused-silica open tubular column (1.0 im film
thickness), and a 30 m x 0.53 mm ID DB-210 fused-silica open tubular column
(1.0 /im film thickness); a 30 m x 0.53 mm ID DB-5 fused-silica open tubular
column (1.5 fun film thickness), a 30 m x 0.53 mm ID RL-5, fused-silica open
tubular column (0.5 urn film thickness), a 30 m x 0.5§ mm ID DB-608 fused-
silica open tubular column (0.83 urn film thickness), and a 30 m x 0.53 mm ID
45
-------
TABLE 20. TYPICAL LEVELS (IN NANOGRAMS PER PIECE OF GLASSWARE) OF PHTHALATE
ESTERS CONTAMINATION REMAINING ON LABORATORY GLASSWARE THAT WAS
SUBJECTED TO COMMON LABORATORY CLEANUP PROCEDURES BEFORE AND AFTER
BAKING AT 400'C TEMPERATURES FOR 4 HRSa
Precleaned amber bottle
Precleaned amber bottle
spiked with 5,000 ng of each
compound and heated for
4 hours at 400°C
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
First
<5.0
37
7.0
115
6.2
<5.0
<5.0
<5.0
<5.0
62
52
<5.0
105
<5.0
5.5
6.8
washing
<5.0
42
6.6
62
<5.0
<5.0
<5.0
<5.0
<5.0
50
30
<5.0
61
<5.0
<5.0
<5.0
Second
<5.0
127
11
26
<5.0
7.5
<5.0
<5.0
<5.0
19
<5.0
<5.0
29
<5.0
<5.0
<5.0
washing
<5.0
25
<5.0
36
<5.0
<5.0
<5.0
<5.0
<5.0
29
16
<5.0
46
6.1
<5.0
<5.0
First
<5.0
18
12
29
<5.0
<5.0
<5.0
<5.0
<5.0
32
13
5.3
41
6.8
<5.0
<5.0
washing
<5.0
41
13
37
<5.0
9.9
<5.0
<5.0
5.6
41
23
<5.0
58
6.6
<5.0
<5.0
Second
<5.0
19
<5.0
25
<5.0
8.8
<5.0
<5.0
<5.0
28
15
<5.0
35
6.5
<5.0
<5.0
washing
<5.0
56
9.9
56
5.5
5.4
12
8.9
11
37
29
<5.0
41
7.0
<5.0
<5.0
'Values given are in nanograms per piece of glassware and they represent
measurements from duplicate experiments (i.e., two different sets of
glassware treated the same).
46
-------
TABLE 21. TYPICAL LEVELS (IN NANOGRAMS PER PIECE OF GLASSWARE) OF PHTHALATE
ESTERS CONTAMINATION ON LABORATORY GLASSWARE SUBJECTED TO COMMON
LABORATORY CLEANUP PROCEDURES, BEFORE AND AFTER STORAGE IN AN
ANALYTICAL LABORATORY ENVIRONMENT
Compound
After storage
separatory funnel
(precleaned,
stored on shelf
1n laboratory)
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
OCP
OOP
DNP
61
28
17
47
5.9
17
<5.0
<5.0
<5.0
51
59
<5.0
140
<5.0
9.0
6.6
37
37
8.3
73
<5.0
29
7.9
5.5
<5.0
52
74
<5.0
100
<5.0
10
<5.0
After storage
continuous
liquid-liquid
extractor
(precleaned,
stored on shelf
in laboratory)
2100
47
75
173
15
54
22
7.0
15
190
230
12
490
9.3
32
<5.0
2100
36
16
155
5.9
3.5
9.3
6.4
<5.0
110
140
<5.0
110
<5.0
7.0
<5.0
Before storage
amber jar
(precleaned)
<5.0
12
15
11
<5.0
17
<5.0
<5.0
<5.0
50
32
<5.0
24
<5.0
<5.0
<5.0
<5.0
<5.0
12
11
<5.0
28
<5.0
<5.0
<5.0
36
11
<5.0
48
<5.0
<5.0
<5.0
After storage
amber jar
(precleaned,
stored on shelf
in laboratory)
<5.0
29
14
27
<5.0
16
<5.0
<5.0
<5.0
38
30
<5.0
67
<5.0
10
22
<5.0
20
7.3
17
<5.0
16
<5.0
<5.0
<5.0
29
9.0
<5.0
32
<5.0
<5.0
<5.0
Before storage
Soxhlet
extractor
(precleaned)
36
42
14
62
8.0
47
35
14
13
53
74
<5.0
77'
<5.0
18*
45'
55
24
23
67
43
64
63
20
22
94
220
<5.0
500*
<5.0
94*
170*
After storage
Soxhlet
extractor
(precleaned,
stored on shelf
in laboratory)
45
15
19
43
19
8.0
26
11
34
130
72
22
90'
45'
22'
8.9*
102
12
10
20
20
5.0
20
9.0
8.0
100
90
22
150'
56'
22'
8.9'
'Value given is approximate since it was estimated from the peak area.
-------
TABLE 22. TYPICAL LEVELS OF PHTHALATE ESTERS CONTAMINATION IN HEXANE
STORED IN SAMPLE VIALS FOR UP TO 61 DAYS AT 4°C
oo
Hexane
Hexane
Hexane
Hexane
Hexane blank
Hexane
Hexane
Hexane
Hexane
Hexane
Hexane
Hexane
Hexane blank
Hexane
Hexane
Hexane
Hexane
Hexane blank
Concentration (ng/mL of uncondensed solvent)
Solvent
Day
Vial
size
(mL)
Type
of
liner
DHP
DEP
DIBP DBP
BHPP BHEP DAP
BEEP
HEHP DHP
BBP
BBEP DEHP DCP
DOP DNP
1.8 metal
1.8 Teflon
15 metal
15 Teflon
12 1.8 metal
12 1.8 Teflon
12 15 metal
12 15 Teflon
20 1.8 metal
20 1.8 Teflon
20 15 metal
20 15 Teflon
61 1.8 metal
61 1.8 Teflon
61 15 metal
61 15 Teflon
31
31
34
38
13
20
19
19
19
37
19
28
22
27
17
16
14
15
16
16
15
16
11
15
14
20
12
29
15
19
17
15
75
39
38
36
42
38
33
36
33
34
29
29
27
25
33
28
30
23
38
15
16
17
14
16
<10a
•Data not available.
Hexane used was a Burdick & Jackson product, Lot No. AQ075.
-------
TABLE 23. TYPICAL LEVELS OF PHTHALATE ESTERS CONTAMINATION
STORED IN SAMPLE VIALS FOR UP TO 61 DAYS AT 4°C
IN ISOOCTANE
Sol vent
Day
Vial Type
size of
(ml) liner
Concentration (ng/mL of uncondensed solvent)
OHP DEP DIBP DBP BMPP BHEP DAP BEEP HEHP DHP BBP BBEP DEHP DCP DOP DNP
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
Isooctane
2
2
2
2
8
8
8
8
12
12
12
12
Isooctane blank 12
Isooctane
Isooctane
Isooctane
Isooctane
20
20
20
20
Isooctane blank 20
Isooctane 61
Isooctane 61
Isooctane 61
Isooctane 61
Isooctane blank
1.8 metal <50 <50
1.8 Teflon <50 <50
15 metal <50 <50
15 Teflon <50 <50
1.8 metal <50 <50
1.8 Teflon <50 <50
15 metal <50 <50
15 Teflon <50 <50
1.8 metal
1.8 Teflon
15
15
15
metal
Teflon
metal
1.8 metal
1.8 Teflon
15
15
15
metal
Teflon
metal
1.8 metal
1.8 Teflon
15 metal
15 Teflon
26
36
25
33
31
23
24
35
24
20
22
19
27
25
17
15
clO
17
44
30
30
34
30
39
21
58
16
14
16
17
16
24
22
29
35
35
82
13
12
13
13
13
17
17
35
33
36
36
36
22
18
24
24
24
26
26
36
28
25
14
14
15
15
15
'Data not available.
Isooctane used was a Fisher Scientific product, Lot No. 864429 pesticide grade.
-------
DB-1701 fused-silica open tubular column (1.0 /wn film thickness). The GC
operating conditions for the first three columns are given in Table 5. The
columns were connected to either an ECD or an FID. The other four columns
were tested in pairs using an injection tee to split the standard or sample
extract introduced into the injector to two different fused-silica open
tubular columns, each connected to an ECD. The single-column approach is
discussed in Section 6.2.1; the dual-column/dual-detector approach is
discussed in Section 6.2.2.
6.2.1 Single-Column Approach
The experimental work began with the evaluation of a DB-5 fused-silica
capillary column. Thirty-eight phthalate esters standards were analyzed
individually to establish their elution order when the DB-5 column was
programmed from 150°C to 275'C at 15eC/min (Tables 24 and 25), or 120°C to
250'C at 15'C/min, and then from 250°C to 300eC at 2eC/min (Table 24).
Some of the compounds (e.g., diisohexyl phthalate, diisooctyl phthalate,
diisononyl phthalate, diisodecyl phthalate, hexyl decyl phthalate, and butyl
decyl phthalate) were found to contain multiple peaks and were eliminated from
further consideration. GC/ECD chromatograms of a composite standard
containing 16 compounds and GC/ECD chromatograms of some of the phthalate
esters found to give multiple peaks are presented in Figures 6 through 9.
Other compounds that had identical retention times on the DB-5 fused-silica
capillary column were not included in the composite standard. For example,
dimethyl cyclohexyl phthalate and dioctyl phthalate and benzyl
2-ethylhexylphthalate had retention times of 12.12 min, 12.18 min, and
12.12 min, respectively, and didecyl phthalate and bis(2-ethylhexyl)phthalate
had retention times of 10.82 min and 10.85 min, respectively. Following this
preliminary work, the group of 16 compounds listed in Table 24 were selected
for further evaluation. The GC conditions were optimized to get the best
separation of the 16 compounds and the total analysis time not to exceed
30 min.
Retention times of the 16 compounds analyzed on the Supelcowax-10 fused-
silica open tubular column are given in Tables 26 and 27. GC/FID
chromatograms obtained during the optimization of the GC analysis are
presented in Figures 10 through 15. Attempts made to connect the
Supelcowax-10 column to an ECD were unsuccessful because of a high detector
background caused by column bleed.
Table 28 presents the results for a DB-210 fused-silica open tubular
column, and Figure 16 shows a GC/ECD chromatogram of a standard containing 11
phthalate esters which was analyzed on the DB-210 fused-silica open tubular
column. This column was found to be the least desirable because of a
significant drift in baseline during column programming. Since this column
was not acceptable for the 11 phthalate esters tested, additional phthalate
esters were not considered.
Our ECD was originally optimized at 285*C, and all experimental work for
the single-column approach was carried out at 285°C. In addition, we also
performed requirements in which the detector temperature was raised to 310°C
or 320°C. Quite often we had to bake the detector at higher temperatures in
50
-------
TABLE 24. RETENTION TIMES (MIN) OF METHOD 8060 COMPOUNDS
ANALYZED ON A DB-5 FUSED-SILICA CAPILLARY COLUMN
Retention time (min)
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Individual
standards'
(10 ng/ML)
3
4
6
7
8
7
8
8
8
9
9
10
10
10
12
14
.01
.10
.34
.06
.02
.47
.40
.25
.69
.62
.70
.60
.85
.76
.18
.08
2.97
4.10
6.34
7.06
8.39
9.63
9.69
10.85
10.75
12.19
14.08
Composite
standard*
(10 ng//iL)
2.97
4.13
6.36
7.08
8.41
9.64
9.71
10.86
10.77
12.20
14.09
3.01
4.09
6.34
7.05
8.39
9.63
9.70
10.85
10.76
12.19
14.08
Composite Composite
standard" standard0
(10 ng//iL) (10 ng//iL)
3.48
4.50
6.52
7.19
8.46
9.68
9.75
11.13
11.01
12.84
15.14
to
to
to
to
to
to
to
to
to
to
to
3.49
4.51
6.53
7.20
8.48
9.70
9.77
11.15
11.02
12.85
15.16
4
5
7
8
9
10
10
12
12
14
17
.22
.37
.50
.18
.47
.81
.89
.57
.39
.69
.56
aGC operating conditions are given in Table 5; the temper
is 150'C to 275°C at 15"C/min; injector temperature 275'
the temperature program
_ __. . . _ . ... ., . ........rature 275'C; detector
temperature 285°C. Duplicate determinations.
bVa1ues given represent the range of retention times for five
consecutive determinations performed with an autosampler. GC
conditions are given in Table 5; the temperature program is 1208C to
260°C at 15°C/min; then to 280°C at 2'C/min; injector temperature
275eC; detector temperature 310'C.
CGC conditions are given in Table 5; the temperature program is 120*C
to 250°C at 15°C/min; then 250*C to 300eC at 2'C/min; injector
temperature 2758C; detector temperature 310°C.
51
-------
TABLE 25. RETENTION TIMES OF THE ADDITIONAL
PHTHALATE ESTER STANDARDS ANALYZED
ON A DB-5 FUSED-SILICA CAPILLARY
COLUMN
Compound
Retention time3 (min)
DIHP )
DIOP >
DINP )
DIDP I
HDP /
BEHHP
DMCP
BEHP
BEEEP
BCP
BDP
BIDP
BOP
DCAP
DUP
EHIDP
I DTP
OOP
OIDP
IHBP
DMIP
DOIP
See Figure 7
See Figure 8
10.70
12.12
11.20 and 12.12
11.16
9.00
See Figure 9
b
9.78 (small peaks at 7.22 and 12.95)
11.18
b
b
9.05 to 9.74 (multiple peaks)
b
b
9.08 and 10.83
b
b
aFor GC operating conditions, refer to Table 5;
the temperature program is 150'C to 275°C at
15"C/min; detector temperature 285"C.
bNo peak was observed at 10 ppm concentration.
52
-------
11 14
r>j
^
IT"
r-- H
ll UOil
-j
o
10
T-tf
\ so
iri
13
15
16
-•1
r-
•
*r
r •!
f
Figure 6. GC/ECD chromatogram of composite phthalate esters standard
analyzed on a 30 m x 0.25 mm ID DB-5 fused-silica capillary
column. For GC operating conditions refer to Table 24,
footnote b. Peak assignments are given 1n Table 24.
53
-------
10
-------
•£>
rl
Figure 8. GC/ECD chromatograms of diisodecyl phthalate (top) and hexyl
decyl phthalate (bottom) analyzed on a 30 m x 0.25 mm ID DB-5
fused-siUca capillary column. For GC operating conditions, refer
to Table 5; the temperature program 1s 150°C to 275°C at 15°C/min;
detector temperature 285°C.
55
-------
r--
iVi
LU
OJ
Tl
CO
in
CO
Figure 9. 6C/ECD chromatogram of butyl decyl phthalate analyzed on a 30 m
x 0.25 mm ID DB-5 fused-silica capillary column. For GC operating
conditions, refer to Table 5; the temperature program is 150°C to
275°C at 15eC/min; detector temperature 285°C.
56
-------
TABLE 26. RETENTION TIMES (MIN) OF METHOD 8060
COMPOUNDS ANALYZED ON A SUPELCOWAX-10
FUSED-SILICA OPEN TUBULAR COLUMN
Retention time (min)*
Compound
no.
1
2
3
4
7
10
11
12
14
15
16
Compound
DMP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
OOP
DNP
Individual
standards
(50 ng//zL)
10.04
10.72
12.25
13.56
15.02
16.51
20.61
17.36
20.80
21.40
25.97
Composite
standard
(50 ng/ML)
10.04
10.73
12.27
13.55
15.04
16.54
20.68
17.36
20.87
21.44
26.00
10.03
10.72
12.26
13.55
15.04
16.51
20.69
17.34
20.87
21.46
26.05
aGC operating conditions are given in Table 5.
Duplicate determinations.
57
-------
TABLE 27. RETENTION TIMES OF 16 PHTHALATE ESTER STANDARDS ANALYZED
ON A SUPELCOWAX-10 FUSED-SILICA OPEN TUBULAR COLUMN
Retention time (min)
Compound
No.
1
2
3
4
5
7
8
9
10
11
12
13
14
15
16
Compound
DMP
DEP
DIBP
DBP
BMPP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Figure 11*
10.02
10.72
12.26
13.54
13.28
15.04
16.50
15.70
16.50
20.74
20.74
17.36
20.74
22.63
25.88
Figure 12* Figure 13a Figure 14" Figure 15"
7.33
7.81
9.17
10.67
10.29
12.85
15.62
14.04
15.37
c
c
16.71
c
20.86
24.70
11.24
12.24
14.46
16.26
15.89
18.43
20.66
19.38
20.47
c
c
21.42
c
24.19
26.48
5.67
6.17
7.33
8.51
8.21
10.24
12.51
11.22
12.31
16.47
17.05
13.41
16.77
18.15
20.86
b
6.11
7.26
8.43
8.14
10.15
12.41
11.13
12.21
16.36
16.94
13.31
16.66
18.28
20.73
aThe GC operating conditions are listed in Figures 11 through 15.
Compound not added to the composite standard.
Compounds were resolved; however, the integrator did not give retention time.
58
-------
347
Pfl
Figure 10. 6C/FID chromatogram of composite phthalate esters standard
analyzed on a 30 m x 0.53 mm ID Supelcowax-10 fused-silica
open tubular column. For 6C operating conditions, refer to
Table 5. Peak assignments are given in Table 26.
59
-------
Figure 11. GC/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular column.
Temperature program: 100°C (hold 2 min) to 240"C (hold 19 mln)
at 10°C/m1n; helium at 33 ps1.
60
-------
12
o Figure 12.
GC/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular column.
Temperature program: 100"C (hold 2 min) to 200°C at 25°C/min,
then to 260'C (hold 13 min) at 4°C/min; helium at 20 psi.
61
-------
Figure 13. 6C/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular column.
Temperature program: 120°C (hold 3 min) to 270°C (hold 10 min)
at 7'C/min; helium at 25 psi.
62
-------
f
H
r-.j
M.
i LI
10
12
Figure 14. 6C/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular column.
Temperature program: 150°C (hold 2 min) to 220°C at 15°C/min,
then to 260°C (hold 15 min) at 4°C/min; helium at 33 psi.
63
-------
r--
in
CO
©
Figure 15. GC/FID chromatogram of composite phthalate esters standard
analyzed on a Supelcowax-10 fused-silica open tubular column.
Temperature program: 150°C (hold 2 min) to 220°C at l5°C/m1n,
then to 260°C (hold 16 min) at 4°C/min; helium at 20 psi.
64
-------
TABLE 28. RETENTION TIMES OF THE PHTHALATE ESTER
STANDARDS ANALYZED ON A DB-210 FUSED-
SILICA OPEN TUBULAR COLUMN
Compound
No.
1
2
3
4
7
10
11
13
14
15
16
Compound Retention time" (min)
DMP
DEP
DIBP
DBP
DAP
DHP
BBP
DEHP
DCP
DOP
DNP
8.38
9.53
13.29
14.66
17.22
19.64
20.63
21.56
22.41
24.09
26.62
a
'The GC operating conditions are given in
Table 5. A GC/ECD chromatogram of the
composite phthalate esters standard is
given in Figure 16.
65
-------
Figure 16. GC/ECD chromatogram of composite phthalate esters standard
analyzed on a DB-210 fused-silica open tubular column. The GC
operating conditions are given in Table 5. For peak assignments
refer to Table 28. Concentration 1s 25 ppm in hexane.
66
-------
TABLE 29. RATIO OF EC DETECTOR ABSOLUTE RESPONSE AT 350'C AND 320'C
Compound
Absolute response (percent RSD)
320°Ca
350'Cb
Ratio0
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB (IS)
DPTP (SU)
76,450
55,630
86,605
75,230
172,490
75,860
79,210
86,820
111,270
63,040
894,010
75,930
76,400
177,600
48,670
36,120
85,780
124,270
(3.1)
(1.1)
(1.7)
(1.7)
(3.0)
(10.8)
(6.3)
(2.8)
(5.5)
(8.1)
(1.7)
(8.0)
(5.3)
(6.8)
(9.3)
(12.5)
(0.9)
(10.8)
82,260
71,370
122,950
100,800
330,810
111,180
109,890
122,630
234,660
79,230
1,029,260
98,100
102,940
351,900
58,750
47,130
119,540
83,730
(2.7)
(2.7)
(8.2)
(6.9)
(6.2)
(4.8)
(4.1)
(2.8)
(5.1)
(6.7)
(2.0)
(10.9)
(8.5)
(8.9)
(16.1)
(22.6)
(1.2)
(14.2)
1.08
1.28
1.42
1.34
1.92
1.47
1.39
1.41
2.11
1.26
1.15
1.29
1.35
1.98
1.21
1.30
1.39
0.67
aThe number of determinations was 4.
bThe number of determinations was 3.
°Ratio of absolute response at 350eC to absolute
response at 320°C at range 1 and attenuation 32. The
GC operating conditions are given in Table 5 under
the DB-5 fused-silica capillary column; the temperature
program is 120°C to 260*C (hold 15.7 min) at 15'C/min;
injector temperature 275*C.
67
-------
order to ensure satisfactory performance. An attempt made to test the
detector at 350°C gave interesting results. Contrary to literature reports
(3,4), we found that when the ECD is operated at 350°C, the detector responses
are between 1.08 and 2.11 times as high as at 320°C, except for diphenyl
terephthalate, for which the response decreased (Table 29). However, because
the DB-1701 fused-silica open tubular column could not withstand temperatures
above 320°C, all experimental work for the dual-column approach was performed
at 320°C (Table 6).
6.2.2 Dual-Column/Dual-Detector Approach
The dual-column/dual-detector approach makes use of the split injection
technique in which two fused-silica open tubular columns bonded with
dissimilar stationary phases are connected to an injection tee and similar
detectors. Four 30 m x 0.53 mm ID columns, DB-5, RT -5, DB-608, and DB-1701,
of different film thickness (Table 6) were paired as shown in Tables 30
through 33. The best separation of the 16 phthalate esters and three of the
surrogates proposed for incorporation in the revised Method 8060 (Method 8061)
seems to be achieved with the DB-5/DB-1701 column pair. Retention times and
relative retentions of the 16 compounds and 3 surrogates are presented in
Tables 30 through 33. GC/ECD chromatograms of a composite phthalate esters
standard are presented in Figures 17 through 20. In addition, the
reproducibility (percent relative standard deviation) of ECD response using
a Supelco 8-in injection tee or a J&W Scientific glass "Y-shape" press fit
inlet splitter was determined and is presented in Tables 34, 35, and 36.
Method linearity was determined only for the DB-5/DB-1701 column pair and is
presented in Table 37. Benzyl benzoate seems to work well as internal
standard. The GC method reproducibility was better than 10.9 percent for 16
consecutive injections of calibration standards, method blanks, and actual
sample extracts (Table 38).
6.3 SAMPLE EXTRACTION
Several extraction techniques were evaluated for extracting phthalate
esters from aqueous and solid matrices. These include the separatory funnel
technique (Method 3510) and the continuous liquid-liquid extraction technique
(Method 3520) for aqueous samples, and Soxhlet extraction (Method 3540) and
sonication (Method 3550) for solid matrices. In addition, two nonconventional
techniques, one using a Mixxor device and the other using the preconcentration
of the phthalate esters onto a Cia-membrane disk followed by elution with
acetonitrile were evaluated. Tables 39, 40, and 41 present the results
obtained for 12 phthalate esters using the separatory funnel technique, the
continuous liquid-liquid extraction technique, and the Mixxor device,
respectively. The spiking levels were 50 ng/L for the experiments performed
with the separatory funnel and the continuous liquid-liquid extractor, and
1 mg/L for the Mixxor device. The extracting solvent was methylene chloride
for the separatory funnel and continuous liquid-liquid extraction, and hexane
for the Mixxor extractor. The extracts from the separatory funnel and
continuous liquid-liquid extractor were analyzed by both GC/ECD and GC/FID.
Recoveries were >70 percent for most compounds and reproducibilities were
better than 10 percent for two-thirds of the compounds when the separatory
funnel extraction was employed. The continuous liquid-liquid extraction
technique gave unacceptable reproduc^ilities for all compounds; the average
68
-------
TABLE 30. RETENTION TIMES AND RELATIVE RETENTION TIMES OF
PHTHALATE ESTERS ON THE OB-5/DB-608 COLUMN PAIR"
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
DB-5
Phthalate
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB(IS)
Surrogates:
DPTP
DPP
DBZP
tr (min)
6.49
8.64
13.67
15.43
17.84
16.15
19.29
18.50
20.06
23.56
23.81
26.53
28.20
27.79
32.48
37.59
11.92
32.27
28.37
33.19
RRT
0.545
0.725
1.146
1.294
1.496
1.355
1.618
1.552
1.682
1.976
1.996
2.225
2.365
2.331
2.724
3.152
1.000
2.707
2.380
2.784
DB-608
tr (min)
6.36
8.25
12.22
14.13
15.21
16.60
17.31
18.26
19.01
20.80
23.87
24.98
24.23
27.55
28.42
32.27
11.45
32.09
30.19
35.38
RRT
0.555
0.721
1.067
1.233
1.328
1.449
1.511
1.594
1.660
1.816
2.084
2.181
2.116
2.405
2.481
2.818
1.000
2.802
2.636
3.089
aThe GC conditions are given in Table 6.
69
-------
TABLE 31. RETENTION TIMES AND RELATIVE RETENTION TIMES OF
PHTHALATE ESTERS ON THE DB-608/DB-1701 COLUMN PAIR8
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
DB-608
Phthalate
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB(IS)
Surrogates:
DPTP
DPP
DBZP
DOIP
DPIP
tr (min)
6.72
8.69
12.74
14.68
15.76
17.24
17.94
18.93
19.70
21.50
24.64
25.71
24.94
28.33
29.14
32.97
12.13
33.99
31.97
37.42
31.13
33.79
RRT
0.554
0.716
1.050
1.210
1.299
1.421
1.479
1.561
1.624
1.772
2.031
2.120
2.056
2.336
2.402
2.718
1.000
2.802
2.636
3.085
2.566
2.786
DB-1701
tr (min)
6.73
8.85
13.36
15.13
16.73
16.96
18.64
18.80
19.56
22.48
23.76
25.96
26.35
27.06
30.57
34.71
11.50
32.85
29.56
34.34
33.26
32.86
RRT
0.585
0.770
1.162
1.316
1.455
1.475
1.621
1.635
1.701
1.955
2.066
2.257
2.291
2.353
2.658
3.018
1.000
2.856
2.570
2.986
2.892
2.857
aThe GC conditions are given in Table 6.
70
-------
TABLE 32. RETENTION TIMES AND RELATIVE RETENTION TIMES OF
PHTHALATE ESTERS ON THE RT -5/DB-608 COLUMN PAIR8
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Phthalate
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB(1S)
Surrogates:
DPTP
DPP
DBZP
DOIP
DPIP
RTx-5
tr (min)
2.94
4.13
7.77
9.25
11.20
9.93
12.81
12.33
13.29
15.53
15.53
17.75
18.42
19.00
22.56
26.44
6.32
21.96
18.81
22.92
24.63
21.90
RRT
0.465
0.653
1.129
1.464
1.772
1.571
2.027
1.951
2.103
2.457
2.457
2.809
2.915
3.006
3.570
4.184
1.000
3.499
2.999
3.643
3.916
3.496
DB-608
tr (min)
6.33
8.48
12.49
14.40
15.50
16.91
17.30
18.59
19.37
21.16
23.45
25.37
24.27
27.98
28.81
32.70
11.86
32.85
30.94
36.27
30.04
32.78
RRT
0.534
0.715
1.053
1.214
1.307
1.426
1.459
1.567
1.633
1.784
1.977
2.139
2.046
2.359
2.429
2.757
1.000
2.770
2.609
3.058
2.533
2.764
'The GC conditions areas given in Table 6.
71
-------
TABLE 33. RETENTION TIMES AND RELATIVE RETENTION TIMES OF
PHTHALATE ESTERS ON THE DB-5/DB-1701 COLUMN PAIR0
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
DB-5
Phthalate
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB(IS)
Surrogates:
DPP(SU-l)
DPIP(SU-2)
DBZP(SU-3)
tr (min)
7.06
9.30
14.44
16.26
18.77
17.02
20.25
19.43
21.07
24.57
24.86
27.56
29.23
28.88
33.33
38.80
12.71
29.46
32.99
34.40
RRT
0.555
0.732
1.136
1.279
1.477
1.339
1.593
1.529
1.657
1.933
1.955
2.168
2.299
2.272
2.622
3.084
1.000
2.318
2.596
2.707
DB-1701
tr (min)
6.37
8.45
12.91
14.66
16.27
16.41
18.08
18.21
18.97
21.85
23.08
25.24
25.67
26.35
29.83
33.84
11.07
28.32
31.37
32.65
RRT
0.575
0.763
1.166
1.324
1.470
1.482
1.633
1.644
1.714
1.973
2.084
2.280
2.318
2.380
2.694
3.056
1.000
2.558
2.834
2.949
'The GC conditions are given in Table 6.
72
-------
DB-608
30 m x 0.53 mm ID
0.83- urn Film
O
ID
DB-5
30 m x 0.53 mm ID
1.5- jim Film
O
LLJ
I
10
20
TIME (min)
30
40
Figure 17. GC/ECD chromatograms of the phthalate esters standard analyzed
on the DB-608/DB-5 column pair. The GC operating conditions are
given in Table 6. For peak assignments, refer to Table 30.
73
-------
O
u.
i A '
•'-IU-
7
MM
npoc
I v
I ttHf
-a
DB-608
30 m x 0.53 mm ID
0.83-pm Film
DB-1701
30 m x 0.53 mm ID
1.0-jimFilm
V
-M
JU
1
(11) 0?)
u
10
20
TIME (min)
30
40
Figure 18. GC/ECD chromatograms of the phthalate esters standard analyzed
on the DB-608/DB-1701 column pair. The GC operating conditions
are given in Table 6. For peak assignments refer to Table 31.
74
-------
DB-608
30 m x 0.53 mm ID
0.83-iim Rim
RTx-5
30 m x 0.53 mm ID
0.5-um Film
COC2J (IS)
O
LLJ
i
J
vj
L
10
20
TIME (min)
30
40
Figure 19. 6C/ECD chromatograms of the phthalate esters standard analyzed
on the DB-eoS/R'T-S column pair. The GC operating conditions are
given in Table o. For peak assignments refer to Table 32.
75
-------
DB-5
30 mx 0.53 mm ID
1.5-^m Film
IS
if"* "i r *- -
11 12 SU-1 SU-2 SU-3
6 8
5
16
O
HI
1 IS
Ju
SU-2 SU-3
12 SU-1 15 T T 16
113
DB-1701
30 mx 0.53 mm ID
1.0-|imFilm
10
11
k
14
JJ
10
20
30
(min)
40
Figure 20. GC/ECD chromatograms of the phthalate esters standard analyzed on
the DB-5/DB-1701 column pair. The GC operating conditions are
given In Table 6. For peak assignments refer to Table 33.
76
-------
TABLE 34. REPRODUCIBILITY OF THE DETECTOR RESPONSE
USING THE DB-5/DB-608 COLUMN SETUP (J&W
SCIENTIFIC INLET SPLITTER)
Percent RSDa
DB-5 DB-608
Phthalate (0.83-^m film) (0.83-/an film)
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB (IS)
3.1
2.5
3.8
1.9
1.6
2.2
1.8
1.9
1.2
1.6
2.2
2.3
2.4
1.5
2.9
3.0
4.8
7.6
10.5
10.0
10.3
8.6
8.7
8.7
12.1
8.8
9.3
10.3
11.1
11.0
14.2
13.6
16.8
10.1
aThe number of determinations was 7. The GC
operating conditions are given in Table 6.
77
-------
TABLE 35. REPRODUCIBILITY OF THE DETECTOR RESPONSE
USING THE DB-608/DB-1701 COLUMN SETUP
(8-IN INJECTION TEE FROM SUPELCO)
Phthalate
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB (IS)
Percent
DB-608
(0.83-/an film)
1.1
1.2
1.4
1.4
7.2
1.4
1.3
6.9
17.1
5.5
0.9
5.5
0.9
1.3
1.9
2.8
1.0
RSDa
DB-1701
(1.0-jfln film)
1.1
1.2
1.2
1.0
1.0
0.9
1.3
1.0
b
1.4
1.2
1.1
1.4
1.1
0.9
1.0
1.4
a The number of determinations was 7. The GC
operating conditions are given in Table 6.
b Data not available.
78
-------
TABLE 36. REPRODUCIBILITY OF THE DETECTOR RESPONSE
USING THE DB-5/DB-1701 COLUMN SETUP
(J&W SCIENTIFIC INLET SPLITTER)
Phthalate
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB (IS)
Percent
DB-5
(1.5-/un film)
5.1
4.6
5.8
2.7
2.0
3.3
3.9
1.5
9.8
3.1
2.9
3.1
2.2
7.1
3.5
1.8
1.8
RSDa
DB-1701
(1.0-/an film)
1.7
2.2
3.7
2.0
3.0
1.9
1.7
1.8
b
2.2
2.8
1.6
2.1
4.2
2.1
3.5
3.5
a The number of determinations was 8. The GC
operating conditions are given in Table 6.
b Data not available.
79
-------
TABLE 37. METHOD LINEARITY USING THE DB-5/DB-1701
SETUP
Correlation Coefficient"
Phthalate
DB-5 DB-1701
5-/an film) (1.0-/zm film)
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
0.99284
0.99246
0.97853
0.99834
0.99806
0.99932
0.98967
0.99595
0.99281
0.99736
0.99783
0.99493
0.99877
0.99885
0.89106
0.93325
0.99960
0.99855
0.99746
0.99890
0.99900
0.99900
0.99832
0.99885
0.98062
0.98142
0.99555
0.99897
0.98771
0.99811
0.99796
0.99563
The concentration range was 20 to 80 ng//iL.
The number of data points was 10. The
phthalate esters could not be detected at
1 ng//iL. The 5-ng//ig and the 100-ng//*L
concentrations did not fit the linear range.
The GC operating conditions are given in
Table 6.
80
-------
TABLE 38. GC METHOD EVALUATION—INTERNAL STANDARD SUMMARY
Sample
identification"
Absolute area
DB-5
DB-1701
Calibration Standard
8113003-19b
8113003-1
8113003-2
8113003-3
8113003-7
8113003-8
8113003-9
8113003-20b
8113003-4
8113003-5
8113003-6
8113003-10
8113003-11
8113003-12
Calibration Standard
Average
Standard deviation
Percent RSD
70,835
51,025
56,274
53,898
56,027
57,869
56,401
60,154
55,121
56,961
59,041
60,010
55,184
54,724
54,672
76.037
58,390
6,385
10.9
73,745
65,399
74,859
71,789
74,392
74,346
69,100
74,342
71,674
75,143
79,203
78,749
69,627
68,981
70,921
76.105
73,023
3,682
5.0
aThe samples listed here are the acetonitrile extracts
of the HPLC-grade water samples and the groundwater
samples spiked with the 16 phthalate esters at
25 /jg/L and 100 jtg/L and extracted using the C.8-
membrane disk method. The GC operating conditions
are given in Table 6 for the DB-5/DB-1701.
"Sample 8113002-19 is an HPLC-water blank; Sample
8113003-20 is a groundwater blank.
81
-------
TABLE 39. AVERAGE RECOVERIES OF METHOD 8060 COMPOUNDS USING SEPARATORY
FUNNEL EXTRACTION (METHOD 3510)"
Compound
GC/ECDb
average recovery
(percent RSD)
GC/FIDC
average recovery
(percent RSD)
DMP
DEP
DIBP
DBP
BMPP
DAP
HEHP
DHP
BBP
DEHP
OOP
DNP
67.6 (18)
73.0 (3.2)
75.0 (2.7)
70.6 (4.5)
58.8 (8.2)
63.0 (3.5)
76.8 (7.4)
60.0 (12)
69.0 (3.1)
63.8 (8.0)
56.6 (24)
54.0 (15)
80.8 (6.6)
80.6 (6.3)
79.7 (5.6)
79.2 (5.5)
71.5 (10)
75.7 (5.6)
77.9 (5.6)
64.9 (17)
72.4 (7.8)
64.0 (17)
59.8 (20)
56.1 (25)
aThe spiking level was 50 /ig/L; the number of
determinations was 5. HPLC-grade water (Fisher
Scientific Lot No. 873266) was used for this
experiment. Extraction was performed with
methylene chloride (American Burdick & Jackson
Lot No. AQ 352).
bThe GC/ECD operating conditions were as
follows: 30 m x 0.25 mm ID DB-5 fused-silica
capillary column (0.25 /«n film thickness); 120°C
to 260°C (hold 15.7 min) at 15eC/min; carrier
gas: helium at 21 psi; injector temperature
275'C; detector temperature 320°C; spitless
injection.
cThe GC/FID operating conditions were as
follows: 30 m x 0.53 mm ID Supelcowax 10
fused-silica open tubular column (1 /*m film
thickness); 150°C (hold 2 min) to 220°C at
15eC/min then to 260°C (hold 13.5 min) at
4'C/min; carrier gas: helium at 20 psi;
injector and detector temperature: 270°C;
spitless injection.
82
-------
TABLE 40. AVERAGE RECOVERIES OF METHOD 8060 COMPOUNDS USING CONTINUOUS
LIQUID-LIQUID EXTRACTION (METHOD 3520)a
Compound
GC/ECDb
average recovery
(percent RSD)
GC/FIDC
average recovery
(percent RSD)
DMP
DEP
DIBP
DBP
BMPP
DAP
HEHP
DHP
BBP
DEHP
OOP
DNP
67.5 (7.4)
72.8 (15)
71.0 (17)
69.7 (17)
51.3 (18)
62.5 (20)
78.0 (22)
44.8 (20)
68.0 (5.6)
62.7 (29)
32.7 (52)
34.2 (41)
89.7 (9.3)
87.6 (11)
81.4 (12)
79.2 (14)
57.1 (17)
68.5 (12)
71.7 (14)
37.9 (31)
15.7 (78)
24.9 (55)
21.1 (69)
20.3 (53)
aThe spiking level was 50 ng/i; the number of
determinations was 6 for GC/ECD and 3 for
GC/FID. HPLC-grade water (Fisher Scientific
Lot No. 873266) was used for this experiment.
Extraction was performed with methylene
chloride (American Burdick & Jackson Lot
No. AQ 352).
bThe GC/ECD operating conditions were as
follows: 30 m x 0.25 mm ID DB-5 fused-silica
capillary column (0.25 tun film thickness);
120°C to 260eC (hold 15.7 min) at 15eC/min;
carrier gas: helium at 21 psi; injector
temperature 275°C; detector temperature
320eC; spitless injection.
°The GC/FID operating conditions were as
follows: 30 m x 0.53 mm ID Supelcowax 10
fused-silica open tubular column (1 /wn film
thickness); 150'C (hold 2 min) to 220'C at
15eC/min then to 260'C (hold 13.5 min) at
4"C/min; carrier gas: helium at 20 psi;
injector and detector temperature: 270*C;
spitless injection.
83
-------
TABLE 41. AVERAGE RECOVERIES OF METHOD 8060
COMPOUNDS USING MIXXOR EXTRACTOR
AND HEXANE
Average recovery8'"
Compound (percent RSD)
DMP
DEP
DIBP
DBP
BMPP
DAP
HEHP
DHP
BBP
DEHP
OOP
DNP
40 (13)
67 (12)
75 (13)
66 (17)
37 (32)
52 (19)
57 (17)
43 (29)
65 (11)
52 (25)
36 (14)
24 (77)
aThe spiking level was 1 mg/L;
the number of determinations
was 5. HPLC-grade water
(Fisher Scientific Lot
No. 873266) was used for this
experiment. Vextrac{ was 1 mL.
Each extract was diluted
500-fold prior to GC/ECD
analysis.
bThe GC/ECD operating
conditions were as follows:
30 m x 0.25 mm ID DB-5
fused-silica capillary column
(0.25 pm film thickness);
120°C to 260*C (hold 15.7 min)
at 15'C/min; carrier gas:
helium at 21 psi; injector
temperature 275"C; detector
temperature 320°C; spitless
injection.
84
-------
recoveries for five of the 12 phthalate esters were 20 to 45 percent.
Extraction with hexane in the Mixxor device also gave unacceptable recoveries
and reproducibilities. Therefore, the separatory funnel extraction method was
further evaluated on samples containing 16 phthalate esters at three
concentrations: 1 /zg/L, 10 M9/L, and 25 M9/L, each in quadruplicate. The
results are presented in Tables 42, 43, and 44. Diphenyl terephthalate was
used as a surrogate; its average recovery in the 12 samples was 98.2 percent,
and the relative standard deviation was 19.2 percent (Table 45). These
results indicate that the separatory funnel extraction method is more
desirable than the continuous liquid-liquid extraction and the Mixxor device
extraction methods.
Three other surrogate compounds were tested for use with Method 3510.
Their average recovery and relative standard deviation are also presented in
Table 45. Excellent recoveries and reproducibilities were achieved for each
surrogate compound.
Tables 46 through 48 present the results of the extraction procedure using
the 3M-Empore membrane disks containing either the C8-silica (Tables 46 and
47) or the C18-silica (Tables 48 and 49). The shorter-chain esters and the
alkoxy-substituted esters had recoveries greater than 65 percent when the
Cg-membrane disks were used for sample filtration. The longer-chain esters
(>C5 alky!) exhibited recoveries <40 percent, and there seems to be a trend
in decreasing recoveries with the increasing number of carbons in the alkyl
chain. In the case of relatively clean samples, the membrane disks can be
reused, provided that they are cleaned and reconditioned after each sample.
The disks were used four consecutive times with HPLC-grade water spiked with
16 target phthalate esters at 24 ng/L. The recovery data for each extraction
are presented in Tables 47 for the C8-membrane disks and 48 for the C18-
membrane disks. Since the C18-membrane disks were found to give higher
recoveries for the longer-chain esters, they were further evaluated with both
HPLC-grade water and groundwater at three concentrations, each in triplicate.
Table 49 presents the data for the 25-/KJ/L and 100-/KJ/L spike level. The
target phthalate esters could not be detected of the 5-/KJ/L spike level. As
expected, method accuracy and precision seem to be a function of concentration
(Table 49). The surrogate recoveries for the samples that were
preconcentrated on the membrane disks and then extracted with acetonitrile are
presented in Table 50.
Extraction of soil samples by sonication with methylene chloride/acetone
(1:1) and Soxhlet extraction with hexane/acetone (1:1) was evaluated using a
sandy loam soil. Tables 51 and 52 summarize the data. In the case of
Method 3550, we prepared two batches of spiked sandy loam soil, and each batch
was split into 8 or 9 portions of material. The average recoveries ranged
from 31.9 to 112 percent for Batch 1 and 32.1 to 108 percent for Batch 2.
The percent relative standard deviations ranged from 4.2 to 24.6 percent for
Batch 1 and 8.1 to 25 percent for Batch 2 (Table 51). The average recoveries
for Method 3540 were slightly higher, ranging from 53.5 to 135 percent;
however, the percent relative standard deviations were much higher than in the
case of Method 3550 ranging from 19.8 to 46.9 percent. Method 3550 was
subsequently evaluated with other solid matrices including an estuarine
sediment and a municipal sludge. The results of these method evaluation
85
-------
TABLE 42. RESULTS OF METHODS 3510 AND 8060 PERFORMANCE (CONCENTRATION l)a
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Spike
level
(MA)
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
Concentration (/ig/L)
Rep.l
1.2
0.85
0.97
0.81
0.84
0.69
0.67
0.61
0.81
0.85
1.2
0.55
1.0
0.75
0.88
0.74
Rep. 2
1.4
0.99
1.0
0.93
0.88
0.73
0.68
0.74
0.74
0.93
1.7
0.65
1.2
0.82
0.95
0.79
Rep. 3
1.3
0.91
0.99
0.86
0.87
0.38
0.66
0.38
0.64
0.89
1.8
0.28
1.3
0.74
0.91
0.76
Average ± SD
Rep. 4 (percent RSD)
1.4
0.91
1.0
0.85
0.87
0.92
0.69
0.74
0.74
0.92
1.4
0.65
1.0
0.79
0.93
0.77
1.3 ± 0.1
(7.3)
0.92 ± 0.06
(6.2)
0.97 ± 0.04
(4.2)
0.86 ± 0.05
(5.8)
0.87 ± 0.02
(2.0)
0.68 ± 0.22
(33)
0.68 ± 0.01
(1-9)
0.62 ± 0.17
(27)
0.73 ± 0.07
(9.6)
0.90 ± 0.04
(4.0)
1.5 ± 0.3
(18)
0.53 ± 0.17
(33)
1.1 ± 0.2
(14)
0.78 ± 0.04
(4.7)
0.92 ± 0.03
(3.2)
0.76 ± 0.02
(2.7)
Average
recovery
(percent RSD)
132 (7.3)
91.5 (6.2)
96.7 (4.2)
86.2 (5.8)
86.5 (2.0)
68.0 (33)
67.5 (1.9)
61.7 (27)
73.2 (9.6)
89.7 (4.0)
152 (18)
53.2 (33)
112 (14)
77.5 (4.7)
91.7 (3.2)
76.5 (2.7)
aEach water sample (1 L reagent water, Fisher Scientific Lot No. 873154)
was spiked with 100 /*L of a 10-ng//iL composite phthalate ester standard
(in hexane) and 100 /jL of 50 ng/jiL diphenyl terephthalate (in hexane).
vext was 10 mL after solvent exchange. The GC/ECD operating conditions
were as follows: 30 m x 0.25 mm ID DB-5 fused-silica capillary column
(0.25 /on film thickness); 120eC to 260°C (hold 15.7 min) at 15eC/min;
carrier gas: helium at 21 psi; injector temperature 275*C; detector
temperature 320°C; spitless injection.
86
-------
TABLE 43. RESULTS OF METHODS 3510 AND 8060 PERFORMANCE (CONCENTRATION 2)a
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Spike
level
(M/L)
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
Concentration (jtg/L)
Rep.l
7.0
9.4
10.4
10.3
10.1
10.7
9.0
9.7
9.8
10.1
12.3
8.4
10.0
9.2
10.2
8.3
Rep. 2
9.8
8.9
11.4
10.9
9.6
11.9
8.4
10.6
9.8
8.2
9.0
6.2
8.6
6.4
7.4
5.5
Rep. 3
10.4
9.4
12.5
11.9
10.8
12.9
9.6
11.7
11.3
9.9
10.9
7.7
9.0
8.1
10.1
7.8
Rep. 4
10.6
9.5
10.3
10.1
9.2
11.3
7.8
9.0
8.8
9.3
10.0
7.8
10.3
8.0
10.0
7.7
Average ± SD
(percent RSD)
9.5 ± 1.7
(18)
9.3 ± 0.3
(2.9)
11.2 ± 1.0
(9.2)
10.8 ± 0.8
(7.5)
9.9 ± 0.7
(7.0)
11.7 ± 0.9
(8.0)
8.7 ± 0.8
(8.9)
10.3 ± 1.2
(11)
9.9 ± 1.0
(10)
9.4 ± 0.8
(9.1)
10.6 ± 1.4
(13)
7.5 ± 0.9
(12)
9.5 ± 0.8
(8.5)
7.9 ± 1.2
(15)
9.4 ± 1.4
(14)
7.3 ± 1.2
(17)
Average
recovery
(percent RSD)
94.5 (18)
93.0 (2.9)
112 (9.2)
108 (7.5)
99 (7.0)
117 (8.0)
87.0 (8.9)
103 (11)
99.2 (10)
93.7 (9.1)
106 (13)
75.2 (12)
94.7 (8.5)
79.2 (15)
94.2 (14)
73.2 (17)
aEach water sample (1 L reagent water, Fisher Scientific Lot No. 873154)
was spiked with 200 /iL of a 50-ng/0L composite phthalate ester standard
(in hexane) and 100 /*L of 50 ng//iL diphenyl terephthalate (in hexane).
Extract was 1° mL after solvent exchange. The GC/ECD operating conditions
were as follows: 30 m x 0.25 mm ID DB-5 fused-silica capillary column
(0.25 urn film thickness); 120*C to 260°C (hold 15.7 min) at 15°C/min;
carrier gas; helium at 21 psi; injector temperature 275°C; detector
temperature 320°C; spitless injection.
87
-------
TABLE 44. RESULTS OF METHODS 3510 AND 8060 PERFORMANCE (CONCENTRATION 3)'
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Spike
level
C/tg/L)
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
Concentration (/*g/L)
Rep
17
23
26
32
28
35
28
33
28
28
32
28
27
25
30
24
.1
.3
.2
.7
.3
.5
.7
.7
.0
.2
.3
.7
.3
.6
.7
.2
.5
Rep.
24.
22.
25.
26.
25.
30.
23.
27.
24.
24.
29.
22.
24.
23.
25.
20.
2
9
5
4
3
9
4
2
8
9
1
6
4
4
6
9
6
Rep. 3
25.0
23.0
26.1
26.9
27.1
32.4
24.8
30.0
26.4
26.0
35.2
25.5
26.6
25.9
29.3
24.2
Rep
26
23
26
26
26
31
23
29
25
26
29
23
24
23
26
20
Average t SD
.4 (percent RSD)
.3
.9
.2
.4
.1
.8
.8
.2
.2
.6
.7
.7
.2
.2
.2
.9
23.4 ± 4.1
(18)
23.2 ± 0.6
(2.5)
26.1 ± 0.5
(2.1)
28.0 ± 2.9
(1.0)
26.9 ± 1.2
(4.4)
32.6 ± 2.2
(6.9)
25.1 ± 2.5
(1.0)
30.0 ± 2.2
(7.3)
26.2 ± 1.5
(5.7)
26.3 ± 1.7
(6.6)
31.8 ± 2.7
(8.4)
25.0 ± 2.6
(10)
25.7 ± 1.7
(6.5)
24.6 ± 1.4
(5.7)
27.9 ± 2.2
(7.8)
22.6 ± 2.1
(9.2)
Average
recovery
(percent RSD)
93
92
104
112
108
130
100
120
105
105
127
100
103
98
112
90
.6 (18)
.8 (2.5)
(2.1)
(1.0)
(4.4)
(6.9)
(1.0)
(7.3)
(5.7)
(6.6)
(8.4)
(10)
(6.5)
.4 (5.7)
(7.8)
.4 (9.2)
aEach water sample (1 L reagent water, Fisher Scientific Lot No. 873154)
was spiked with 500 pi of a 50-ng/^L composite phthalate ester standard
(in hexane) and 100 pL of 50 ng/pL diphenyl terephthalate (in hexane).
Extract was *° mL a^ter solvent exchange. The GC/ECD operating conditions
were as follows: 30 m x 0.25 mm ID DB-5 fused-silica capillary column
(0.25 (jm film thickness); 120*C to 260*C (hold 15.7 min) at 15'C/min;
carrier gas: helium at 21 psi; injector temperature 275°C; detector
temperature 320'C; spitless injection.
-------
TABLE 45. RECOVERY OF SURROGATE COMPOUNDS SPIKED INTO REAGENT HATER SAMPLES (METHOD 3510)
Compound
DPTP
DPP
DPIP
DBZP
Spike
level
(MA)
5
10
10
10
Percent recovery
Rep
81.
78.
79.
.1
8
3
0
Rep. 2
83.8
85.9
75.0
Rep. 3
98.3
90.2
90.2
Rep
100
94
96
.4
.1
.3
Rep. 5
96.2
93.1
94.3
Average
98
92
88
86
.2a
.0
.3
.9
RSD
(percent)
19.2s
9.3
7.3
10.9
'The number of determinations was 12.
00
-------
TABLE 46. EVALUATION OF C8-MENBRANE
DISKS FOR PRECONCENTRATION
OF PHTHALATE ESTERS
Phthalate
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Percent
Vol ume
(500 mL)
94.7
91.9
71.3
64.5
37.3
111
39.7
112
64.3
35.2
63.7
84.4
23.8
71.9
19.4
20.4
Recovery*
Vol ume
(1000 mL)
101
122
101
97.5
48.3
138
57.5
145
102
52.5
110
121
41.2
103
34.8
33.8
a500 mL or 1000 mL of HPLC-grade
water were spiked with the
target compounds at 24 /jg/L and
were passed through a pre-
conditioned 3M-Empore Cg-
membrane disk in approximately
26 min. The target compounds
were eluted with 10 mL of
acetonitrite. A second elution
with an additional 10 mL of
acetonitrile was performed. No
compounds were detected in the
second extract. The GC/ECD
opera-ting conditions are given
in Table 6 under the
DB-608/DB-1701 column pair.
Single determination.
90
-------
TABLE 47. EVALUATION OF C8-MEMBRANE DISKS FOR
PRECONCENTRATION OF PHTHALATE ESTERS
(MULTIPLE USE OF DISKS)
Percent Recovery*
Phthalate 1st 2nd 3rd 4th
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
92.5
85.8
69.4
68.5
57.6
119
57.2
131
97.5
56.6
90.8
97.5
50.7
90.8
47.3
45.3
116
106
86.7
92.5
99.2
123
85.0
136
115
84.2
116
113
83.3
104
78.0
79.2
103
98.3
84.2
92.5
115
120
57.4
137
112
56.2
102
123
51.2
96.7
45.4
47.9
105
91.7
78.1
85.0
136
120
67.8
131
108
65.3
104
109
62.0
92.5
56.1
60.5
a500 mL of HPLC-grade water were spiked with
the target compounds at 24 ug/L and were
passed through a preconditioned 3M-Empore C8-
membrane disk in approximately 26 min. The
target compounds were eluted with 10 mL of
acetonitrile. A second elution with an
additional 10 mL of acetonitrile was
performed. No compounds were detected in the
second extract. The membrane disk was
reconditioned with 10 mL of methanol followed
by 10 mL of water and reused three consecutive
times. The GC operating conditions are given
in Table 6 for the DB-608/DB-1701 column pair.
91
-------
TABLE 48. EVALUATION OF C1B-HEMBRANE DISKS
FOR PRECONCENTRATION OF PHTHALATE
ESTERS (MULTIPLE USE OF DISKS)
Percent Recovery8
Phthalate 1st 2nd 3rd 4th
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
75.5
73.8
76.2
78.7
70.5
94.2
67.1
97.6
82.7
73.6
84.4
85.4
73.5
82.4
75.1
84.4
56,3
64.9
65.1
65.8
63.7
68.0
63.3
82.0
72.6
62.9
80.9
84.6
70.0
83.0
75.9
84.5
58.9
55.1
53.1
56.3
49.3
67.2
46.2
75.2
63.4
46.7
62.9
60.8
51.9
60.8
56.3
71.3
63.6
66.5
63.3
63.3
57.9
76.7
55.3
92.4
65.7
57.2
73.0
69.9
61.2
67.5
55.2
55.9
a500 ml of HPLC-grade water were spiked with
the target compounds at 24 ug/L and were
passed through a preconditioned 3M-Empore
membrane disk in approximately 20 min. The
target compounds were eluted with 10 mL of
acetonitrile. A second elution with an
additional 10 mL of acetonitrile was per-
formed. No compounds were detected in the
second extract. The membrane disk was
reconditioned with 10 mL of methanol
followed by 10 mL of water and reused three
consecutive times. The GC/ECD operating
conditions are given in Table 6 under the
DB-5/DB-608 column pair.
92
-------
TABLE 49.
C18-MEMBRANE DISKS METHOD EVALUATION8
HPLC-Grade Water
Phthalate
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP'
DEHP
DCP
OOP
DNP
25 /zg/L
81.
76.
64.
69.
62.
83.
54.
90.
47.
66.
79.
57.
78.
47.
44.
38.
7
6
2
9
5
3
3
9
7
5
1
9
6
7
5
0
(5.
(1.
(16.
(17.
(16.
(3.
(34.
(9.
(40.
(26.
(17.
(37.
(13.
(23.
(39.
(52.
9)
6)
7)
2)
7)
1)
1)
0)
0)
3)
1)
2)
5)
7)
6)
4)
100
88.6
92.3
87.6
90.3
87.2
107
93.6
108
93.9
98.4
97.3
91.3
94.8
106
84.9
96.9
/*g/L
(17
(10
(16
(13
(9
(13
(21
(8
(22
(5
(2
(7
(6
(19
(3
(11
.7)
•3)
.2)
.2)
•5)
.6)
.0)
.9)
•4)
.0)
.6)
.4)
•3)
•9)
•8)
•1)
25 /
67.7
71.9
80.0
73.5
59.6
72.3
55.4
72.5
92.6
73.3
c
79.4
90.0
75.9
78.1
71.2
Groundwater
in 71 ^
75.6
69.3
57.6
63.1
58.9
69.5
54.0
99.8
24.0
54.5
c
50.0
72.7
41.4
38.8
27.4
100
86.6
92.6
89.3
95.0
86.7
113
78.9
102
83.4
97.7
66.0
96.3
98.7
108
90.1
95.2
M9/L
(14.3)
(7.2)
(1.6)
(1.5)
(4.9)
(2.8)
(5.8)
(4.0)
(8.8)
(14.8)
(39.3)
(7.9)
(6.0)
(13.3)
(6.1)
(12.7)
aThe number of determinations was 3. 500 mL of HPLC-grade water or
groundwater were spiked with the target compounds at the levels indicated
and were passed through a preconditioned 3M-Empore C18-membrane disk in
approximately 20 min. The target compounds were eluted with 10 mL
acetonitrile. The GC/ECD operating conditions are given in Table 6 under
the DB-5/DB-1701 column pair. Value given is the average recovery; number
in parentheses is the relative standard deviation.
bOne of the replicates was discarded because all of the recoveries were
biased low.
°Not able to calculate recovery because compound was present in the sample
at 28 iig/l.
93
-------
TABLE 50.
C18-MEMBRANE DISKS METHOD EVALUATION—SURROGATE RECOVERIES"
Percent recovery
Sample
identification
8113003-1
8113003-2
8113003-3
8113003-4
8113003-5
8113003-6
8113003-7
8113003-8
8113003-9
8113003-10
8113003-11
8113003-12
8113003-13
8113003-14
8113003-15
8113003-16
8113003-17
8113003-18
Diphenyl
Sample description phthalate
HPLC-grade water spiked
with phthalate esters
at 5 ng/l
Groundwater sample spiked
with phthalate esters
at 5 fig/I
HPLC-grade water spiked
with phthalate esters
at 25 pg/L
Groundwater sample spiked
with phthalate esters
at 25 ng/L
HPLC-grade water spiked
with phthalate esters
at 100 fig/I
Groundwater sample spiked
with phthalate esters
at 100 fig/I
98.4
97.6
96.8
103
102
115
63.2
96.8
106
99.2
81.6
47.2
79.3
78.9
93.8
81.7
81.9
83.3
Di phenyl
isophthalate
99.1
99.2
95.4
106
103
120
61.1
99.1
107
99.5
82.1
49.9
82.9
86.7
102
78.2
89.8
89.7
Di benzyl
phthalate
83.8
85.5
84.2
91.9
89.1
109
56.2
89.1
99.6
87.1
73.0
39.1
82.8
83.9
99.3
80.3
88.0
90.9
8113003-19
8113003-20
HPLC-grade water
method blank
Groundwater sample
method blank
Average
Standard deviation
Percent RSD
101
70.3
88.9
16.3
18.3
104
70.7
91.3
16.8
18.4
106
63.2
84.1
16.4
19.5
aThe spike levels of the surrogates were 25 ng/L for Samples -1 through -12,
-19, and -20, and 50 ng/l for Samples -13 through -18.
94
-------
TABLE 51. RESULTS OF METHODS 3550 AND 8060 PERFORMANCE
Batch la
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
DOP
DNP
Average
recovery
112
96.0
83.4
89.1
31.9
93.1
102
74.9
62.9
103
94.9
103
72.0
69.1
84.0
72.0
RSD
(percent)
17.6
8.2
12.4
13.3
24.6
9.3
9.9
11.1
8.4
4.2
9.1
21.3
12.3
10.6
7.7
9.4
Batch 2b
Average
recovery
79.4
73.1
65.1
73.1
32.1
61.7
108
68.0
58.3
87.4
85.7
74.9
98.3
63.4
61.7
65.7
RSD
(percent)
25.0
16.9
8.1
13.3
16.0
21.5
19.7
9.4
9.3
8.7
8.5
9.7
11.9
10.1
15.9
11.4
aThe number of determinations was 9. Compounds were spiked
at 1 ppm.
bThe number of determinations is 8. Compounds were spiked
at 1 ppm.
95
-------
TABLE 52. RESULTS OF METHODS 3540 AND 8060 PERFORMANCE
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Average
Recovery3
73.7
129
70.3
73.7
75.4
74.9
87.4
54.3
53.5
135
95.4
89.7
107
66.3
58.3
62.3
RSD
(percent)
19.8
28.0
23.3
27.7
34.5
31.8
26.2
31.6
25.8
37.2
25.5
22.4
46.9
24.0
22.1
22.4
aThe number of determinations was 8.
Compounds were spiked at 1 ppm.
96
-------
studies are presented in Section 6.6. Surrogate recoveries for Method 3550
are summarized in Table 53.
6.4 EXTRACT CLEANUP
The current Method 8060 recommends cleanup of sample extracts by either
alumina (Method 3610) or Florisil (Method 3620) chromatography. Both methods
use large volumes of solvents that may increase the contamination of sample
extracts because of impurities present in the eluting solvent. For example,
Method 3610 recommends 140 mL of 20 percent diethyl ether in hexane, and
Method 3620 recommends 100 ml of 20 percent diethyl ether in hexane. Neither
method indicates whether or not the phthalate esters are recovered
quantitatively.
We have evaluated both methods with standards in hexane containing
16 phthalate esters. The percent recoveries of the 16 compounds are presented
in Table 54.
Alumina cleanup is preferred over the Florisil cleanup since it allows
recovery of all target compounds by elution with 20 percent diethyl ether in
hexane. Using Florisil cleanup, three of the 16 target compounds could not
be recovered at all, and dimethyl and diethyl phthalates gave recoveries of
only 40 and 57 percent, respectively.
To improve the recoveries of the five phthalate esters mentioned above,
we have taken Florisil and alumina SPE cartridges of 0.5-g, 1.0-g,.and 2-g
size, charged them with our target compounds and interferents, and eluted
them with 10 percent acetone in hexane (for Florisil) and 20 percent acetone
in hexane (for alumina). We first attempted the elution of the phthalate
esters from the alumina cartridge with 20 percent diethyl ether in hexane.
Since none of the phthalate esters was recovered after 10 mi solvent passed
through the cartridge, we changed the eluting solvent to 10 percent acetone
in hexane and later to 20 percent acetone in hexane to improve the recovery
of bis(2-methoxyethyl) phthalate, bis(2-ethoxyethyl) phthalate and
bis(2-n-butoxyethyl) phthalate. The results of these experiments are
summarized in Tables 55, 56, and 57.
The data shown in Table 55 indicate that all but two phthalate esters can
be recovered from a 0.5-g or a 1.0-g Florisil cartridge with 5 mL 10-percent
acetone in hexane (Fraction 1) and from a 2.0-g cartridge with 10 mL
10-percent acetone in hexane (no phthalate esters were recovered in
Fraction 1, therefore an additional fraction had to be collected). The two
phthalate esters that could not be recovered are bis(2-methoxyethyl) phthalate
and bis(2-ethoxyethyl) phthalate. When working with the 0.5-g Florisil
cartridge, these two phthalate esters were recovered almost quantitatively by
eluting the cartridge with an additional 5 mL 10-percent acetone in hexane;
however, they could not be recovered from either the 1.0-g or the 2.0-g
Florisil cartridge under similar conditions. The alumina cartridge procedure
(Table 57) allowed recovery of all 16 phthalate esters except for one
compound, bis(2-methoxyethyl) phthalate, from the 2.0-g cartridge.
97
-------
TABLE 53. RECOVERY OF SURROGATE COMPOUNDS SPIKED INTO SOIL SAMPLES
(METHOD 3550)
Percent recovery
Spike
level RSD
Compound (ng/g) Rep.l Rep.2 Rep.3 Rep.4 Rep.5 Average (percent)
DPTP
DPP
DPIP
DBZP
330
330
330
330
90.2
91.5
113
79.1
83.9
85.0
90.5
97.1
93.9
110
111
105
86.9
87.9
81.4
95.4
91.3
94.3
95.7
12.8
12.5
11.2
13.9
98
-------
TABLE 54. PERCENT RECOVERIES OF METHOD 8060 COMPOUNDS USING ALUMINA
(METHOD 3610) AND FLORISIL (METHOD 3620) COLUMN CHROMATOGRAPHY
Percent Recovery3
Adsorbent
Alumina
Alumina
Florisil
Florlsil
DMP
63
66
43
37
DEP
63
62
58
56
OIBP
82
72
83
77
DBP
73
80
98
72
BMPP
91
88
88
81
BMEP
74
67
0
0
DAP
77
73
84
79
BEEP
67
67
0
0
HEHP
92
89
110
100
DHP
73
67
0
0
BBP
87
87
92
88
BBEP
62
63
0
0
DEHP
76
106
200
64
DCP
83
86
86
81
OOP
96
120
120
110
DNP
73
69
76
69
"Alumina and Florisil chromatography were done according to Methods 3610 and 3620, respectively.
to
-------
TABLE 55. ELUTION PATTERNS AND PERCENT RECOVERIES OF THE METHOD 8060
COMPOUNDS FROM THE FLORISIL CARTRIDGES USING 10 PERCENT ACETONE IN
HEXANE"
0.5-g cartridge
40-/*g spike
Compound Fraction 1
DMP
DEP
DIBP
DBF
BMPP
BMEP"
DAP
BEEP
HEHP
DHP
BBP
BBEP
OEHP
OCP
OOP
DNP
98.9
108
109
104
106
16.0
107
96.6
99.6
105
108
113
111
91.4
120
118
101
113
106
107
114
15.6
111
97.6
98.4
109
109
117
111
81.6
114
118
80-/ig spike
Fraction 1
81.2
91.3
92.8
87.4
98.2
11.9
94.8
74.7
75.6
91.0
84.1
98.5
92.1
62.9
102
103
80.8
93.1
91.7
88.5
95.9
14.8
91.5
69.3
75.9
87.6
82.3
94.3
87.6
61.7
93.2
92.2
120-/ig spike 40-0g spike
Fraction 1 Fraction 1
73.9
83.7
87.8
80.1
86.4
10.7
82.7
65.8
65.3
78.6
67.5
83.2
76.5
53.5
83.3
83.1
76.4
85.3
91.8
83.1
90.2
15.9
87.9
71.6
68.9
86.0
71.3
91.4
84.5
57.0
95.4
95.9
91.1
99.5
94.9
100
112
c
96.8
96.0
101
99.4
99.8
105
95.6
97.0
99.9
111
87.6
95.1
88.6
105
99.1
C
92.1
91.1
90.8
94.1
97.3
89.9
87.4
84.1
94.3
98.8
1.0-g cartridge
80-09 spike
Fraction 1
42.1
81.2
88.7
89.1
95.4
C
89.4
d
70.6
88.6
80.4
95.3
68.0
62.0
98.1
98.1
57.7
87.3
92.1
87.4
94.1
c
92.5
d
70.2
87.9
82.8
102
74.7
53.7
97.9
100
120-0g spike 40-0g spike
Fraction 1 Fraction 1
28.5
78.6
90.6
87.9
92.5
c
88.1
d
66.1
85.1
70.1
97.4
47.1
59.0
98.5
98.3
35.7
76.5
88.1
85.8
89.2
c
85.6
d
63.9
82.5
69.5
93.5
50.2
48.0
93.0
94.8
70.9
111
114
108
115
C
113
d
83.9
106
109
118
111
69.6
114
118
95.3
94.0
103
108
116
c
106
d
93.8
103
104
112
65.8
72.0
115
115
2.0-g cartridge
80-09 spike
Fraction 1
37.1
100
92.2
91.4
94.8
c
91.8
d
68.9
86.4
83.2
98.3
82.9
59.9
103
101
39.5
101
92.8
92.4
95.8
c
92.3
d
84.3
90.9
93.9
95.9
84.4
57.3
99.4
99.3
120-0g spike
Fraction 1
63.6
107
86.9
82.5
105
c
83.4
d
77.8
80.6
101
94.1
81.5
72.2
97.3
92.8
72.9
107
86.0
80.9
98.5
c
82.6
d
77.5
78.3
97.2
87.0
77.6
59.9
86.9
88.5
'Each cartridge was preconditioned with 4 mL hexane prior to use. Each experiment was performed in duplicate. Fraction 1 was eluted
with 5 mL of 10 percent acetone in hexane; Fraction 2 with 5 ml of 10 percent acetone in hexane. A third fraction was collected from
the 2-g cartridge by elution with 5 ml of 10 percent acetone in hexane.
bAdditional bis(2-methoxyethyl) phthalate was recovered from the 0.5-g Florisil cartridge by eluting the cartridge with an additional
5 mL of 10 percent acetone in hexane. The recoveries in Fraction 2 were 70.3 and 71.3 percent for the 40-0g spike, 55.4 and
54.3 percent for the 80-0g spike, 53.4 and 54.1 percent for the 120-0g spike.
'Compound not recovered even when the cartridge was eluted with an additional 5 mL of JO percent acetone in hexane.
dBis(2-ethoxyethyl) phthalate was recovered by eluting the cartridge with an additional 5 mL of 10 percent acetone in hexane. Total
recoveries were 75.1 and 79.3 percent for the 80-0g spike (1.0-g cartridge), 57.3 and 56.5 percent for the 120-0g spike (1.0-g
cartridge), 94.6 percent for the 400-0g spike (2-g cartridge), 55.4 and 62.0 percent for the 80-0g spike (2.0-g cartridge), 70.2 and
79.0 percent for the 120-0g spike (2.0-g cartridge).
-------
TABLE 56. ELUTION PATTERNS AND PERCENT RECOVERIES OF THE METHOD 8060 COMPOUNDS
USING 10 PERCENT ACETONE IN HEXANE AND 20 PERCENT ACETONE IN HEXANE'
10 Percent acetone in hexane
Compound
DMP
DEP
DIBP
DBP
BMPP
BHEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
ONP
Fraction 1
78.6
101
94.0
101
86.4
0
63.6
16.6
68.5
99.9
93.8
75.4
99.1
73.3
97.9
89.5
(5 ml)
65.6
84.5
75.8
88.1
37.0
0
57.3
19.6
56.9
89.3
89.6
66.3
88.3
67.0
120
113
Fraction 2
47.9
0
0
0
0
31.5
0
109
0
0
10.3
50.2
0
0
0
0
(5 ml)
28.8
0
0
0
0
40.3
0
102
0
0
5.4
47.6
0
0
0
0
20 Percent acetone in hexane
Fraction 1
101
103
105
109
104
66.6
106
121
103
112
106
114
102
99.0
118
93.5
(5 ml)
102
103
104
107
102
61.6
100
102
99.3
105
100
103
83.1
90.5
107
101
Fraction 2
0
0
0
0
0
38.8
0
0
0
0
0
0
0
0
0
0
(5 ml)
0
0
0
0
0
34.8
0
0
0
0
0
0
0
0
0
0
M-g Alumina cartridges (Supelco, lot SP 0214) were used; each cartridge was preconditioned
with 4 ml hexane prior to use. Each experiment was performed in duplicate; amount spiked was
40 ng per component per cartridge (2 ml of 20 /ig/mL in hexane).
-------
o
ro
TABLE 57. ELUTION PATTERNS AND PERCENT RECOVERIES OF THE METHOD 8060
COMPOUNDS FROM ALUMINA CARTRIDGES OF VARIOUS SIZES BY ELUTION WITH
20 PERCENT ACETONE IN HEXANE"
0.5-g cartridge
Compound
OHP
DEP
OIBP
OBP
BMPP
BNEP
DAP
BEEP
HEHP
OHP
BBP
BBEP
DEHP
OCP
OOP
DNP
40 -jig spike
Fraction 1
106
111
92.5
101
98
98.8
103
107
102
101
113
86.4
112
100
84.3
77.1
108
112
94.3
103
96.4
104
104
110
104
100
116
78.5
112
102
74.3
66.2
80-pg spike
Fraction 1
67.8
76.2
79.8
83.8
81.4
76.0
83.6
80.9
68.3
81.5
59.2
83.0
77.3
63.9
79.6
80.7
72.6
78.9
80.6
86.7
82.6
79.4
83.2
84.6
72.0
87.4
61.9
91.1
87.8
62.3
90.5
89.5
120-pg spike
Fraction 1
69.5
73.1
116
91.3
100
87.7
99.3
95.2
78.1
101
62.0
117
96.6
73.5
103
96.4
71.3
75.3
115
89.6
109
88.8
98.7
94.5
69.8
85.4
60.5
113
91.3
72.6
101
94.7
40-/ig spike
Fraction 1
108
139
77.5
101
75.1
87.1
92.4
96.5
81.1
116
103
88.1
99.4
99.3
93.3
106
117
148
85.4
109
90.4
93.8
101
105
104
126
110
92.5
107
105
108
114
1.0-g cartridge
60- tig spike
Fraction 1
100
106
93.8
109
92.3
92.2
103
97.7
90.3
94.9
99.1
99.4
100
92.4
101
103
92.1
125
108
121
107
101
116
109
110
106
106
110
113
106
113
114
120- tig spike
Fraction 1
115
126
96.9
103
83.3
90.3
95.0
103
87.4
118
104
104
93.5
90.5
103
113
113
127
94.0
101
81.5
88.5
93.0
96.6
86.2
116
103
103
92.0
89.9
102
110
2.0-g cartridge
40-pg spike
Fraction 1
98.6
106
84.6
95.4
81.8
b
82.4
81. 6C
74.9
95.5
89.3
73.3
85.6
57.4
85.6
88.9
93.8
108
86.5
94.6
80.3
b
84.4
67. 9e
78.8
97.8
91.0
73.9
87.0
59.1
86.5
91.5
80- ng spike
Fraction 1
92.3
105
119
108
101
b
98.6
76.9'
94.1
87.8
96.8
101
99.1
82.1
101
100
73.2
102
110
103
98.1
b
94.9
55. 8C
87.9
81.6
92.8
96.5
93.1
71.1
97.1
96.1
120 -ng spike
Fraction 1
84.1
112
99.8
98.2
81.5
b
87.7
45.6'
78.5
101
94.5
87.3
90.2
60.7
93.1
102
93.0
104
100
92.8
78.8
b
83.3
70.3'
74.2
93.7
88.4
79.5
85.6
57.5
85.9
92.9
"Each cartridge was preconditioned with 4 mL hexane prior to use. Each experiment was performed in duplicate. Fraction 1 was eluted with 5 mL of
20 percent acetone In hexane; Fraction 2 with 5 raL of 20 percent acetone in hexane. A third fraction was collected from the 2-g cartridge by elution
with 5 ml of 20 percent acetone in hexane.
"BMEP was recovered from the 2.0-g alumina cartridge by eluting the cartridge with two additional 5-mL portions of 20 percent acetone in hexane
(Fractions 2 and 3). The recoveries In Fraction 2 were 77.6 and 50.9 percent (40-pg spike), 57.8 and 46.2 percent (80-/*g spike), 26.3 and 61.5 percent
(120-fig spike). The recoveries In Fraction 3 were 38.1 percent (40-/ig spike), 37.9 and 31.9 percent (120-/ig spike).
'Additional BEEP was recovered from the 2.0-g alumina cartridge by eluting the cartridge with an additional 5 ml of 20 percent acetone in hexane.
The recoveries In Fraction 2 were 13.3 and 30.9 percent (40-pg spike), 28.6 and 55.9 percent (80-^g spike), 62.7 and 28.9 percent (120-/ig spike).
-------
Matrix interferents such as corn oil, diesel hydrocarbons, elemental
sulfur, and the organochlorine pesticides listed in SW-846 Method 8081 were
added to hexane solutions containing the target analytes at known
concentrations, and the hexane solutions were then subjected to the Florisil
or alumina cartridge procedure to establish if there are any changes in the
compound elution pattern and in the target analyte recovery when matrix
interferents are present (Table 58). Such interferents were selected because
they mimic typical background contamination in certain environmental sample
matrices that could also be contaminated with the target compounds. For
example, corn oil would be representative of fatty acid triglycerides, diesel
hydrocarbons of petroleum hydrocarbons, and organochlorine pesticides of
compounds of environmental significance that would be expected to behave in
the same way as the target analytes investigated in this study. The data
presented in Table 58 indicate that neither the corn oil nor the diesel
hydrocarbons affected the elution patterns of the 16 phthalate esters. Corn
oil was also removed from the Florisil cartridge with 10 percent acetone in
hexane. Fortunately, its presence does not seem to affect the determination
of the 16 phthalate esters. This statement is true only for corn oil
concentrations below 0.2 mg/mL of solvent (or 1 mg per cartridge) because this
is the maximum concentration we used. Diesel hydrocarbons do not seem to
cause problems with the quantification of the phthalate esters because the
detector is transparent to aliphatic hydrocarbons. Elemental sulfur, if
present, is eluted from the Florisil cartridge with 10 percent acetone in
hexane. Therefore, extracts that are known to contain elemental sulfur should
be subject to sulfur cleanup (Method 3660) prior to Florisil cartridge
cleanup.
The effect of interferents when the Florisil cartridges were eluted with
hexane/diethyl ether (1:1) is presented in Table 59.
Corn oil was also removed from the Florisil cartridge by hexane/diethyl
ether; however, it did not seem to affect the determination of the 16
phthalate esters if the corn oil levels did not exceed 1 mg/cartridge
(Table 59).
Recoveries of the test compounds in the presence of diesel hydrocarbons
were quantitative except for DAP which could not be quantified because of
interference from the diesel fuel hydrocarbons (Table 59).
The organochlorine pesticides overlapped with the phthalate esters when
the GC analysis was performed on the DB-5 fused-silica capillary column and
had to be separated prior to the gas chromatographic analysis. The
hexane/diethyl ether (1:1) combination did not give adequate recoveries for
half of the target compounds (Table 59) but use of 20 percent methylene
chloride in hexane followed by elution with hexane/acetone (9:1) was found to
remove most of the organochlorine pesticides and gave quantitative recoveries
for 14 of the 16 phthalate esters (Table 60).
Additional experiments were performed in order to develop a procedure
that will allow the determination of the phthalate esters in the presence of
the organochlorine pesticides. The experimental design is presented in
Table 61. Tables 62 and 63 present the recovery data and elution patterns of
the 16 target phthalate esters when the cartridges were eluted with methy1<*ne
103
-------
TABLE 58. PERCENT RECOVERIES OF METHOD 8060 COMPOUNDS FROM
ALUMINA CARTRIDGES WHEN INTERFERENTS ARE PRESENT"
FLORISIL AND
Compound
Florisil cartridge
Alumina cartridge
Corn oil
(1000 fig per cartridge)
Diesel
hydrocarbons
(2000 fig per cartridge)
Corn oil
(1000 ng per cartridge)
Diesel
hydrocarbons
(2000 ng per cartridge)
DMP
DEP
DIBP
DBP
BMP
BMEP"
DAP
BEEP0
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
119
133
101
111
104
b
96.6
53.3
89.8
108
106
104
99.9
81.4
109
114
123
133
104
111
104
b
96.8
64.6
91.2
106
107
104
99.4
81.2
108
114
106
123
111
110
93.2
b
98.8
43.7
87.1
103
102
98.8
92.1
68.2
102
107
111
129
107
114
95.7
b
98.8
32.3
86.6
104
104
100
94.6
68.2
103
111
105
120
88.8
92.4
61.2
81.4
82.7
70.9
74.3
99.8
93.8
87.8
83.3
81.8
93.1
98.5
104
119
87.7
91.1
63.1
81.8
83.1
71.8
82.9
98.9
92.6
87.8
83.1
81.3
92.7
99.2
92.5
92.5
82.8
88.7
69.8
74.1
74.9
66.0
71.1
90.3
84.6
88.3
72.6
72.0
80.9
86.4
94.4
94.4
85.8
90.4
71.0
75.8
76.9
67.9
73.1
91.5
87.3
81.6
74.6
73.8
82.7
88.3
*l-g cartridges were used for these experiments. Each cartridge was preconditioned with 4 ml hexane. Each
experiment was performed in duplicate. The Florisil cartridge was eluted with 5 ml of 10 percent acetone in
hexane. The alumina cartridge was eluted with 5 ml of 20 percent acetone in hexane.
"BMEP was recovered from the Florisil cartridge by eluting the cartridge with an additional 5 ml of 10 percent
acetone in hexane. The recoveries were 81.9 and 95.6 percent when corn oil was present as interferent and
71.5 and 62.3 percent when diesel hydrocarbons were the interferents.
'Additional BEEP was recovered from the Florisil cartridge by eluting the cartridge with an additional 5 ml of
10 percent acetone in hexane. The recoveries in Fraction 2 were 41.6 and 31.7 percent when corn oil was present
as interferent, and 56.8 and 63.4 percent when diesel hydrocarbons were the interferents.
-------
o
en
TABLE 59. ELUTION PATTERNS AND PERCENT RECOVERIES OF METHOD 8060 COMPOUNDS
FROM FLORISIL CARTRIDGES WITH HEXANE/DIETHYL ETHER (l:l)a
Fraction 1
Compound
OMP
DEP
OIBP
DBP
BMPP
BNEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
No
(5 nL of hexane/di ethyl ether 1:1)
With corn
Interferents
94
96
122
116
108
2.6
95
8.9
112
98
111
65
95
98
106
90
94
95
119
114
109
96
101
99
104
98
117
97
103
101
109
91
oil
78
84
122
b
103
38
b
7.8
127
128
122
78
89
118
114
80
82
93
115
105
87
19
103
44
116
106
117
90
83
86
80
72
With dlesel
hydrocarbons
116
74
105
124
109
128
b
55
88
107
118
103
85
114
114
104
108
70
97
113
96
109
b
48
81
103
112
112
78
104
107
95
With
organochlorine
pesticides
93
103
118
b
b
b
b
14
86
b
b
b
91
100
108
94
91
98
111
b
b
b
b
58
83
b
b
b
86
96
102
90
Fraction 2 (5 mL of hexane/di ethyl ether (1:1)
No
Interferents
<5
<5
<5
<5
<5
32
<5
51
<5
<5
<5
27
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
<5
With corn
oil
<5
10
<5
<5
<5
<5
<5
66
11
6.0
<5
25
<5
<5
<5
<5
<5
<5
<5
<5
<5
20
<5
54
8.2
<5
<5
10
<5
<5
<5
<5
With diesel
hydrocarbons
<5
7.6
<5
<5
<5
32
6.1
62
8.2
<5
<5
9.6
<5
<5
<5
<5
<5
<5
<5
<5
<5
22
7.2
58
6.0
<5
<5
10
<5
<5
<5
<5
With
organochlorine
pesticides
<5
6.2
19
<5
60
103
74
53
<5
17
6.8
58
<5
<5
<5
<5
<5
6.4
22
<5
72
128
87
54
<5
20
7.4
69
<5
<5
<5
<5
"Florisil cartridges (1 g) were used. Fraction 1 was eluted with 5 ml hexane/diethyl ether (1:1) and Fraction 2 with an additional 5 ml
hexane/diethyl ether (1:1). Final volume of each fraction was 5 mL. Method 8060 compounds were spiked at 2,500 ng per Florisil
cartridge.
"Not able to determine because of interference from organochlorine pesticides.
-------
TABLE 60. RESULTS OF THE FLORISIL CARTRIDGE CLEANUP
EVALUATION STUDY (PHTHALATE ESTER
STANDARDS ONLY; ELUTION WITH 20 PERCENT
NETHYLENE CHLORIDE IN HEXANE, AND
HEXANE/ACETONE (9:1))
Percent
8712-013-10
Fraction
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
I
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
II
68.0
87.7
107
103
116
36.0
75.0
78.3
118
72.0
91.0
113
107a
106
125
100
recovery
8712-013-11
Fraction
I
0
0
0
23.5
0
0
0
0
0
0
0
0
0
0
0
0
II
121
90.7
140
129
123
39.1
130
99.0
134
61.0
102
188
113a
109
131
112
8712-013-12
Fraction
I
0
0
0
0
0
0
6.6
0
0
0
0
0
0
0
0
0
II
201
86.3
108
131
130
20.7
76.0
69.0
127
53.0
102
103
110a
102
114
94.7
aThe value not corrected for method blank; if
corrected, recovery becomes 0.
106
-------
TABLE 61. EXPERIMENTAL DESIGN FOR FLORISIL CARTRIDGE CLEANUP METHOD
DEVELOPMENT"
Amount
spiked (ng)
Sample
identification
8712-013-1
8712-013-2
8712-013-3
8712-013-4
8712-013-5
8712-013-6
8712-013-7
8712-013-8
8712-013-9
8712-013-10
8712-013-11
8712-013-12
8712-013-13
8712-013-14
8712-013-15
8712-013-16
8712-013-17
8712-013-18
8712-013-22
Phthalate
esters
500
500
500
500
500
500
500
500
500
500
500
500
0
0
0
500
500
500
0
OCPs
500
500
500
500
500
500
500
500
500
0
0
0
500
500
500
500
500
500
0
Fraction I
(5mL)
Hexane
Hexane
Hexane
Hexane
Hexane
Hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
Fraction II
(5 mL)
Hexane/di ethyl ether
(1:1)
Hexane/di ethyl ether
(1:1)
Hexane/di ethyl ether
(1:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
20% methyl ene
chloride in hexane
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
Hexane/acetone
(9:1)
(continued)
107
-------
TABLE 61 (continued)
Amount
spiked (ng)
Sample
identification
8712-013-23
8712-013-24
8712-013-25
8712-013-26
8712-013-27
8712-013-28
8712-013-29
8712-013-30
8712-013-31
8712-013-32
8712-013-33
8712-013-34
8712-013-35
8712-013-36
8712-013-37
8712-013-38
8712-013-39
8712-013-40
8712-013-41
8712-013-42
Phthalate
esters
0
500
500
500
500
500
500
500
500
500
0
0
0
500
500
500
500
500
500
500
OCPs
0
0
0
0
0
0
0
0
0
0
500
500
500
500
500
500
500
500
500
500
Fraction I
(5 ml)
20% methyl ene
chloride in hexane
15% methyl ene
chloride in hexane
15% methyl ene
chloride in hexane
15% methyl ene
chloride in hexane
25% methyl ene
chloride in hexane
25% methyl ene
chloride in hexane
25% methyl ene
chloride in hexane
30% methyl ene
chloride in hexane
30% methyl ene
chloride in hexane
30% methyl ene
chloride in hexane
15% methyl ene
chloride in hexane
25% methyl ene
chloride in hexane
30% methyl ene
chloride in hexane
15% methyl ene
chloride in hexane
15% methyl ene
chloride in hexane
15% methyl ene
chloride in hexane
25% methyl ene
chloride in hexane
25% methyl ene
chloride in hexane
25% methyl ene
chloride in hexane
30% methyl ene
chloride in hexane
Fraction II
(5 ml)
Hexane/acetone
(9:1)
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
(continued)
108
-------
TABLE 61 (concluded)
Amount
spiked (ng)
Sample Phthalate
identification esters
8712-013-43
8712-013-44
8712-013-45
8712-013-46
8712-013-47
500
500
0
0
0
OCPs
500
500
0
0
0
Fraction I
(5 ml)
30% methyl ene
chloride in hexane
30% methyl ene
chloride in hexane
15% methyl ene
chloride in hexane
25% methyl ene
chloride in hexane
30% methyl ene
chloride in hexane
Fraction II
(5 ml)
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
10% ethyl
acetate in hexane
al-g Florisil cartridges were used. The 16 phthalate esters were spiked at
500 ng per component. The organochlorine pesticides (OCPs) were also spiked
at 500 ng per component.
109
-------
TABLE 62.
RECOVERY OF PHTHALATE ESTERS FROM THE 1-G FLORISIL CARTRIDGE BY
ELUTION WITH 15 PERCENT, 25 PERCENT, AND 30 PERCENT METHYLENE
CHLORIDE IN HEXANE (FRACTION I) AND 10 PERCENT ETHYL ACETATE IN
HEXANE (FRACTION
Percent recovery
8712-013-24
Fraction
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
I
7.5
0
0
13.1
0
0
0
0
13.7
49.0
4.0
0
0
0
0
0
II
7.0
34.5
98.0
65.0
101
0
156
6.0
91.0
40.1
73.0
0
54.0
125
94.2
80.3
8712-013-25
Fraction
I
5.7
0
0
13.5
0
0
0
0
0
40.2
0
0
0
0
0
0
II
6.6
37.9
60.4
80.0
102
21.0
161
6.2
90.0
42.5
81.0
0
46.0
97.5
95.3
75.4
8712-013-26
Fraction
I
4.1
0
0
0
0
0
0
0
0
43.3
0
0
0
0
0
0
II
0
40.1
64.2
85.0
105
32.8
166
5.0
101
44.3
89.0
0
56.0
110
107
82.9
8712-013-27
Fraction
I
0
0
0
0
0
0
0
0
0
46.2
27.3
0
0
0
0
0
11
13.0
65.8
64.6
112
102
23.4
143
6.8
110
50.9
95.4
0
31.3
110
108
93.4
8712-013-28
Fraction
I
12.3
0
0
0
0
0
0
0
0
46.2
24.7
0
0
0
0
0
II
0
27.3
60.7
83.7
90.7
24.2
179
0
120
44.9
84.4
0
36.2
100
99.8
81.1
8712-013-29
Fraction
I
12.6
12.9
0
0
0
0
0
0
14.7
80.5
29.2
0
0
0
0
0
II
2.2
38.6
89.8
69.8
96.6
24.2
184
17.6
89.3
51.3
93.4
0
36.0
108
108
78.8
8712-013-30
Fraction
I
4.4
0
0
0
0
0
0
0
0
0
3.7
0
0
0
0
0
II
4.3
26.6
58.6
49.8
79.4
0
159
0
115
50.7
101
0
50.0
142
104
105
8712-013-31
Fraction
I
13.6
0
0
0
0
0
0
0
0
0
6.5
0
0
0
0
0
II
13.2
27.2
53.6
52.4
78.7
18.9
108
0
120
47.2
106
0
43.9
142
100
115
8712-013-32
Fraction
I
12.8
0
0
0
0
0
0
0
0
1.5
0
0
0
0
0
II
12.4
24.8
56.1
51.9
79.4
18.2
103
0
101
49.2
103
0
44.9
148
106
94.3
'The spiking level was 500 ng of each phthalate ester per cartridge.
-------
TABLE 63. RECOVERY OF PHTHALATE ESTERS FROM THE 1-6 FLORISIL CARTRIDGE IN THE
PRESENCE OF THE OCPs BY ELUTION WITH 15 PERCENT, 20 PERCENT, AND
30 PERCENT NETHYLENE CHLORIDE IN HEXANE (FRACTION I) AND 10 PERCENT
ETHYL ACETATE IN HEXANE (FRACTION II)a
Percent Recovery
Compound
DMP
DEP
OIBP
OBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
8712-013-36
Fraction
II
160"
49.5
150"
123
1560"
138"
5,550"
ND
112
137"
123
560"
135"
112
117
43.0
8712-013-37
Fraction
II
172"
57.5
105
121
74.9
645"
5,620"
ND
102
123
124
520"
119
104
118
40.6
8712-013-38
Fraction
II
174"
60.0
108
115
72.7
224"
5,530"
ND
103
129"
117
530"
112
97.8
109
38.4
8712-013-39
Fraction
II
118
63.2
98.2
118
78.8
456"
1,420"
28.3
108
146"
127"
338"
151"
110
126"
42.0
8712-013-40
Fraction
II
112
56.3
108
121
116
162"
1,410"
26.3
113
120
125
362"
128"
110
123
43.6
8712-013-41
Fraction
II
101
56.7
99.0
108
110
185"
850"
29.2
110
113
129"
274"
119
112
126"
43.8
8712-013-42
Fraction
II
120
40.2
92.5
109
109
124
1,640"
29.0
110
126"
117
335"
220"
103
112
45.2
8712-013-43
Fraction
II
113
51.1
95.8
106
102
210"
454"
24.2
88.6
92.7
106
171"
181"
94.3
90.7
38.3
8712-013-44
Fraction
II
114
34.9
97.6
102
102
367"
881"
25.7
99.7
110
121
227"
249"
107
111
44.3
4The spiking level was 500 ng of phthalate ester per cartridge. The OCPs were also spiked at 500 ng per component per
cartridge.
"High recovery due to interferents eluted from the cartridge by 10 percent ethyl acetate in hexane and/or contaminants in ethyl
acetate.
-------
chloride/hexane and then ethyl acetate/hexane. Interferences in the
determination of phthalate esters caused by organochlorine pesticides (OCRs)
and method blank analyses are presented in Tables 64 and 65.
The Florisil procedure was further evaluated using extracts of
environmental samples spiked with 16 phthalate esters at known concentration.
The results presented in Table 66 indicate that recoveries were greater than
74 percent except for bis(2-ethoxyethyl) phthalate (recovery ranges from 24
to 62 percent) and bis(methoxyethyl) phthalate which could not be recovered
in Fraction 2.
6.5 PRESERVATION STUDY
This section summarizes the results of the preservation studies for
reagent water samples and soil samples spiked with the test compounds at
5 /Kj/L and 1 ng/g, respectively. Tables 67, 68, and 69 summarize results for
the water samples. Table 70 summarizes results for the soil samples. The
same results given in Tables 67 through 69 are presented in Figures 21, 22,
and 23.
Spiked water samples preserved at neutral and acidic pH and 4'C give
comparable recoveries which are above 60 percent for most compounds, even
after 21 days of storage. Preservation of water samples at pH 9 and 4"C
should be avoided since most compounds show a significant decrease in
concentration after 14 days of storage.
The recovery data presented in Table 70 and Figures 24 and 25 indicate
that the average recoveries of the phthalate esters except BMPP from soil are
>60 percent. Storage of spiked soil samples at -10"C is preferred over
refrigeration at 4'C, since it allows higher recoveries of the lower-
molecular-weight esters, which are more easily degradable, to be recovered.
6.6 REVISED METHOD 8060
The revised Method 8060 was evaluated in terms of reproducibility of the
injection technique, the linearity of response over several orders of
magnitude in concentration, how precise the identification and the measurement
of the gas chromatographic technique is, and in terms of what the minimum
detectable levels for the phthalate esters are. Subsequent sections address
the reproducibility of the injection technique, the instrument calibration,
the method accuracy, and the method detection limit.
6.6.1 Reproducibility of the GC Technique
To establish the reproducibility of the GC technique, 11 consecutive
injections of hexane blanks spiked with 10 /*L of a benzyl benzoate solution
at 50 ng/nl (nominal concentration of benzyl benzoate that was analyzed is
500 ng//nL) were performed by an autosampler.
The results are presented in Table 71 as the average retention time of
the 11 replicates and the RSD, and the average detector response and the RSD.
The reproducibility of the retention time was 0.072 percent and the
reproducibility of the detector response was 2.3 percent. When actual samples
112
-------
TABLE 64. INTERFERENCES IN THE DETERMINATION OF PHTHALATE ESTERS CAUSED BY
OCPs
Concentration (ng/mL)*
8712-013-13
Fraction
Compound
DMP
OEP
OIBP
DBP
BMPP
BHEP
OAP
BEEP
HEHP
OHP
BBP
BBEP
DEHP
OCP
OOP
DNP
I
NDe
ND
23.2
64.1
1,810
6,160
370
NO
3,070
42.2
33.2
NO
48.4
NO
ND
ND
II
260
ND
24.5
34.0
660
23.8
4,220
ND
30.9
122
36.3
586
70.4
ND
ND
ND
8712-013-14
Fraction
I
ND
ND
57.3
26.2
2,250
6,420
2,070
ND
3,270
52.6
37.2
31.6
53.0
ND
ND
NO
II
227
ND
70.4
30.0
386
325
2,870
32.7
69.9
103
31.7
583
129
ND
ND
ND
8712-014-15
Fraction
I
45.2
ND
50.9
27.8
2,020
6,274
857
38.2
3,120
63.8
34.5
NO
69.3
NO
ND
ND
II
205
37.7
66.6
26.7
19.8
544
3,960
32.1
251
89.4
30.5
606
112
ND
ND
ND
8712-013-33
Fraction
I
ND
ND
45
28
1,260
7,510
557
60
3,540
65
35
ND
65
ND
ND
ND
.6
.6
.4
.1
.8
.9
II
208
NO
20.4
20.8
18.0
585
6,200
22.1
21.7
107
36.5
629
219
ND
ND
ND
8712-013-34 8712-013-35
Fraction Fraction
I" II I"
137
ND
30.3
ND
130
313
1,110
21.7
19.9
87.2
33.2
346
124
ND
ND
ND
II
143
ND
36.3
ND
ND
293
491
22.1
150
63.0
28.5
99.1
76.6
ND
ND
ND
The Florisil cartridges were charged only with OCPs. The various peaks detected in these fractions as phthalate esters are
not contaminants in the OCP standards but rather OCP peaks eluting at the same retention times as the target analytes. If
the OCPs would not interfere with the quantification of the phthalate esters, the sum of the concentrations in Fractions I
band II should be 100.
cAnalysis did not pass QC criteria, results are therefore not reported.
Not detected; detection limit was approximately 10 ng/mL.
-------
TABLE 65. RESULTS OF METHOD BLANK ANALYSES FOR THE FLORISIL CARTRIDGE
Concentration (ng/mL)a
8712-013-22
Fraction
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
I
40.7
NDb
ND
ND
ND
ND
63.9
ND
33.9
50.6
24.9
ND
68.8
ND
52.2
ND
II
151
ND
ND
26.8
ND
ND
137
16.3
18.6
49.8
30.2
ND
121
ND
ND
ND
8712-013-23
Fraction
I
51.9
ND
22.8
ND
ND
ND
ND
18.8
18.3
ND
35.9
ND
100
ND
64.1
ND
II
200
ND
22.1
30.1
ND
ND
139
18.2
ND
54.4
37.5
ND
114
ND
ND
ND
8712-013-45
Fraction
I
12.4
ND
ND
ND
ND
ND
ND
ND
ND
ND
35.7
ND
60.2
ND
ND
ND
II
55.4
ND
ND
36.8
ND
18.5
ND
14.9
16.7
40.0
35.9
ND
56.1
ND
ND
ND
8712-013-46
Fraction
I
17.0
ND
ND
36.6
ND
ND
46.8
ND
ND
ND
ND
59.1
81.1
ND
ND
ND
II
33.2
ND
ND
13.9
ND
ND
ND
14.8
ND
38.2
35.6
ND
61.1
ND
ND
ND
8712-013-47
Fraction
I
24.2
ND
ND
ND
ND
ND
61.8
ND
ND
49.0
27.1
ND
58.2
ND
ND
ND
II
32.4
ND
ND
33.1
16.4
17.9
ND
20.7
ND
35.1
24.7
56.7
58.1
ND
ND
ND
aThe Florisil cartridge (1 g) was eluted with 5 mL of 20 percent methylene
chloride in hexane and 5 mL of hexane/acetone (9:1) in the case of samples
8712-013-22, -23, and 5 mL of 15 percent, 25 percent, and 30 percent
methylene chloride in hexane for samples -45, -46, and -47. Fraction II for
samples -45, -46, -47 was eluted with 5 mL of 10 percent ethyl acetate in
hexane. Final volume of each fraction was 5 mL.
bNot detected; detection limit was approximately 10 ng/mL.
114
-------
TABLE 66. PERCENT RECOVERIES OF PHTHALATE ESTERS FROM VARIOUS MATRICES BY
FLORISIL CARTRIDGE CLEANUP WITH HEXANE/METHYLENE CHLORIDE (4:1)
AND HEXANE/ACETONE (9:1) AS ELUANTS*
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Sandy loam
soil"
78
79
79
74
77
0
82
37
80
78
82
86
74
91
80
84
Sediment
of unknown
origin
75
79
82
78
84
0
86
24
88
88
99
94
85
96
92
96
NBS
SRM-1572
80
89
90
84
102
0
100
62
95
86
114
98
108
106
104
106
NBS
SRM-1632a
76
79
108
83
91
0
76
32
93
92
102
106
88
98
95
111
NBS
SRM-1633a
82
84
86
83
86
0
89
33
81
80
98
98
112
95
88
92
"The spiking level was 50 ng/mL of extract for each compound. Data shown
are for Fraction 2 which was eluted with 5 mL hexane/acetone (9:1).
"Sandy loam soil from Puyallup, Washington, with the following physico-
chemical characteristics: pH 5.9 to 6.0; 89 percent sand, 7 percent
silt, 4 percent clay; cation exchange capacity 7 meg/100 g; total organic
carbon content 1,290 ± 185 mg/Kg.
115
-------
TABLE 67. CONCENTRATION OF THE 16 PHTHALATE ESTERS AS A
OF TIME AT pH 6.8 TO 6.9
FUNCTION
Concentration (pg//iL of extract)6
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
OAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
DPTP"
t
3,180
3,970
,620
,840
b
,460
,100
,420
5,210
4,680
6,470
4,860
4,590
4,660
5,240
4,100
135
=0
2,790
3,100
3,970
4,190
b
4,530
3,490
3,970
4,220
3,780
5,610
4,540
4,040
4,010
4,170
3,350
106
t=l
2,180
3,010
3,920
4,100
b
4,540
3,520
3,460
3,830
3,110
5,450
3,970
3,910
3,930
4,150
3,180
112
day
1,930
2,910
3,840
3,790
b
4,140
2,990
3,350
3,810
2,980
5,590
3,750
3,750
3,830
4,150
3,180
118
t=3
3,100
2,620
3,170
3,030
b
5,220
2,340
2,300
2,990
2,560
4,440
3,510
3,240
2,730
2,870
1,800
115
days
3,690
3,700
4,100
4,010
b
5,910
3,310
3,410
4,240
3,240
5,920
4,580
3,780
3,880
4,140
3,140
138
t=7
2,900
3,560
4,420
4,400
b
4,970
3,230
4,020
3,970
4,280
4,590
4,240
3,800
3,810
4,290
3,430
78.5
days
2,970
3,430
4,390
4,440
b
5,770
3,960
3,580
3,950
3,890
4,880
4,200
3,760
3,810
4,130
3,360
87.5
t=10
2,750
3,350
4,110
3,960
b
4,610
2,770
3,610
3,640
3,600
4,720
3,990
3,450
3,470
3,630
2,890
76.2
days
2,580
3,080
3,660
3,450
b
4,200
2,350
3,090
3,060
3,200
4,220
3,330
2,980
3,000
3,130
2,580
72.7
t=14
2,760
3,330
3,880
4,190
b
4,240
3,180
2,990
3,080
3,000
4,190
4,060
2,630
2,890
2,520
1,820
74.8
days
2,710
3,280
3,340
3,190
b
4,600
2,570
2,200
2,460
2,720
3,220
4,060
2,240
2,120
2,050
1,450
72.6
t=21
3,870
3,840
3,790
3,230
b
4,480
3,080
2,470
2,800
3,060
3,590
3,700
2,990
2,970
2,920
2,390
77.6
days
3,440
3,550
3,700
3,300
b
5,370
2,870
1,370
2,670
2,730
3,350
3,570
2,860
2,830
3,510
2,180
76.9
'The spike level was equivalent to 5,000 pg/pL of extract or 5 /zg/L of water (1 L reagent water was spiked
with 5 /ig of each phthalate ester; extraction by Method 3510; Vextract was 1 ml). Duplicate determinations
were performed at each time event.
bNot able to quantitate because of interference.
°The value given Is the percent recovery for the extraction step.
-------
TABLE 68. CONCENTRATION OF THE 16 PHTHALATE ESTERS AS A FUNCTION
OF TIME AT pH 9
Concentration (pg//iL of extract)*
Compound t=0 t»7 days t»14 days
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
DPTPe
2,320
2,870
3,790
4,090
b
4,950
3,650
3,870
4,400
3,560
7,080
4,980
4,120
4,250
4,370
3,230
155
1,910
2,730
3,130
3,100
b
4,480
2,860
3,030
3,250
2,990
5,860
4,170
3,440
3,390
3,500
2,660
124
3,180
3,850
4,390
4,240
b
5,440
4,080
3,770
3,860
4,020
5,140
4,250
3,670
3,660
3,650
2,990
90.1
750
1,070
3,760
2,980
b
1,430
3,070
3,220
2,690
3,360
1,510
1,390
3,660
3,040
3,030
2,480
94.8
380°
450°
1,810
1,500
b
756
1,270
1,930
1,560
2,000
226
d
2,670
1,890
2,290
1,290
70.9
380°
450°
1,870
1,290
b
819
1,180
2,430
1,420
1,960
265
d
2,600
1,720
2,200
1,410
73.1
aThe spike level was equivalent to 5,000 pg//iL of extract or
5 ng/L of water (1 L reagent water was spiked with 5 ng of
each phthalate ester, adjusted to pH 9 with 6N NaOH;
extraction by Method 3510; Ve_a(? was 1 ml). Duplicate
determinations were performea at each time event.
bNot able to quantitate because of interference.
cManual quantitation of the peak.
dNot detected.
"The value given for the DPTP surrogate is the percent
recovery for the extraction step.
117
-------
TABLE 69. CONCENTRATION OF THE 16 PHTHALATE ESTERS AS A FUNCTION
OF TIME AT pH 2
Concentration (pg//iL of extract)0
Compound t=0 t=7 days t=14 days
DMP
DEP .
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
DPTPC
2,690
5,710
3,600
3,450
b
4,610
3,110
3,240
3,900
3,130
5,510
3,990
3,330
3,350
3,220
2,630
105
2,920
5,920
3,740
3,700
b
5,180
3,510
3,370
4,150
3,540
6,220
5,040
4,160
3,870
3,660
2,960
128
3,150
3,350
3,890
4,070
b
5,000
4,490
3,490
3,660
3,950
4,400
4,520
3,680
3,380
3,600
2,940
84.5
3,050
3,500
3,720
3,850
b
5,150
3,380
3,130
3,470
3,550
4,480
3,680
2,990
3,080
3,130
2,660
83.3
2,780
3,100
3,550
4,060
b
4,320
3,520
2,730
3,080
2,900
4,240
3,640
2,760
2,790
2,540
1,840
74.3
2,900
3,270
3,390
3,680
b
3,830
3,360
2,770
2,700
2,720
4,060
3,400
2,600
2,540
2,620
1,900
72.1
aThe spike level was equivalent to 5,000 pg//iL of extract or
5 ng/l of water (1 L reagent water was spiked with 5 jtg of
each phthalate ester, adjusted to pH 2 with 6N H3S04 (3 mL);
extraction by Method 3510; Ve^ra^ was 1 ml). Duplicate
determinations were performed at each time event.
bNot able to quantitate because of interference.
°The value given for the DPTP surrogate is the percent
recovery for the extraction step.
118
-------
LJ
O
O
LJ
o:
z
UJ
O
o:
u
Q.
140
120 -
100 -
80 -
60 -•
40 -
20 -'
0
DMP DEP DIBP DBP BMED BEEP DAP HEHP DHP BBP BBEP DCHP DEHP OOP DNP
ZZL'~T-0'•
T=7
T=10
T=14
T = 21
Figure 21. Average recovery of Method 8060 compounds from HPLC-grade water
as a function of time at pH 7.
-------
ro
o
LJ
O
U
Ul
o:
i-
z
LJ
O
01
LJ
Q.
140
120 -
100 -
80 -
60 -
40 -
20 -
0
/\
x\
/\
/\
/\
/\
x\
/\
x\
/\
/\
/\
x\
\
x\
\
\
\
\
\
x\
\
\
\
xl\
x\
7"
\
\/J
\/
X
xK
x
\
•\1
\
x\
x\
/\
\7
x
DMP DEP DIBP DBP BMEP BEEP DAP HEHP DHP BBP BBEP DCHP DEHP OOP DNP
T = 0
T=7
T-14
Figure 22. Average recovery of Method 8060 compounds from HPLC-grade water
as a function of time at pH 9.
-------
120
ro
100 ^
QL
U
O
O
LJ
UJ
U
a:
LJ
Q.
80 -
60 -
40 ~
20 -
0
\
M/
/
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[71
y\
\yj
/
y
y
y
y\
y
\
\
y\
y\
\
\
\
\
y\
\
/\
\
\
\
\
y\
y
\
\
\
\
\
\
\
\
\
y\
y\
y\
\
y\
y\
y\
y\
l/\
y\
y\
y\
y\
y\
y\
\
y\
DMP DEP DIBP DBP BMEP BEEP DAP HEHP DHP BBP BBEP DCHP DEHP OOP DNP
T-14
Figure 23. Average recovery of Method 8060 compounds from HPLC-grade water
as a function of time at pH 2.
-------
TABLE 70. RECOVERY OF THE 16 PHTHALATE ESTERS AS A
FUNCTION OF TIME AT -10eC (SOIL MATRIX)
Percent Recovery*
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
t=0
77.3
61.3
75.8
111
83.8
68.4
76.2
41.4
68.2
191
96.7
77.7
94.4
73.2
86.9
112
t=3 days
60.8
69.2
110
91.1
145
25.9
73.6
11.9
82.4
90.4
73.3
47.9
159
66.7
85.1
94.9
t=7 days
51.4
52.1
60.4
70.2
74.9
59.2
56.1
26.8
49.2
79.8
80.9
65.4
106
73.4
87.1
128
t» 14 days
47.4
58.2
108
87.4
20.5
57.4
63.7
0
23.8
110
75.4
62.9
91.4
48.0
95.4
124
t=28 days
59.0
60.7
102
99.1
29.9
52.4
71.3
0
19.6
105
95.4
75.8
89.2
48.0
109
164
aThe spike level was 1,000 ng/g; samples were extacted by Method 3550;
volume of extract was 3 ml; 2 ml were subjected to Florisil
chromatography. GC analyses were performed on the DB-5/DB-1701 column
pair (Table 6).
122
-------
ro
4\J\J
190 -
180 -
170 -
160 -
150 -
140 -
& 130 -
Ul
§ 120-
a 110-
K 100 -
8 80-,
B 70-
60 -
50 -
40 -
30 -
20 -
10 -
A _ -
_
,'.
<• ' \
t ' i
i
< 1
> l
1
'" . '
i l ' i
,
•
j
' \
..
,
,,
!,
>
n
,
•
(
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;
t • i
•V
p
• •
h. •-
* i
:, ..
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'-<
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i * t
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i
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11 • \
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i "
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;;, ,.
«
n
1
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l,
l,
,,
;,
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'•'
.',,
'• •
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'• •
DMP DEP DIBP DBP BMPPBMEP DAP BEEP HEHP DHP BBP BBEP DEHP DCP OOP DNP
1-3
t-7
t-14
t-28
Figure 24. Recovery of Method 8060 compounds from sandy loam soil as a
function of time at -10°C.
-------
ro
^.O\J
260 -
240 -
220 -
200 -
£ 18°-
8 160-
u
K 140 -
u 120 -
tt
U 100 -
O. IUW
80 -
60 -
40 -
20 -
A —
q
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'«
j-
!
i
, ..
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1
l 1 > l
i 1 i >
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r
/:.
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•
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l
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•
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1 '
1
s
s
s ,
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s
«
n
, ( r
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f ,
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pi
••
3
!,
s,
•
(
:« ,,
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-
q
::
, s
' •
"
;
«
,
:, ,.
:, ,,
(
r
s
s
'
,
:, ,.
:, .,
•
i
!,
;,
!,
;
'•
•
M,
'
DMP DEP DIBP DBF BMPPBMEP DAP BEEP HEHP DHP BBP BBEP DEHP DCP OOP DNP
1771 to
t1T1
t1T2
*2T1
IXXl t2T2
Figure 25. Recovery of Method 8060 compounds from sandy loam soil as a
function of time at -10°C and +4°C (t0 - initial times; tt -
1 month; t2 - 1.5 months; T, - temperature of -10°; T2 -
temperature of +4°C).
-------
TABLE 71. REPRODUCIBILITY OF THE ABSOLUTE AREA
AND RETENTION TIME OF BENZYL BENZOATE*
Experiment
No.
1
2
3
4
5
6
7
8
9
10
11
Average
SD
RSD (percent)
Retention time
(min)
5.821
5.808
5.807
5.808
5.809
5.806
5.809
5.808
5.806
5.807
5.810
5.809
0.004
0.072
Absolute area
59,682
58,450
58,429
58,897
58,331
60,242
61,203
57,292
56,784
57,912
57,132
58,578
1,360
2.3
aThe reproducibility was determined for 11
consecutive injections. The nominal spike level
of benzyl benzoate was 500 pg//*L. The GC/ECD
operating conditions were as follows: 30 m x
0.25 mm ID DB-5 fused-silica capillary column
(0.25 urn film thickness); 120'C to 260eC (hold
15.7 min) at 15*C/min; carrier gas: helium at
21 psi; injector temperature 275*C; detector
temperature 320"C; spitless injection.
125
-------
were analyzed over a 24-hr period, the reproducibility of detector response
was 6.2 percent (Table 72); nonetheless, this value is quite acceptable.
Reproducibility of detector response for each of the 16 phthalate esters is
given in Table 73. The data indicate that excellent reproducibilities were
achieved at concentrations of 100 pg//iL.
6.6.2 Instrument Calibration
Quantification of compounds is typically performed using two types of
calibration: external standard calibration and internal standard calibration.
In the former case, working solutions containing the test compound are
analyzed prior to samples to determine the linear dynamic range of the
instrument. Quantification of compounds in an unknown sample is performed by
comparing the detector response obtained for the unknown sample to that
measured for a calibration standard within the linear range of the instrument.
In the latter case, the linear dynamic range of the instrument needs to be
established in the same way. In addition, an internal standard is spiked into
every calibration standard and unknown sample, and the ratio of detector
responses of the test compound to the internal standard for the calibration
standard and the unknown sample are compared.
During the course of this project we have performed five sets of
multilevel calibrations using the DB-5 fused-silica capillary column.
Initially, we analyzed standards ranging in concentration from 50 to 5,000
pg/jiL. From these data, we concluded that the linear range extends from 50
to 3,000 pg. The data presented in Tables 74 through 76 give the average
response factors (RF), slopes, intercepts, and correlation coefficients for
the five sets of multilevel calibrations. It can be seen that the RFs were
for the most part low for the first two calibrations, reached a maximum for
the November 17, 1987, calibration, and began decreasing for the last two
calibrations. The explanation is that the detector was so contaminated in
August, 1987, that we had to send it to the manufacturer for cleaning. When
the cleaned detector was in operation, the response increased and then began
decreasing steadily with use. To exemplify that, the daily response factors,
measured at three concentrations over a period of 5 days, are shown in
Tables 77 through 85. It is quite obvious, from comparing the percent
differences between the daily response and the average response, that the
instrument performance changes significantly from day to day.
One way to compensate for that change is to use an internal standard.
Table 86 presents the average relative response factors (RRFs), slopes,
intercepts, and correlation coefficients for two multilevel calibrations
performed with benzyl benzoate as internal standard. It can be seen now that
the values for the slopes of the calibration lines are much closer (ideally
they should be identical).
The following comments are made based on observations gathered during the
course of this project:
• Establishment of the instrument dynamic range should be considered;
however, because the ECD is such a sensitive detector and its
performance changes so drastically even during the course of the day,
we recommend that a three-level calibration bracketing the sample
126
-------
TABLE 72. ABSOLUTE AREAS OF THE INTERNAL STANDARD
Sample identification Absolute area'
Phthalate std (2,500 pgM)
Phthalate std (2,500 pgM)
Phthalate std (2,000 pgM)
Phthalate std (1,500 pgM)
8711-049-9 (Rep.l)
8711-049-10 (Rep. 2)
8711-049-11 (Rep. 3)
8711-049-12 (Rep. 4)
8711-049-15 (spiking solution)
Phthalate std (2,500 pgM)
Phthalate std (1,000 pgM)
Phthalate std (1,000 pgM)
Phthalate std (500 pgM)
8711-049-5 (Rep.l)
8711-049-6 (Rep. 2)
8711-049-7 (Rep. 3)
8711-049-8 (Rep. 4)
8711-049-14 (spiking solution)
Phthalate std (1,000 pgM)
Phthalate std (500 pgM)
Phthalate std (100 pgM)
Phthalate std (50 pgM)
8711-049-1 (Rep.l)
8711-049-2 (Rep. 2)
8711-049-3 (Rep. 3)
8711-049-4 (Rep. 4)
8711-049-13 (spiking solution)
Phthalate std (50 pgM)
Average
SO
RSD (percent)
n
50,512
59,728
54,883
53,074
55,670
48,495
48,342
45,572
52,162
52,900
53,481
57,261
53,231
51,325
45,797
51,206
51,323
53,405
53,200
50,911
48,125
51,826
50,359
50,951
50,159
48,668
51,436
47,477
51,481
3,193
6.2
28
"The internal standard was benzyl benzoate;
nominal spike level was 500 pgM. The
analyses are listed in the order in which they
were performed over a 24-hr period. The GC/ECD
operating conditions were as follows: 30 m x
0.25 mm ID DB-5 fused-silica capillary column
(0.25 fun film thickness); 120*C to 260°C (hold
15.7 min) at 15*C/min; carrier gas: helium at
21 psi; injector temperature 275*C; detector
temperature 320*C; splitless injection.
127
-------
TABLE 73. REPRODUCIBILITY OF THE GC/ECD RESPONSE FACTORS
USED IN QUANTITATION"
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Average
response
factor"
395
418
376
423
443
253
404
376
355
358
1,080
348
499
564
394
397
RSDb
(percent)
2.3
1.3
1.1
5.0
2.9
1.3
2.4
3.1
2.8
2.6
3.7
4.2
3.0
3.3
3.0
1.9
Average
response
factor6
322
351
373
370
441
244
385
371
310
346
1,140
378
470
582
393
369
RSD°
(percent)
6.2
5.9
4.7
5.1
1.7
6.7
9.3
7.3
11
2.5
4.3
8.5
2.0
2.6
2.0
7.3
aThe GC/ECD operating conditions were as follows:
30 m x 0.25 mm ID DB-5 fused-silica capillary
column (0.25 pin film thickness); 120'C to 260eC
(hold 15.7 min) at 15*C/min; carrier gas: helium
at 21 psi; injector temperature 275*C; detector
temperature 320eC; splitless injection.
bDate: November 10, 1987, and November 11, 1987;
time of analysis for the three calibration
standards was: 3:59 p.m.; 10:04 p.m.; 2:06 a.m.
Concentration was 100 pg//iL.
°Date: November 12, 1987; time of analysis for the
three calibration standards was: 10:36 a.m.;
11:09 a.m.; and 8:21 p.m. Concentration was
100 pg/^L.
128
-------
TABLE 74. COMPARISON OF THE MULTILEVEL CALIBRATIONS PERFORMED ON
AUGUST 18, 1987, AND AUGUST 28, 1987
August 18, 1987
Average
Compound RF" Slope Intercept
CC6
August 28, 1987
Average
RF' Slope Intercept CCb
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
64.7
78.2
90.6
129
c
c
143
c
c
c
1,180
c
328
338
169
183
52.5
56.8
54.5
57.4
c
c
86.3
c
c
c
797
c
186
246
97.6
126
8,552
13,917
24,921
71,079
c
c
23,064
c
c
c
269,700
c
105,200
70,350
56,680
39,520
0.9986
0.9962
0.9911
0.9822
c
c
0.9423
c
c
c
0.9904
c
0.9803
0.9931
0.9817
0.9927
32.9
46.4
65.7
79.9
c
c
84.3
c
c
68.3
1,000
c
218
308
112
121
45.4
44.6
45.7
52.4
c
c
60.8
c
c
30.4
726
c
154
199
72.8
86.0
-8,566
1,980
10,130
25,300
c
c
8,254
c
c
14,170
137,500
c
31,720
47,950
22,170
16,700
0.9921
0.9990
0.9954
0.9853
c
c
0.9911
c
c
0.9892
0.9969
c
0.9905
0.9958
0.9861
0.9880
aNumber of determinations was 10 for August 18, 1987, and 11 for August 28,
1987. The GC/ECD operating conditions were as follows: 30 m x 0.25 mm ID
DB-5 fused-silica capillary column (0.25 /«n film thickness); 120°C to 260°C
(hold 15.7 min) at 15eC/min; carrier gas: helium at 21 psi; injector
temperature 2758C; detector temperature 320eC; splitless injection.
bCC - Linear regression correlation coefficient.
Compound was not available for testing.
129
-------
TABLE 75. COMPARISON OF THE MULTILEVEL CALIBRATIONS PERFORMED ON
NOVEMBER 17, 1987, AND NOVEMBER 25, 1987
November 17, 1987
November 25, 1987
Average
Compound RFa Slope Intercept
CCB
Average
RF* Slope Intercept CCfc
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
183.6
171.4
162.2
176.9
304.6
151.0
178.0
185.1
225.7
138.5
661.9
202.6
198.6
398.9
196.5
195.9
24,418
30,299
37,851
35,716
22,387
27,730
40,534
40,415
22,535
33,899
91,943
43,765
59,356
49,510
30,271
31,541
0.9954
0.9950
0.9924
0.9931
0.9985
0.9933
0.9920
0.9920
0.9973
0.9918
0.9963
0.9898
0.9739
0.9976
0.9951
0.9933
157
172
193
198
335
174
214
233
244
179
861
240
250
472
215
233
110.3
97.0
98.7
104.7
228.6
96.0
104.1
111.3
164.8
75.2
566.4
127.3
131.6
307.4
126.9
144.5
29,685
37,012
45,580
45,414
57,890
39,003
55,675
61,906
46,205
47,015
15,344
57,774
57,217
85,690
48,591
49,159
0.9829
0.9800
0.9755
0.9743
0.9878
0.9731
0.9621
0.9591
0.9837
0.9600
0.9854
0.9688
0.9727
0.9858
0.9718
0.9770
aThe number of determinations was 11. The GC/ECD operating conditions were as
specified in Table 74.
bCC - Linear regression correlation coefficient.
130
-------
TABLE 76. MULTILEVEL CALIBRATION PERFORMED ON DECEMBER 8, 1987
Average
Compound RFa Slope Intercept CCb
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
162
164
178
163
289
119
159
173
206
132
656
141
190
357
131
151
134.2
116.1
110.5
106.0
227.2
89.99
105.9
106.2
160.3
71.08
487.6
103.2
112.5
272.9
94.17
110.7
17,331
23,203
27,970
25,875
27,798
18,582
27,207
32,387
21,293
23,445
63,386
17,575
26,970
37,954
15,630
16,825
0.9974
0.9954
0.9950
0.9946
0.9956
0.9961
0.9902
0.9874
0.9947
0.9920
0.9933
0.9924
0.9931
0.9927
0.9917
0.9915
aThe number of determinations was 11. The GC/ECD
operating conditions were as specified in
Table 74.
bCC - Linear regression correlation coefficient.
131
-------
TABLE 77. DAILY RESPONSE FACTOR AT 500 pg//iL
Average response
factor^ RF*
Compound November 17, 1987 NovemberlS, 1987 Percent Dc
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DCP
DEHP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
505
309
253
251
188
184
210
210
310
181
233
240
243
191
815
271
461
328
239
244
-18
-23
-14
-16
-12
-8.6
-10
-9.4
-8.7
-11
-3.4
-4.9
-8.7
+6.1
-5.5
-2.8
aMultilevel calibration; 11 data points at concentrations
between 50 pg//iL and 3,000 pg//tL. The GC/ECD conditions
were as specified in Table 74.
bRF500 is the compound's response factor.
cPercent difference.
132
-------
TABLE 78. DAILY RESPONSE FACTOR AT 1,000 pg/^L
Average response
factor' RF b
Compound November 17, 1987 November 18, 1987 Percent 0
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
184
182
187
204
307
168
215
222
233
168
769
241
231
423
224
230
-20
-24
-23
-18
-13
-15
-17
-16
-12
-22
-8.9
-15
-25
-16
-12
-8.4
aMultilevel calibration; 11 data points at concentrations
between 50 pg//*L and 3,000 pg/jU.. The GC/ECD conditions
were as specified in Table 74.
bRF1000 is the compound's response factor.
cPercent difference.
133
-------
TABLE 79. DAILY RESPONSE FACTOR AT 1,500 pg//iL
Average response
factor1* RF >
Compound November 17, 1987 November 18, 1987 Percent Dc
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
188
181
178
191
293
159
201
207
221
160
716
229
224
402
209
204
-18
-24
-27
-24
-16
-20
-23
-22
-17
-26
-15
-18
-28
-20
-17
-19
aMultilevel calibration; 11 data points at concentrations
between 50 pg//iL and 3,000 pg//iL. The GC/ECD conditions
were as specified in Table 74. '
bRF1500 is the compound's response factor.
°Percent difference.
134
-------
TABLE 80. DAILY RESPONSE FACTOR AT 500 pg//iL
Average response
factor*
RF.
i uwvwi m JQQ
Compound November 17, 1987 November 19, 1987 Percent D
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
185
173
201
210
337
187
245
253
262
190
870
290
278
488
248
250
-19
-28
-18
-16
-4.0
-5.6
-5.8
-4.5
-1.5
-12
+3.1
+1.8
-10
-3.4
-2.0
-0.4
aMultilevel calibration; 11 data points at concentrations
between 50 pg//iL and 3,000 pg//iL. The GC/ECD conditions
were as specified in Table 74.
bRF500 is the compound's response factor.
cPercent difference.
135
-------
TABLE 81. DAILY RESPONSE FACTOR AT 1,000 pg//iL
Average response
factor*
RF,
Compound November 17, 1987 November 19, 1987 Percent Dc
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
186
183
185
206
323
175
222
229
245
170
780
286
256
440
245
228
-19
-23
-24
-18
-8.0
-12
-15
-14
-7.9
-21
-7.6
+0.4
-17
-13
-3.2
-9.2
aMultilevel calibration; 11 data points at concentrations
between 50 pg/jiL and 3,000 pg//*L. The GC/ECD conditions
were as specified in Table 74.
'RF
1000
is the compound's response factor.
°Percent difference.
136
-------
TABLE 82. DAILY RESPONSE FACTOR AT 1,500 pg//iL
Average response
factor* RF150pb
Compound November 17, 1987 November 19, 1987 Percent Dc
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
202
198
184
207
320
170
217
223
240
166
744
264
239
425
216
198
-12
-17
-25
-17
-8.8
-14
-17
-16
-9.8
-23
-12
-7.4
-23
-16
-15
-21
"Multilevel calibration; 11 data points at concentrations
between 50 pg/^L and 3,000 pg//iL. The GC/ECD conditions
were as specified in Table 74.
'RF
1500
is the compound's response factor.
Percent difference.
137
-------
TABLE 83. DAILY RESPONSE FACTOR AT 500 pg//iL
Average response
factor1* RF*
Compound November 17, 1987 November23, 1987 Percent D°
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
206
201
225
235
339
209
269
274
269
220
892
294
283
490
244
214
-10
-16
-7.8
-6.0
-3.4
+5.6
+3.5
+3.4
+1.1
+2.3
+5.7
+3.2
-8.4
+3.0
-3.6
-15
aMultilevel calibration; 11 data points at concentrations
between 50 pg//iL and 3,000 pg/jjL. The GC/ECD conditions
were as specified in Table 74.
bRF500 is the compound's response factor,
cPercent difference.
138
-------
TABLE 84. DAILY RESPONSE FACTOR AT 1,000
Average response
factor* RF b
Compound November 17, 1987 November 23, 1987 Percent Dc
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
150
144
157
172
291
153
185
193
216
147
746
217
207
391
185
163
-35
-40
-36
-31
-17
-23
-29
-28
-19
-32
-12
-24
-33
-23
-27
-35
aMultilevel calibration; 11 data points at concentrations
between 50 pg/ML and 3,000 pg/jtL. The GC/ECD conditions
were as specified in Table 74.
bRF1000 is the compound's response factor.
°Percent difference.
139
-------
TABLE 85. DAILY RESPONSE FACTOR AT 1,500 pg//iL
Average response
factor8 RF >
Compound November 17, 1987 NovemberzS, 1987 Percent D°
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
229
239
244
250
351
198
260
265
266
215
844
285
309
505
253
251
161
151
155
173
287
147
184
189
215
149
737
217
208
394
190
170
-30
-37
-37
-31
-18
-26
-29
-29
-19
-31
-13
-24
-33
-22
-25
-32
aMultilevel calibration; 11 data points at concentrations
between 50 pg//uL and 3,000 pg//iL. The GC/ECD conditions
were as specified in Table 74.
bRF1500 is the compound's response factor.
°Percent difference.
140
-------
TABLE 86. COMPARISON OF THE MULTILEVEL CALCULATIONS PERFORMED ON
NOVEMBER 25, 1987, AND DECEMBER 8, 1987
Compound
November 25, 1987
December 8, 1987
Average
RRFa Slope Intercept CCb
Average
RRF* Slope
Intercept CCb
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
OCP
OOP
DNP
0.0028
0.0031
0.0035
0.0036
0.0060
0.0032
0.0039
0.0042
0.0044
0.0032
0.0154
0.0043
0.0045
0.0085
0.0039
0.0042
0.0040
0.0017
0.0017
0.0018
0.0040
0.0017
0.0018
0.0019
0.0029
0.0013
0.0099
0.0022
0.0023
0.0053
0.0022
0.0025
0.5511
0.6823
0.7834
0.8543
1.0793
0.7178
1.0174
1.1302
0.8572
0.8585
2.8515
1.0587
1.0494
1.5932
0.8934
0.9077
0.9806
0.9770
0.9720
0.9701
0.9872
0.9696
0.9594
0.9561
0.9711
0.9570
0.9848
0.9658
0.9711
0.9852
0.9697
0.9731
0.0036
0.0031
0.0034
0.0031
0.0055
0.0025
0.0030
0.0033
0.0039
0.0026
0.0125
0.0027
0.0037
0.0066
0.0025
0.0029
0.0022
0.0019
0.0018
0.0017
0.0038
0.0015
0.0017
0.0017
0.0027
0.0012
0.0080
0.0017
0.0018
0.0045
0.0016
0.0018
0.5197
0.6122
0.7024
0.6556
0.8597
0.4943
0.6718
0.7775
0.6386
0.5631
1.9222
0.4880
0.6899
1.1257
0.4930
0.4874
0.9877
0.9850
0.9848
0.9853
0.9938
0.9879
0.9804
0.9954
0.9926
0.9856
0.9938
0.9899
0.9904
0.9929
0.9913
0.9423
aThe number of determinations was 4. The GC/ECD operating conditions were as
follows: 30 m x 0.25 mm ID DB-5 fused-silica capillary column (0.25 urn film
thickness); 120°C to 260°C (hold 15.7 min) at 15'C/min; carrier gas: helium at
21 psi; injector temperature 275"C; detector temperature 320°C; splitless
injection. RRF is the compound's response factor relative to benzyl benzoate
(internal standard).
bCC - Linear regression correlation coefficient.
141
-------
concentration be performed at least at 8- to 10-hour intervals or
every 10 samples. If the expected sample concentrations are, for
example, 100 pg/jiL of extract, then the three levels should be 50,
100, and 150 pg//xL. If the expected concentrations are not known,
then the sample should be analyzed twice, before and after the
calibration standards that bracket the unknown sample were analyzed.
It may appear that such an approach would double the effort required
in the analysis; however, if the analyses are performed with an
autosampler, then such an effort will not be so burdensome.
• Whenever routine analyses are performed with an autosampler it may be
necessary to begin the daily analysis sequence with four or five
calibration standards of the same concentration. We observed that the
instrument response usually stabilizes after the fourth injection.
• After changing the septum in the injector, we have observed that the
detector requires as much as 48 hrs to stabilize. Therefore, septa
should always be preheated in the oven at 250eC prior to use and they
should be replaced at the end of the working day, or preferably before
the weekend, thus allowing the detector enough time to stabilize and
without affecting the sample throughput during the working day.
6.6.3 Method Accuracy
Method performance data are presented in this report as method accuracy,
method precision, and method detection limits. Table 87 and 88 and Figures 26
through 29 summarized the method accuracy and precision for the aqueous and
solid samples, respectively. In the case of aqueous samples, method accuracy,
given as percent recovery of the 16 phthalate esters spiked into estuarine
water, leachate, and groundwater, at two concentrations (20 jtg/L and 60 jugA)
(Figure 26) ranged from 59.5 to 117. In the case of solid samples, the
percent recoveries were distributed over a much wider range (Figure 28),
demonstrating that method accuracy is indeed a function of both matrix and
analyte concentration. Method precision for aqueous samples (Figure 27) was
better than 27.5 percent. Method precision for solid samples (Figure 29)
varied from matrix to matrix.
In addition to the real samples, which were used during the entire method
protocol evaluation, we also analyzed six EPA performance evaluation samples
on a DB-5 fused-silica capillary column. Four of these samples contained
phthalate esters and two did not. The results of the GC/ECD analyses are
presented in Tables 89 through 94 and in Figures 30 through 33. Percent
biases range from -26.2 and +15.5. All phthalate esters that were reported
by EPA to be present in those samples were identified correctly. Because
these samples contained many other compounds and the analyses were carried out
only on one capillary column, we reported false positives. Confirmation of
the target phthalate esters on another capillary column probably would have
eliminated the false positives.
6.6.4 Method Detection Limits
The determination of the method detection limits was performed according
to the following procedure:
142
-------
TABLE 87. ACCURACY AND PRECISION DATA FOR METHOD 3510 AND METHOD 8061"
Spike level
(20 M9A)
Compound
DMP
DEP
OIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Surrogates :
DPP
DPIP
DBZP
Estuarine
water
84.0
71.2
76.0
83.2
78.6
73.8
78.2
75.6
84.7
79.8
84.1
78.5
81.4
77.4
74.9
59.5
98.5
95.8
93.9
(4.1)
(3.8)
(6.5)
(6.5)
(2.6)
(1.0)
(7.3)
(3.3)
(5.3)
(7.2)
(6.4)
(3.5)
(4.1)
(6.5)
(4.9)
(6.1)
(2.6)
(1.9)
(4.4)
Leachate
98.9
82.8
95.3
97.5
87.3
87.2
92.1
90.8
91.1
102
105
92.3
93.0
88.2
87.5
77.3
113
112
112
(19.6)
(19.3)
(16.9)
(22.3)
(18.2)
(21.7)
(21.5)
(22.4)
(27.5)
(21.5)
(20.5)
(16.1)
(15.0)
(13.2)
(18.7)
(4.2)
(14.9)
(11-7)
(14.0)
Groundwater
87.1
88.5
92.7
91.0
92.6
82.4
88.8
86.4
81.4
90.9
89.6
89.3
90.5
91.7
87.2
67.2
110
109
106
(8.1)
(15.3)
(17.1)
(10.7)
(13.7)
(4.4)
(7.5)
(5.8)
(17.6)
(7.6)
(6.1)
(3.6)
(4.9)
(15.2)
(3.7)
(8.0)
(3.3)
(3.3)
(3.8)
Estuarine
water
87.1
71.0
99.1
87.0
97.4
82.5
89.2
88.7
107
90.1
92.7
86.1
86.5
87.7
85.1
97.2
110
104
111
(7.5)
(7.7)
(19.0)
(8.0)
(15.0)
(5.5)
(2.8)
(4.9)
(16.8)
(2.4)
(5.6)
(6.2)
(6.9)
(9.6)
(8.3)
(7.0)
(12.4)
(5.9)
(5.9)
Spike level
(60 /zg/L)
Leachate
112
88.5
100
106
107
99.0
112
109
117
109
117
107
108
102
105
108
95.1
97.1
93.3
(17.5)
(17.9)
(9.6)
(17.4)
(13.3)
(13.7)
(14.2)
(14-6)
(11.4)
(20.7)
(24.7)
(15.3)
(15.1)
(14.3)
(17.7)
(17.9)
(7.2)
(7.1)
(9.5)
Groundwater
90.9 (4.5)
75.3 (3.5)
83.2 (3.3)
87.7 (2.7)
87.6 (2.9)
76.9 (6.6)
92.5 (1.8)
84.8 (5.9)
80.1 (4.1)
88.9 (2.4)
93.0 (2.0)
92.4 (0.6)
91.1 (3.0)
71.9 (2.4)
90.4 (2.0)
90.1 (1.1)
107 (2.4)
106 (2.8)
105 (2.4)
'The number of determinations was 3. Values given in parentheses are the percent relative standard
deviations of the average recoveries. The sample extracts were not subjected to Florisil cleanup.
-------
TABLE 88. ACCURACY AND PRECISION DATA FOR METHOD 3550 AND METHOD 8061*
Spike level
(i /*g/g)
EstuaHne
Compound sediment"
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
77.9
68.4
103
121
108
26.6
95.0
c
c
103
113
114
c
36.6
c
c
(42.8)
(1.7)
(3.1)
(25.8)
(57.4)
(26.8)
(10.2)
(3.6).
(12.8)
(21.1)
(48.8)
Municipal
sludge
52.1
68.6
106
86.3
97.3
72.7
81.9
66.6
114
96.4
82.8
74.0
76.6
65.8
93.3
80.0
(35.5)
(9.1)
(5.3)
(17.7)
(7.4)
(8.3)
(7.1)
(4.9)
(10.5)
(10.7)
(7.8)
(15.6)
(10.6)
(15.7)
(H.6)
(41.1)
Sandy loam
soil
c
54.7
70.3
72.6
e
0
81.9
c
57.7
77.9
56.5
c
99.2
92.8
84.7
64.2
(6.2)
(3.7)
(3-7)
(15.9)
(2.8)
(2.4)
(5.1)
(25.3)
(35.9)
(9.3)
(17.2)
Estuarine
sediment"
136
60.2
74.8
74.6
104
19.5
77.3
21.7
72.7
75.5
72.9
38.3
59.5
33.9
36.8
c
(9.6)
(12.5)
(6.0)
(3.9)
(1.5)
(14.8)
(4.0)
(22.8)
(11.3)
(6.8)
(3.4)
(25.1)
(18.3)
(66.1)
(16.4)
Spike level
(3 /ig/g)
Municipal
sludge
64.8
72.8
84.0
113
150
59.9
116
57.5
26.6
80.3
76.8
98.0
85.8
68.5
88.4
156
(11.5)
(10.0)
(4.6)
(5.8)
(6.1)
(5.4)
(3.7)
(9.2)
(47.6)
(4.7)
(10.3)
(6.4)
(6.4)
(9.6)
(7.4)
(8.6)
Sandy loam
soil
70.2 (2.0)
67.0 (15.1)
79.2 (0.1)
70.9 (5.5)
83.9 (11.8)
0
82.1 (15.5)
84.7 (8.5)
28.4 (4.3)
79.5 (2.7)
67.3 (3.8)
62.0 (3.4)
65.4 (2.8)
62.2 (19.1)
115 (29.2)
115 (13.2)
'The number of determinations was 3. Values given in parentheses are the percent relative standard
deviations of the average recoveries. All samples were subjected to Florisil cartridge cleanup.
"The estuarine sediment extract (Florisil, Fraction 1) was subjected to sulfur cleanup (Method 3660
with tetrabutylammonium sulfite reagent).
cNot able to determine because of matrix interferent.
-------
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Figure 26. Method accuracy for aqueous matrices (N, - estuarine water;
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Figure 27. Method precision for aqueous matrices (N, - estuarine water;
M2 - leachate; M3 - groundwater; C, - concentration at 20 /ig/L per
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Figure 29. Method precision for solid matrices (M1 - estuarine sediment;
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at 1 /ig/g per component; C - concentration at 3 /ig/g per
component)
-------
TABLE 89. RESULTS OF METHOD 8060 ANALYSIS FOR EPA HP-482 SAMPLE 4
Compounds known to be present
in WP-482 sample 4 at /ig/L
Phthalates found by the
revised EPA Method 8060 Percent
at /ig/L bias
1,4-Dichlorobenzene 251
Bis(2-chloroisopropyl)ether 204
Hexachloroethane 303
Nitrobenzene 373
Naphthalene 250
DMP 404
Acenaphthene 197
Fluorene 250
4-Chlorophenyl phenyl ether 374
4-Bromophenyl phenyl ether 373
Anthracene 200
Fluoranthene 301
BBP 250
Chrysene 209
Ethyl hexyl phthalate 153
Benzo(b)fluoranthene 203
Benzo(a)pyrene 224
Dibenzo(a,h)anthracene 204
Benzo(g,h,i)perylene 300
DMP
298
BBP
227
False Positives:
-26.8
-9.2
DBP
BMEP
BEEP
DHP
DEHP
8.8
10.2
20.8
27.6
78.6
"The concentrations reported are based on spiking 1 mL of the stock
solution WP-482 into 1 L HPLC-grade water. 100 pi of the stock
solution WP-482 were spiked into 1 L HPLC-grade water and extracted
with methylene chloride using EPA Method 3510 for Florisil cleanup.
The GC/ECD operating conditions were as follows: 30 m x 0.25 mm ID
DB-5 fused-silica capillary column (0.25 jwi film thickness); 120°C
to 260*C (hold 15.7 min) at 15'C/min; carrier gas: helium at 21
psi; injector temperature 275°C; detector temperature 320°C;
splitless injection. Since we used only 100 0L of the stock
solution to spike, our results have been multiplied by a factor of
10 in order to compare them with the true values reported by EPA.
149
-------
TABLE 90. RESULTS OF METHOD 8060 ANALYSIS FOR EPA WP-482 SAMPLE 1
Compounds known to be present
in WP-482 sample 1 at
Compounds found by EPA Percent
Method 8060a at ng//iLa bias
Bis(2-chloroethyl)ether 48.2
1,3-Dichlorobenzene 52.0
1,2-Dichlorobenzene 24.7
Nitrosodipropylamine 34.8
Isophorone 76.6
Bis(2-chloroethoxy)methane 48.5
1,2,4-Trichlorobenzene 25.3
Hexachlorobutadiene 49.6
2-Chloronaphthalene 25.4
2,6-Dinitrotoluene 76.5
2,4-Dinitrotoluene 73.8
DEP 25.1
Hexachlorobenzene 35.7
Phenanthrene 40.2
DBP 24.9
Pyrene 60.2
Benzo(a)anthracene 73.9
OOP 43.9
Benzo(k)fluoranthene 45.7
DEP
DBP
OOP
29.0
23.6
39.2
False Positives:
+15.5
- 5.2
-10.7
BMEP
BMPP
BEEP
DAP
HEHP
DHP
BBP
DCP
DEHP
2.8
0.35
2.9
1.9
0.94
0.50
1.2
0.74
3.2
"The EPA spiking solution WP-482 was diluted with hexane and analyzed
by GC/ECD. The GC/ECD operating conditions were as follows: 30 m x
0.25 mm ID DB-5 fused-silica capillary column (0.25 (an film
thickness); 120°C to 260'C (hold 15.7 min) at 15'C/min; carrier gas:
helium at 21 psi; injector temperature 275"C; detector temperature
320"C; splitless injection.
150
-------
TABLE 91. RESULTS OF METHOD 8060 ANALYSIS FOR EPA WP-482 SAMPLE 2
Compounds known to be present in
EPA Method WP-482 sample 2 at ng//iL
Compounds found by EPA Percent
Method 8060 at ng//zLa bias
Bis(2-chloroethyl)ether 253
1,3-Dichlorobenzene 148
1,2-Dichlorobenzene 250
Nitrosodipropylamine 352
Isophorone 149
Bis(2-ch1oroethoxy)methane 255
1,2,4-Trichlorobenzene 256
Hexachlorobutadiene 157
2-Chloronaphthalene 251
2,6-Dinitrotoluene 229
2,4-Dinitrotoluene 277
DEP 254
Hexachlorobenzene 350
Phenanthrene 202
DBP 252
Pyrene 298
Benzo(a)anthracene 315
OOP 230
Benzo(k)fluoranthene 246
DEP
DBP
OOP
216
192
200
False Positives:
-15.0
-23.8
-13.0
DMP
DIBP
BMEP
BEEP
DAP
HEHP
DHP
BBP
BBEP
DCP
DEHP
10.7
6.6
29.4
19.5
15.8
11.3
46.4
9.2
7.7
4.6
29.9
aA 6C/ECD chromatogram is shown in Figure 30. The EPA spiking
solution was diluted with hexane and analyzed by GC/ECD. The GC/ECD
operating conditions were as follows: 30 m x 0.25 mm ID DB-5
fused-silica capillary column (0.25 ion film thickness); 120*C to
260°C (hold 15.7 min) at 15'C/min; carrier gas: helium at 21 psi;
injector temperature 275eC; detector temperature 320eC; splitless
injection.
151
-------
TABLE 92. RESULTS OF METHOD 8060 ANALYSIS FOR EPA WP-482 SAMPLE 3
Compound known to be present In
WP-482 sample 3 at ng//iL
Compounds found by EPA Percent
Method 8060 at ng//iLa bias
1,4-Dichlorobenzene 24.8
Bis(2-chloroisopropyl)ether 38.8
Hexachloroethane 30.0
Nitrobenzene 76.5
Naphthalene 24.8
DMP 40.0
Acenaphthene 19.5
Fluorene 51.2
4-Chlorophenyl phenyl ether 76.7
4-Bromophenyl phenyl ether 41.5
Anthracene 40.0
Fluoranthene 29.8
BBP 51.3
Chrysene 69.9
Ethyl hexyl phthalate 29.1
Benzo(b)fluoranthene 40.0
Benzo(a)pyrene 24.9
Dibenzo(a,h)anthracene 40.7
Benzo(g,h,i)perylene 80.4
DMP
33.5
BBP
43.4
False positives:
-16.3
-15.4
DEP
BMPP
DHP
BBEP
DCP
DEHP
16.2
0.35
3.4
0.94
0.20
24.3
aA GC/ECD chromatogram is shown in Figure 31. The EPA spiking
solution was diluted with hexane and analyzed by GC/ECD. The GC/ECD
operating conditions were as follows: 30 m x 0.25 mm ID DB-5
fused-silica capillary column (0.25 /an film thickness); 120°C to
260°C (hold 15.7 min) at 15eC/min; carrier gas: helium at 21 psi;
injector temperature 275eC; detector temperature 320*C; splitless
injection.
152
-------
TABLE 93. RESULTS OF METHOD 8060 ANALYSIS FOR EPA WP-485
Compounds found by EPA
Compounds known to be present Method 8060 at ng//iLa
In the sample at ng/pl (false positives)
Acenaphthylene
Phenanthrene
Fluoranthene
Benzo(a)anthracene
Benzo(a)pyrene
Benzo(b)fluoranthene
Dibenzo( a, n) anthracene
Benzo(g,h,i)perylene
100
100
10.0
10.0
10.0
10.0
10.0
10.0
DMP
BMPP
DAP
DHP
3.6
0.14
0.09
6.4
aA GC/ECD chromatogram is shown in Figure 32. The EPA spiking
solution was diluted with hexane and analyzed by GC/ECD. The
GC/ECD operating conditions were as follows: 30 m x 0.25 mm
ID DB-5 fused-silica capillary column (0.25 /im film
thickness); 120°C to 260°C (hold 15.7 min) at 15eC/min;
carrier gas: helium at 21 psi; injector temperature 275'C;
detector temperature 320'C; splitless injection.
153
-------
TABLE 94. RESULTS OF METHOD 8060 ANALYSIS FOR WP-281 SAMPLE 2
Compounds known to be present
in WP-281 sample 2 at ng//iL
Compounds found by EPA
Method 8060 at ng//iLa
(false positives)
Phenol
2,4-Dimethylphenol
2-Chlorophenol
2-Chloro-3-methyl phenol
2,4-Dichlorophenol
2,4,6-Trichlorophenol
Pentachlorphenol
2-Nitrophenol
4-Nitrophenol
15.0
12.5
8.3
20.0
10.0
12.5
10.0
20.0
15.0
DMP
BMPP
DAP
DHP
BBP
OEHP
5.2
0.50
0.90
1.6
0.36
2.4
a A GC/ECD chromatogram is shown in Figure 33. The EPA
spiking solution was diluted with hexane and analyzed by
GC/ECD. The GC/ECD operating conditions were as follows:
30 m x 0.25 mm ID DB-5 fused-silica capillary column
(0.25 pm film thickness); 120'C to 260°C (hold 15.7 min)
at 15"C/min; carrier gas: helium at 21 psi; injector
temperature 275°C; detector temperature 320'C; splitless
injection.
154
-------
OJ
CO
Figure 30. 6C/ECD chromatogram of HP-482 Sample 2.
155
-------
en
o»
(M
ni
Figure 31. GC/ECD chromatogram of WP-482 Sample 3.
-------
in
to
CD rvi
CO
MOJ
*T O-
T in
— ro
in
Figure 32. GC/ECD chromatogram of WP-485.
-------
GO
o-oi
en
00
r
Figure 33. GC/ECD chromatogram of WP-281 Sample 2.
-------
• Make an estimate of the detection limit from the concentration value
that corresponds to an instrument signal-to-noise ratio in the range
of 5 to 10. The data in Table 95 indicate that all 16 phthalate
esters are detected in the 25-pg//iL standard. The GC/ECD chromatogram
in Figure 39 indicates that BBP is significantly larger than the other
phthalate esters. Consequently, this compound should be spiked at 10
times lower levels for MDL determination. Furthermore, DMP and DEP
should be spiked at higher concentrations because the detector
response to these compounds is low compared to DBP or DAP.
• Spike reagent water at a concentration that is in the same
concentration range as the estimated method detection limit; in this
case, the concentration is 200 ng/L for all compounds except BBP at
20 and DMP and DEP at 1,000 ng/L.
• Take seven aliquots of the spiked matrix and process each aliquot
through the entire analytical process. Make all computations
according to the protocol written for the revised Method 8060. Seven
blank measurements are also performed to determine the concentrations
of the 16 phthalate esters in the sample matrix (these values are then
subtracted from the respective sample measurements).
• Calculate the standard deviation of the seven replicate measurements
and compute the MDL as follows:
MDL ' Vl.0.99) X SD
where t,n.., ^^ is the student's t value appropriate for a 99 percent
confidence level and a standard deviation with n-1 degrees of freedom, and SD
is the standard deviation of the seven replicate measurements. The value of
t for 6 degrees of freedom is 3.143.
Table 95 presents the values found for each of the 16 phthalate esters
in a hexane blank and six calibration standards at 0.5, 1.0, 5.0, 10, 25, and
50 pg//iL. The GC/ECD chromatograms of these analyses are shown in Figures 34
through 40. The results of method blank analyses with and without Florisil
cleanup are shown in Tables 96 and 97, and the MDL data for water samples are
summarized in Tables 98 and 99.
Approximate MDLs for soil samples can be derived from these data assuming
that a 30 g soil sample is taken for extraction and the final extract volume
is 10 ml.
6.6.5 Ruggedness Testing
A ruggedness test was performed for Method 8060 to determine how
sensitive the method is to changes of the seven conditions identified in
Table 100. They include: injector temperature, detector temperature,
injection volume, time set up for the injector during which all sample is
transferred to the fused-silica capillary column (splitless time), type of
solvent, and matrix interferences from diesel hydrocarbons and chlorinated
organochlorine pesticides. The seven conditions are assigned the letters A
through G, and the altered values are assigned the same letters in the lower
159
-------
TABLE 95. ESTIMATION OF THE INSTRUMENT DETECTION LIMIT
Concentration (pg//zL)a
Retention
time
Compound (min)
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
3
4
6
7
7
7
8
8
8
9
9
10
11
11
13
16
.422
.465
.496
.161
.980
.417
.433
.195
.656
.638
.715
.560
.157
.011
.067
.049
Hexane
blank
--
--
--
--
--
--
35
--
22
7.0
--
6.2
3.4
--
• -
Std
0.5
--
--
4.
--
--
--
33
--
25
8.
--
9.
4.
.-
- *
2
6
6
4
Std
1.0
--
--
13
9.2
19
--
50
--
39
10.3
--
14
16
-.
• ~
Std
5.0
--
2.3
10
8.1
7.8
15
38
8.1
31
14
8.8
14
10
..
• ~
Std
10
20
22
29
20
30
15
42
21
30
19
15
24
18
26
- "
Std
25
54
54
22
29
12
28
20
40
30
43
32
40
35
30
31
23
Std
50
137
84
69
58
52
59
41
48
47
53
53
61
60
53
67
75
aThe value given was obtained by calibrating against a
50 pg//iL standard analyzed prior to the seven analyses shown
in this table. Reproducibility of the injection technique
is 2.87 percent as determined from the absolute response of
the internal standard. The five standards (1 to 50 pg//xL)
were obtained from a 100-pg//*L standard by appropriate
dilutions; the 0.5-pg/juL standard was obtained from the
50-pg/jiL standard. The GC/ECD operating conditions were as
follows: 30 m x 0.25 mm ID DB-5 fused-silica capillary
column (0.25 pm film thickness); 120°C to 260eC (hold
15.7 min) at 15°C/min; carrier gas: helium at 21 psi;
injector temperature 275°C; detector temperature 320*C;
splitless injection.
160
-------
-------
CD
0>
oo
in
to
ru
111 CO
111
0.
I/I
ecu
Figure 35. 6C/ECD chromatogram of composite phthalate esters standard at
0.5 pg/ML analyzed on a 30 m x 0.25 mm ID DB-5 fused-silica
capillary column; helium at 20 ps1; temperature program: 120°C
to 260°C (15.7 m1n hold) at 15°C/min, Injector temperature
275°C; detector temperature 320°C; range 1; attn 8.
162
-------
ID
00
(M
ru
o
blO
Ul
O.
I/I
ecu)
«l-
If-'
Figure 36. GC/ECD chromatogram of composite phthalate esters standard at
1 pg//iL analyzed on a 30 m x 0.25 mm ID DB-5 fused-sllica
capillary column; helium at 20 ps1; temperature program: 120°C
to 260°C (15.7 min hold) at 15°C/m1n, Injector temperature
275°C; detector temperature 320°C; range 1; attn 8.
163
-------
00
CD
o-
(M
CD
o
bid)
Ul
0.
UI
Kill
Figure 37. GC/ECD chromatogram of composite phthalate esters standard at
5 pg//*L analyzed on a 30 m x 0.25 mm ID DB-5 fused-silica
capillary column; helium at 20 psi; temperature program: 120°C
to 260°C (15.7 min hold) at 15°C/min, Injector temperature
275°C; detector temperature 320°C; range 1; attn 8.
164
-------
Kill
Figure 38. GC/ECD chromatogram of composite phthalate esters standard at
10 pg//iL analyzed on a 30 m x 0.25 mm ID DB-5 fused-sillca
capillary column; helium at 20 ps1; temperature program: 120°C
to 260°C (15.7 m1n hold) at 15°C/min, Injector temperature
275°C; detector temperature 320°C; range 1; attn 8.
165
-------
•o
CD
r-
VI
CD
10
Oil
Figure 39. GC/ECD chromatogram of composite phthalate esters standard at
25 pg//iL analyzed on a 30 m x 0.25 mm ID DB-5 fused-silica
capillary column; helium at 20 psi; temperature program: 120°C
to 260°C (15.7 min hold) at 15eC/min, injector temperature
275°C; detector temperature 320°C; range 1; attn 8.
166
-------
K)
CO
in
(M
m
m
o
UJOD
UJ
0.
tn
(Kbl
Figure 40. GC/ECD chromatogram of composite phthalate esters standard at
50 pg/^L analyzed on a 30 m x 0.25 mn ID DB-5 fused-silica
capillary column; helium at 20 psi; temperature program: 120°C
to 260°C (15.7 min hold) at 15°C/min, injector temperature
275°C; detector temperature 320°C; range 1; attn 8.
167
-------
TABLE 96. RESULTS OF THE METHOD BLANK ANALYSES FOR THE MDL STUDY
FOR WATER SAMPLES'
Concentration (ng/L)
Compound Rep.l Rep.2 Rep.4 Rep.5 Rep.6 Rep.7 Average ± SD
DMP
DEP
DIBP
DBP
BMPP
BMEP
BMPP
DAP
BEEP
DAP
HEHP
DHP
BBP
BBEP
DEHP
DCP
DOP
DNP
DPTP (percent
recovery)
130 140 120 56 150 119 ± 37
400 160 110 220 57 170 186 ± 119
110 130 110 110 120 95 113 ± 12
170 100 75 82 82 84 99 ± 36
93 86 70 85 82 75
aHPLC-grade water lot 880930 from Fisher Scientific. One-liter aliquots
were extracted 3 times with 60 mL methylene chloride (American Burdick
& Jackson, lot AR 348), and solvent-exchanged with hexane (American
Burdick & Jackson, lot AQ 406). Vextract was 10 mL. If no value is
given, then the compound was not detected by GC/ECD. No Florisil
cleanup was used. The GC/ECD operating conditions were as follows:
30 m x 0.25 mm ID DB-5 fused-silica capillary column (0.25 tun film
thickness); 120'C to 260°C (hold 15.7 min) at 15eC/min; carrier gas:
helium at 21 psi; injector temperature 275°C; detector temperature
320'C; splitless injection.
168
-------
TABLE 97. METHOD DETECTION LIMIT STUDY -- FLORISIL DISPOSABLE
CARTRIDGES METHOD BLANKS
Concentration (ng/cartridge)a
Compound Rep.l Rep.2 Rep.3 Rep.4 Rep.5 Average SD
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
412
192
94
129
60
303
28
69
43
32
16
30
295
<10
<10
<10
.7
.0
.4
.2
.9
.9
.6
.4
411
76.2
96.6
116
73.3
134
40.3
61.9
16.2
29.3
16.1
29.8
267
<10
<10
<10
466
155
107
142
106
302
140
65
40
31
16
42
332
<10
<10
<10
.3
.3
.6
.6
.2
495
228
119
162
152
291
33.6
64.2
12.9
35.8
24.3
59.0
320
18.2
13.8
<10
496
144
107
132
51
282
22
71
4
33
18
43
243
<10
<10
<10
.9
.0
.2
.7
.2
.2
.1
456
159
105
136
88
262
52
66
23
32
18
40
291
<10
<10
<10
.6
.9
.4
.6
.6
.4
.9
42
66
10
17
41
72
49
4
17
2
3
12
37
--
--
™ ~
8 Each Florisil disposable cartridge (Supelco, Inc.) was
eluted with 5 mL of 20 percent methylene chloride in
hexane (Fraction 1) which was discarded, followed by S ml
of hexane/acetone (9:1) which was concentrated to 1 mL and
analyzed by GC/ECD. The GC/ECD operating conditions were
as follows: 30 m x 0.25 mm ID DB-5 fused-silica capillary
column (0.25 pm film thickness); 120eC to 260'C (hold
15.7 min) at 15°C/min; carrier gas: helium at 21 psi;
injector temperature 275'C; detector temperature 320°C;
splitless injection.
169
-------
TABLE 98. METHOD DETECTION LIMITS FOR MATER SAMPLES NOT SUBJECTED TO
FLORISIL CARTRIDGE CLEANUP
Compound
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
Spike
level
(ng/L)
1,000
1,000
200
200
200
200
200
200
200
200
20
200
200
200
200
200
Concentration
Rep.l
164
113
63.0
155
b
154
7.3
234
262
188
67.4
101
90.0
37.0
c
14.4
Rep. 2
143
107
61.0
163
b
127
33.2
256
220
173
60.8
72.4
97.2
37.2
c
16.0
Rep. 3
230
167
53.4
382
b
216
41.6
181
182
178
82.8
204
a
61.2
78.8
b
Rep. 4
140
94.4
42.2
89.4
b
50.2
10.7
63.8
230
145
58.0
106
114
34.4
C
C
(ng/L)a
Rep. 5
262
145
42.8
90.8
b
113
13.1
148
187
149
73.8
158
134
33.8
c
c
Rep. 6
210
111
39.8
216
b
220
25.0
80.0
200
150
92.4
142
b
44.2
c
b
Rep. 7
310
138
57.0
246
b
228
47.0
130
197
212
166
137
164
95.8
67.0
64.4
Average
208
135
51.3
192
b
158
25.4
156
183
171
85.9
131
120
49.1
72.9
31.6
SD
64
25.6
9.6
102
--
66.8
15.7
72.7
76.9
24.7
37.5
43.0
30.1
22.7
8.3
28.4
MDL
200
80
30
320
--
210
49
230
240
78
120
140
95
71
26
89
aThe values given were not corrected for the blank. The GC/ECD operating conditions
were as follows: 30 m x 0.25 nun ID DB-5 fused-silica capillary column (0.25 urn film
thickness); 120*C to 260eC (hold 15.7 min) at 15°C/min; carrier gas: helium at 21 psi;
injector temperature 275°C; detector temperature 320*C; splitless injection.
bCompound not quantitated because of interference.
°Not detected.
-------
TABLE 99. METHOD DETECTION LIMITS FOR HATER SAMPLES SUBJECTED TO FLORISIL CARTRIDGE CLEANUP
Concentration (ng/L)a
Compound
DMP
DEP
DIBP
OBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
DOP
DNP
level
(ng/L)
1,000
1,000
200
200
200
200
200
200
200
200
20
200
200
200
200
200
Rep.l
650
450
310
54
65.6
696
21.6
44.2
125
141
16.0
174
622
25.7
c
8.82
Rep. 2
496
416
294
44
111
448
62.6
61.0
119
130
13.6
172
512
20.5
c
c
Rep. 3
458
370
284
296
226
508
87.2
209
211
96.4
50.4
198
b
42.5
44.4
b
Rep. 4
292
334
276
74
406
692
127
32.2
100
94.4
14.0
152
390
25.3
c
13.96
Rep. 5
800
498
200
84
154
930
80.2
51.2
167
124
30.0
222
518
28.7
c
12.5
Rep. 6
776
384
314
276
67.0
558
98.6
213
140
121
33.8
154
b
25.9
20.4
b
Rep. 7
836
250
292
132
188
550
115
189
90.2
83
21.0
152
554
25.9
15.4
25.2
Average
615
387
282
137
174
626
84.6
114
136
113
25.5
175
520
27.8
26.7
15.1
SD
205
79.6
38.4
106
118
162
35
84.5
41.5
21.5
13.5
26.6
84.4
6.93
15.5
7.06
MDL
640
250
120
330
370
510
110
270
130
68
42
84
270
22
49
22
aThe values given were not corrected for the blank. The GC/ECD operating conditions
were as follows: 30 m x 0.25 mm ID DB-5 fused-silica capillary column (0.25 /im film
thickness); 120eC to 260°C (hold 15.7 min) at 15°C/min; carrier gas: helium at 21 psi;
injector temperature 275°C; detector temperature 320°C; splitless injection.
bNot able to quantitate because of interference.
cNot detected.
-------
TABLE 100. CONDITIONS VARIED AND ASSIGNED VALUES FOR GAS CHROMATOGRAPHIC
ANALYSIS (METHOD 8060) FOR RUGGEDNESS TEST
Condition
Injector temperature (°C)
Detector temperature (°C)
Injection volume (/iL)
Splitless time (sec)
Solvent
No.
1
2
3
4
5
Letter
A, a
B,b
C,c
D,d
E,e
Value for
capital
letter
300
350
3
60
Hexane
Value for
lower case
letter
250
300
1
30
Hexane/ethyl ether
Interferences from matrix
(diesel hydrocarbons)
Interferences from matrix
(chlorinated pesticides)
F,f
G,g
(80:20)
With diesel Without
hydrocarbons
at 20 mg/mL
With
chlorinated
pesticides
at 0.5 ng//iL
Without
172
-------
case (a through g). For example, the injector temperature is 300°C in the
experiments 1 through 4 and 250°C in the experiments 5 through 8. Table 101
presents the design for the test of experimental conditions for the eight
experiments.
Tables 102 and 103 present the analytical results. The group differences
VA through VG were calculated using equations 1 through 7.
VA = l/4(s + t + u + v) -l/4(w + x + y + z) = A-a (1)
VB = l/4(s + t + w + x) -l/4(u + v + y + z) = B-b (2)
Vc = l/4(s + u + w + y) -l/4(t + v + x + z) = C-c (3)
VD = l/4(s + t + y + z) -l/4(u + v + w + x) = D-d (4)
VE = l/4(s + u + x + z) -l/4(t + v + w + y) = E-e . (5)
VF = l/4(s + v + w + z) -l/4(t + u + x + y) = F-f (6)
VG = l/4(s + v + x + y) -l/4(t + u + w + z) = G-g (7)
It appears from the data presented in Table 103 that only certain
compounds are affected by changes of the various parameters in the GC
procedure. DAP seems to be affected the most, followed by BMEP and the
internal standard. This is not unexpected since diesel fuel components and
the organochlorine pesticides overlap with these compounds, making their
identification difficult, if not impossible. We have noted during the
performance of the ruggedness test that the detector sensitivity decreased
drastically after the diesel- or the organochlorine pesticide-containing
working solutions were analyzed. It is therefore imperative that such
interferents be eliminated prior to the gas chromatographic analysis,
otherwise the data obtained for samples containing such interferences are
unreliable.
6.6.6 Confirmation by GC/MS
Table 104 gives the retention times (as scan numbers) for 16 phthalate
esters, benzyl benzoate, and 5 other compounds proposed as surrogate compounds
in the revised Method 8060. A GC/MS chromatogram of a composite standard
containing 16 phthalate esters is shown in Figure 41. Mass spectra in both
the plot and list format are included 1n Appendix C of this final report. The
concentration of the composite standard is 10 ng//iL per compound. The
sensitivity of the GC/MS instrument is estimated to approximately 1 to 2 ng
per compound.
173
-------
TABLE 101. DESIGN FOR RUGGEDNESS TEST OF
EXPERIMENTAL CONDITIONS
Values of conditions
in determination
No.
Experimental
condition 12345678
1 AAAAaaaa
2 BBbbBBbb
3 CcCcCcCc
4 DDddddDD
5 EeEeeEeE
6 F f f F F f f F
7 GggGgGGg
174
-------
TABLE 102. RUGGEDNESS TEST FOR METHOD 8060 -- CONCENTRATIONS OF
TEST COMPOUNDS FOUND FOR EACH EXPERIMENT
Concentration (/ig/mL)
Compound
DMP
DEP
BB-IS
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
DPTP
OOP
DNP
Exp
1
2
0
9
5
49
0
0
0
0
4
4
57
2
2
5
2
2
1
.77
.81
.93
.07
.99
.8
.773
.534
.247
.927
.11
.39
.3
.98
.63
.79
.22
.81
Exp 2
6.24
5.24
10.5
8.40
7.86
8.66
8.58
7.88
7.16
8.76
9.30
9.56
7.86
7.26
7.46
10.3
7.52
6.58
Exp 3
2.47
2.16
3.74
2.61
2.08
3.40
3.97
2.11
3.26
3.69
2.60
4.06
3.04
3.32
4.15
4.61
3.33
3.26
Exp
1
1
224
5
1
96
300
0
4
0
0
6
88
3
2
7
2
2
4
.87
.04
.18
.92
.2
.062
.30
.054
.052
.60
.2
.38
.28
.90
.62
.42
Exp 5
4.18
4.14
1.61
0.483
0.654
0.068
0.798
0.268
2.74
2.97
2.42
4.27
2.72
3.12
3.40
4.68
2.57
2.75
Exp 6
7.
9.
258
12.
17.
172
360
828
7.
5.
17.
6.
162
7.
7.
3.
5.
5.
86
58
6
1
20
54
3
38
22
82
56
50
88
Exp 7
3.49
5.81
185
3.06
4.40
67.3
174
285
3.84
1.83
5.33
4.09
68.0
4.46
5.21
5.06
4.72
4.81
Exp 8
0.266
2.56
6.52
0.218
2.92
20.2
6.52
3.06
2.46
0.136
1.54
6.30
3.18
3.20
2.16
7.14
3.78
2.86
175
-------
TABLE 103. RUGGEDNESS TEST FOR METHOD 8060 -- GROUP DIFFERENCES FOR
THE TEST COMPOUNDS
Condition
Compound
DMP
DEP
BB-IS
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
DPTP
OOP
DNP
(1)
-0.86
-2.71
-52.97
2.22
-1.81
-25.38
-57.00
-276.44
-0.32
0.74
-2.63
0.89
-19.88
-0.27
-0.52
2.04
-0.22
-0.31
(2)
2.99
2.55
-37.03
4.87
5.07
10.86
-28.58
136.61
0.87
3.12
5.90
0.89
16.87
1.56
1.88
-0.09-
0.84
1.17
(3)
-1.08
-0.88
-76.91
-2.79
-4.17
-44.12
-123.89
-137.77
-2.76
-1.27
-3.43
-3.01
-32.55
-1.80
-1.08
-2.19
-1.65
-1.03
(4)
-1.15
-0.13
-71.13
-0.03
-0.15
-31.43
-118.72
-133.49
-0.95
-0.15
-0.52
0.76
-29.91
0.22
-0.05
1.89
1.06
0.69
(5)
-0.85
0.22
-38.01
1.84
3.31
18.29
-28.03
135.12
-1.22
-0.83
2.11
-0.85
14.69
-0.38
-0.40
-1.71
-0.65
-0.44
(6)
-2.99
-3.06
-56.07
-2.93
-4.99
-21.27
-59.61
-279.77
-2.93
-3.93
-6.60
-0.63
-22.38
-2.40
-3.54
0.49
-2.47
-2.42
(7)
0.46
1.29
161.42
4.55
3.97
88.24
203.73
275.07
-0.01
-1.80
2.73
-0.68
89.68
0.28
0.19
-1.11
-0.53
0.12
176
-------
TABLE 104. RETENTION TIMES (SCAN NUMBERS) AND THREE MOST INTENSE PEAKS IN
THE MASS SPECTRA OF METHOD 8060 COMPOUNDS"
Compound Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
name
DMP
DEP
DIBP
DBP
BMPP
BMEP
DAP
BEEP
HEHP
DHP
BBP
BBEP
DEHP
DCP
OOP
DNP
BB (IS)
DPP (SU)
DPIP (SU)
DPTP (SU)
DBZP (SU)
DOIP (SU)
Scan
number
1128
1264
1499
1573
1662
1602
1714
1687
1737
1844
1851
1928
1973
1962
2082
2191
1397
1959
2057
2061
2087
2127
Mass spectrum
m/z (relative intensity)
163 (100),
149 (100),
149 (100),
149 (100),
149 (100),
59 (100),
149 (100),
45 (100),
149 (100),
149 (100),
149 (100),
57 (100),
149 (100),
149 (100),
149 (100),
149 (100),
105 (100),
225 (100),
225 (100),
225 (100),
107 (100),
167 (100),
77 (21),
177 (23),
57 (28),
41 (9.2),
85 (66),
58 (73),
43 (19),
72 (94),
57 (19),
43 (21),
91 (69),
56 (82),
57 (39),
167 (34),
43 (14),
43 (18),
91 (43),
77 (39),
76 (27),
104 (44),
91 (81),
279 (41),
164 (10)
150 (12)
41 (13)
150 (8.9)
43 (66)
149 (13)
150 (8.8)
73 (92)
55 (12)
41 (8.4)
206 (26)
45 (80)
167 (36)
55 (24)
57 (12)
150 (9.8)
77 (28)
226 (15)
104 (23)
76 (20)
149 (70)
149 (21)
aThe GC/MS operating conditions are given in Section 5.7.
177
-------
DATA: E3TERSTD »78
CALIi ESTERSTD #2
RIC DATA: ESTERSTD »78 SCANS 300 TO 2S80
82/02/88 23:84:00
S APPLE:
COHDS.:
MCE: G 1,2600 LABEUl N 8, 4.9 QUAN: A 0, 1.0 J 8 BASE: U 20, 3
RIC
9424.
590
8:29
1990
16:40
1509
23:09
2960
33:20
2589
41:40
SCON
Tins
Figure 41. GC/NS chromatogram of composite standard containing the 16
phthalate esters listed in Table 104. The GC/MS operating
conditions are given in Section 5.7.
178
-------
REFERENCES
1. Test Methods for Evaluating Solid Waste, Vol. IB: Laboratory Manual
Physical/Chemical Methods, SW-846, 3rd Edition, U.S. Environmental
Protection Agency, Washington, DC, November 1986.
2. Michael, P. R., W. J. Adams, A. F. Werner, and 0. Hicks," Surveillance
of Phthalate Esters in Surface Waters and Sediments in the United
States," Environ. Toxicol. Chem. 3:377-389, 1984.
3. Russell, D. J., and B. McDuffie, "Analysis of Phthalate Esters in
Environmental Samples: Separation from PCBs and Pesticides Using Dual
Column Liquid Chromatography," Intern. J. Environ. Anal. Chem.
15:165-183, 1983.
4. Thuren, A., "Determination of Phthalates in Aquatic Environmentals,"
Bull. Environ. Contam. Toxicol. 36:33-40, 1986.
179
-------
APPENDIX A
SINGLE-LABORATORY EVALUATION OF
METHOD 8060, "PHTHALATE ESTERS"
LITERATURE REVIEW
A-l
-------
CONTENTS
1. Introduction 1
2. Phthalate Ester Structures, Physico-Chemical Properties,
Production and Uses 2
2.1 Phthalate Ester Structures and Physico-Chemical Properties 2
2.2 Production and Uses 2
3. Analytical Methodologies for Phthalate Ester Determination . . 8
3.1 Sample Preservation 8
3.2 Extraction 10
3.2.1 Extraction of Water Samples 10
3.2.2 Extraction of Sediment and Soil Samples 15
3.3 Sample Extract Cleanup 18
3.3.1 Liquid-Liquid Partitioning 18
3.3.2 Gel Permeation Chromatography 18
3.3.3 Sulfur Removal 22
3.3.4 Liquid-Solid Chromatography 22
3.3.5 Selection of Cleanup Technique 26
3.4 GC Analysis 26
3.4.1 Gas Chromatographic Columns 27
3.4.2 Problems With Gas Chromatography 34
3.5 High-Performance Liquid Chromatography 39
3.5.1 HPLC Methods 39
3.6 Confirmation of Compound Identity 46
3.7 Background Contamination 48
3.8 Stability of Phthalate Esters 61
References 62
-------
FIGURES
Number Page
1 Combined alumina-silica gel chromatographic cleanup
technique 25
2 GC/FID chromatogram of phthalate esters analyzed on a
15 m x 0.53 mm ID DB-1 fused-silica capillary column . . 31
3 GC/FID chromatogram of phthalate esters analyzed on a
15 m x 0.53 mm ID SPB-5 fused-silica capillary column . . 32
4 GC/FID chromatogram of phthalate esters analyzed on a
30 m x 0.25 mm ID DB-1301 fused-silica capillary
column 33
5 ECD response as a function of phthalate ester molecular
weight 37
6 Normal-phase HPLC separation of phthalate esters .... 41
7 Reversed-phase HPLC separation of phthalate esters ... 42
8 High-performance GPC separation of phthalate esters ... 43
9 C18-reversed-phase HPLC separation of phthalate esters
using methanol-water 45
10 Retention time of the phthalate esters as a function of
alkyl chain length 45
11 Phthalate ester fragmentation scheme 49
12 Electron impact mass spectra of common phthalate
esters 49
iv
-------
TABLES
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
The chemical structures and nomenclatures of the phthalate
esters
Physical properties of phthalate esters
Plasticizer production in the United States
Summary of analytical methodologies for phthalate
esters
Summary of extraction techniques for water samples . . .
Summary of spike recovery data for the microextraction
technique
Summary of duplicate analyses for the microextraction
technique
Summary of the extraction techniques for soils and
sediments
Liquid-Liquid solvent partitioning systems
Summary of GPC techniques
GPC elution volumes of the phthalate esters
Summary of cleanup procedures with Florisil
Instrumental detection limits for phthalate esters . . .
GC columns and conditions reported for the analysis of
phthalate esters
GC conditions and retention data for the phthalate
esters
ECD responses at 320°C
ECD responses at 255°C
HPLC conditions and retention data for the phthalate
esters
Pac
3
6
7
9
11
16
16
17
19
20
21
23
27
28
35
38
38
40
-------
TABLES (CONCLUDED)
Number Page
19 Summary of conditions for normal-phase HPLC separations
of phthalate esters 44
20 Identities and concentrations of organic compounds in
solvents by GC/MS 51
21 Identities and concentrations of organic compounds in
solvents by SIM 52
22 Concentrations of DBP and DEHP in organic solvents
and waters 53
23 Concentrations of DBP and DEHP in solid reagents .... 54
24 Concentrations of DBP and DEHP in various materials ... 55
25 Cleanup procedures used on reagents, glassware, and other
laboratory items 57
26 Component losses after evaporation-reconstitution of
100-ng//iL and 10-ng//tL standard solutions 60
27 Bis(2-ethylhexyl) phthalate loss as a function of storage
conditions and storage time 61
VI
-------
SECTION 1
INTRODUCTION
The Resource Conservation and Recovery Act (RCRA) of 1976 and its
elements require the Environmental Protection Agency (EPA) to regulate
hazardous waste activities. Implementation and enforcement of RCRA requires
analytical methodologies that will provide reliable data. The document "Test
Methods for Evaluating Solid Waste, Office of Solid Waste Manual SW-846" (1),
revised recently, provides a compilation of methods for evaluating RCRA solid
wastes for environmental and human health hazards. The SW-846 Method 8060
addresses the determination of phthalate esters.
This report presents a literature review pertinent to this study.
This literature review was performed using the computerized Chemical Abstracts
search and several EPA reports dealing with the analysis of organic compounds
in water. Furthermore, recent issues of Analytical Chemistry, the Journal of
Chromatography. the Journal of Chromatographic Science, the Association of
Official Analytical Chemists Journal. and Environmental Science and Technology
were searched for information that had not yet been entered in the computer
data bases.
The computer searches were performed using DIALOG. Chemical Abstracts
files were searched back to 1977 for all references containing "phthalate
esters," "gas chromatography," "extraction," and "cleanup." Eighty articles
that were judged to be scientifically relevant to the objectives of this study
were retrieved for the literature review.
The literature review summary that is presented in this report
addresses the following:
• Sample preservation techniques
• Extraction techniques for water, soil, and sediment samples
• Sample extract cleanup techniques
• Gas Chromatographic analysis (columns, retention time information,
Chromatographic problems)
• Compound confirmation
• Background contamination by phthalate esters.
-------
SECTION 2
PHTHALATE ESTER STRUCTURES, PHYSICO-CHEMICAL PROPERTIES,
PRODUCTION AND USES
2.1 PHTHALATE ESTER STRUCTURES AND PHYSICO-CHEMICAL PROPERTIES
Phthalate esters or phthalic acid esters are esters of phthalic acid
or 1,2-benzene dicarboxylic acid.
The chemical structures, common names, Chemical Abstracts Registry
Numbers (CAS), and alternate names for the 17 phthalate esters proposed for
evaluation in this study are given in Table 1. Their physico-chemical
properties are listed in Table 2.
In general, these compounds have low vapor pressures, low water
solubilities, and high boiling points. They are colorless, odorless, viscous
liquids under ambient conditions. As the lengths of the ester chains
increase, the phthalate ester volatility decreases. The branching of the
ester chain has the opposite effect on volatility.
Chemical hydrolysis of the esters to the carboxylic acid and the
alcohol in the presence of acid or base has been reported to occur (2). Acid
hydrolysis is usually reversible, whereas alkaline hydrolysis is irreversible.
Hydrolytic half-lives of 80 to 4000 days for a series of dialkyl phthalates
at pH 8 and 30"C were reported (3). Photolytic degradation of the phthalate
esters does not seem to be a major degradation pathway. Gledhill et al. (4)
reported no significant decrease in BBP concentrations in water after 28 days
for samples exposed to sunlight, as opposed to controls kept in the dark.
2.2 PRODUCTION AND USES
Phthalate esters are synthesized from phthalic anhydride which is made
either from naphthalene or ortho-xylene by oxidation in the presence of
vanadium pentoxide. The anhydride is further esterified with an alcohol at
high temperature or in the presence of sulfuric acid.
The annual production of phthalate esters in the United States amounted
to more than 2 x 109 Ib in 1978 (5). Although an accurate estimate of their
uses is not available, most of the phthalate esters produced are used as
plasticizers for polymers. Among the various plasticizers produced in the
United States, the phthalate esters represent approximately 60 percent of the
total amount produced (Table 3). The main uses of the plastics containing
phthalates are in building and construction (e.g., wire and cable, flooring,
swimming pool liners), home furnishings (e.g., furniture upholstery, wall
coverings, housewares), cars, wearing apparel, food wrapping and closures,
-------
TABLE 1. THE CHEMICAL STRUCTURES AND NOMENCLATURES OF THE PHTHALATE ESTERS
Compound name
Chemical structure
Chemical
Abstracts
Registry no.
Alternate names
Dimethyl phthalate
O
II
C
II
o
O —CH3
O —CH3
131-11-3 DMP
l,2-Benzened1carboxy!1c add,
dimethyl ester
Phthallc add dimethyl ester
Methyl phthalate
Diethyl phthalate
O
II
C
. 0 — CH2CH3
— C — O — CH2CH3
O
84-66-2 DEP
l,2-Benzened1carboxy!1c add,
d1ethyl ester
Ethyl phthalate
D1-n-butyl
phthalate
C
II
O
O—(CH2)3CH3
O — (CH2) CH3
84-74-2 DBP
o-Benzenedlcarboxyllc acid,
d1butyl ester
01 butyl phthalate
Benzene-o-d1carboxyl1c add,
d1-n-butyl ester
n-Butyl phthalate
Dlamyl phthalate
O
II
C
C
o
131-18-0 DAP
o-Benzened1carboxyl1c add,
O—(CH2)4CH3 d1-n-penty1 ester
D1-n-pentyl phthalate
O—(CH2)4CH3 Any 1 phthalate
Pentyl phthalate
Dihexyl phthalate
O
II
C
C
II
o
84-75-3 DHP
o-Benzened1carboxyl1c add,
O — (CH2)5CH3 d1 -n-hexyl ester
D1-n-hexyl phthalate
•O—
-------
TABLE 1. (continued)
Compound name
Chemical structure
Chemical
Abstracts
Registry no.
Alternate names
D1-n-octyl
phthalate
— C — O — (CH2)?CH3
117-84-0 OOP
o-Benzened1carboxy!1c add,
dloctyl ester
n-01octyl phthalate
Octyl phthalate
Dloctyl-o-benzened1-
carboxylate
Dlnonyl phthalate
t
I
Dldecyl phthalate
1
1
Butyl benzyl
phthalate
Hexyldecyl
phthalate
Olcyclohexyl
phthalate
DUsobutyl
phthalate
0
^ it
^^ — c
f » 1
lk_^J — c
^•"^ II
o
0
rJ^]-c
M-s
o
o
0-c
— c
II
o
o
0 — c
— c
II
0
o
\
o
\
°*°\
<
o
— c
II
o
O — (CH laCH
2>8 3
— 0 (CH2)8CH3
— 0 — (CH2)gCH3
— 0
-------
TABLE 1. (concluded)
Compound name
DUsohexyl
phthalate
Chemical
O
0-c -
— c —
II
o
structure
a
- O CHCH3
- O — CHCH3
1
(CH2)3CH3
Chemical
Abstracts
Registry no. Alternate names
DIHP
o-Benzened1carboxy!1c add,
d1-(2-n-hexyl) ester
D1-1sohexyl phthalate
Isohexyl phthalate
Dlisooctyl
phthalate
O
0-c
— c
II
0
(CH2)5 CH3
— O — CHCH3
— O — CHCH3
1
(CH2)5CH3
27554-26-3 DIOP
o-Benzened1carboxy!1c add.
d1-(2-n-octyl) ester
01-1sooctyl phthalate
Isooctyl phthalate
Olisononyl
phthalate
O
II
C
c
II
o
(CH2)eCH3
O —CHCH3
O CHCH3
I
28553-12-0 OINP
o-Benzened1carboxy11c add,
d1(2-n-nonyl) ester
01-1sononyl phthalate
Isononyl phthalate
Ollsodecyl
phthalate
O
II
— C — O —CHCH3
c
II
o
O —CHCH3
I
26761-40-0 DIOP
o-Benzened1carboxy!1c add,
d1-(2-n-decyl) ester
D1-1sodecyl phthalate
Isodecyl phthalate
-------
TABLE 2. PHYSICAL PROPERTIES OF PHTHALATE ESTERS"
Phthalate
Dimethyl
Diethyl
Oi-n-butyl
D i amy 1
Dihexyl
Bis(2-ethylhexyl)
Oi-n-octyl
Dinonyl
Didecyl
Butyl benzyl
Hexyldecyl
Dicyclohexyl
Diisobutyl
Diisohexyl
Diisooctyl
Diisononyl
Diisodecyl
Melting
Molecular point
weight "C
194b
222b
278C
306
334
390b
390b
418
446
312d
390
330
278
334
390
418
446
Oe
-40. 5b
-35"
<-55m
-50b
-25b
-35'
58 to 65m
<-50m
-48m
Boiling
point
'C (torr)
282 (760)e
298 (760)e
340 (760)b
342m
386.9 (5)b
220 (4)b
377 (760)'
212 to 218m
295 to 298m
228 to 239m
255 (5)m
Log
octanol /water
Vapor pressure Water solubility partition
(torr) (mg/L) coefficient
<0.01 (20'C)b 5000 (20'C)d 2.12i'k
4000 (32'C)h
4320 (25'C)1
0.05 (70'C)b 1000 (32'C)h 3.22''k
896 (25'C)1
0.1 (115'C)b 13 (25'C)1 5.2''"'1
1.9 x 10-3" 0.24" 5.65 to 5.93"
<0.01 (20T)b 0.4 (25'C)1 6.7^
2 x 10-7 (20'C)9 1.38 5.31
<0.2 (150'C)b 3 (25'C)1 3.2^
2.98 5. si."
4.8'
7.5 x 10-4 0.09"
7.2 x 10-5" 0.2"
1.19"
•Data taken from References 7,8,9.
"Patty 1963 cited in Reference 7.
eWeast 1977 cited in Reference 7.
dFishbein and Albro 1972 cited in Reference 7.
dVerschueren 1977 cited in Reference 7.
' EPA-OHMIADS cited in Reference 7.
siilrzy et al. 1978 cited in Reference 7.
hPeakall 1975 cited in Reference 7.
1 Determined by Wolfe et al. 1979 cited in Reference 7.
i Calculated as per Leo et al. 1971 cited in Reference 7.
"Calculated by Wolfe et al. 1979 cited in Reference 7.
1 Value does not include a correction for the influence of molecular folding because data are not
available; cited in Reference 7.
mData taken from Reference 8.
"Data taken from Reference 9.
-------
TABLE 3. PLASTICIZER PRODUCTION IN THE UNITED STATES3
Production
Plasticizer (x 106 Ib)
Phthalic acid esters -- total 1,259
Dioctyl phthalateb 409
Diisodecyl phthalate 171
Diethyl phthalate 22
Dibutyl phthalate 17
n-Hexyl-n-decyl phthalate 16
Dimethyl phthalate 10
Phosphoric acid esters (cyclic) -- total 99
Trimellitic acid esters -- total 33
Trioctyl trimellitate 16
Adipic acid esters -- total 68
Di-(2-ethylhexyl)adipate 45
Epoxidizer soya esters -- total 114
All others 513
Total production -- all plasticizers 2,086
aData taken from Reference 5.
bMostly as bis(2-ethylhexyl) phthalate.
-------
medical tubing, etc. Phthalate esters used otherwise are found in pesticide
carriers, oils, and insect repellent formulation (6).
-------
SECTION 3
ANALYTICAL METHODOLOGIES FOR PHTHALATE ESTER DETERMINATION
A summary of published methods for the determination of phthalate
esters in water, wastewater, soils, etc. is presented in Table 4. The
detection methods in this table, gas chromatography with flame ionization or
electron capture detection, gas chromatography/mass spectrometry, and stable
isotope dilution gas chromatography/mass spectrometry vary in sensitivity,
selectivity, complexity, degree of evaluation, ease of operation, etc. A
detailed discussion of each of these techniques follows. Sample preservation
and. the isolation of phthalate esters from water, soil, and sediment samples,
are discussed first. Sample extract cleanup and analysis techniques are
discussed next, followed by a discussion of the extent of background
contamination by phthalate esters.
3.1 SAMPLE PRESERVATION
The importance of proper sample preservation must be emphasized. The
choice of the preservation method depends on the type of sample, the types of
phthalate esters that need to be determined, the duration of sample storage
prior to analysis, and the analytical procedure to be used (10). The sample
preservation method chosen must not impair the analytical procedure to be
used. Amber glass bottles are the containers of choice because of the
protection they offer from photodegradation. The bottles must not be
prerinsed with sample before collection (11). Composite samples should be
collected in refrigerated glass containers and, if automatic sampling
equipment is to be used, it must be free of any Tygon tubing (11). Most
investigators report that samples should be refrigerated (15,16) or frozen
(15,16,17,18,19,20) until analysis.
The use of aluminum foil either to wrap whole samples or to line the
container caps prior to freezing has been reported (16,17,19). Contamination
by phthalate esters present on the aluminum foil is not a problem for whole
biological samples (such as fish) where the exterior skin is not utilized for
analysis (17). However, use of aluminum foil to line container caps has been
shown to cause severe contamination problems due to extraction of the
phthalate esters from the cork or the rubber backing of the cap by the solvent
vapor in the vial. Container caps lined with Teflon were found to be superior
to those lined with aluminum-foil-lined caps (21,22,23).
Sample preservation by the addition of solvents such as methylene
chloride (24) or hexane (25) immediately after sample collection has been
reported. The addition of small amounts of organic solvents acts as inhibitor
-------
TABLE 4. SUMMARY OF ANALYTICAL METHODOLOGIES FOR PHTHALATE ESTERS
Method Matrix
EPA Method 606 Hater
EPA Method 625 Hater
EPA Method 1625 Hater
EPA Method 8060 Liquids
Solids
Eastman-Kodak Hater
Method
Compounds
DMP
DEP
OBP
BBP
DEHP
OOP
CMP
DEP
DBP
BBP
DEHP
OOP
DMP
DEP
DBP
BBP
DEHP
OOP
DMP
DEP
DBP
BBP
DEHP
OOP
DMP
DEP
DBP
BBP
DHP
DEHP
DIOP
DINP
DIDP
Other
Extraction
procedure
Separatory funnel
Methyl ene chloride
Separatory funnel
Continuous liquid-
liquid extractor
Methylene chloride
Separatory funnel
Continuous liquid-
liquid extractor
Methylene chloride
EPA Method 3510
EPA Method 3520
EPA Method 3540
EPA Method 3550
Separatory funnel
Methylene chloride
Cleanup procedure
10 g Florisil (3% water-
deactivated); 100 ml
20X ethyl ether in
hexane
10 g alumina (3X Mater-
deactivated); 140 ml
20X ethyl ether in
hexane
None
None
EPA Method 3610
EPA Method 3620
None
Analysis procedure
GC/ECO
1.5X SP 2250/1. 95%
SP 2401 on Supelcoport
(100/120 mesh)
3X OV-1 on Supelcoport
(100/120 mesh)
GC/MS
3X SP 2250 on Supelcoport
(100/120 mesh)
Isotope dilution GC/MS
30 a x 0.25 am ID
fused-silica capillary
column, 94X oethyl, 4X
phenyl, IX vinyl
polysiloxane bonded
GC/FID or GC/ECD (GC/FID)
1.5X SP 2250/1. 95X
SP 2401 on Supelcoport
(100/120 mesh)
GC/ECD
3X OV-101 on Gas Chrom Q
(100/120 mesh)
MDL
(fjg/L or fig/kg) Reference
0.29 to 3 11
1.9 to 2.5 12
10 13
14 to 31* (GC/FID) 1
0.29 to 3* (GC/ECD)
14
"Values given are for clean waters.
"MDL not specified.
-------
to bacterial degradation of the phthalate esters (16,17,18,24,25,26) and also
initiates sample extraction. Addition of a single crystal of thymol as
bacterial inhibitor prior to freezing was reported by Glazer et al. (18).
Aqueous solutions containing DEP decreased in DEP-concentration by
20 percent in 4 days and by 70 percent in 12 days when kept under fluorescent
light, and no DEP was detected after 14 days. In the absence of light and
under refrigeration, no loss in DEP was detected after 60 days (27). Shouten
et al. (25) reported that DBP and DEHP concentrations decreased with time, and
the losses were attributed to biodegradation. At the 50-ppb level, a
50-percent loss of DEHP occurred in 5 days. In the case of DBP, over
90 percent was lost in about 3 days. Sullivan et al. (28,29) reported
significant losses of phthalate esters from seawater during storage and
attributed the losses to adsorption onto glassware. A kinetic study showed
that most of the adsorption of DEHP occurred within the first hour and that
after 12 hrs the solid adsorbent and the aqueous phase reached equilibrium
(28). The adsorbed phthalates were only partially recovered by subsequent
water rinses. A solvent rinse of the containers recovered most of the
compounds adsorbed onto the glassware (29).
The stability of the phthalate esters in solid matrices has not been
systematically investigated. Storage of soil and sediment samples at room
temperature should be avoided since bacterial degradation does occur
(16,18,24,26). Freezing at -10 to -30°C appears to be the most suitable
method for preservation of solid samples since it has the widest range of
applications and causes the least changes in the sample matrix.
3.2 EXTRACTION
A number of methods have been reported for the isolation of the
phthalate esters from water, soil, sediment, and particulate material
collected on Hi-Vol glass fiber filters. The extraction techniques that have
been used on water samples include stirring, separatory funnel partitioning,
and liquid-liquid extraction; adsorption onto XAD-resins or other liquid
chromatographic bonded stationary phases has also been reported. The methods
that have been reported for soil and sediment samples include solvent
partitioning, liquid-solid extraction, Soxhlet extraction, blending, and
ultrasonication.
3.2.1 Extraction of Water Samples
Table 5 presents a summary of extraction methods for water samples as
published in the literature. The following describes in detail some of the
procedures for the extraction of phthalate esters from water samples.
3.2.1.1 Liquid-Liquid Extraction
Liquid-liquid extraction is the simplest and the most widely used
technique for the extraction of organic compounds from water. This technique
consists of mixing the sample with a water-Immiscible solvent in the original
sample container, a separatory funnel, or a continuous liquid-liquid
extractor. An important parameter for the liquid-liquid extraction is the
11
-------
TABLE 5. SUMMARY OF EXTRACTION TECHNIQUES FOR WATER SAMPLES
Nethod
Solvent
Phthalate
esters
Recovery
percent
Spiking
level
(ppb)
Reference
Solvent (separatory
funnel)
Solvent (separatory
funnel)
Solvent (separatory
funnel)
Solvent (separatory
funnel)
Solvent (separatory
funnel)
Solvent (homogenlzer)
Solvent (separatory
funnel)
Solvent (separatory
funnel)
Solvent (separatory
funnel)
Solvent (separatory
funnel, continuous
liquid-liquid
extractor)
Solvent (separatory
funnel, continuous
liquid-liquid
extractor)
Methylene chloride DMP, DEP, DALP,
DBP, DHP, BBP,
DPP, OOP
Petroleum ether
DMP, DBP, BBP, 25 to 130
DEHP
Methylene chloride BBP, TPMBP
C7,9benzyl-P
CrA11DAKP
53 to 89 0.1 to 20
Methylene chloride DMP, DEP, DBP, 85 to 110
DHEP, DIBP,
DEHP, DNP
Methylene chloride DMP, DEP, DBP,
BBP, DHP, DEHP,
0.1
110-
DAKP,
, DNP, DDP,
,,,,-DAKP,
Hexane
Hexane
D
DEP
DBP, DEHP
94 to 96
88 to 1001
Methylene chloride DMP, DEP, DBP
OOP, BBP, DEHP
Methylene chloride DMP, DEP, DBP,
OOP, BBP, DEHP
Methylene chloride DMP, DEP, DBP,
OOP, BBP, DEHP
Methylene chloride DMP, DEP, DBP,
OOP, BBP, DEHP
10 to 100
100
0.7 to 106
16
17
24
30
14
27
31
11
12
13
(continued)
'Extraction efficiency was a function of the shaking tines; 5 »1n (88 percent); 10 m1n
(99 percent); 20 rain (100 percent).
"Refer to methods for data.
°Data obtained with type A polyurethane foam plugs (density 0.030 g/ml).
"Data obtained with type B polyurethane foam plugs (density 0.019 to 0.022 g/ml).
'Value given 1s for preconcentratlon from 500 to 1,000 ml water sample.
NOTE: DALP - dlallyl phthalate
DPP - dlphenyl phthalate
DAKP - dlalkyl phthalate
DHEP - diheptyl phthalate
TPMBM - 2,2,4-tr1methyl-l,3-pentaned1ol nonolsobutyrate phthalate
12
-------
TABLE 5. (concluded)
Method
Solvent
Phthalate
esters
Recovery
percent
Spiking
level
(ppb)
Reference
Solvent (separatory
funnel, bottle
stirrbar, continuous
liquid-liquid
extractor)
Methylene chloride DMP, DEP, DBP,
BBP, OOP, DEHP
25 to 129
*»«o p* uuiiuapap^
(10 cm x 4 mm ID,
10 im particles)
Polyurethane Foam
LiChrosorb RP-18
(2mm plug, 5 im
particles)
Supelclean
LC-18 SPE
Gradient elution
from 100 percent
water to 100
percent methanol
Extract with 2 mL
acetone and 3 mL
hexane in a glass
syringe (dynamic
conditions)
Extract with 50 mL
solvent (acetone-
hexane 1:4); time
varies from 1 hr
to 72 hrs except
for DMP from
10 min to 1 hr
Elute with
85 percent
methanol in water
Elute with three
1-mL aliquots of
acetonitrile
DHP, DBP, DEHP,
DNP
DHP, DEP, DBP,
DIBP, DAM, DHP
DEHP, OOP,
DIDP, BBP
DMP, DHP, DBP,
OOP, DHEP
20 to 102e
8 to 101d
25 to 100
1.0
DBP, DEHP
DMP, DEP, DBP,
BBP
99*
85.9 to 103 2,000
32
33
34
34
35
36
XAD-4
(1.2-1.8 mm
XAD-2
XAD-4
XAD-8
(1.2 cm x 6
Filtrasorb
(1.2 cm x 6
x
.5
300
.5
25 mm)
cm)
cm)
Elute with
50 to 100 ML
acetone
Acetone and
chloroform
Chloroform or
benzene
DEP, DBP 85 to 99 2 to 100 37
DMP, DBP, DEHP 19 to 116 50 38
DMP, DBP, DEHP 0 to 45 50 38
'Extraction efficiency was a function of the shaking tine; 5 min (88 percent); 10 min
(99 percent); 20 min (100 percent).
"Refer to methods for data.
'Data obtained with type A polyurethane foam plugs (density 0.030 g/mL).
"Data obtained with type B polyurethane foam plugs (density 0.019 to 0.022 g/mL).
'Value given Is for preconcentration from 500 to 1,000 mL water sample.
NOTE: DALP - diallyl phthalate
DPP - diphenyl phthalate
DAKP - dialkyl phthalate
DHEP - diheptyl phthalate
TPMBM - 2,2,4-tr1methyl-l,3-pentanediol monoisobutyrate phthalate
13
-------
solvent. It must be immiscible with water, extract the organics of interest
from the sample, and be compatible with the analysis procedure.In the case of
the separatory funnel technique, up to 1 liter of an aqueous sample is poured
into a separatory funnel and extracted by shaking with an organic solvent.
The layers are allowed to separate, and the organic fraction is drawn off.
Solvents with a specific gravity greater than that of water are preferred
since the lower layer can be removed more easily.
Disadvantages of separatory funnel partitioning are:
• Limited sample size
• Emulsion formation with many wastewaters.
To break emulsions, it has been recommended that the extract be passed through
a 25-mm-thick glass wool pad (39,40).
Several reports (14,16,17,24,27,30,31) dealing with the determination
of phthalate esters in water samples involved separatory funnel partitioning;
however, there appears to be little consensus as to the best organic solvent,
the extracting conditions (e.g., sol vent-to-liquid ratios, time, degree of
agitation), or the use of salts to enhance partitioning into the organic
layer. Table 5 lists the various systems used, although no data on the
percent recovery for each procedure have been reported.
One of the more recent devices introduced specifically for
liquid-liquid extractions is the Mixxor device. These devices come in various
sizes to handle sample volumes up to 50 ml. The mixing is accomplished by
moving the piston up and down in the mixing chamber 5 or 6 times (equivalent
to 40 or more shakes in a separatory funnel). The system is fast, precise,
and safe (41). The main disadvantage is the ease with which emulsions are
generated (41).
3.2.1.2 Adsorption
Several techniques have been used to adsorb phthalate esters onto solid
supports. The materials used include uncoated polyurethane foams (PUT) or PUF
coated with a viscous-liquid stationary phase (34), and support-bonded
polymers such as HPLC column materials (33,35). The polyurethane foams seem
to work well in removing some phthalate esters from water at the part-per-
million level; however, the foams do not retain the larger-molecular-weight
compounds such as DEHP and OOP (34). Once adsorbed from water, the phthalate
esters may be desorbed by either elution using a high-pressure liquid
chromatograph (HPLC), if the preconcentration was performed on support-bonded
HPLC columns (33,35), or Soxhlet extraction with organic solvents such as
acetone or hexane if the preconcentration was done on PUFs (34). Table 5
lists the solid supports, compounds, and their recoveries using the eluting
solvent specified in the table.
A solid-phase extraction method for the preconcentration of phthalate
esters from water samples has been reported recently (36). Although there
14
-------
are no solid-phase extraction methods currently approved for use by the EPA,
a solid-phase extraction method for the determination of endrin, lindane,
methoxychlor, and toxaphene has been recently proposed as an alternative to
the current EPA method (42). Major factors influencing the reversed-phase
solid-phase retention are sample pH, sample volume, sorbent mass, and analyte
concentration (43). These factors are first selected a priori, based on
experience, in order to optimize the elution scheme (solvent strength and
solvent volume). Once the elution conditions are optimized, then the choices
made initially for sample pH, sample volume, and sorbent mass are varied to
maximize compound retention.
Recoveries of phthalate esters using solid-phase extraction on
Supelclean LC-18 cartridges (Supelco Inc.) range from 91.6 to 102 percent
(36). The experiments were conducted as follows: each tube was conditioned
with 2 mL methanol followed by 4 ml purified water. The preconcentration
step was done by passing 50 mL of purified water spiked with selected
phthalate esters at 2000 ppb to which 3 ml of acetonitrile was added.
Following sample preconcentration, each tube was washed twice with deionized
water (1 ml) and then eluted with three 1-mL portions of acetonitrile (36).
The major drawback of this technique is the fact that with environmen-
tal samples a large number of contaminants will be sorbed on and eluted with
the compounds of interest, which will inevitably lead to broad solvent peaks
and capacity problems. To avoid this, three alternatives have been recom-
mended (35):
• Use of gradient elution to achieve a stepwise elution of the sorbed
components
• Proper selection of the stationary phase
• Use of a selective detection system (e.g., fluorescence detection
or postcolumn derivatization).
Other solid adsorbents that are used as alternatives for isolation of
phthalate esters from water samples include the XAD resins (37,38) and carbon
(38). The recovery efficiencies of XAD-2, XAD-4, XAD-7, and XAD-8 resins and
resin mixtures were reported by Tateda and Fritz (37) and Van Rosum and Webb
(38). The results indicate that the resin performs well for the shorter-chain
esters; the longer-chain esters such as DEHP are not retained efficiently by
the resin. Activated carbon was reported to be effective in extracting
phthalate esters from aqueous samples; however, the recovery of the adsorbed
compounds is not quantitative when the carbon is extracted with chloroform
or benzene in a Soxhlet extractor (38).
D. Blevins (45) reported recently a new procedure for preconcentrating
phthalate esters and other organics from aqueous samples. The procedure
involves filtration of the aqueous sample through a PTFE membrane disk
impregnated with Cs-bonded silica. The membrane, known as the 3M-Empore
membrane disk, consists of chemically modified 8-/un silica particles tightly
bound in a densely woven "web" of micro-PTFE fibrils. The silica particles
15
-------
are individually suspended, with the surface of each particle free to interact
with the aqueous liquid that is filtered through or the organic solvent used
in extracting the organics retained by the membrane disk. Aqueous samples
containing 5 percent methanol are passed through at approximately 25 mL/min,
and the compounds are extracted from the membrane disk with 10 mi
acetonitrile. Typical recoveries for DMP, DEP, DBP, and OOP are in the range
of 82 to 117 percent.
3.2.1.3 Nicroextractlon
Rhoades and Nulton (44) developed a technique for the extraction of
industrial wastewaters using microextraction (a one-step extraction in which
the solvent-to-sample ratio is 1:40 to 1:100). Use of their method for
extracting relatively nonpolar analytes is justified from both theoretical and
practical viewpoints. Comparison of both the recovery and the variability of
the microextraction technique with other methods is presented in Tables 6
and 7. Microextraction compares favorably with other techniques and offers
several practical advantages:
• Being a single-step extraction, it offers the possibility to
extract selectively the analyte of interest
• Contamination is minimized by sample preparation in a single
container
• It is less labor intensive, since extract concentration is not
necessary
• The need for extract cleanup is minimized.
3.2.2 Extraction of Sediment and Soil Samples
This section summarizes the extraction techniques reported in the
literature for soil and sediment samples. Examples of solvent and solvent
mixtures used for extraction, type of extraction, compounds investigated, and
recovery data are presented in Table 8.
Techniques used for the extraction of phthalate esters from soil and
sediment samples can be classified into three categories: 1) blending the
sample with solvents, 2) sonication, and 3) Soxhlet extraction. Each of these
techniques will be described in more detail below.
Blending consists of the extraction of 10 to 20 g soil or sediment
material on a wrist-action shaker (16) or in an extraction jar (17) with
various solvents. Acetone-hexane and acetonitrile have been recommended for
blending (16,17). After extraction with acetonitrile, the soil was extracted
twice with methylene chloride-petroleum ether (1:9), and all extracts were
combined (17).
Soxhlet extraction seems to be preferred because of the ease and
neatness in handling. Because of the nature of the system, freeze-dried soil
16
-------
TABLE 6. SUMMARY OF SPIKE RECOVERY DATA FOR THE MICROEXTRACTION TECHNIQUE
Microextraction
Other methods*
Phthalate ester
Average No. of Average No. of
percent Std. spiked percent Std. spiked
recovery Dev. samples recovery Dev. samples
Bis(2-ethylhexyl)
Di-n-butyl
Di-n-octyl
85
61
82
25
29
21
50
13
13
56
66
92
56
66
_ —
92
55
2
aData supplied by William F. Cowen, Catalytic, Inc. Data obtained by
Southwest Research Institute and several other laboratories taking
part in an EPA-Effluent Guidelines verification program are also
included (44).
TABLE 7. SUMMARY OF DUPLICATE ANALYSES FOR THE MICROEXTRACTION TECHNIQUE
Microextraction
Variability"
Phthalate ester
Bis(2-ethylhexyl)
Di-n-butyl
Di-n-octyl
Average
39.8
31.9
21.6
Range
0 to 143
0 to 67
0 to 35
No. of
duplicates
27
11
8
Other methods"
Variability"
No. of
Average Range duplicates
110 5 to 200 31
86 4 to 200 14
"Data supplied by William F. Cowen, Catalytic, Inc. Data obtained by Southwest
Research Institute and several other laboratories taking part in an EPA-Effluent
Guidelines verification program are also Included (44).
"All values in absolute difference between duplicates expressed as the percent of
- mean value: lOOJx, -x2|/x.
17
-------
TABLE 8. SUMMARY OF THE EXTRACTION TECHNIQUES FOR SOILS AND SEDIMENTS
Spiking
Type of Recovery level
Solvent extraction Phthalate ester (percent) (ng/g) Reference
Acetone/hexane Blending DMP, DEP, DAP, -- 35-1,000 16
DBP, DHP, BBP,
DEHP, DPP, OOP
Acetonitrile, Blending DMP, DBP, BBP, 61-81 300-700 17
methylene chloride/ DEHP
petroleum ether
(1:9)
Water/cyclohexane Sonication BBP, TPMBP, DAKP 91-160 50-20,000 24
(1:1)
Methylene chloride Sonication DMP, DEP, DBP, 53-99 20,000 32
or DEHP, OOP, BBP
Acetone/hexane (1:1)
Benzene Sonication DMP, DEP, DBP, -- -- 47
OOP
Hexane/acetone/
methanol (8:1:1)
Acetonitrile
Methylene chloride
Methylene chloride,
toluene/methanol
(10:1) or Acetone/
hexane (1:1)
Chi orof orm/methanol
(1:1)
Soxhlet
Soxhlet
Soxhlet
Soxhlet
Soxhlet
DBP,
DEP
DMP,
DHP,
DMP,
DEHP,
DBP,
DEHP
DEP, DBP,
DEHP, DNP
DEP, DBP,
DOP, BBP
DEHP
90-110 -- 46
90-94 200-2,000 27
85-110 -- 30
1
1-5 48
NOTE: TPMBP - 2,2,4-tr1methyl-l,3-pentanediol monoisobutyrate phthalate
DAKP - diallcyl phthalate
18
-------
is preferred (46). The extraction time varies from solvent to solvent.
Soxhlet extraction with acetonitrile gave DEP recoveries of 90 to 94 percent
after 4 hours (27), while 90 to 100 percent recoveries for DBP and DEHP were
achieved after 17 hours extraction with 8:1:1 hexane-acetone-methanol (46).
A comparison of the extraction efficiencies obtained by blending,
sonication, and Soxhlet has not been reported. Furthermore, the choice of
solvent appears to be a matter of personal preference at the present time.
3.3 SAMPLE EXTRACT CLEANUP
Several types of cleanup techniques are available for the removal of
coextractants from a sample matrix. They are:
• Liquid-liquid partitioning
• Gel permeation chromatograpy
• Sulfur removal
• Liquid-solid chromatography (Florisil, silica gel, alumina).
3.3.1 Liquid-Liquid Partitioning
This technique is widely used for removing fats, waxes, and polar
materials. It consists of partitioning the sample extract with an immiscible
solvent of different polarity. For example, an extract in hexane, petroleum
ether, or benzene can be partitioned with a polar solvent such as
acetonitrile. The waxes and fatty materials will remain in the nonpolar phase
(hexane, petroleum ether, or benzene) while the phthalate esters will
partition into the acetonitrile. The polar phase containing the phthalate
esters is then diluted with water, and a sodium sulfate or sodium chloride
solution is added to "salt out" the phthalate esters from the polar phase back
into another nonpolar solvent. Giam et al. (15) reported that partitioning
of extracts containing lipids and DEHP with acetonitrile and dimethyl
formamide did not selectively remove the lipids and DEHP coextracted with the
lipids when the hexane extract was partitioned with acetonitrile.
Solvent systems used for liquid-liquid partitioning of the phthalate
esters are listed in Table 9.
Poole and Wibberley (49) have devised a very simple 3-step
liquid-liquid partitioning procedure. After sample homogenization with
acetonitrile and filtration, hexane-water is used to partition the phthalate
esters from polar coextractants. Dimethyl formamide-water (94:6) and hexane
is then used to further partition the moderately polar species. Finally,
hexane-acetonitrile is used to partition the nonpolar species into the hexane
phase, leaving the phthalate esters in the acetonitrile.
19
-------
TABLE 9. LIQUID-LIQUID SOLVENT PARTITIONING SYSTEMS
Initial solvent
system
Partitioning
system
Secondary
partitioning
Reference
Acetonitrile
Acetonitrile
and methylene
chloride/
petroleum ether
Ethylene glycol
Acetonitrile
Acetonitrile
Acetonitrile
Methylene chloride/
petroleum ether
(1:5); water
(5 percent NaCl)
Hexane/acetone Water
Water (5 percent
NaCl)
Hexane
Hexane
Hexane/water
Hexane/water
Methylene chloride/
petroleum ether
phase partitioned
with water
Water/acetone phase
partitioned with
hexane
Aceton i tri1e/water
phase partitioned
with methylene
chloride/petroleum
ether
Hexane phase parti-
tioned with water
Dimethyl formamide/
water (94:6);
hexane; hexane/
acetonitrile
15
16
17
50
51
27
49
20
-------
3.3.2 Gel Permeation Chromatography (GPC)
In gel permeation chromatography the separation mechanism is based on
differences in molecular size. Large molecules are excluded from the pores
of the gel and elute first, while smaller molecules which can diffuse into the
pores are eluted last.
The GPC techniques reported for the phthalate esters are summarized in
Table 10. The elution volume of DEHP from Biobeads SX-3 and 1:1 methylene
chloride/cyclohexane is reported in Table 11. The elution volumes of other
phthalates, isophthalates, and terephthalates also have been reported (52).
GPC with Biobeads SX-2 as a preparative technique prior to GC analysis
of phthalates has been investigated by Baker (53). It was demonstrated that
small amounts of phthalate esters can be separated from an excess of lipids,
providing sufficiently clean sample extracts. Burns et al. (20) separated
lipids and phthalate esters using Biobeads SX-3 and methylene
chloride/cyclohexane (1:1).
Problems with GPC stem mainly from samples with high lipid content,
where broad elution of the lipid fraction overlaps the elution of the
phthalate esters. The presence of triglycerides affects the performance of
TABLE 10. SUMMARY OF GPC TECHNIQUES
Column/solvent
system
Phthalate
esters
Detector/level Reference
HP-GPC Shodex A801
Styrene-di vi nylbenzene
resin (exclusion limit
1,000 as polystyrene)/
chloroform
LP-GA Biobeads SX-3/1:!
methylene chloride-
cyclohexane
HP-GPC Shodex A801
Styrene-divinyl benzene
resin (exclusion limit
1,000 as polystyrene)/
chloroform
Biobeads SX-2/benzene
DMP.DEP.DBP, Ultraviolet at
DHEP, DEHP, 243 nm/20 ppb
DDP
DBP,DEP,
DEHP,DHEP
Cl to CIS
DAKP
31
Ultraviolet at
254 nm; preparatory
column
Ultraviolet at
254 nm
20
52
DEHP,DNP,DMP Ultraviolet at
281 nm/20 pg
53
NOTE: DHEP - diheptyl phthalate
DDP - didecyl phthalate
DAKP - dialkyl phthalate
21
-------
TABLE 11. 6PC ELUTION VOLUMES OF THE PHTHALATE ESTERS8
Eluate
(mL)
0 to 50
50 to 60
60 to 70
70 to 80
80 to 90
90 to 100
100 to 110
110 to 120
120 to 130
130 to 140
Total
Lipld
(percent
removal )
..
0.6
41.1
44.7
5.3
1.3
0.6
--
--
--
93.6
Phthalate
ester
(percent
recovery)
..
..
-.
36.4
62.5
--
--
--
--
98.9
aData taken from Reference 52.
22
-------
electron-capture detectors, generating negative peaks in that portion of the
chromatogram where the phthalate esters elute. Additionally, gradual
deterioration of both the column and the detector may occur due to lipid
materials (20).
3.3.3 Sulfur Removal
Sulfur and other organosulfur compounds, if present, may give large
solvent peaks which could mask the region from the solvent peak to the
earliest eluting phthalate ester peaks.
Sulfur may be removed as a discrete fraction by GPC (54). Alterna-
tively, several chemical methods are available for the removal of sulfur:
reaction with metallic mercury (55), activated copper (56), Raney Nickel (57),
tetrabutyl ammonium sulfite (TBA+)2 S032" (58), or potassium cyanide (59).
Suzuki et al. (51) used a silver-nitrate-coated Florisil column for the
cleanup of interfering coextractants (carotenes) and organic sulfides derived
from some vegetables.
3.3.4 Liquid-Solid Chromatography
The following discussion will demonstrate the application of Florisil,
silica gel, and alumina to the cleanup of sample extracts prior to the
determination of phthalate esters.
3.3.4.1 Florisil
Florisil is a synthetic magnesium silicate manufactured by the Floridin
Company from magnesium sulfate and sodium silicate. Following precipitation
from solution, the magnesium silicate is filtered, dried, and calcinated at
650eC. Examples of cleanup methods for phthalate esters using Florisil are
listed in Table 12.
Giam et al. (15) in their study of the determination of trace phthalate
esters in open-ocean biota samples noted that DDTs and PCBs were present in
virtually all biological samples in the ppb range, and that this required a
separation scheme for the phthalate esters, since some peaks overlap (Aroclor
1260/DEHP, Aroclor 1254/DBP). They found that 3-percent water-deactivated
Florisil gave the best recoveries for the phthalate esters, and that elution
followed the pattern determined previously: 6 percent diethyl ether in
petroleum ether for elution of the PCBs, then 15 percent diethyl ether in
petroleum ether for DEHP, and 20 percent diethyl ether in petroleum ether for
DBP. Spiked sample recoveries ranged from 70 to 100 percent depending on the
type and amount of tissue used. Giam et al. (15) encountered an interference
from lipid and other tissue components which eluted with DEHP in the
15-percent ether fraction. Solvent partitioning of the lipid prior to column
cleanup was attempted; however, that resulted in loss of phthalates. It was
further noted that, if the sample size was adjusted to maintain lipid levels
to 3 g per sample, then the interference was reduced to minimum.
23
-------
TABLE 12. SUMMARY OF CLEANUP PROCEDURES WITH FLORISIL
Material
Eluting solvents
Percent
recovery Reference
3 percent water-
deactivated
FTorisil, 35 g
3 percent water-
deactivated
Florisil, 3 g
Florisil, 25 cm x
1 cm ID (2 cm
anhydrous sodium
sulfate on top)
AgNO.-coated
Flonsil, 3 g
(top with 3 cm
anhydrous sodium
sulfate)
Florisil, 7 g
2 percent water-
deactivated
Florisil
3 percent water-
deactivated Florisil
30 g (top with
2.5 cm layer of
anhydrous sodium
sulfate)
Florisil, 10 g (top
with 1 cm anhydrous
sodium sulfate)
6 percent diethyl ether in petroleum
ether (DDT, PCBs); 200 ml
15 percent diethyl ether in petroleum
ether (DEHP); 200 ml
20 percent diethyl ether in petroleum
ether (DBP); 200 ml
Petroleum ether (PCBs, DDT); 10 ml
20 percent diethyl ether in petroleum
ether (phthalates) 25 ml
Hexane (80 ml)
25 percent diethyl ether in hexane
(80 ml)
Diethyl ether (phthalates); 200 ml
Hexane
2 percent ethyl acetate in hexane
(chlorinated pesticides and
phthalates)
3 percent benzene in hexane
(organochlorine pesticides); 40 ml
2 percent ethyl acetate in hexane
(phthalates); 60 ml
100 ml hexane/diethyl ether (50:3);
discard
100 ml hexane/diethyl ether (17:3)
(DEHP, DPB)
30 percent methylene chloride in
hexane (PCBs, pesticides); 250 ml
10 percent ethyl acetate in hexane
(phthalates); 100 ml
6 percent diethyl ether in petroleum
ether
15 percent diethyl ether in petroleum
ether (DEHP DBP)
20 percent diethyl ether 1n petroleum
ether (DBP, DEP, DMP)
Hexane, 40 mL, discard
20 percent diethyl ether in hexane,
100 ml
70 to 100 15
100
17
50
51
46
19, 60
61
1, 11
24
-------
Similar elution schemes involving diethyl ether and petroleum ether
have been reported by others (17,46,50,61). Other eluting solvents, such as
2 percent ethyl acetate in hexane (51) and 10 percent ethyl acetate in hexane,
have also been used (19,60). In both cases, however, the organochlorine
pesticides interfere and must be removed from the phthalate esters either with
3 percent benzene in hexane (51) or 30 percent methylene chloride in hexane
(19,60).
Suzuki et al. (51) separated the phthalate esters from the
organochlorine pesticides using AgN03-coated Florisil; the pesticides were
eluted with 40 ml of 3 percent benzene in hexane, while the phthalate esters
were eluted with 60 mL of 2 percent ethyl acetate in hexane (51).
A method for the standardization of the Florisil activity by measure-
ment of its adsorptive capacity has been developed by Mills (62). Laurie acid
(present in excess) is adsorbed from hexane solution onto the Florisil column;
the amount not adsorbed is determined by alkali titration of the eluant.
Calculation of the Florisil activity (equivalents per gram) allows comparison
of different batches of activated Florisil.
3.3.4.2 Silica Gel
Andersen and Lam (23) reported the use of a Celite column for the
separation of phthalate esters from edible oils. The Celite column was first
coated with an edible oil (triolein), and DEHP was eluted with propylene
carbonate. Experiments were carried out using different amounts of stationary
phase. To determine the elution of DEHP, the propylene carbonate eluant was
collected in 1-mL fractions and analyzed by gas chromatography.
3.3.4.3 Alumina
Persson et al. (63) and Webster and Nickless (64) reported sample
cleanup using alumina and deactivated alumina, respectively. The sediment or
water extract was applied to a 20-g column of deactivated alumina and eluted
with 40 mL hexane (Fraction 1), 100 mL of 6 percent diethyl ether in hexane
(Fraction 2), 100 mL of 15 percent diethyl ether in hexane (Fraction 3), 100
mL of 25 percent diethyl ether in hexane (Fraction 4), and 100 mL of
50 percent diethyl ether in hexane (Fraction 5). Most phthalate esters elute
in Fraction 3, the more oxygenated esters elute in Fraction 4, and bis(methoxy
ethyl) phthalate was recovered in Fraction 5 (64). A simple separation of
neutral lipids and phthalate esters from polar lipids was carried out by
Waldock (30) using 5-percent water-deactivated alumina. Recoveries ranged
from 98 to 101 percent when phthalate ester standards dissolved in fish oil
were eluted from the alumina column with methylene chloride.
Phthalate esters were separated from organochlorine pesticides and PCBs
using combined alumina-silica gel chromatography (16) as shown in Figure 1.
The separation of the organochlorine pesticides, PCBs, and phthalate esters
is first achieved on an alumina column (16). The organochlorine pesticides
and PCBs are then eluted with hexane while the phthalate esters are eluted
with benzene. Further fractionation of the benzene fraction is achieved on
25
-------
CO
Hexane extract from sample
evap. to 0.5 ml
Alumina column
Elute with 40 ml hexane,
followed by 20 ml benzene*
0-20 ml (A-.l fraction)
Treat with Hg to remove S;
evap. to 0.5 ml
Silica gel column
20-35 mL (A-2 fraction)
Dieldrin
Endrin
Heptachlor epoxide
35-55 ml (A-3 fraction)
Silica qel column
ro
o»
Elute with 30 ml hexane,
followed bv 20 ml benzene*
20 ml eluent (A-3),
followed by 20 ml of
10 percent acetone in
benzene*
0-25 mL (S-l fraction)
Aldrin
PCBs
PCNs
Ml rex
25-45 ml (S-2 fraction) 0-20 ml (AS-1 fraction)
Chlordane Discard
DDD
DDE
DDT
Heptachlor
Lindane
Toxaphene
20-35 ml (AS-2 fraction)
Phthalate Esters
Dimethyl
Diethyl
Diallyl
Di-n-butyl
Di-n-hexyl
Butyl benzyl
Di-2-ethylhexyl
Diphenyl
Di-n-octyl
*Approximately 5 ml of eluting liquid is retained by column.
Figure 1.
Combined alumina-silica gel chromatographic cleanup technique.
Figure taken from Reference 16.
-------
a silica gel column; the phthalate esters are eluted with 10 percent acetone
in benzene. Benzene was chosen since it is compatible with GC/ECD. Such a
fractionation scheme was needed because of some interfering polar substances
that could not be retained on the alumina column.
Burns et al. (20) and Hazelton Laboratories (65) utilized sulfuric-
acid-impregnated alumina to increase the separation of lipid materials from
the phthalate esters. Ordinary alumina removed only about one-half of the
lipid which remained after GPC, and it was, therefore, inefficient if used
alone. Elution with 2 percent diethyl ether in hexane removed early
coextractants, while elution with 10 percent diethyl ether in hexane removed
the phthalate esters. Degradation of the phthalate esters was minimal on the
acid-impregnated alumina. However, it was recommended that fresh
acid-impregnated alumina be prepared every 2 weeks.
Several difficulties with the above method were noted (65):
• The capacity of the alumina column was low
• Because of the ubiquitous nature of DEHP, the blank levels for DEHP
were in the range of 50 to 100 ppb, with occasional values as high
as 1,000 ppb
• GC/MS analysis indicated a distinct change in retention times, and
broad peak shapes for DEHP were attributable to a buildup of
extraneous high-boiling sample materials.
3.3.5 Selection of a Cleanup Technique
The choice of the adsorbent depends upon many factors, but the type of
matrix and the interfering coextractants present seem to be the most important
considerations. Alumina has been found to be more efficient than silica gel
and Florisil in the removal of fats (16,20). Silica gel can remove edible
oils efficiently when a suitable solvent such as propylene carbonate is used
(23). Florisil cleanup has had the most attention. Though not quantitatively
compared with other methods for specific sample types, Florisil appears to
enable the separation of phthalate esters from a greater number of extracts
of different matrices than any other method presently used. Lopez-Avila
et al. (66) reported elution patterns and recovery data for 16 phthalate
esters in the presence of interferents such as corn oil, diesel hydrocarbons,
organochlorine pesticides, and chlorinated phenols using 1-g Florisil
disposable cartridges. Hexane, hexane/diethyl ether (1:1), hexane/acetone
(9:1), and various combinations of hexane/methylene chloride were used as
eluants. The authors reported that organochlorine pesticides can be removed
efficiently with hexane/methylene chloride (4:1); under these conditions, the
phthalate esters are retained on the Florisil cartridge and can be recovered
by elution with hexane/acetone (9:1).
When dealing with multicomponent analyses, and especially in those
cases where the GC analysis does not allow complete separation of all the
sample components, a combination of cleanup techniques may be required.
27
-------
Combined techniques may offer the advantage of fractionating several sample
constituents (phthalates, PCBs, pesticides, etc.) for analysis in the same
sample with a minimum amount of preparation time and materials.
3.4 GC ANALYSIS
This section will address only the GC analysis techniques, including
gas chromatography/mass spectrometry (GC/MS). Basically, the phthalate esters
are separated on the chromatographic column, either packed or capillary, at
elevated temperatures, and the compounds are detected with an electron capture
detector (ECD), a flame ionization detector (FID), or a mass spectrometer
(MS). Section 3.5 addresses the high performance liquid chromatographic
(HPLC) techniques. A comparison of GC and HPLC detection limits is presented
in Table 13.
3.4.1 Gas Chromatographic Columns
Several factors must be considered when selecting columns for the GC
analysis of phthalate esters.
• Operating parameters should yield maximum resolution and detector
response at a minimal retention time
• The stationary phases should be stable and not bleed during the GC
analysis
TABLE 13. INSTRUMENTAL DETECTION LIMITS FOR PHTHALATE ESTERS8
Detection limit
Method
HPLC
GC
Detector
U.V. (254 nm)
U.V. (224 nm)
FID
ECD
MS(SIM)
MS
Amount
injected
(ng)
10
2
1
0.02
0.01
1
Concentration/
volume injected
(ng/mL)
100 (100 nl)
20 (100 ML)
1,000 (1 ML)
20 (1 Hi)
10 (1 ML)
1,000 (1 /zL)
aData taken from Reference 5.
SIM -- selected ion monitoring.
28
-------
• The columns should be durable enough to allow analysis of very
complex matrices.
Single and mixed stationary phases, both nonpolar and polar, in packed
and capillary columns have been used for the analysis of phthalate esters.
Some of the more commonly used columns and GC conditions are listed in Table
14. Representative chromatograms of phthalates on various columns are shown
in Figures 2 through 4.
The majority of the reports in the literature discusses packed columns
for the separation of phthalate esters. Capillary columns are beginning to
emerge as suitable alternatives to packed columns. Though the resolution of
capillary columns is superior to that of packed columns, the small loading
factors of a capillary column have limited its sensitivity unless coupled
with a very sensitive detector such as BCD. By using large-bore capillary
columns with a relatively thick layer of stationary phase, sample loadings,
and therefore sensitivity, can be greatly increased without significant losses
in resolution.
Gas chromatographic retention data for 13 phthalate esters and mixtures
have been compiled by Banerjee et al. (82). Table 15 contains operating
conditions and retention data from the GC portion of the study. It should be
noted from Table 15 that in GC analysis of phthalate esters (especially
commercial preparations) problems may be encountered arising from the
distribution of alkyl isomers, which overlap to a great extent. The GC
profile of a particular phthalate ester is influenced by the branching of
alkyl groups and also by the inclusion of asymmetric phthalate esters. High-
resolution capillary columns may be required to completely characterize
specialty phthalate ester plasticizers in commercial use.
Compound identification is usually done by retention time match on two
dissimilar columns, or by mass spectrometry. Quantitation is accomplished
by use of an internal standard such as di-n-nonyl phthalate (48),
bis(2-ethylhexyl) sebacate (23), or a stable-isotope internal standard if
GC/MS is used in the analysis (12,13,14,65,72).
3.4.2 Problems With Gas Chromatography
Three separate problems with the analysis of phthalate esters by GC
have been identified:
• Column and detector performance degradation caused by insufficient
sample cleanup (15,18,30,67)
• Loss of sensitivity at high ECD temperatures (16,17,73)
• Interferences by overlapping peaks (e.g., PCBs, organochlorine
pesticides, sulfur, or triglycerides).
Mass spectrometry appears to be the solution to interference problems
(30,73). Losses in sensitivity of the ECD can be minimized by operation of
29
-------
TABLE 14. GC COLUMNS AND CONDITIONS REPORTED
FOR THE ANALYSIS OF PHTHALATE ESTERS
Colimn
3% SE-30 on Gas Chrom Q (100/200 mesh)
1.5% SP-2250 and 1.95% SP-2401
on Supelcon AW-OMCS (100/200 mesh)
1.5% SP-2250 and 1.95% SP-2401 on
Supelcoport (100/120 mesh)
15 m x 0.25 mm ID Capillary SE-52 (on-
column injection)
8% Diethylene glycol succinate
polyester (DEGS)
3% SE-30
2% OV-17
1.5% SE-52
3% QF-1
10% Polyethylene glycol — 20 M
10% Apiezon M
3% SE-30 (Ultraphase) on Chromosorb W
(AW-OMCS, 60/80 mesh)
10% OV-25 Gas Chrom Q (100/120 mesh)
10% OV-225 HP Chromosorb W
(100/120 mesh)
50 m x 0.3 mm ID Capillary (0.35 )m
film thickness) OV-101
42 m x 0.3 mm ID Capillary (0.39 )m
film thickness) SE-30
Aue packing
Aue packing (100/120 mesh)
1.5% SE-52 Chromosorb W (60/80 mesh)
1.5% OV-17 Shimalite W (80/100 mesh)
3% Silar-lOC Gas Chrom Q
(100/120 mesh)
GC conditions
220'C iso
195'C iso
180'C (4 min hold)
at 4'C/min to
240'C (10 min
hold)
70'C to 150'C at
40'C/min, 150'C
260'C at 6'C/min
180'C iso
230'C iso
230'C iso
180'C iso
230'C iso
230'C iso
250'C iso
210'C iso
l&O'C iso
175'C iso
230'C iso
250'C iso
80'C to 250'C at
4'C/min
100'C (4 min hold)
to 240'C (20 min
hold) at 4'C/min
230'C iso
240'C iso
150'C to 250'C
(7 min hold) at
16*C/min
Detection
Detector limit Reference
ECD 1 ppb 15
ECD 15
ECD 1-100 ppb 16
ECD 0.05-0.1 ppb 17
FID 67
ECD 50
FID 0.5 nmoL/mL 68
FID 69
FID 0.5 ng/ml 21
FID 22
FID 10 ppb 48
MS 0.1-8 ppb 24
(continued)
30
-------
TABLE 14. (continued)
Co limn
2% DEGS -0.5% phosphoric acid
3% SE-30 Gas Chrom Q (100/120 mesh)
3% SE-30 Chromosorb WHP (100/120 mesh)
3% SE-30 Gas Chrom Q (80/100 mesh)
5% OV-101 AW-Chromosorb W
(80/100 mesh)
3% OV-17 on Chromosorb G (80/100 mesh)
4% OV-101 Chromosorb WHP (80/100 mesh)
6% Dexsil 300 on Chromosorb W
3% OV-1 Chromosorb Q (100/120 mesh)
24 m x 0.3 mm ID Capillary SE-54
30 m x 0.32 mm ID Capillary SE-52
3% XE-60 Gas Chrom Q (80/100 mesh)
3% OV-101 Supelcoport
(80/100 mesh)
Outlet end 3% SP-2100/3% OV-210
Inlet end 1% SP-2100/2% SP-2401
Supelcoport (100/120 mesh)
3% OV-17 Chromsorb W DMCS (60/80 mesh)
Aue packing
3% SE-30 Gas Chrom Q (100/200 mesh)
6% OV-210 + 4% SE-30 Chromosorb W (AW)
(60/80 mesh)
GC conditions
—
200'C iso
200'C iso
200'C iso
230'C iso
280-C iso
220-C iso
255'C iso
150-C (2 min hold)
to 240-C, (5 min
hold) at 5'C/min
25'C initial;
100'C to 250'C at
10'C/min
50'C to 320'C at
6"C/min
199'C iso
210'C Iso
200'C iso
--
90'C to 250'C
(15 min hold) at
4'C/min
180'C iso
202'C iso
Detection
Detector limit
—
ECD 1 ng
ECD
ECD 0.5 ng
FID
FID
ECD 10 ng
FID
MS
MS
MS 30 pg
MS
ECD 1.4 ppb
ECD
ECD 1 ng/g
ECD
FID
ECD 30 ng
ECD
Reference
51
76
27
23
71
46
72
73
30
74
26
20
34
47
75
60
(continued)
31
-------
TABLE 14. (concluded)
Colum
GC conditions
Detection
Detector limit
Reference
2% OV-225 Gas Chrom Q (80/100 mesh)
6% OV-210 + 4% SE-30 Chromosorb W (AW)
Aue packing (AP) (ultrathin, thermally
treated layer of Carbowax 20M
deposited as a 0.2% layer on acid-
washed 100/120 mesh Chromosorb W)
3% OV-17 Gas Chrom Q (100/120 mesh)
SE-30
OV-1
2.5% silicone gum E-301 Chromosorb
(BDH)
3% SE-30 Gas Chrom Q (80/100 mesh)
1% DC-560 Gas Chrom Q (100/120 mesh)
3% OV-101 Gas Chrom Q (100/120 mesh)
30 m x 0.32 trni ID Capillary SPB-5
180'C iso
203'C iso
100-C (4 min hold)
to 240'C (50 min
hold) a 4'C/min
230-C iso
223'C iso
125'C (2 min hold)
to 275'C, (7 min
hold) at 15'C/min
ECD
ECD
FID
MS
0.1 pg
ECD 0.01 ng/m3
FID
FID
FID
70'C (1 min hold) MS
to 250'C (hold) at
20'C/min
20-50 ppb
26
19
77
78
79
53
63
14
65
32
-------
8min
1. Dimethyl phthalate
2. Diethyl phthalate
3. Di-n-butyl phthalate
4. Butyl benzyl phthalate
5. Bis(2-ethylhexyl) phthalate
6. Di-n-octyl phthalate
Figure 2. GC/FID chromatogram of phthalate esters analyzed on a 15 m x
0.53 mm ID DB-1 fused-silica capillary column (1.5 im film
thickness); temperature program 150°C to 275°C at 15°C/m1n;
helium at 21 ml/mln. Figure taken from Reference 80.
33
-------
1. Dimethyl phthalate
2. Diethyl phthalate
3. Di-n-butyl phthalate
4. Butyl benzyl phthalate
5. Bis(2-ethylhexyl) phthalate
6. Di-n-octyl phthalate
I I I I I I I
0 2 4 8 8 10 12 14
Figure 3. GC/FID chromatogram of phthalate esters analyzed on a 15 m x
0.53 mm ID SPB-5 fused-silica capillary column (1.5 /an film
thickness); temperature program 115°C (4 min hold) to 250°C
(15 min hold) at 16"C/min; helium at 30 mL/min. Figure taken
from Reference 81.
34
-------
1. Dimethyl phthalate
2. Diethyl phthalate
3. Di-n-butyl phthalate
4. Butyl benzyl phthalate
5. Bis(2-ethylhexyl) phthalate
6. Di-n-octyl phthalate
lOmin
Figure 4. GC/FID chromatogram of phthalate esters analyzed on a 30 m x
0.25 ran ID DB-1301 fused-silica capillary column (0.25 ion film
thickness); temperature program 180eC (1 min hold) to 270°C at
158C/m1n; hydrogen at 50 cm/sec. Figure taken from
Reference 80.
35
-------
TABLE 15. GC CONDITIONS AND RETENTION DATA FOR THE PHTHALATE ESTERS3'5
Ester
GC conditions
Retention
time (rain)
Percent
area
Alkyl chain
length
Dimethyl
Oiethyl
Di-n-butyl
Butylbenzyl
Oihexyl
Butyl-2-ethylhexyl
Bis(2-ethylhexyl)
160'C iso 1.11
Injector temperature: 225'C
Detector temperature: 260'C
Flowrate: 42 mL/min
160'C iso 1.88
Injector temperature: 225'C
Detector temperature: 260'C
Flowrate: 42 mL/min
160'C iso 8.32
Injector temperature: 225*C
Detector temperature: 260'C
Flowrate: 42 mL/min
200'C iso 6.74
Injector temperature: 225'C
Detector temperature: 260'C
Flowrate: 40 mL/min
190'C iso 7.11
Injector temperature: 225'C 7.66
Detector temperature: 260"C 8.58
Flowrate: 40 mL/min 9.31
10.51
160'C (1 min hold) to 220'C at 30'C/min 3.63
Injector temperature: 225'C 5.54
Detector temperature: 260'C 9.20
Flowrate: 42 mL/min
200'C (1 min hold) to 250'C at 30'C/min 3.68
Injector temperature: 250'C 4.75
Detector temperature: 300'C 6.26
Flowrate: 41 mL/min 8.61
160'C (1 min hold) to 220'C at 30'C/min 9.64
Injector temperature: 250'C
Detector temperature: 300'C
Flowrate: 41 mL/min
<5
17
30
31
22
<8
34
66
1
14
33
38
12
12
12
12
12
12
8
12
16
12
14
16
18
16
"Data taken from Reference 82.
GC column 6' x 1/8" glass column packed with 3% OV-1 on Supelcoport 100/120.
(continued)
36
-------
TABLE 15. (concluded)
. Ester
Di isooctyl
01 isononyl
Oi(heptyl,nonyl,
undecyl)
GC conditions
220'C iso
Injector temperature: 225'C
Detector temperature: 260'C
Flowrate: 30 mL/min
180'C (2 min hold) to 250'C at 30'C/min
Injector temperature: 250'C
Detector temperature: 300'C
Flowrate: 28 mL/min
180'C (2 min hold) to 250'C at 30'C/min
Injector temperature: 250'C
Detector temperature: 300'C
Flowrate: 39 mL/min
Retention
time (min)
6.13
7.18
8.42
7.74C
8.05C
8.23C
8.74C
5.97
6.37
7.26
7.88
9.19
10.26
12.32
14.09
17.42
20.56
Percent
area
7
39
54
26
8
28
38
7.2
11.2
11.6
19.2
13.1
21.1
6.01
10.7
<3.0
<3.0
Alkyl chain
length
15, 16
16
16
18
18
18
19
14
14
16
16
18
18
20
20
22
22
Diisodecyl
200'C (1 min hold) to 250'C at 30'C/min 8.26
Injector temperature: 250'C Unresolved
Detector temperature: 300'C envelope
Flowrate: 41 mL/min
19, 20
Diundecyl
Ditridecyl
180"C (2 min hold) to
Injector temperature:
Detector temperature:
Flowrate: 39 mL/min
250'C iso
Injector temperature:
Detector temperature:
Flowrate: 41 mL/min
250'C at 30'C/min
250'C
300'C
250'C
300'C
13.13
15.06
16.36
17.48
20.50
16.7
Unresolved
envelope
3
14
8
28
43
—
22
22
22
22
22
24, 25.
26. 27
"Data taken from Reference 82.
bGC column 6 ft x 1/8 in ID glass column packed with 3% OV-1 on Supelcoport 100/120.
cPoor peak resolution.
37
-------
the detector below 300°C. The only requirement is maintaining the detector
above the highest column temperature, which is usually 240 to 260°C. The
decrease in sensitivity with temperature has been attributed to a nondissocia-
tive electron capture mechanism (16). Figure 5 shows ECD response as a
function of molecular weight, and Tables 16 and 17 list the ECD responses at
320 and 255eC for various phthalate esters, respectively.
The "reaction ECD," reported recently by Hall (83) enhances the per-
formance of the ECD by altering the molecular structure of the compound being
analyzed as it passes through a high-temperature, flowthrough reactor mounted
at the inlet of the ECD. The ECD response in the case of the phthalate esters
increases at moderately high temperatures because the phthalate esters are
converted to the phthalic anhydride, which gives a greater response than the
ester (83).
3.5 HIGH-PERFORMANCE LIQUID CHRONAT06RAPHY
3.5.1 HPLC Methods
High-performance liquid chromatography can be performed in normal
phase, reversed phase, and gel permeation. Normal-phase operation utilizes
a polar stationary phase such as silica or a bonded phase such as cyanopropyl
silane, and elution from this stationary phase by a progressively more polar
solvent gradient. Reversed-phase operation makes use of a nonpolar bonded
phase such as octadecyl silane, and elution from this stationary phase by a
progressively less polar solvent. Gel permeation uses a porous polymer phase
similar to that used in preparative work, but with a smaller particle size.
High-molecular-weight compounds elute first because of their exclusion from
the gel pores, and low-molecular-weight compounds elute later depending on
their diffusion into and out of the pores of the gel. All three techniques
have been investigated for the separation of phthalate esters
(27,31,33,35,46,53,84).
Banerjee et al. (82) studied the retention of 13 phthalate esters and
mixtures using reversed-phase HPLC and reported retention at several mobile
phase compositions (Table 18).
Mori (31) utilized a Shodex HP-255 porous polymer bead column for
adsorption (normal- and reversed-phase) chromatography with n-hexane for
normal-phase operation (Figure 6) or methanol for reversed-phase operation
(Figure 7). Also, a Shodex A801 styrene-divinyl benzene resin column was used
for gel permeation chromatography, with an exclusion limit of 1,000 as
polystyrene, with chloroform as eluant (Figure 8). UV detection at 224 nm for
hexane and methanol, and 243 nm for chloroform were reported (31). Other
chromatographic methods operating in the normal-phase mode are listed in
Table 19.
Otsuki (33) investigated the use of hyperbolic gradients for the
separation of phthalate esters by reversed-phase HPLC on a C-18 column using
a methanol-water gradient. Figure 9 shows the separation obtained, while
Figure 10 shows the relationship between retention time and linear alky! chain
38
-------
8.0
7.S
< 7.0
I
6.5
6.0
150 200
250 300
MW
350 400
Figure 5. ECD response as a function of phthalate ester molecular weight.
Figure taken from Reference 16.
39
-------
TABLE 16. ECD RESPONSES AT 320°C
Compound
Retention time
(min)
Detector response
area/ng
(x 10-3)
Dimethyl phthalate
Dibutyl phthalate
Butyl benzyl phthalate
Bis(2-ethylhexyl) phthalate
6.0
12.1
17.1
20.6
43
29
334
52
aData taken from Reference 17.
TABLE 17. ECD RESPONSES AT 255°C
Compound
GC retention
time
(min)
Detector response9
area/nmol
(xlO-6)
Dimethyl phthalate
Diethyl phthalate
Diallyl phthalate
Di butyl phthalate
Dihexyl phthalate
Butyl benzyl phthalate
Bis(2-ethylhexyl) phthalate
Diphenyl phthalate
Dioctyl phthalate
2.5
3.7
6.4
10.5
18.0
18.8
21.0
25.5
26.5
74
51
67
13
8
90
16
78
5
aData taken from Reference 16.
GC: 1.5% SP-2250 and 1.95% SP-2401 on 100/120 mesh Supelcoport.
40
-------
TABLE 18. HPLC CONDITIONS AND RETENTION DATA FOR THE PHTHALATE ESTERSa'b
HPLC conditions
mobile phase
Acetonitrile
Phthalate (percent)
Dimethyl
Diethyl
Di-n-butyl
Butyl benzyl
Dihexyl
Butyl -2-ethylhexyl
Di(n-hexyl,n-octyl,
n-decyl)
Bis(Z-ethylhexyl)
Diisooctyl
Diisononyl
Di(heptyl, nonyl
undecyl )
Diisodecyl
Di undecyl
Oitridecyl
70
50
80
70
60
50
80
70
60
70
60
70
90
80
70
60
80
70
65
70
60
80
70
65
80
70
65
85
80
70
65
85
70
95
90
80
Mater
(percent)
30
50
20
30
40
50
20
30
40
30
40
30
10
20
30
40
20
30
35
30
40
20
30
35
20
30
35
15
20
30
35
15
30
5
10
20
No. of
peaks
1
1
1
1
1
1
1
1
1
1
2
3
1
4
5
5
1
1
1
1
1
2
3
3
5
6
6
1
3
3
3
1
1
2
2
4
Retention volume
(mL)
5.2
7.6
4.4
7.2
10.4
22.0
5.5
5.9
8.7
10.1
16.8, 19.3
6.4, 8.7, 13.3
4.6
5.5, 6.4, 7.8, 9.7
8.3, 10.6, 13.3, 17.9, 23.9
17.9, 25.8, 38.2, 58.9, 89.2
8.3
15.2
19.3
14.3
29.4
6.4, 7.4
9.7, 11.0, 12.0
26.2, 31.7, 38.6
6.9, 9.2, 11.0, 11.7, 13.3
11.0, 12.4, 15.6, 19.8, 23.9,
34.5
12.0, 15.6, 21.2, 28.5, 38.2,
53.3
8.3
9.2, 10.1, 11.5
18.4, 22.1, 26.7
27.1, 33.1, 40.5
10.1
32.7
9.7, 10.1
9.7, 10.6
20.2, 24.8, 27.6, 32.7
'Data taken from Reference 82.
bHPLC column 10 tan RP-2, UV 200 nm.
41
-------
o
in
i
0)
a
u
-------
-------
a
n
I
ta
<0
•o
u
o
o
a
a:
OHP
DBP
DLP
20
30
Elution Volume (ml)
Figure 8. High-performance GPC separation of phthalate esters (Shodex
A801, chloroform). Figure taken from Reference 31.
44
-------
TABLE 19. SUMMARY OF CONDITIONS FOR NORMAL-PHASE HPLC
SEPARATIONS OF PHTHALATE ESTERS
Detection
Column Eluant Detector limit Reference
Porasil 10 /zm 1:1 Methylene chloride/ UV-254 nm 0.05 /ig 84
isooctane
Shodex Porous n-hexane UV-254 nm 2 nga 31
Polymer Beads HP-255 UV-224 nm
Pellicular Peri sorb A 2% diisopropyl ether in UV-281 nm 53
n-hexane
Silica gel LiChrosorb 1:2 n-hexane/methylene UV-233 nm 10 ng 46
SI 60 (5 /im) chloride with 0.2% UV-224 nm
ethanol
Zorbax-CN 2% isopropanol/hexane UV-224 nm 20 ng 27
aValue given is for OOP at 224 nm.
45
-------
rtoo
i. 5
z
-so 3
-0
10
20 min
Figure 9. Cig Reversed-phase HPLC separation of phthalate esters using
methanol-water. Figure taken from Reference 33.
20
t-e
Z°
_| L III I I 1 I I
45 7 9 10
LINEAR ALKYL CARBON NUMBER
Figure 10. Retention time of phthalate esters as a function of alkyl chain
length (A - gradient elution with a 10-min holding time;
B - gradient elution without the holding process). Figure taken
from Reference 33.
46
-------
number. The phthalate esters were found to be completely adsorbed onto the
bonded phase with 100-percent water eluant. The hyperbolic gradient with the
introduction of a holding time during the gradient allowed elution of other
organic compounds prior to that of the phthalate esters.
Van Vliet et al. (35) combined sample collection, preconcentration,
and sample preparation into a single step to decrease the amount of contamina-
tion introduced by.handling during each step in the process. Water samples
were passed through a CJ8 precolumn where phthalates were completely adsorbed
onto the stationary phase. Later, elution, using a methanol gradient,
separated the phthalate esters. Experiments indicated that the passage of too
large a sample volume resulted in breakthrough of those sample components that
had very small capacity ratios. Three solutions to the problem were
offered (35):
• Carefully designed gradient profile to achieve separation of the
compounds of interest from the contaminants
• Selection of more selective stationary phases for the trace
enrichment step
• More selective detection (e.g., fluorescence, MS, post-column
derivatization).
Post-column derivatization is particularly attractive since, after the
separation step, the phthalate esters could be detected as a fluorescent
derivative of phthalic acid. This derivative could be independent of the
particular ester, and the added selectivity of the derivatization step
combined with a highly sensitive detection method could greatly increase the
utility and scope of method.
3.6 CONFIRMATION OF COMPOUND IDENTITY
Three basic techniques have been used for the confirmation of the
identities of phthalate esters:
• Comparison of the retention times on two or more stationary phases
• Formation of a derivative and either analysis of the derivative or
observation of the loss of the suspected phthalate peak in the
chromatogram
• Identification by gas chromatography/mass spectrometry.
Confirmation of phthalate esters by hydrolysis and peak disappearance
has been reported (20). The samples are analyzed both before and after
treatment with alkaline methanol, which saponifies the esters to the cor-
responding alkanols and phthalic acid salts.
47
-------
Conversion of phthalate esters to the corresponding phthalic acid
followed by reaction to produce a very ECD-sensitive ester derivative has
been reported by several investigators (18,26,75,76,85).
Giam et al. (75) confirmed phthalate esters in environmental samples
by derivatization involving: (a) hydrolysis to the acid; (b) fusion with
2-chloroethylamine hydrochloride to yield N-(2-chloroethylJphthalimide, (c)
back extraction of the reaction mixture into isooctane, and (d) analysis by
GC/ECD. The method is simple, fast, inexpensive, and sensitive. It produces
the derivative in high yield (90 percent), free from most other contaminants,
and requires minimal amounts of glassware and reagents.
Takashita et al. (76) utilized a more complicated procedure in which
the phthalate esters were also hydrolyzed, esterified with
BF3-2,2,2-trifluorp-ethanol solution, and extracted into hexane before GC/ECD
analysis. Derivatization yields of 100 percent were achieved, and the overall
recoveries ranged from 70 to 110 percent (76). The detection limit of the
derivative was 0.1 pg; this represents approximately a 100-fold increase in
sensitivity over that generally obtained when the phthalate esters are
directly analyzed by GC/ECD. No impurities from byproducts were noted.
Glazer et al. (18) developed a procedure using BF3-methanol for the
esterification reaction and 4-chlorophthalate as internal standard. The
method determines phthalates, isophthalates, and terephthalates
simultaneously.
Arbin and Ostelius (26) performed derivatization by a solid-liquid
phase-transfer catalysis, utilizing hexyl iodide to form a di-ester for GC/ECD
analysis. This approach allowed the determination of both phthalate diesters
and metabolite monoesters to be analyzed simultaneously. Cleanup of the
reaction mixture by extraction with 1M H2S04, followed by addition of silver
sulfate to the organic phase, was necessary to prevent tailing caused by
degradation products from quaternary ammonium halides (byproducts of the
alkylation reaction). The efficiency of the reaction was >99 percent.
Mass spectrometry has been used for compound confirmation by Michael
et al. (24) and Safe and Hutzinger (86). Mes et al. (60) confirmed the
presence and identity of phthalate esters by MS after separation by alumina
thin-layer chromatography.
Because of the high phthalate ester concentrations relative to other
trace organics, Cautreels and van Cauwenberghe (87) suggest using secondary
ions since the fragment ions at mass 149 may overload the collector.
Dziwinski et al. (71) reported that the identification of long-chain
dialkyl phthalates by electron impact MS is made difficult because the
molecular ions M+ are not detectable in the MS. Better results are obtained
with field ionization or chemical ionization. Addison (73) noted a decrease
in [M+H]+ at higher reaction temperatures. Comparison of electron impact and
chemical ionization for the identification of phthalate demonstrated that
isobutane is the preferred gas, since isobutane produces simpler spectra,
48
-------
gives greater abundance of the quasimolecular >[M+H]+ ions, and forms no
adduct ions.
Hunt et al. (88) have reported the use of an MS/MS technique to avoid
the preparation and separation steps usually involved in environmental
analysis. They identify several disadvantages of the current analytical
methods involving GC and GC/MS:
• Inability to detect highly polar, nonvolatile, or thermally labile
compounds
• High labor costs associated with sample preparation
• Large amount of time required per analysis.
MS/MS with collision-activated dissociation (CAD) utilizing a triple
quadrupole offers several advantages:
• Direct analysis
• Rapid qualitative and semi-qualitative analysis
• Applicable to both liquid and solid matrices
• Analysis at the 10 to 100 ppb level
• Elimination of most sample preparation and chromatographic steps
• Detection of both knowns and unknowns by molecular weight and
functional group
• Total analysis time of under 30 minutes per sample.
Figure 11 shows the phthalate ester fragmentation scheme upon
electron-impact. The mass 149 fragment, corresponding to the protonated
phthalic anhydride, is produced in high abundance for all but the dimethyl
phthalate. Electron impact mass spectra of 5 phthalate esters are shown in
Figure 12.
3.7 BACKGROUND CONTAMINATION
Interferences in the determination of phthalate esters are common,
though few detailed studies have been undertaken. Background contamination
from the ubiquitous nature of phthalate esters (DEHP and DBF) has become a
greater concern since much lower levels of phthalate esters are being traced
in the environment. Giam et al. (15) stated that the major problem in the
phthalate ester analysis is the reduction of background contamination to
levels less than the levels generally present in samples. Phthalate esters
are present on almost all laboratory equipment. Giam et al. (15) also
indicated that nonplastic materials (cork, glass wool, Teflon sheets, and
aluminum foil) contain DEHP. When reported, the procedure blank levels tended
49
-------
0
II
COST
II CHS
COET
0
^
m/x 177
•COET
CAO
^
m/z223
^COET CAD,
|f - ETON,
SH"
O
+O'H
149
Figure 11. Phthalate ester fragmentation scheme. Figure taken from
Reference 88.
77
... i.t.>....!- .
163
OMETHW PHTHAUtTE
J_LJ_
oi-0-airrrt. PHTHALATE
223
OUTYiaSNZYt. PMTHALATE
91
.1 I.L
sr
M9
«7
Ot-2-CTMYUiEXVI. PHTHAkATS
279
I
OI-A-OCm. PMTHALATJ
2S2
279
M 00 BO 200 2SO 300 030 «»
Figure 12. Electron impact mass spectra of common phthalate esters.
taken from Reference 5.
50
Figure
-------
to be very high (1,500 ng for DEHP, 100 ng for DBF), regardless of careful
preparation. The problem stems from four areas related to most analytical
techniques used to determine phthalate esters:
• Contamination by traces of phthalate esters in high-purity solvents
which becomes appreciable when solvent is concentrated to a few
milliliters
• Contamination from laboratory items containing parts comprised of
phthalates, or processed with equipment containing phthalate esters
• Contamination from adsorption of phthalate esters from the
laboratory onto equipment and glassware
• Losses incurred in the concentration step when the sample extract
is evaporated to dryness.
Each of these background sources will be discussed in detail below.
Bowers et al. (21) have undertaken a detailed study of high-purity
solvents used in trace organic analysis. Distilled-in-glass and
pesticide-grade solvents commonly used in the analyses of environmental
samples for organic compounds at trace levels were concentrated 200-fold and
analyzed by GC and GC/MS to determine contaminants and contaminant levels.
Table 20 lists the total amount of impurities found in the different solvents.
Lower levels were determined by selected ion monitoring (SIM) and are
presented in Table 21.
Distilled-in-glass cyclohexane, methylene chloride, and methanol
solvents contained an estimated total of 1 to 150 ng of organic impurities per
mL of uncondensed solvent. At a 200-fold concentration, this corresponds to
0.2 to 3 ng/mi of impurities. Included were phthalate esters, n-hydrocarbons,
and chlorinated hydrocarbons (21). Up to 21 compounds were identified in
pesticide-grade solvents, with a maximum single component concentration of 30
to 50 ng/mL of uncondensed solvent. It was found that distilled-in-glass
grade solvents contained fewer and lesser amounts of similar impurities, and
therefore are most suitable for trace organic analysis (21).
Ishida et al. (48) detected phthalate esters in plastic laboratory
apparatus and in benzene, chloroform, n-hexane, and light petroleum ether.
They studied this contamination problem in greater detail, examining solvents,
water, solids, and laboratory apparatus. Table 22 lists the concentrations
of DBF and DEHP in solvents and water. Table 23 shows the levels of phthalate
esters present in solids commonly used in the laboratory, and Table 24 lists
the concentrations found in various materials. Removal techniques for these
phthalate esters contaminants were also investigated. It was found that the
phthalate esters could be removed from organic solvents by distillation.
Solids could be cleaned by either heating to 300°C for 12 hours (where
applicable) or by extraction with chloroform-methanol.
51
-------
TABLE 20. IDENTITIES AND CONCENTRATIONS OF ORGANIC COMPOUNDS IN SOLVENTS BY GC/MS1
01
ro
Caledon Labs Burdick & Jackson
Fisher Scientific
Concentration Concentration
Solvent RIb Compound (ng/mL) RI Compound (ng/mL)
Cyclohexane 1150 C6H120 49 2180 Diethyl phthalate 3.5
2878 Dioctyl phthalate 1.8
Methylene 2200 Diethyl phthalate 0.52
chloride 2350 Dibutyl phthalate 7.7
Methanol
RI
1085
1705
1828
1027
1038
1155
1655
2760
1265
1475
Compound
n-Butyl-n-butyrate
Tributyl phosphate isomer
Tributyl phosphate isomer
1.1,1 -Tr ich loropropane
1,2,3-Trichloropropane
1,1,1, 2-Tetrachloropropane
Tetrachloroethane
Phthalic anhydride
Dioctyl phthalate
C8H12; l,4-Bis(methylene)-
cyclohexane
C8H100; 2,6-Dimethylphenol
Concentration
(ng/mL)
29
2.9
0.89
16
21
1.9
0.42
14
1.6
2.6
"Data taken from Reference 21.
RI — retention index.
-------
TABLE 21. IDENTITIES AND CONCENTRATIONS OF ORGANIC COMPOUNDS IN SOLVENTS BY SIM*
en
to
Solvent
Cyc lohexane
Hethylene
chloride
Hethanol
Caledon
Compound
Dibutyl phthalate
Dioctyl phthalate
Dibutyl phthalate
Dioctyl phthalate
Dibutyl phthalate
Dioctyl phthalate
Labs
Concentration
(ng/mL)
0.1
<0.1
0.2
0.2
0.4
0.2
Burdick & Jackson
Compound
Dibutyl phthalate
Oioctyl phthalate
Tetracosane
Pentacosane
Hexacosane
Heptacosane
Octacosane
Nonacosane
Triacontane
Dibutyl phthalate
Dioctyl phthalate
Dibutyl phthalate
Dioctyl phthalate
Concentration
(ng/mL)
0.5
0.2
0.1
0.2
0.2
0.2
0.2
0.1
0.1
0.9
0.2
0.1
0.1
Fisher Scientific
Compound
Dimethyl phthalate
Diethyl phthalate
Dibutyl phthalate
Dioctyl phthalate
Dimethyl phthalate
Diethyl phthalate
Dibutyl phthalate
Dioctyl phthalate
Dimethyl phthalate
Diethyl phthalate
Dibutyl phthalate
Dioctyl phthalate
Concentration
(ng/mL)
<0.1
0.4
7
1
<0.1
0.1
0.7
11
<0.1
0.1
1
0.6
*Data taken from Reference 21.
-------
TABLE 22. CONCENTRATIONS OF DBP AND DEHP IN
ORGANIC SOLVENTS AND WATERS9
Concentration
Sample
DBP
DEHP
City water
Well water0
Tap water (well water)
Ion-exchanged water
Benzene
Acetone
n-Hexane
Chloroform
Diethyl ether
Methanol
Ethanol
Light petroleum
Dichloromethane
Ethyl acetate
Acetonitrile
2.04 ppb
2.49 ppb
1.93 ppb
0.83 ppb
0.17 ppm
Trace
4.82 ppb
3.85 ppb
1.31 ppb
1.96 ppm
43.6 ppb
78.7 ppb
69.3 ppb 61.7 ppb
0.18 ppm
aData taken from Reference 48.
bFrom Tohoku University.
°Less than 10 ppb (109).
54
-------
TABLE 23. CONCENTRATIONS OF DBP AND DEHP IN SOLID REAGENTS9
Concentration
Reagent
Silicic acidb
Silicic acicf
Wakogel S-l
Floridil (PR)d
Floridil (EP)e
Alumina powder
Celite (GR)f
Activated charcoal powder
Na2S04 (EP)
Na2S04 (GR)
Na2S04 (PR)
Dnerite
CaC03 (GR)
NaCl (EP)
Nad (GR)
DEAE-cellulose
CM-cellulose
Sephadex G-100
Amberlite IR-45
Dowex L-X8
DBP
0.59 ppm
0.23 ppm
1.61 ppm
23.8 ppb
91.9 ppb
41.8 ppb
0.12 ppm
95.8 ppb
0.09 ppm
24.3 ppb
24.3 ppb
16.7 ppb
18.7 ppb
Trace
Trace
0.25 ppm
9.89 ppm
4.32 ppm
2.27 ppm
9.76 ppm
DEHP
2.36 ppm
0.97 ppm
0.84 ppm
63.5 ppb
87.5 ppb
44.8 ppb
Trace9
47.9 ppb
0.04 ppm
16.9 ppb
14.5 ppb
Trace
21.6 ppb
0.11 ppm
11.8 ppb
0.41 ppm
4.12 ppm
Trace
0.25 ppm
0.19 ppm
Company
E. Merck
Mallinkcrodt
Wako
Floridin
Wako
Wako
Wako
Wako
Wako
Wako
Wako
W. A. Hammond
Wako
Wako
Wako
Pharmacia
Pharmacia
Pharmacia
Rohm and Haas
Dow Chemical
aData taken from Reference 48.
bFor thin layer chromatography.
cFor column chromatography.
dChemical for analysis of pesticide residues.
eExtra-pure reagent.
Guaranteed reagent.
9Less than 10 ppb.
55
-------
TABLE 24. CONCENTRATIONS OF DBP AND DEHP IN VARIOUS MATERIALS8
Concentration
Material
DBP
DEHP
Company
Heavy-walled tubing
(elicon tubing)
Polyvinyl tubing
Black rubber tubing
PTFE tubing
Black rubber stopper
Red rubber stopper
Injection packing0
Vial cap
Wash bottle
Cork stopper
Thimble filter-paper
Filter-paper No. 2
Chromatography paper
Glass-wool
Kimwipe
Parafilm
Saranwrap
Aluminum foil A
Aluminum foil B
Aluminum foil C
--
23.3%
--
Traceb
--
Trace
22.5 ppm
--
--
0.05%
5.28 ppm
0.01%
0.83 ppm
41.9 ppm
--
8.28 ppm
0.13 ppm
1.62 ppm
0.97 ppm
67.2%
11.6%
0.2%
0.3%
--
0.25 ppm
0.65 ppm
--
--
1.34 ppm
0.62 ppm
2.28 ppm
4.15 ppm
--
--
8.35 ppm
4.85 ppm
1.01 ppm
0.60 ppm
Seimi
Plastec
Seimi
Niplon Products
Seimi
Seimi
Hitachi
Seimi
Seimi
Seimi
Toyo Roshi
Toyo Roshi
Whatmann
Wako
Kimberly-Clark
American Can
Asahidow
Toyo Alumi
Showa Alumi
Mitsubishi Alumi
aData taken from Reference 48.
bLess than 10 ppb.
clnjection packing for gas chromatograph.
56
-------
Giam et al. (15) found that normally all reagents were pure enough
except for diethyl ether, which had to be freshly distilled because of
contamination problems during storage. Materials for their work were either
extracted with petroleum ether, or heated to 320°C for 10 hours, and covered
with purified aluminum foil. A check was made of all solvents and reagents
prior to use by 200-fold concentration and GC/ECD analysis. Giam et al. (15)
found filter paper to be a major problem; the use of glass fiber filters,
which can be heated, was suggested. Previously cleaned Teflon sheets were
used in place of rubber stoppers to hold Buchner funnels to minimize
extraction of phthalate esters from the rubber. Also reported was a possible
problem posed by airborne contaminants; levels of DEHP at air conditioning
vents were found to be about 35 ng/m3.
Table 25 lists the various cleanup and testing methods reported in the
literature to determine and control the extent of phthalate ester background
contamination.
Glass sample vials with foil-lined screw caps are generally considered
to be sufficiently inert to produce no sample contamination. Bowers et al.
(21) and Denney et al. (22) noted that storage of samples in a methanol
solvent for several weeks caused substantial changes in composition. These
samples had been stored in glass sample vials with Sn-Pb alloy foil-lined
screw caps at room temperature. They concluded that the contaminants
originated from the cork backing of the metal foil. This, they postulated,
occurred through permeation by the methanol solvent vapor. To determine the
cause, extent, and prevention of the contamination, Denney et al. (22) tested
sample vials with foil or PTFE liners. The vials were filled with methanol
and sealed for 8, 24, 48, and 72 hours, and the solvent was then analyzed by
GC/MS. The analysis after only 8 hours indicated the presence of three
compounds in the vial with the foil-lined screw cap, while the PTFE-lined
samples showed no contamination. After 72 hours, the vials with the
foil-liner showed nine contaminants while the PTFE-lined vials still showed
no contamination. Solvents with vapor pressures different from that of
methanol may produce different degrees of contamination. It was recommended
(22) that PTFE-rubber laminated discs, or their equivalent, be used in
conjunction with screw cap vials to insure sample integrity. The effect of
sample storage temperature was not investigated.
Burke et al. (89) reported losses of phthalate esters whenever extracts
were concentrated to below 500 /*L, independent of the concentration technique.
Chiba reported that the use of petroleum ether as the extraction solvent
resulted in greater losses upon condensation of the solvent as compared to
benzene (90). Detectable sample loss was observed even when a viscous
solvent, such as ethylene glycol, was used. Bowers et al. (47) examined the
problems which could occur when samples or sample extracts are taken to
dryness and then reconstituted as part of the analytical preparative scheme.
Surface-active agents were used to prevent losses occurring by irreversible
adsorption onto glass surfaces, since silylation only masks silanol groups
leaving active metal sites accessible. The surface coating technique appeared
to make little difference in the case of phthalate esters (47). Greater
losses were noted for samples with low concentrations (10 ng//*L versus
57
-------
TABLE 25. CLEANUP PROCEDURES USED ON REAGENTS, GLASSWARE,
AND OTHER LABORATORY ITEMS
Ite
Procedure
Comments
Reference
Water
Liquid-liquid extraction with petroleum ether
Ooubly-distillation in all glass still
Adsorption on a series of Sep Pak columns
Adsorption onto charcoal filters
Extraction with n-hexane
Extraction with methylene chloride
Redistillation from potassium permanganate/potassium
hydroxide
Redistillation
Distillation/hexane extraction
Phthalates not removed by
distillation
15.27,70
16
17
68
48.76
30
33
34
35
Diethyl ether
cn
00
Distillation
Extraction with methanol/sodlum hydroxide after reflux,
washing with water, then drying
Check by concentration
and GC/ECD analysis
15.70
68
23
15.19.27.30.48,
50.70.76
Sodium chloride
Sodium sulfate
Florisil
Alumina
Silica gel
Celite
Heating at 320'C
Heating at 400'C for 10 hours
Heating at 320'C
Heating at 210'C for several hours
Heating at 400'C for 10 hours
Heating at 650'C for 3 hours
Heating at 500'C for 3 hours
Extraction with methylene chloride
Washing with methylene chloride, acetone, then hexane,
drying at 120'C
Hashing with acetone, then hexane
Heating at 320'C
Soxhlet extraction for 12 hours with hexane, storage in
n-hexane
• Heating at 210'C for several hours
• Extraction with methylene chloride
• 800'C for 4 hours
• Heating at 210'C for several hours
• Extraction with methylene chloride
• Heating at 500'C for 3 hours
15.27,70
17
15,27.70
16
17
51
23
30
20
34
15,27.70
46
16
30
20
16
30
26
-------
TABLE 25. (continued)
Ue
Procedure
Contents
Reference
Polyurethane foam (PUF)
Paper filters
Soxhlet thimbles
Glass fiber filters
Soxhlet extraction for 14 hours with acetone
Soxhlet extraction sequentially with: methano). toluene,
methylene chloride, acetone, and petroleum ether
Soxhlet extraction for 12 hours with hexane
Soxhlet extraction for 12 hours with hexane
Heating at 320'C
34
46
46
15.27.70.78
Glassware
cn
Acid washing, rinsing with acetone, then hexane
Washing with Alconox, sonication. rinsing with tap water.
then delon(zed water. Heating at 300'C for 1 hour.
Washing In 5% "Decon 90," soaking in 10X nitric acid.
rinsing with tap water, soaking in methylene chloride-
methanol (1:1) 24 hours, rinsing with methylene chloride
before use
Rinsing with acetone, then hexane, drying at 150'C
Detergent washing, rinsing with hot tap water, acetone.
then hexane
Keeping in chromic acid for >3 hours before use
Washing in acetone and n-hexane before use
Washing in water, heating at 250'C for 10 hours, or rinsing
with acetone (4x) and hexane (2x)
Rinsing with acetone
Cleaning with Extran solution, washing with deionized
water, heating at 400"C for 10 hours, and storing in
aluminum foil
Detergent washing rinsing in: tap water, reagent grade
acetone, heating at 210'C overnight, hexane rinsing before
use
Washing with cleaning solution (Micro), rinsing with
deiontzed water, distilled acetone, nanopure acetone
(distilled over 5-ft column and potassium permanganate).
Heating at 320'C for 10 hours; covering with aluminum foil
(treated); rinsing with nanograde petroleum ether before
Use only glass or stainless-steel
labware
19.34.60
47
30
34
46
23
51
48.51
68
17
16
15,27.70
(continueoj
-------
TABLE 25. (concluded)
Ite
Procedure
Contents
Reference
Aluminum foil
Heating at 350'C to 400'C for 14 hours
Washing with cleaning solution (Micro), rinsing with
deionized water, distilled acetone, nanopure acetone
(distilled over 5-ft column and potassium permanganate).
Heating at 320'C for 10 hours; covering with aluminum foil
(treated), rinsing with nanograde petroleum ether before
use.
Cleaning with Extran solution, washing with deionized
water, heating at 400T for 10 hrs.
48
15.27.70
17
en
o
Stopcocks
Spoons,
Porcelain ware.
Blender,
Spatulas,
Forceps,
Dissecting knives
Glass wool
GLC septa
Volumetric flasks
• Washing and rinsing as above for glassware
• Washing with petroleum ether
• Washing with cleaning solution (Micro), rinsing with
deionized water, distilled acetone, nanopure acetone
.(distilled over 5 ft column and potassium permanganate).
Heating at 320"C for 10 hours; covering with aluminum foil
(treated); rinsing with nanograde petroleum ether before
use.
• Cleaning with Extran solution, washing with deionized
water, heating at 400'C for 10 hours, and storing in
aluminum foil
• Rinsing with methylene chloride and drying at 180"C
• Heating at 500'C for 3 hours
• Extraction with methylene chloride
• Baking at 250'C for several weeks before using, then
backing with Teflon discs
• Washing In acetone, hexane, drying in vacuum
15.27.70
17
Check prior to use by screening:
rinse with methylene chloride or
hexane and analyze by GC/MS
24
23
30
30
34
-------
100 ng/jzL, or 1 percent versus 10 percent). Slight concentration increases,
as well as losses, were noted for the phthalate esters (Table 26).
Van Vliet et al. (35) developed a preconcentration method using an
HPLC precolumn for HPLC analysis of phthalate esters which eliminates much
of the contamination stemming from laboratory apparatus and solvents.
Preconcentration methods based on the precolumn collection technique cannot
be recommended for complex matrices and heavily contaminated samples, because
resolution of the phthalate esters from matrix components may be insufficient.
3.8 STABILITY OF PHTHALATE ESTERS
Little information was obtained from the literature on the stability
of the phthalate esters in various matrices. However, storage of samples at
room temperature should be avoided since bacterial degradation and
base-catalyzed hydrolysis do occur (16,18,24,26).
Payne and Benner (27) studied the losses of phthalate esters as a
function of time at room temperature and at 4°C. The data are presented in
Table 27. Losses incurred in samples kept under fluorescent light were higher
than in samples kept in the dark. The best storage method was found to be
refrigeration (no loss in 60 days). However, environmental samples must be
analyzed for phthalates immediately, since sample losses of 64 percent in 1
day and 89 percent in 2 days were noted for DEHP (27).
Freezing at -10 to -30°C appears to be the most suitable method for
storage of solid matrices since it has the widest range of application, causes
the least changes in the samples, and makes the addition of preservatives
unnecessary.
61
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TABLE 26. COMPONENT LOSSES AFTER EVAPORATION-RECONSTITUTION OF 100-ng//iL AND 10-ng//il
STANDARD SOLUTIONS*
o>
Loss (ng//iL)
Phthalate ester
Dimethyl
Diethyl
Di butyl
Dioctyl
Dimethyl
Diethyl
Di butyl
Dioctyl
Retention
time
(min)
8.4
10.6
18.0
28.9
8.4
10.6
18.0
28.9
Original
solution
(ngM)
134
101
102
101
13.4
10.1
10.2
10.1
Deactivated
No. 1
36
17
8
6
13.4
5.0
+1.2
+1.0
Deactivated
No. 2
40
21
14
13
13.4
6.9
0.8
0.5
Untreated
No. 1
42
24
19
17
13.4
5.1
0.4
0.5
Untreated
No. 2
31
15
12
10
13.4
4.8
1.1
1.0
"Data taken from Reference 47.
-------
TABLE 27. BIS(2-ETHYLHEXYL) PHTHALATE LOSS AS A FUNCTION OF
STORAGE CONDITIONS AND STORAGE TIME'
Conditions
Time Losses
interval (%)
Evaporation to dryness in an air stream
(solvent methylene chloride)
(Solvent: water) Ambient temperature
(fluorescent lighting)
(Solvent: water) Refrigeration at 4°C
Environmental water sample
0.5 hours 10 to 25
4 Days
12 Days
14 Days
60 Days
24 Hours
48 Hours
20
70
>99
<1
64
89
aData taken from Reference 27.
63
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70
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APPENDIX B
METHOD 8061'
PHTHALATE ESTERS
"Method 8061 Is the revised version of Method 8060.
-------
METHOD 8061
PHTHALATE ESTERS
1.0 SCOPE AND APPLICATION
1.1 Method 8061 is used to determine the identities and concentrations
of various phthalate esters in liquid and solid matrices. Table 1 lists
compounds that may be determined by this method, their CAS Registry Numbers,
and their method detection limits (MDL) in a water matrix. The MDLs for the
components of a specific sample may differ from those listed in Table 1
because MDLs depend on the nature of interferences in the sample matrix.
Table 2 lists the practical quantitation limits (PQL) for other matrices.
1.2 When this method is used to analyze for any or all of the compounds
listed in Table 1, compound identification should be supported by at least one
additional qualitative technique. This method describes conditions for
parallel column, dual electron capture detector analysis which fulfills the
above requirement. Retention time information obtained on two megabore fused-
silica open tubular columns is given in Table 1. Alternatively, gas
chromatography/mass spectrometry could be used for compound confirmation.
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of a gas chromatograph and in the
interpretation of gas chromatograms.
2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 liter for
liquids, 10 to 30 g for solids) is extracted by using the appropriate sample
extraction technique specified in Methods 3510, 3540, and 3550. Method 3520
is not recommended for the extraction of aqueous samples because the longer
chain esters dihexyl phthalate, bis(2-ethylhexyl) phthalate, di-n-octyl
phthalate, and dinonyl phthalate tend to adsorb to the glassware and
consequently, their extraction recoveries are <40 percent. Aqueous samples
are extracted at a pH of 5 to 7 with methylene chloride in a separatory funnel
(Method 3510). Alternatively, aqueous samples could be filtered through 3M-
Empore membrane disks that contain C^-bonded silica. The phthalate esters are
retained by the silica and later eluted with, acetonitrile. Solid samples are
extracted with hexane/acetone (1:1) or methylene chloride/acetone (1:1) in a
Soxhlet extractor (Method 3540) or with a sonicator (Method 3550). After
cleanup, the extract is analyzed by gas chromatography with electron capture
detection (GC/ECD).
2.2 The sensitivity of Method 8061 usually depends on the level of
interferences rather than on instrumental limitations. If interferences
prevent detection of the analytes, cleanup of the sample extracts is
necessary. Either Method 3610 or 3620 alone or followed by Method 3660,
Sulfur Cleanup, may be used to eliminate interferences in the analysis.
Method 3640, gel-permeation cleanup, is applicable for samples that contain
high amounts of lipids and waxes.
8061-1 Revision 2
September 1989
****September 1989****
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3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Interferences coextracted from the samples will vary considerably
from waste to waste. While general cleanup techniques are referenced or
provided as part of this method, unique samples may require additional cleanup
approaches to achieve desired sensitivities for the target analytes.
3.3 Glassware must be scrupulously clean. All glassware require
treatment in a muffle furnace at 400*C for 2 to 4 hrs or thorough rinsing with
pesticide-grade solvent prior to use. Refer to Chapter 4, Step 4.1.4 for
further details regarding the cleaning of glassware. Volumetric glassware
should not be heated in a muffle furnace.
Soxhlet extractors cannot be baked in the muffle furnace, and
thorough rinsing with hot tap water, followed by deionized water and acetone
is not adequate. Even after a Soxhlet extractor was refluxed with acetone
for three days, with daily solvent changes, the levels of bis(2-ethylhexyl)
phthalate were as high as 500 ng per washing. Storage of glassware in the
laboratory introduces contamination even if the glassware is wrapped in
aluminum foil. Therefore, any glassware used in Method 8061 should be cleaned
immediately prior to use.
3.4 Florisil and alumina may be contaminated with phthalate esters and,
therefore, use of these materials in sample cleanup should be employed
cautiously. Washing of these materials prior to use with the solvent(s) that
is/are used for elution during extract cleanup was found helpful, however,
heating at 320eC for Florisil and 210°C for alumina is recommended. Phthalate
esters were detected in Florisil cartridge method blanks at levels ranging
from 10 to 460 ng, with 5 phthalate esters in the 105 to 460 ng range.
Complete removal of the phthalate esters from Florisil cartridges does not
seem possible, and it is therefore desirable to keep the steps involved in
sample preparation to a minimum.
3.5 Paper thimbles and filter paper must be exhaustively washed with the
solvent that will be used in the sample extraction. Soxhlet extraction of
paper thimbles and filter paper for 12 hrs with fresh solvent should be
repeated for a minimum of three times. Method blanks should be obtained
before any of the precleaned thimbles or filter papers are used. Storage of
precleaned thimbles and filter paper in precleaned glass jars covered with
aluminum foil is recommended.
3.6 Glasswool used in any step of sample preparation should be a
specially treated pyrex wool, pesticide grade, and must be baked at 400°C for
4 hrs immediately prior to use.
3.7 Sodium sulfate must be purified by heating at 400°C for 4 hrs in a
shallow tray. To avoid recontamination, the precleaned material must be
stored in glass bottles, covered with precleaned aluminum foil. The storage
period should not exceed two weeks. To minimize contamination, extracts
8061-2 Revision 2
September 1989
****September 1989****
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should be dried directly in the glassware in which they are collected by
adding small amounts of precleaned sodium sulfate until excess of free flowing
material is noted.
3.8 The presence of elemental sulfur will result in large peaks which
often mask the region of the compounds eluting before dicyclohexyl phthalate
(Compound No. 14) in the gas chromatograms shown in Figure 1. Method 3660 is
suggested for removal of sulfur.
3.9 Waxes and lipids can be removed by gel-permeation chromatography
(Method 3640). Extracts containing high amounts of lipids are viscous and
may even solidify at room temperature.
4.0 APPARATUS AND MATERIALS
4.1 Glassware, see Methods 3510, 3540, 3550, 3610, 3620, 3640, and 3660
for specifications.
4.2 Kuderna-Danish (K-D) apparatus, standard taper 19/22 ground-glass
joints (Kontes K-570025-0500):
4.2.1 Concentrator tube, 10 ml graduated (Kontes K-570050-1025
or equivalent). A ground-glass stopper is used to prevent evaporation
of solvents or analytes after removal from the concentration apparatus.
4.2.2 Evaporation flask, 500 ml (Kontes K-570001-50 or
equivalent). Attach to concentrator tube with springs.
4.2.3 Snyder column, three-ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Springs, 1/2 in (Kontes K-662750).
4.2.5 Boiling chips, approximately 10/40 mesh. Heat to 400eC for
30 min or Soxhlet-extract with methylene chloride prior to use.
4.3 Gas chromatograph:
4.3.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for on-column and split/splitless injection, 8-in
injection tee (Supelco, Inc., Catalog No. 2-3665) or J&W Scientific
"press-fit" glass Y splitter for megabore columns (Catalog No. 705-0733)
and all required accessories, including syringes, analytical columns,
gases, electron capture detectors, and data system.
4.3.2 Columns:
4.3.2.1 Column 1, 30 m x 0.53 mm i.d. DB-5 fused-silica
open tubular column or equivalent, 1.5 fun film thickness.
8061-3 Revision 2
September 1989
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4.3.2.2 Column 2, 30 m x 0.53 mm i.d. DB-1701 fused-silica
open tubular column or equivalent, 1.0 urn film thickness.
4.4 Vacuum system for eluting disposable solid-phase extraction
cartridges.
4.4.1 Vacuum manifold consisting of individually adjustable,
easily accessible flow-control valves for up to 24 cartridges, sample
rack, chemically resistant cover and seals, heavy-duty glass basin,
removable stainless steel solvent guides, built-in vacuum gauge and
valve.
4.4.2 Vacuum trap made of 500-mL side arm flask fitted with a
one-hole stopper and glass tubing.
4.4.3 6-mL, 1-g solid-phase extraction cartridges, LC-Florisil
or equivalent, prepackaged, ready to use.
4.5 Water bath, heated, with concentric ring cover, capable of
temperature control (±2"C).
4.6 Apparatus for filtration of aqueous samples through 3M-Empore
extraction disks (optional).
4.6.1 Vacuum apparatus:
4.6.1.1 1-L suction flask.
4.6.1.2 Disk base.
4.6.1.3 Graduated funnel.
4.6.1.4 Clamp.
4.6.1.5 Vacuum gauge.
4.6.1.6 Pinch clamp.
4.6.1.7 25 x 200 mm test tube.
4.6.2 47-mm 3M-Empore Ci8-extraction disks (Analytichem
International, Catalog No. 1214-5004, or equivalent).
5.0 REAGENTS
5.1 Reagent-grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to
specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may
be used, provided it is first ascertained that the reagent is of sufficiently
8061'4 Revision 2
September 1989
****September 1989****
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high purity to permit its use without lessening the accuracy of the
determination.
5.2 Reagent-grade water. All references to water in this method refer
to HPLC-grade water unless otherwise specified.
5.3 Sodium sulfate (ACS certified), granular, anhydrous. Purify by
heating at 400"C for 4 hrs in a shallow tray.
5.4 Florisil -- J. T. Baker, Lot No. 442707, 60/80 mesh, activated at
400eC for 16 hrs, then deactivated with water (3 percent by weight).
5.5 Florisil disposable cartridges -- Supelclean™ SPE tubes or
equivalent consisting of serological-grade 6-mL polypropylene tubes, packed
each with 1 g LC-Florisil (40-/im particles, 60-A pores) held between
polyethylene frits.
5.6 Alumina -- Alumina Woelm N Super I, activated/deactivated as
described for Florisil.
5.7 Hexane, methylene chloride, acetone, acetonitrile, and methanol.
Pesticide quality or equivalent for hexane, methylene chloride, and acetone
and HPLC grade for methanol and acetonitrile.
5.8 Stock standard solutions:
5.8.1 Prepare stock standard solutions at a concentration of
1 ng/nl by dissolving 0.0100 g of assayed reference material in hexane
and diluting to volume in a 10-mL volumetric flask. When compound purity
is assayed to be 96 percent or greater, the weight can be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standard solutions can be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.8.2 Transfer the stock standard solutions into Teflon-lined
screw-cap bottles. Store at 4°C and protect from light. Stock standard
solutions should be checked periodically by gas chromatography for signs
of degradation or evaporation, especially just prior to preparing
calibration standards from them.
5.8.3 Stock standard solutions must be replaced after 6 months,
or sooner, if comparison with check standards indicates a problem.
5.9 Calibration standards: Calibration standards at a minimum of five
concentration levels for each parameter of interest are prepared through
dilution of the stock standard solutions with hexane. One of the
concentration levels should be at a concentration near, but above, the method
detection limit. The remaining concentration levels should correspond to the
expected range of concentrations found in real samples or should define the
working range of the GC. Calibration solutions must be replaced after 1 to
8061'5 Revision 2
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2 months, or sooner, if comparison with calibration verification standards
indicates a problem.
5.10 Internal standards (if internal standard calibration is used): To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Benzyl benzoate has been tested
and found appropriate for Method 8061.
5.10.1 Prepare a spiking solution of benzyl benzoate in hexane at
5 ng/nl. Addition of 10 (iL of this solution to 1 ml of sample extract
is recommended. The spiking level of the internal standard should be
kept constant for all samples and calibration standards. Store the
internal standard spiking solution at 4°C in a Teflon-sealed container.
Standard solution should be replaced when ongoing QC (Section 8)
indicates a problem.
5.11 Surrogate standards: The analyst should monitor the performance
of the extraction, cleanup (when used), analytical system, and the
effectiveness of the method in dealing with each sample matrix by spiking
each sample, standard, and blank with surrogates. Three surrogates are
suggested for Method 8061: diphenyl phthalate, diphenyl isophthalate, and
dibenzyl phthalate.
5.11.1 Prepare a surrogate standard spiking solution in acetone
which contains 50 ng//iL of each compound. Addition of 500 fil of this
solution to 1 L of water or 30 g solid sample is equivalent to 25 /ig/L
of water or 830 ng/g of solid sample. The spiking level of the surrogate
standards may be adjusted accordingly if the final volume of extract is
reduced below 2 mL for water samples or 10 mL for solid samples. Store
the surrogate spiking solution at 4'C in a Teflon-sealed container. The
solution must be replaced after 6 months or sooner if ongoing QC
(Section 8) indicates problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See introductory material to this chapter, Organic Analytes,
Section 4.1.
6.2 The stability of the phthalate esters in soil or sediment samples
has not been systematically investigated. Storage of soil samples at room
temperature should be avoided since degradation of some phthalate esters has
been reported to occur. Deep-freezing at -10"C or -20°C appears to be the
most suitable method for storage of solid matrices since it has the widest
range of application, causes the least changes in the samples, and makes the
addition of preservatives unnecessary.
6.3 All aqueous samples must be extracted within 7 days of sample
collection; all soil and sediment samples must be extracted within 30 days
8061'6 Revision 2
September 1989
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of sample collection. Extracts must be stored at <4°C and must be analyzed
within 30 days of extraction.
7.0 PROCEDURE
7.1 Extraction:
7.1.1 Refer to Chapter Two for guidance on choosing the
appropriate extraction procedure. In general, water samples are
extracted at a pH of 5 to 7 with methylene chloride in a separatory
funnel (Method 3510). Method 3520 is not recommended for the extraction
of aqueous samples because the longer chain esters dihexyl phthalate
bis(2-ethylhexyl) phthalate, di-n-octyl phthalate, and dinonyl phthalate
tend to adsorb to the glassware and consequently, their extraction
recoveries are <40 percent. Solid samples are extracted with
hexane/acetone (1:1) or methylene chloride/acetone (1:1) in a Soxhlet
extractor (Method 3540) or with a sonicator (Method 3550). Immediately
prior to extraction, spike 500 /iL of the surrogate standard spiking
solution (concentration 50 ng/jiL) to 1 L aqueous sample or 30 g solid
sample.
7.1.2 Spiked samples are used to verify the applicability of the
chosen extraction technique for each new sample type. Each matrix must
be spiked with the compounds of interest to determine the percent
recovery and the limit of detection for that matrix. Spiking of water
samples should be performed by adding appropriate amounts of phthalate
esters, dissolved in methanol or acetone, to the water sample immediately
prior to extraction. After addition of the spike, mix the samples
manually for 1 to 2 minutes. Typical spiking levels for water samples
are 5 to 10 nq/l for samples in which phthalate esters were not detected
and 2 to 5 times the background level in those cases where phthalate
esters are present. Spiking of solid samples should be performed by
adding appropriate amounts of phthalate esters which are dissolved in
methanol or acetone to the solid samples. The solid sample should be wet
prior to the addition of the spike (at least 40 percent moisture) and
should be mixed thoroughly with a blender for 10 minutes to homogenize
the material. Transfer the whole portion that was spiked with the test
compounds to the extraction thimble for Soxhlet extraction (Method 3540)
or proceed with the sonication extraction (Method 3550).
7.1.3 Extraction of aqueous samples using the 3M-Empore C18-
extraction disks (optional):
7.1.3.1 Disk preconditioning: Place the 3M-Empore C18-
extraction disk into the filtration apparatus and prewash the disk
with 10 to 20 ml acetonitrile. Apply vacuum to pull the solvent
through the disk. Maintain vacuum to pull air through for 5 min.
Follow with 10 ml of methanol. Apply vacuum and pull most of the
methanol through the disk. Release vacuum before the disk gets dry.
Follow with 10 mL HPLC-grade water. Apply vacuum and pull most of
8061-7 Revision 2
September 1989
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the water through the disk. Release the vacuum before the disk gets
dry.
7.1.3.2 Sample preconcentration: Add 2.5 ml methanol to the
500 ml aqueous sample in order to get reproducible results. Pour
the sample into the filtration apparatus. Adjust vacuum so that it
takes approximately 20 min to process 502.5 ml of sample. After all
sample has passed through the membrane disk, pull air through the
disk for 5 to 10 min to remove any residual water.
7.1.3.3 Sample elution: Break the vacuum and place the tip
of the filter base into the test tube that is contained inside the
suction flask. Add 10 ml acetonitrile to the graduated funnel
making sure to rinse the walls of the graduated funnel with the
solvent. Apply vacuum to pass the acetonitrile through the membrane
disk.
7.1.3.4 Extract concentration: Concentrate the extract to
2 ml or less using a gentle stream of pure nitrogen.
7.2 Prior to Florisil cleanup or gas chromatographic analysis, the
extraction solvent must be exchanged to hexane. The exchange is performed as
follows:
7.2.1 Following K-D concentration of the methylene chloride or
methylene chloride/acetone extracts obtained according to Section 7.1.2
to 1 ml using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 min.
7.2.2 Momentarily remove the Snyder column, add 50 ml hexane, a
new glass bead, and attach the macro-Snyder column. Concentrate the
extract using 1 ml hexane to prewet the Snyder column. Place the K-D
apparatus on the water bath so that the concentrator tube is partially
immersed in the hot water. Adjust the vertical position of the apparatus
and the water temperature, as required, to complete concentration in 5-10
min. At the proper rate of distillation the balls 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 and allow it to
drain and cool for at least 10 min.
7.2.3 Remove the Snyder column and rinse the flask and its lower
joint into the concentrator tube with 1 to 2 ml hexane. A 5-mL syringe
is recommended for this operation. Adjust the extract volume to 2 ml for
water samples or 10 mL for solid samples. Stopper the concentrator tube
and store at 4°C if further processing will bfr performed immediately.
If the extract will be stored for two days or longer, it should be
transferred to a Teflon-lined screw-cap vial. Proceed with the gas
chromatographic analysis.
8061'8 Revision 2
September 1989
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7.3 Cleanup/Fractionation:
7.3.1 Cleanup may not be necessary for extracts from a relatively
clean sample matrix. If polychlorinated biphenyls (PCBs) and
organochlorine pesticides are known to be present in the sample, proceed
with the procedure outlined in Methods 3610 or 3620. Collect Fraction
1 by eluting with 140 ml (Method 3610) or 100 ml (Method 3620) of
20-percent diethyl ether in hexane. Note that, under these conditions,
bis(2-methoxyethyl) phthalate, bis(2-ethoxyethyl) phthalate, and bis(2-
n-butoxyethyl) phthalate are not recovered from the Florisil column. The
elution patterns and compound recoveries are given in Table 3.
7.3.2 As an alternative to Method 3620, the following Florisil
cartridge procedure can be used for extract cleanup. With this method,
bis(2-methoxyethyl) phthalate, bis(2-ethoxyethyl) phthalate, and bis(2-
n-butoxyethyl) phthalate are recovered quantitatively.
7.3.2.1 Every lot of Florisil cartridges must be checked
prior to use as follows. Install 1-g cartridges in the vacuum
manifold. Wash each cartridge with 4 ml of pesticide-grade hexane
and discard the eluate. Add to each cartridge 2 ml of a composite
standard containing the test compounds at 5 to 10 /ig/mL and elute
each cartridge with 5 ml of 10-percent acetone in hexane.
Concentrate the eluate to a final volume of 2 ml and analyze by
GC/ECD. The lot of Florisil cartridges is acceptable if all 16
target compound recoveries, except bis(2-methoxyethyl) phthalate,
are between 80 and 120 percent and if no other interferences are
detected.
7.3.2.2 Prior to cleanup of sample extracts, the cartridges
must be washed with hexane. This is accomplished by placing 10, 12,
or 24 cartridges in the vacuum manifold (the number depends on the
type of vacuum manifold; for example, Vac Elut SPS24 from
Analytichem International can accommodate 24 cartridges) and passing
at least 4 ml of pesticide-grade hexane through each cartridge.
While washing the cartridges, adjust the vacuum applied to each
cartridge so that the flows through the cartridges are approximately
equal. Do not allow the cartridges to go dry after they have been
washed.
7.3.2.3 After the cartridges have been washed, release the
vacuum and replace the collecting vials with 5-mL volumetric flasks
or culture tubes. Care must be taken to ensure that the solvent
line from each cartridge is placed inside the correct volumetric
flask or culture tube.
7.3.2.4 After the volumetric flasks or the culture tubes
have been set in the vacuum manifold, the vacuum is restored and the
sample extracts (the whole extract for the aqueous sample and 2 ml
of the 10-ml extract for the solid samples) are added to the
8061~9 . Revision 2
September 1989
****September 1989****
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appropriate cartridges. Use a syringe or a volumetric pipet for
transferring the extracts.
7.3.2.5 Elute each cartridge with 5 ml of 10-percent acetone
in hexane and collect the eluate in a 5-mL volumetric flask or
culture tube held inside the vacuum manifold. Adjust to the 5-mL
mark if not all solvent is recovered. Transfer the eluates to clean
sample vials for further concentration using nitrogen blow-down
evaporation with a gentle stream of pure nitrogen. The elution
patterns and compound recoveries are given in Table 3.
7.3.3 If PCBs and organochlorine pesticides are known to be
present in the sample, and if the Florisil cartridge procedure is
considered, then two fractions are collected: Fraction 1 is eluted with
5 ml of 20-percent methylene chloride in hexane and Fraction 2 is eluted
with 5 ml of 10-percent acetone in hexane. The elution patterns and
compound recoveries are given in Table 4. Fraction 1 contains the
organochlorine pesticides and PCBS and can be discarded. Fraction 2
contains the phthalate esters and is analyzed by GC/ECD.
7.4 Gas chromatography conditions (recommended):
7.4.1 Column 1 (DB-5) and Column 2 (DB-1701): Temperature program
150'C (0.5 min hold) to 220'C at 5eC/min, then to 2758C (13 min hold) at
3°C/min; carrier gas helium at 6 mL/min; makeup gas nitrogen at
20 mL/min; injector temperature 250"C; detector temperature 320'C.
7.4.2 Table 1 gives the retention times and MDLs that can be
achieved by this method for the 16 phthalate esters. An example of the
separations achieved with the DB-5 and DB-1701 fused-silica open tubular
columns is shown in Figure 1.
7.5 Calibration:
7.5.1 Refer to Method 8000 for proper calibration techniques.
Use Tables 1 and 2 for guidance on selecting the lowest point on the
calibration curve.
7.5.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for the description of each of these
procedures.
7.6 Gas chromatographic analysis:
7.6.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 fiL of internal standard solution at 5 /ig//iL
to the sample prior to injection.
7.6.2 Follow step 7.6 in Method 8000 for instructions on the
analysis sequence, appropriate dilutions, establishing daily retention
time windows, and identification criteria.
8061-10 Revision 2
September Iy89
****September 1989****
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7.6.3 Record the sample volume injected and the resulting peak
areas.
7.6.4 Using either the internal or the external calibration
procedure (Method 8000), determine the identity and the quantity of each
component peak in the sample chromatogram which corresponds to the
compounds used for calibration purposes.
7.6.5 If the response of a peak exceeds the working range of the
system, dilute the extract and reanalyze.
7.6.6 Identify compounds in the sample by comparing the retention
times of the peaks in the sample chromatogram with those of the peaks in
standard chromatograms. The retention time window used to make
identifications is based upon measurements of actual retention time
variations over the course of 10 consecutive injections. Three times the
standard deviation of the retention time can be used to calculate a
suggested window size.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and
in the extraction method utilized. If extract cleanup was performed, follow
the QC specified in Method 3600 and in the specific cleanup method.
8.2 Mandatory quality control to evaluate the GC system operation is
found in Method 8000, Section 8.6.
8.2.1 The quality control check sample concentrate (Method 8000,
Section 8.6) should contain the test compounds at 5 to 10 ng//iL.
8.3 Calculate the recoveries of the surrogate compounds for all samples,
method blanks, and method spikes. Determine if the recoveries are within
limits established by performing QC procedures outlined in Method 8000
Step 8.10.
8.3.1 If the recoveries are not within limits, the following are
required:
• Make sure there are no errors in calculations, surrogate solutions
and internal standards. Also check instrument performance.
• Recalculate the data and/or reanalyze the extract if any of the
above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are a
problem, or flag the data as "estimated concentration."
8.4 An internal standard peak area check must be performed on all
samples. The internal standard must be evaluated for acceptance by
8061-n Revision 2
September 1989
****September 1989****
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determining whether the measured area for the internal standard deviates by
more than 50 percent from the average area for the internal standard in the
calibration standards. When the internal standard peak area is outside that
limit, all samples that fall outside the QC criteria must be reanalyzed.
8.5 GC/MS confirmation: Any compounds confirmed by two columns may also
be confirmed by GC/MS if the concentration is sufficient for detection by
GC/MS as determined by the laboratory-generated detection limits.
8.5.1 The GC/MS would normally require a minimum concentration of
10 ng//zL in the final extract for each single-component compound.
8.5.2 The sample extract and associated blank should be analyzed
by GC/MS as per Section 7.0 of Method 8270.
8.5.3 A reference standard of the compound must also be analyzed
by GC/MS. The concentration of the reference standard must be at a level
that would demonstrate the ability to confirm the phthalate esters
identified by GC/ECD.
8.6 Include a mid-level calibration standard after each group of
20 samples in the analysis sequence. The response factors for the mid-level
calibration must be within ±30 percent of the average values for the
multilevel calibration.
8.7 Demonstrate through the analyses of standards that the Florisil
fractionation scheme is reproducible. When using the fractionation schemes
given in Methods 3610 or 3620, batch-to-batch variations in the composition
of the alumina or Florisil material may cause variations in the recoveries of
the phthalate esters.
9.0 METHOD PERFORMANCE
9.1 The MDL is defined as the minimum concentration of the test compound
that can be measured and reported with 99 percent confidence as being greater
than zero. The MDL concentrations listed in Table 1 were obtained by using
reagent water. Details on how to determine MDLs are given in Reference 1. The
MDL actually achieved in a given analysis will vary as it is dependent on
instrument sensitivity and matrix effects.
9.2 This method has been tested in a single laboratory by using
different types of aqueous samples and solid samples which were fortified with
the test compounds at two concentrations. Single-operator precision, overall
precision, and method accuracy were found to be related to the concentration
of the compounds and the type of matrix. For exemplification, results of the
single-laboratory method evaluation are given in Tables 5, 6, and 7.
9.3 The accuracy and precision obtained will be determined by the sample
matrix, sample preparation technique, cleanup techniques, and calibration
procedures used.
8061-12 Revision 2
September 1989
****September 1989****
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10.0 REFERENCES
(1) Glazer, J.A.; Foerst, G.D.; McKee, G.D.; Quave, S.A., and Budde, W.L.
"Trace Analyses for Wastewaters," Environ. Sci. and Techno!. 15: 1426,
1981.
8061'13 Revision 2
September 1989
****September 1989****
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TABLE 1. GAS CHROMATOGRAPHIC RETENTION TINES AND METHOD DETECTION LIMITS FOR
THE PHTHALATE ESTERS'
*
*
GO
re
re
3
CT
re
-s
00
to
*
*
*
00
o
at
GO
re
•o
c*
re 73
3 re
cr <
re -••
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
IS
SU-1
SU-2
SU-3
Compound name
Dimethyl phthalate (DMP)
Di ethyl phthalate (DEP)
Ollsobutyl phthalate (OIBP)
Oi-n-butyl phthalate (DBF)
B1s(4-methyl-2-pentyl) phthalate (BMPP)
Bis(2-methoxyethyl) phthalate (BHEP)
Diamyl phthalate (DAP)
B1s(2-ethoxyethy1) phthalate (BEEP)
Hexyl 2-ethylhexyl phthalate (HEHP)
Dlhexyl phthalate (DHP)
Benzyl butyl phthalate (BBP)
B1s(2-n-butoxyetjiy1) phthalate (BBEP)
B1s(2-ethylhexyl) phthalate (DEHP)
Dicyclohexyl phthalate (DCP)
Dl-n-octyl phthalate (OOP)
Dinonyl phthalate
Benzyl benzoate
Dlphenyl phthalate (DPP)
Diphenyl isophthalate (DPIP)
DI benzyl phthalate (OBZP)
Chemical
Abstract
Registry
No.
131-11-3
84-66-2
84-69-5
84-74-2
146-50-9
117-82-8
131-18-0
605-54-9
75673-16-4
84-75-3
85-68-7
117-83-9
117-81-7
84-61-7
117-84-0
84-76-4
120-51-4
84-62-8
744-45-6
523-31-9
Retention time
(mln)
Column 1
7.06
9.30
14.44
16.26
18.77
17.02
20.25
19.43
21.07
24.57
24.86
27.56
29.23
28.88
33.33
38.80
12.71
29.46
32.99
34.40
Column 2
6.37
8.45
12.91
14.66
16.27
16.41
18.08
18.21
18.97
21.85
23.08
25.24
25.67
26.35
29.83
33.84
11.07
28.32
31.37
32.65
MDL"
Liquid
(ng/L)
640
250
120
330
370
510
110
270
130
68
42
84
270
22
49
22
C
c
C
c
VO
00
ro
•Column 1 Is a 30 m x 0.53 mm ID 08-5 fused-sillca open tubular column (1.5-/MI film thickness).
Column 2 Is a 30 m 0.53 mm ID DB-1701 fused-silica open tubular column (1.0-/im film thickness).
Temperature program Is 150*C (0.5 mln hold) to 220'C at 5'C/min, then to 275'C (13 mln hold) at
3'C/min. An 8-1n Supelco injection tee or a J&W Scientific press fit glass inlet splitter is used to
connect the two columns to the injection port of a gas chromatograph. Carrier gas helium at
6 raL/min; makeup gas nitrogen at 20 mL/min; injector temperature 250'C; detector temperature 320*C.
"MDL Is the method detection limit. The HDL was determined from the analysis of seven replicate
aliquots of reagent water processed through the entire analytical method (extraction, Florisil
cartridge cleanup, and GC/ECD analysis using the single column approach: DB-5 fused-sillca capillary
column). NDL • t(n-l, 0.99) x SD where t(n-l, 0.99) is the student's t value appropriate for a 99-
percent confidence Interval and a standard deviation with n-1 degrees of freedom, and SD is the
standard deviation of the seven replicate measurements. Values measured were not corrected for
method blanks.
'Not applicable.
-------
TABLE 2. PRACTICAL QUANTITATION LIMITS (PQL) FOR VARIOUS MATRICES3
Matrix Factor"
Groundwater 10
Low-level soil by sonication with GPC cleanup 670
High-level soil and sludges by sonication 10,000
Nonwater miscible waste 100,000
aSample PQLs are highly matrix-dependent. The PQLs
listed herein are provided for guidance and may not
always be achievable.
bPQL = [Method detection limit (Table 1)] X [Factor
(Table 2)]. For nonaqueous samples, the factor is
on a wet-weight basis.
8061-15 Revision 2
September 1989
****September 1989****
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TABLE 3. AVERAGE RECOVERIES OF METHOD 8061 COMPOUNDS USING METHODS 3610,
3620, AND THE ALUMINA AND FLORISIL DISPOSABLE CARTRIDGE PROCEDURE
Compound
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis (2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Method
3610
alumina*
64.5
62.5
77.0
76.5
89.5
70.5
75.0
67.0
90.5
73.0
87.0
62.5
91.0
84.5
108
71.0
Method
3620
Florisil"
40.0
57.0
80.0
85.0
84.5
0
81.5
0
105
74.5
90.0
0
82.0
83.5
115
72.5
Alumina
cartridge"
101
103
104
108
103
64.1°
103
111
101
108
103
108
97.6
97.5
112
97.3
Florisil
cartridge*1
89.4
97.3
91.8
102
105
78. 3e
94.5
93.6
96.0
96.8
98.6
91.5
97.5
90.5
97.1
105
aThe number of determinations was 2; alumina and Florisil chromatography
were done according to Methods 3610, and 3620, respectively.
bl-g alumina cartridges were used; Fraction 1 was eluted with 5 ml of
20-percent acetone in hexane. The number of determinations was 2. The
amount spiked was 40 /ig per component per cartridge.
°The amount recovered by elution with an additional 5 mL of 20-percent
acetone in hexane was 36.8 percent.
dl-g Florisil cartridges were used; Fraction 1 was eluted with 5 ml of
10-percent acetone in hexane. The number of determinations was 2. The
amount spiked was 40 /ig per component per cartridge.
6The amount recovered by elution with an additional 5 mL of 10-percent
acetone in hexane was 14.4 percent.
8061-16
****September 1989****
Revision 2
September 1989
-------
TABLE 4. ELUTION PATTERNS AND AVERAGE RECOVERIES OF METHOD 8061 COMPOUNDS
USING THE FLORISIL DISPOSABLE CARTRIDGES
Percent recovery8
Compound
Fraction 1
Fraction 2
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis(2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
0
0
0
12
0
0
3.3
0
0
0
0
0
0
0
0
0
130
88
118
121
123
32
94
82
126
62
98
135
110
106
123
102
(52)
(2.8)
(16)
(13)
(5.7)
(31)
(8.3)
(19)
(6.4)
(15)
(6.5)
(34)
(2.7)
(3.3)
(7.0)
(8.7)
aThe number of determinations was 3. The values given in
parentheses are the percent relative standard deviations of
the average recoveries.
8061-17
****
September 1989
Revision 2
September 1989
****
-------
TABLE 5. ACCURACY AND PRECISION DATA FOR EXTRACTION USING THE 3M EMPORE DISKS
AND METHOD 8061
HPLC-grade water
Compound
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis(2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Average
recovery
(%)
88.6
92.3
87.6
90.3
87.2
107
93.6
108
93.9
98.4
97.3
94.8
91.3
106
84.9
96.9
Precision
(% RSD)
17.7
10.3
16.2
13.2
9.5
13.6
21.0
8.9
22.4
5.0
2.6
6.3
7.4
19.9
3.8
11.1
Groundwater
Average
recovery
(%)
86.6
92.6
89.3
95.0
86.7
113
78.9
102
83.4
97.7
66.0
98.7
96.3
108
90.1
95.2
Precision
(% RSD)
14.3
7.2
1.6
1.5
4.9
2.8
5.8
4.0
8.8
14.8
39.3
6.0
7.9
13.3
6.1
12.7
aThe number of determinations was 4.
component.
The spiking level was 100 /zg/L per
8061-18
****September 1989****
Revision 2
September 1989
-------
TABLE 6. ACCURACY AND PRECISION DATA FOR METHOD 3510 AND METHOD 8061'
*
*
*
*
CO
O>
3
O"
to
CD
O
I- *°
IO
00
to
*
*
*
*
Compound
Dimethyl phthalate
Oiethyl phthalate
DUsobutyl phthalate
Di-n-butyl phthalate
B1s(4-methyl-2-pentyl) phthalate
B1s(2-methoxyethyl) phthalate
Dlamyl phthalate
B1s(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Olhexyl phthalate
Benzyl butyl phthalate
B1s(2-n-butoxyethyl) phthalate
B1s(2-ethylhexyl) phthalate
Olcyclohexyl phthalate
Dl-n-octyl phthalate
Dlnonyl phthalate
Surrogates :
Diphenyl phthalate
Dlphenyl isophthalate
Dlbenzyl phthalate
Estuarlne
water
84.0
71.2
76.0
83.2
78.6
73.8
78.2
75.6
84.7
79.8
84.1
78.5
81.4
77.4
74.9
59.5
98.5
95.8
93.9
(4.1)
(3.8)
(6.5)
(6.5)
(2-6)
(1.0)
(7.3)
(3.3)
(5.3)
(7.2)
(6.4)
(3.5)
(4.1)
(6.5)
(4.9)
(6.1)
(2.6)
(1.9)
(4.4)
Spike level
(20 M9/L)
Leachate
98.9 (19.6)
82.8 (19.3)
95.3 (16.9)
97.5 (22.3)
87.3 (18.2)
87.2 (21.7)
92.1 (21.5)
90.8 (22.4)
91.1 (27.5)
102 (21.5)
105 (20.5)
92.3 (16.1)
93.0 (15.0)
88.2 (13.2)
87.5 (18.7)
77.3 (4.2)
113 (14.9)
112 (11.7)
112 (14.0)
Groundwater
87.1
88.5
92.7
91.0
92.6
82.4
88.8
86.4
81.4
90.9
89.6
89.3
90.5
91.7
87.2
67.2
110
109
106
(8.1)
(15.3)
(17.1)
(10.7)
(13.7)
(4.4)
(7.5)
(5.8)
(17.6)
(7.6)
(6.1)
(3-6)
(4.9)
(15.2)
(3.7)
(8.0)
(3.3)
(3.3)
(3.8)
Estuarlne
water
87.
71.
99.
87.
97.
82.
89.
88.
107
90.
92.
86.
86.
87.
85.
97.
110
104
111
1
0
1
0
4
5
2
7
1
7
1
5
7
1
2
(7.5)
(7.7)
(19.0)
(8.0)
(15.0)
(5.5)
(2.8)
(4.9)
(16.8)
(2.4)
(5.6)
(6.2)
(6.9)
(9.6)
(8.3)
(7.0)
(12.4)
(5.9)
(5.9)
Spike
(60
level
M9/L)
Leachate
112
88.5
100
106
107
99.0
112
109
117
109
117
107
108
102
105
108
95.1
97.1
93.3
(17.5)
(17.9)
(9.6)
(17.4)
(13.3)
(13.7)
(H.2)
(14.6)
(11.4)
(20.7)
(24.7)
(15.3)
(15.1)
(14.3)
(17.7)
(17.9)
(7.2)
(7.1)
(9.5)
Groundwater
90.9 (4.5)
75.3 (3.5)
83.2 (3.3)
87.7 (2.7)
87.6 (2.9)
76.9 (6.6)
92.5 (1.8)
84.8 (5.9)
80.1 (4.1)
88.9 (2.4)
93.0 (2.0)
92.4 (0.6)
91.1 (3.0)
71.9 (2.4)
90.4 (2.0)
90.1 (1.1)
107 (2.4)
106 (2.8)
105 (2.4)
'The number of determinations was 3.
the average recoveries.
The values given in parentheses are the percent relative standard deviations of
TJ
c*
n> 70
3 n>
o- <
o> -'•
-$ to
•— o
ID 3
00
to ro
-------
TABLE 7. ACCURACY AND PRECISION DATA FOR METHOD 3550 AND NETHOD 8061*
*
*
*
CD
3
or
(D
-i
U3
00
U3
*
If
*
CD
O
O>
ro
o
Estuarine
Compound
Dimethyl phthalate
Diethyl phthalate
Olisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Oiamyl phthalate
Bls(Z-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis (2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Dl-n-octyl phthalate
Oinonyl phthalate
sediment
77.9
68.4
103
121
108
26.6
95.0
C
c
103
113
114
c
36.6
C
C
(42.8)
(1.7)
(3.1)
(25.8)
(57.4)
(26.8)
(10.2)
(3.6)
(12.8)
(21.1)
(48.8)
Spike level
(1 M9/9)
Municipal
sludge
52.1 (35.5)
68.6 (9.1)
106 (5.3)
86.3 (17.7)
97.3 (7.4)
72.7 (8.3)
81.9 (7.1)
66.6 (4.9)
114 (10.5)
96.4 (10.7)
82.8 (7.8)
74.0 (15.6)
76.6 (10.6)
65.8 (15.7)
93.3 (14.6)
80.0 (41.1)
Sandy loam
soil
c
54.7 (6.2)
70.3 (3.7)
72.6 (3.7)
C
0
81.9 (15.9)
C
57.7 (2.8)
77.9 (2.4)
56.5 (5.1)
C
99.2 (25.3)
92.8 (35.9)
84.7 (9.3)
64.2 (17.2)
Estuarine
sediment
136 (9.6)
60.2 (12.5)
74.8 (6.0)
74.6 (3.9)
104 (1.5)
19.5 (14.8)
77.3 (4.0)
21.7 (22.8)
72.7 (11.3)
75.5 (6.8)
72.9 (3.4)
38.3 (25.1)
59.5 (18.3)
33.9 (66.1)
36.8 (16.4)
C
Spike level
(3 /*g/g)
Municipal
sludge
64.8 (11.5)
72.8 (10.0)
84.0 (4.6)
113 (5.8)
150 (6.1)
59.9 (5.4)
116 (3.7)
57.5 (9.2)
26.6 (47.6)
80.3 (4.7)
76.8 (10.3)
98.0 (6.4)
85.8 (6.4)
68.5 (9.6)
88.4 (7.4)
156 (8.6)
Sandy loam
soil
70.2 (2.0)
67.0 (15.1)
79.2 (0.1)
70.9 (5.5)
83.9 (11.8)
0
82.1 (15.5)
84.7 (8.5)
28.4 (4.3)
79.5 (2.7)
67.3 (3.8)
62.0 (3.4)
65.4 (2.8)
62.2 (19.1)
115 (29.2)
115 (13.2)
aThe number of determinations was 3. The values given in parentheses are the percent relative standard deviations of the
average recoveries. All samples were subjected to Florisil cartridge cleanup.
bThe estuarine sediment extract (Florisil, Fraction 1) was subjected to sulfur cleanup (Method 3660 with tetrabutylammonium
sulfite reagent).
"Not able to determine because of matrix interferent.
to
o>
-a
r*
(D TO
3 (D
CT <
It -*•
-$ (/>
!-• O
to =3
CD
10 ro
-------
DB-5
30 m x 0.53 mm ID
1.5-jim Film
11 12
O
LU
jJ
(\
16
O
LU
10
11
I
10
20
TIME (min)
16
30
DB-1701
30 m x 0.53 mm ID
1.0-pjn Film
40
Figure 1. GC/ECD chromatograms of a composite phthalate esters standard
(concentration 10 ng//iL per compound) analyzed on a DB-5 and a
DB-1701 fused-silica open tubular column. Temperature program:
150°C (0.5 min hold) to 220°C at 5°C/min, then to 275°C (13 min
hold) at 3'C/min.
8061-21
****September 1989****
Revision 2
September 1989
-------
METHOD 8061—PHTHALATE ESTERS (FLOW CHART)
S
CM
T
Q
7.1.1 Choose appropriate
extraction procedure
I
7.1.2 Add appropriate surro-
gates and spiking compounds
to aqueous samples prior to
extraction; extract by either
Method 3510 or with
&-membrane disks
I
7.1.2 Add appropriate surro-
gates and spiking compounds
to solid samples prior to
extraction; extract by Method
3540 or 3550
7.2 Exchange extraction
solvent to hexane as necessary
during K-D procedures
I
7.2.1 Concentrate, methylene
chloride extract, allow K-D
apparatus to drain and cool
I
7.2.2 Add hexane; attach Snyder
column; place apparatus on
water bath; concentrate; remove
from water bath; cool
I
7.2.3 Remove column; rinse
flask and joints with hexane;
adjust extract volume
7.3.1 Choose appropriate cleanup
technique, as necessary; Florisil
cleanup is recommended (Refer to
Method 3620 or to procedures given
in 7.3.2 and 7.3.3)
7.2.3 Will
further processing
be performed
within 2
days?
7.2.3 Transfer extract to Teflon
sealed screw-cap vials;
refrigerate
8061-22
****September 1989****
Revision 2
September 1989
-------
METHOD 8061—PHTHALATE ESTERS (FLOW CHART) (CONTINTUED)
to
CM
TI-
Q
W
LU
7.4.1 Set conditions for Columns
1 and 2
i
7.4.2 Refer to Table 1 for retention
times and MDLs; refer to Figure 1
for example of chromatograms
I
7.5.1 Refer to Method 8000 for
calibration techniques; select
lowest point on calibration curve
I
7.5.2 Choose and execute
internal calibration (refer to
Method 8000)
7.6.1 Add internal standard if
necessary
7.6.2 Establish daily retention
time windows, analysis sequence,
dilutions, and identification criteria
8061-23
****
September 1989
Revision 2
September 1989
****
-------
METHOD 8061—PHTHALATE ESTERS (FLOW CHART) (CONCLUDED)
7.6.3 Record sample volume
injected and resulting peak sizes
I
7.6.4 Determine identity and
quantity of each component peak
that corresponds to compounds
used for calibration
7.6.5
Does peak
exceed working
range of
system?
7.6.5 Dilute extract
reanalyze
7.6.6 Compare standard and
sample retention times; identify
compounds
I
Stop
8061-24
****September 1989****
Revision 2
September 1989
-------
APPENDIX C
GC/MS CHROMATOGRAMS AND MASS SPECTRA
(PLOT AND LIST)* FOR METHOD 8060 COMPOUNDS
aTo identify a particular mass spectrum in this appendix, refer to Table 1
in this appendix which gives compound name, scan number, three most
intense ions and their relative intensities. The GC/MS operating
conditions are given in footnote "a" of Table 1.
C-l
-------
TABLE 1. RETENTION TIMES (SCAN NUMBER) AND THREE MOST INTENSE PEAKS IN THE
MASS SPECTRA OF METHOD 8060 COMPOUNDS3
Compound Compound
no.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
name
DMP
DEP
DIBP
DBP
BMEP
BMPP
BEEP
DAP
HEHP
DHP
BBP
BBEP
DCP
DEHP
OOP
DNP
Benzyl benzoate (IS)
Diphenyl
Diphenyl
Diphenyl
Dibenzyl
Dioctyl
phthalate (SU)
isophthalate (SU)
terephthalate (SU)
phthalate (SU)
isophthalate (SU)
Scan
number
1128
1264
1499
1573
1602
1662
1687
1714
1737
1844
1851
1928
1962
1973
2082
2191
1397
1959
2057
2061
2087
2127
Mass spectrum
m/z (relative intensity)
163
149
149
149
59
149
45
149
149
149
149
57
149
149
149
149
105
225
225
225
107
167
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
(100),
77
177
57
41
58
85
72
43
57
43
91
56
167
57
43
43
91
77
76
104
91
279
(21),
(23),
(28),
(9.2),
(73),
(66),
(94),
(19),
(19),
(21),
(69),
(82) ,
(34),
(39),
(14),
(18),
(43),
(39),
(27),
(44),
(81),
(41),
164
150
41
150
149
43
73
150
55
41
206
45
55
167
57
150
77
226
104
76
149
149
(10)
(12)
(13)
(8.9)
(13)
(66)
(92)
(8.8)
(12)
(8.4)
(26)
(80)
(24)
(36)
(12)
(9.8)
(28)
(15)
(23)
(20)
(70)
(21)
aThe GC operating conditions are as follows: 30 m x 0.25 mm ID
DB-5 fused-silica capillary column, 45 to 300°C at 8°C/min.
C-3
-------
MASS SPECTRUM
02/02/88 23:04t88 + 18:48
SAMPLEc \MMKI3
CONOS.i
ENHANCED
188.01
DATAi ESTERSTD #1128
CALIs ESTERSTD 12
163
BASE M/Zj 163
RICi 7688.
Dimethyl phthalate (DMP)
o
I
50.0-1
77
50
39
4U-
92
104
I
128
M/Z
135
i
149
60
128
140
-ry
160
r 3464.
194
180
200
-------
Class List
O2/O2/88 23: 04: OO + 18: 48
Samp 1e:
I*
*
*<*3
Dimethyl phthalate (DMP)
Data: ESTERSTD *112B
Call: ESTERSTD » 2
Base at/1
RIC:
163
7688.
Conds. :
Enhanced
38
195
Mass
38?
39?
50
51
52
53
59
63
64
65
66
73
74
75
76
77
78
79
91
92
93
1O4
1O5
119
120
132
133
134
135
136
149
162
163
164
165
194
195
(S 15B 2N
0. OO
X RA
1. 53
2. OS
7. 94
3. 2O
1. 79
0. 4O
0. 92
1. 65
2. 80
1. 21
1. 67
0. 38
2. 42
2. 66
9. 56
21. 39
1. 79
2. 25
1. 1O
7. 82
0. 43
4. 76
3. 2O
0. 49
2. 54
0. 43
6. O9
0.64
7. O2
O. 87
0. 43
0. 92
1OO. OO
9. 82
1. 15
6. 87
O. 95
OT)
O.
Inten.
53.
71.
275.
111.
62.
14.
32.
57.
97.
42.
58.
13.
84.
92.
331.
741.
62.
78.
38.
271.
15.
165.
111.
17.
88.
15.
211.
22.
243.
3O.
15.
32.
3464.
34O.
40.
238.
33.
Minima Min Inten:
Maxima * 0
O.
C-5
-------
iee.e-i
o.
I
o>
50.0-
MASS SPECTRUM
82/82/88 23:84:88 + 21:84
SAMPLE: \###<#3
CONOS.i
ENHANCED
-------
Mass List Data: ESTERSTD «1264 Base m/z: 149
O2/02/88 23:04:00 + 21: O4 Call: ESTERSTD * 2 RIC: 5836.
Sample:
1*
*
*<*3
Diethyl phthalate (DEP)
Conds. :
Enhanced (S 15B 2N OT)
36
222
Mass
36?
38?
39?
43
44
45
50
51
52
64
65
66
74
75
76
77
91
93
104
105
1O6
121
122
132
147
149
150
151
176
177
178
222
0. 00
X RA
0. 51
0. 68
3. 9O
0. 38
1. 10
1. 27
6. 19
3. 39
1. 36
0. 93
12. 29
4. 03
1. 40
1. 74
9. 32
4. 7O
0. 72
8. 26
6. 95
8. 94
1. 86
7. 16
3. 69
1.78
0. 89
1OO. OO
12. 46
1. 48
1O. 42
23. 43
4. 49
2. 12
0. Minima Min Inten:
Maxima f O
Inten.
12.
16.
92.
9.
26.
30.
146.
80.
32.
22.
29O.
95.
33.
41.
220.
111.
17.
195.
164.
211.
44.
169.
87.
42.
21.
2360.
294.
35.
246.
553.
106.
SO.
C-7
-------
MASS SPECTRUM
02/92/88 23:04:08 + 24:39
SAMPLES \MKI3
CONDS.i
ENHANCED (S 15B 2N 0T)
188.8-1
DATA: ESTERSTD 11499
CALh ESTERSTD 12
149
BASE M/Zi 149
RIC: 7256.
Diisobutyl phthalate (DIBP)
00
50.0-
57
41
58
76
1
r 3440.
223
167
oe«
283
1
M/Z
48
60
88
120
148
168
188
20
8
228
-------
Mass List Data: ESTERSTO HI499 Base tn/z: 149
02/02/88 23:04:00 + 24:99 Call: ESTERSTD * 2 RIG: 7256.
Sample:
i*
*
*(*3
Diisobutyl phthalate (DIBP)
Conds. :
Enhanced (S 15B 2N OT)
Min Inten: 0.
« O
39
224
Mass
39?
4O
41
42
43
5O
51
55
56
57
58
65
75
76
77
93
104
105
121
122
123
132
135
149
150
151
167
2O5
223
224
0. 00
X RA
3. 11
O. 67
12. 88
0. 99
2. 01
1. 69
0. 78
1. 54
6. 92
28. 37
1. 31
3. 17
0. 38
5. 00
1. 19
2. 97
5. 78
2. O3
2. 38
1. 16
0. 87
1. 31
O. 41
100. 00
8. 98
0. 9O
3. 72
2. 21
6.69
0.81
O. Minima
Maxima
Inten.
107.
23.
443.
34.
69.
58.
27.
53.
238.
976.
45.
1O9.
13.
172.
41.
102.
199.
70.
82.
4O.
3O.
45.
14.
344O.
3O9.
31.
128.
76.
23O.
28.
C-9
-------
190.9-
0
58.8
41
361
T"
40
MASS SPECTRUM
92/82/88 23:84(88 + 26:13
SAMPLE: \tt**<«3
CONOS.s
ENHANCED
-------
Mass List Data: ESTERSTD #1573 Base m/z: 149
O2/02/88 23:O4: OO + 26:13 Call: ESTERSTD # 2 RIC: 5888.
Sample:
!#
*
»<*3
Di-n-butyl phthalate (DBF)
Conds. :
Enhanced (S 15B 2N OT)
36
224
Mass
36?
39?
41
43
44
50
51
55
56
57
65
76
77
93
1O4
1O5
121
122
123
132
135
147
149
ISO
151
160
205
206
223
224
0. 00
y. RA
0. 18
2. 25
9. 18
0. 85
0. 58
1. 35
0. 79
1. 9O
5. O9
5. 50
3. 30
3. 77
1. 20
2. 81
4. 09
2. 54
2. 54
1. 55
1. 02
0. 38
O. 41
0. 38
100. OO
8. 89
0. 76
O. 38
4. 36
0. 44
4. 59
0. 44
O. Minima Min Inten:
Maxima # O
Inten.
6.
77.
314.
29.
20.
46.
27.
65.
174.
188.
113.
129.
41.
96.
14O.
87.
87.
53.
35.
13.
14.
13.
342O.
3O4.
26.
13.
149.
15.
157.
15.
C-ll
-------
o
I
58.8-
MASS SPECTRUM
82/82/88 23:84(88 + 26:42
SAMPLEi MMMKtt
CONOS.s
ENHANCED
-------
Mass List Data: ESTERSTD «1602 Base ffl/z: 99
O2/O2/88 23:O4:OO + 26:42 Call: ESTERSTD * 2 RIC: 3316.
Samp 1e:
*
*(*3
Bis(2-methoxyethy1)
Conds. :
Enhanced
41
2O7
Mass
41
43
44
45
5O
58
59
6O
65
75
76
77
93
1O4
105
132
133
149
163
167
176
194
2O6
2O7
phthalate (BMEP)
(S 156 2N OT)
0. OO
% RA
1. 79
7.39
1. 48
11. 2O
2. 95
73. O9
100. OO
3. 89
1. 56
2. 57
7. 85
3. 58
1. 01
9. 18
1. O9
1.32
1. 01
12. 91
1. 09
2. 10
2. 8O
1. 01
1. 17
4. 82
O. Minima
Maxima
Inten.
23.
95.
19.
144.
38.
940.
1286.
50.
20.
33.
101.
46.
13.
118.
14.
17.
13.
166.
14.
27.
36.
13.
15.
62.
Min Inten:
« O
C-13
-------
198.0
o
58.0-
MASS SPECTRUM
82/82/88 23j84t08 + 27:42
SAMPLE: \W<#3
CONDS.i
ENHANCED (S 15B 2N 8T)
DATA: ESTERSTD #1662
CALI: ESTERSTD #2
BASE M/Zs 149
RIC: 3292.
149
Bis(4-methyl-2-pentyl) phthalate (BMPP)
43
69
57
65
76
93
184
u
121
M/H
748.
167
251
58
158
• i
258
-------
Mass List Data: ESTERSTD «1662 Base m/z: 149
02/02/88 23:04:00 -«• 27:42 Call: ESTERSTD # 2 RIC: 3292.
Sample:
!#
«
«(*3
Bis(4-methyl-2-pentyl) phthalate (BMPP)
Conds. :
Enhanced (S 15B 2N OT)
Minima Hin Znten: O.
Maxima * 0
39
251
Mass
39?
41
42
43
44
55
56
57
65
69
76
83
84
85
86
93
104
105
121
122
149
150
167
168
251
O. OO
X RA
3. 74
2O. 99
4. 81
66. 04
5. 61
7. 89
2. 81
13. 1O
2. 81
22. 99
6. 15
2. 14
20. 59
66. 18
5. 75
2. 67
6. 95
2. 27
2. 54
2. 27
10O. OO
13. 24
46. 12
4. 41
7. 49
O.
Inten.
28.
157.
36.
494.
42.
59.
21.
98.
21.
172.
46.
16.
154.
495.
43.
20.
52.
17.
19.
17.
748.
99.
345.
33.
56.
C-15
-------
188.8-1
o
I
I—•
CT>
58.8-1
fl/Z
45
MASS SPECTRUM
82/92/88 23184:99 + 28:87
SAMPLE: \ft*t<«3
CONDS.r
ENHANCED (S 15B 2N 8T)
DATA: ESTERSTD 11687
CALI: ESTERSTD 12
BASE M/Z: 45
RICs 5392.
r 1838.
72
59
58
65
Bis(2-ethoxyethyl) phthalate (BEEP)
149
184
76
89
93
121
176
'P
T
48
68
88
128
148
168
188
. 228
-------
Mass List
O2/02/88 23:04:00 + 28: O7
Sample:
If
*
«(f3
.Bis(2-ethoxyethyl) phthalate (BEEP)
Conds. :
Enhanced (S 15B 2N OT)
Data: ESTERSTD *1687
Call: ESTERSTD * 2
Base A/Z:
RIG:
45
3392.
39
221
Mass
39?
41
43
44
43
46
50
35
57
59
65
66
72
73
74
75
76
77
89
91
93
104
1O5
121
132
133
148
149
130
163
176
177
193
194
221
0. 00
X RA
1. 36
2. 72
21. 36
26. 41
1OO. OO
2. 72
3. 98
1. 26
2. 33
13. 59
6. 99
4. 95
93. 98
92. 04
5. 44
1. 36
11. 26
3. 11
10. 19
2.62
5. 15
15.73
5. 53
5. 05
5.24
6. 80
3. 50
36. 12
3. 4O
1. 46
9. 13
1.65
10. 19
1.55
3. 39
O.
Inten.
14.
28.
220.
272.
1030.
28.
41.
13.
24.
140.
72.
51.
968.
948.
56.
14.
116.
32.
105.
27.
53.
162.
57.
52.
54.
7O.
36.
372.
33.
15.
94.
17.
105.
16.
37.
Minima
Maxima
Min Inten:
* 0
C-17
-------
HASS SPECTRUM
62/62/88 23s64i66 + 28i34
SAMPLE! \MKi3
CONOS.i
ENHANCED (S 159 2N 6T)
DATA! ESTERSTD #1714
CALI: ESTERSTD 12
BASE M/Zs 149
RIC: 5456.
166.6-1
149
Diamyl phthalate (DAP)
o
I
I—I
00
56.6-
43
55
M/Z
8 T. .'ft..
' I * i • 'i • i
r 2836.
167
219
237
156
268
-------
Mass List Data: ESTERSTD 41714 Base «/z: 149
O2/02/88 23:04:OO * 28:34 Call: ESTERSTD * 2 RIC: 9456.
Sample:
:*
Diamyl phthalate (DAP)
Conds. :
Enhanced (S 15B 2N OT)
39 0. 00 O. Minima Min Inten:
238 Maxima * O
Mass X RA Inten.
39?
40
41
42
43
44
50
55
65
69
70
71
76
77
93
104
105
121
122
123
149
15O
151
167
219
237
238
2. 5O
0. 49
8. 64
5. 50
18. 86
1. 38
1.09
4. 94
2. 79
1. 20
3. 60
3. 67
3. 24
1. 2O
2. 64
3. 21
2. 40
2. 15
1. 52
1. 27
100. 00
8. 78
1. 20
0. 92
2.72
5.22
O. 95
71.
14.
245.
156.
535.
39.
31.
140.
79.
34.
102.
104.
92.
34.
75.
91.
68.
61.
43.
36.
2836.
249.
34.
26.
77.
148.
27.
C-19
-------
MASS SPECTRUM
82/02/88 23i84s88 + 28s57
SAMPLE! \m
-------
Mass List
02/02/88 23:04:00 + 28:57
Samp 1e:
!«
*
«<*3
Hexyl 2-ethylhexyl phthalate (HEHP)
Conds. :
Enhanced (S 15B 2N OT)
Data: ESTERSTO *1737
Call: ESTERSTD * 2
Base m/z:
RIC:
149
3108.
39
223
Mass
39?
41
43
44
5O
54
55
56
57
65
67
76
77
81
82
83
93
104
105
121
122
123
149
150
167
223
0.00
'4 RA
4. 03
13. 44
2. 33
1. 34
1. 13
3. 11
11. 53
3. 32
18. 95
3. 61
5. 37
4. 17
1. 06
0. 99
2. 19
3. 82
2. 19
5. 37
1.34
2. 19
1. 20
1. 13
100. 00
11.24
6. 93
7. 07
0.
Inten.
57.
190.
33.
19.
16.
44.
163.
47.
268.
51.
76.
59.
15.
14.
31.
54.
31.
76.
19.
31.
17.
16.
1414.
159.
98.
1OO.
Minima Min Inten:
Maxima # 0
O.
C-21
-------
188.8-1
o
I
ro
ro
58.0-
M/2
MASS SPECTRUM
82/02/88 23:84i 88 + 38:44
SAMPLE: MMHKI3
CONOS.i
ENHANCED (S 156 2N 8T>
DATA: ESTERSTD 11844
CALI: ESTERSTO 12
BASE M/Z: 149
RIC: 5272.
149
Dihexyl phthalate (DHP)
43
55
I
58
167
159
251
i
238
r 2788.
-------
Mass List Data: ESTERSTD #1844 Base «/z- 149
02/02/88 23:04:00 + 3O:44 Call: ESTERSTD * 2 RIC 3272
Sample:
!*
*
*(*3
Dihexyl phthalate (DHP)
Conds. :
Enhanced (S 15B 2N OT)
O.
39
252
Mass
39?
41
42
43
44
55
56
57
65
67
69
76
77
84
85
93
104
105
121
122
123
147
149
150
151
167
233
251
252
0. OO
% RA
1. 4O
8. 43
3. 19
20. 95
0. 82
4.73
4. 56
2. 40
1. 79
0. 57
2. 08
2. 19
0. 25
1. 79
2. 91
1. 54
2. 08
1. 47
1. 43
1. 22
1. 15
0.47
1OO. 00
9. 18
1. 15
1. 33
2. 51
6. 28
0. 68
O. Minima Min Inten:
Maxima * 0
Inten.
39.
235.
89.
584.
23.
132.
127.
67.
50.
16.
58.
61.
7.
5O.
81.
43.
58.
41.
40.
34.
32.
13.
2788.
256.
32.
37.
70.
173.
19.
C-23
-------
188.8
o
i
ro
58.e H
M/Z
41
MASS SPECTRUM
82/82/88 23:84:88 + 38:51
SAMPlEi \##»<#3
CONDS.:
ENHANCED (S 158 2N 8T)
Butyl benzyl phthalate (BBP)
65
50
II
57
76
i Ih
91
184
123
132
DATA: ESTERSTO #1851
CALI: ESTERSTO #2
BASE n/2: 149
RIC: 4752.
149
58
158
286
178
238
r 1286.
-------
(lass List
O2/02/88 23: 04: 00
Sample:
I*
30: 51
Data:
Call:
ESTERSTD »1891
ESTERSTD « 2
Base
RIC:
149
4752.
*<*3
Butyl benzyl phthalate (BBP)
Conds. :
Enhanced (S 15B 2N OT)
39
238
Mass
0. OO
f. RA
0.
Inten.
Minima
Maxima
Min Inten:
ft O
O.
39?
41
44
50
51
55
56
57
63
65
73
76
77
79
89
90
91
92
93
104
105
1O6
107
12O
121
122
123
132
133
135
136
149
15O
178
2O5
2O6
207
238
5. 39
9. 95
2. 16
3. 57
2. 99
2. 90
5. 14
5. 31
1. 58
14. 34
3. 32
5. 97
5. 47
2. 90
2. 82
2. 40
68. 91
6. 97
2. 57
13. 18
9. 54
1. 33
4. 98
1. 66
2. 24
7. 79
13. 43
15.09
3. 48
7. 38
1. 16
1OO. OO
11.94
4. 39
4.89
26. 20
5.22
4. 15
65.
12O.
26.
43.
36.
35.
62.
64.
19.
173.
4O.
72.
66.
35.
34.
29.
831.
84.
31.
159.
115.
16.
60.
2O.
27.
94.
162.
182.
42.
89.
14.
1206.
144.
53.
59.
316.
63.
SO.
C-25
-------
188.8
MASS SPECTRUM
02/82/88 23:94:99 + 32;08
SAMPLEi st##<#3
CONDS.i
ENHANCED
57
DATA: ESTERSTD 11928
CALI: ESTERSTD 12
BASE H/Zs 57
RICs 4888.
45
o
I
ro
a*
se. eH
41
647.
Bis(2-rKbutoxyethyl) phthalate (BBEP)
149
181
85
76
93
W/Z
5.8
133
II
176
193
163
188
T"
158
249
258
-------
Mass List Data. ESTERSTD #1928 Base */z: 57
O2/O2/88 23:04:OO + 32:OS Call: ESTERSTD # 2 RIC: 4880.
Sample:
i«
f
«(*3
Bis(2-n-butoxyethyl) phthalate (BBEP)
Minima Min Inten:
Maxima # O
Conds. :
Enhanced
36
249
Mace
36?
39?
41
43
44
45
50
55
56
57
58
63
65
66
71
72
73
76
77
83
85
86
88
93
100
101
102
104
1O5
117
119
121
132
133
148
149
150
163
176
177
193
194
249
(S 15B 2N
O. OO
% RA
2. Ol
3. 4O
44. 05
8. 35
13. 60
79. 60
2. 01
16.07
82. 23
1OO. OO
5. 56
5. 56
4. 95
4. 17
4. 02
2.01
3. 25
10. 97
2. 63
3.40
51. 31
2. 32
3. 09
4. 64
28. 59
55. 18
3. 40
20.25
7.73
8. 19
5. 41
4.95
12.98
13. 6O
7.88
63.21
6. 18
3.86
18.39
4. 48
2O. 87
4. O2
4. 17
OT)
O.
Inten.
13.
22.
285.
54.
88.
515.
13.
1O4.
532.
647.
36.
36.
32.
27.
26.
13.
21.
71.
17.
22.
332.
15.
2O.
3O.
185.
357.
22.
131.
5O.
53.
35.
32.
84.
88.
51.
409.
40.
25.
119.
29.
135.
26.
27.
C-27
-------
188.8-1
o
INJ
00
50.0-
41
MASS SPECTRUM
62/92/88 23:84»98 + 32i42
SAMPLE! M»«(tt3
CONDS.s
ENHANCED (S 15B 2N 8T)
DATAi ESTERSTO 11962
CALI: ESTERSTD #2
BASE M/Z: 149
RICi 5832.
1 9
Dicyclohexyl phthalate (DCP)
55
I. -.1
67
83
T
In
93
J_
M/Z
.50
249
297
r 2172.
1 ' ^ I ' I
150
250
-------
Mass List Data: ESTERSTD 41962 Base m/z: 149
02/02/88 23:04:00 + 32:42 Cali: ESTERSTD « 2 RIC: 5832.
Samp 1e:
!#
*(*3
Dicyclohexyl phthalate (DCP)
Conds. :
Enhanced (S 15B 2N OT)
39
249
Mass
39?
41
42
43
44
SO
53
54
55
56
57
65
67
76
77
79
81
82
83
93
99
1OO
1O4
105
121
122
149
ISO
151
167
168
2O7
249
0. 00
X RA
3. 96
14. 87
1. 43
2. 62
1. 70
0. 87
1. 52
4. 28
24.03
1. 61
2. 03
3. 36
7. 41
3. 78
1. 29
1. 70
3. 22
4. 83
9.85
2. 76
2. 62
0. 6O
3. 73
1. 70
2. 16
2. 81
100. 00
14. 36
1. 57
33. 70
3. 36
0.28
4. Ol
O. Minima Min Inten:
Maxima « O
Inten.
86.
323.
31.
57.
37.
19.
33.
93.
522.
35.
44.
73.
161.
82.
28.
37.
70.
105.
214.
60.
57.
13.
81.
37.
47.
61.
2172.
312.
34.
732.
73.
6.
87.
C-29
-------
Mass List Data: ESTERSTD 41962 Base m/z: 149
02/02/88 23:04:00 + 32:42 Cali: ESTERSTD « 2 RIC: 5832.
Samp 1e:
!#
*(*3
Dicyclohexyl phthalate (DCP)
Conds. :
Enhanced (S 15B 2N OT)
39
249
Mass
39?
41
42
43
44
SO
53
54
55
56
57
65
67
76
77
79
81
82
83
93
99
1OO
1O4
105
121
122
149
ISO
151
167
168
2O7
249
0. 00
X RA
3. 96
14. 87
1. 43
2. 62
1. 70
0. 87
1. 52
4. 28
24.03
1. 61
2. 03
3. 36
7. 41
3. 78
1. 29
1. 70
3. 22
4. 83
9.85
2. 76
2. 62
0. 6O
3. 73
1. 70
2. 16
2. 81
100. 00
14. 36
1. 57
33. 70
3. 36
0.28
4. Ol
O. Minima Min Inten:
Maxima « O
Inten.
86.
323.
31.
57.
37.
19.
33.
93.
522.
35.
44.
73.
161.
82.
28.
37.
70.
105.
214.
60.
57.
13.
81.
37.
47.
61.
2172.
312.
34.
732.
73.
6.
87.
C-29
-------
MASS SPECTRUM
02/02/88 23:04100 + 32j53
SAMPLE: \*ft«<*3
CONOS.i
ENHANCED (S 158 2N 0T)
DATA: ESTERSTD #1973
CALh ESTERSTD #2
BASE M/Z: 149
RICs 5248.
100.0
149
Bis(2-ethylhexyl) phthalate (DEHP)
o
I
co
o
50.0-
57
43
71
65
Hi I.
83
76
104
,22.32
167
M/Z
279
50
150
r 1288.
258
-------
Mass List Data: ESTERSTD 91973 Base a/z: 149
02/02/88 23.04.00 •»• 32:53 Call: ESTERSTD # 2 RIC: 5248.
Sample:
;*
«
*(f3
Bis(2-ethylhexyl) phthalate (DEHP)
Conds. :
Enhanced
39
280
Mass
39?
41
42
43
44
55
56
57
58
65
67
69
70
71
72
76
77
82
83
84
93
1O4
1O5
112
113
114
121
122
132
149
150
151
163
167
168
279
28O
(5 15B 2N
0. 00
X RA
3. O5
21. 41
4. O6
28. 44
1. 33
15.00
8. 91
39. 30
2. 42
2.97
0. 62
6. 33
24. 06
28. 36
1. 95
3. 75
0. 70
2. 19
8. 98
5. 00
2. 11
6. 25
2.03
8. 52
13. 12
0.70
2. O3
2. 11
2.42
1OO. OO
11. 25
1. 8O
1. O9
36. O9
3.75
6.48
0. 70
OT)
O. Minima Min Inten.
Maxima * 0
Inten.
39.
274.
52.
364.
17.
192.
114.
5O3.
31.
38.
8.
81.
308.
363.
25.
48.
9.
28.
115.
64.
27.
80.
26.
1O9.
168.
9.
26.
27.
31.
1280.
144.
23.
14.
462.
48.
83.
9.
C-31
-------
MASS SPECTRUM
02/82/88 23i84i88 + 34:42
SAMPLE: \W<*3
COWS*:
ENHANCED
-------
Mass List Data:
02/02/88 23:04:00 + 34:42 Cali:
Samp 1e:
ESTERSTD «2O82
ESTERSTD « 2
Base m/z:
RIC:
149
3744.
ft<*3
Di-n-octyl phthalate (OOP)
Conds. :
Enhanced (S 15B 2N OT)
39
281
Mass
39?
41
42
43
44
55
56
57
65
67
69
70
71
76
83
84
93
104
105
106
112
121
123
135
149
150
151
167
253
261
279
280
281
0. 00
% RA
1. 38
1O. 94
3. 87
13. 76
1. 49
7. 07
4. 09
11. 66
0. 88
0. 88
4. 25
3. 37
7. 73
1. 44
1. 82
1. 38
0. 77
1. 88
0. 50
1. 55
1. 66
1.05
0.88
0. 50
100. 00
9.28
0.83
1.82
0. 39
1. 10
6. 57
O. 77
0. 61
0. Minima Min Inten:
Maxima * 0
Inten.
25.
198.
70.
249.
27.
128.
74.
211.
16.
16.
77.
61.
140.
26.
33.
25.
14.
34.
9.
28.
30.
19.
16.
9.
1810.
168.
15.
33.
7.
20.
119.
14.
11.
O.
C-33
-------
188.8
o
I
U>
58.8-
43
MASS SPECTRUM
92/82/88 23(84:88 + 36i31
SAMPLEi \4MMKt3
CONOS.!
ENHANCED (S 158 2N 8T)
DATA: ESTERSTD 12191
CALI: ESTERSTO »2
BASE M/Z: 149
RICj 3428.
149
Dinonyl phthalate (DNP)
37
i
5ft
293
191
275
r 1566.
158
288
258
-------
Mass List Data: ESTERSTD #2191 Base m/z: 149
02/02/88 23:04:OO + 36:31 Call: ESTERSTD * 2 RIC: 3428.
Sample:
!*
«
*(*3
Dinonyl phthalate (DNP)
Conds. :
Enhanced (S 15B 2N OT)
Minima Min Inten: O.
Maxima • 0
39
294
Mass
39?
41
42
43
44
54
55
56
57
67
69
70
71
76
81
83
84
85
97
98
1O4
105
121
123
126
127
149
ISO
167
177
191
275
293
294
0. 00
X RA
1. 60
10. 54
2. 94
17. 94
2. 04
0. 57
8. 30
4. 60
9. 26
0. 57
4. 53
2. 62
7. 60
1. 53
0. 83
2. 23
1. 53
4. 15
1. 79
1. 34
2. 11
1. 66
0. 57
1. 53
1. 47
1. 53
1OO. OO
9. 83
2. 36
0. 57
1. 15
0. 96
6. 96
1.02
0.
Inten.
25.
165.
46.
281.
32.
9.
130.
72.
145.
9.
71.
41.
119.
24.
13.
35.
24.
65.
28.
21.
33.
26.
9.
24.
23.
24.
1566.
154.
37.
9.
18.
15.
109.
16.
C-35
-------
108.0-1
MASS SPECTRUH
02/04/88 0:23108 + 23:17
SAMPLE! PHTHALATE STD A 100NG/UL
CONOS.t FINN4500
ENHANCED (S 156 2N 0T)
105
DATA: R8203STDA 11397
CALIi R8283STOA #2
BASE M/Z: 105
RICr 16640.
o
I
58.0-
91
77
51
39
65
M/H
40
.,
60
-44
r 5584.
Benzyl benzoate
2 2
:.r
194
100
120
140
160
180
220
-------
Mass List Data:
O2/04/88 0:23:OO + 23:17 Cali:
Sample: PHTHALATE STD A 10ONG/UL
Conds.: FINN4500
Enhanced (S 15B 2N OT)
R0203STDA f!397
R0203STOA « 2
Base oi/z
RIC:
105
16640.
38
214
Mass
38?
39?
40
41
90
51
52
53
62
63
64
65
66
74
75
76
77
78
79
80
89
9O
91
92
1OS
1O6
1O7
1O8
152
153
165
166
167
168
194
195
212
213
214
0. OO
X RA
1. O2
4. 92
0. 43
1. 41
3. 64
12. 46
1. 59
0. 16
0. 61
2. 53
0. 88
10. 3O
0. 64
0. 75
0. 64
1. 61
27. 79
2. 97
5. 07
0. 43
4. 94
7. 4O
42. 91
3. 38
100. 00
8. 70
6. 23
0. 48
0. 56
0. 27
1. 40
O. 59
5.21
1. 13
8.86
1. 47
2O. 59
3.22
0. 25
0.
Inten.
57.
275.
24.
79.
203.
696.
89.
9.
34.
141.
49.
575.
36.
42.
36.
90.
1552.
166.
283.
24.
276.
413.
2396.
189.
5584.
486.
348.
27.
31.
15.
78.
33.
291.
63.
495.
82.
115O.
180.
14.
Minima Min Inten:
Maxima « O
0.
Benzyl benzoate
C-37
-------
MASS SPECTRUM
82/64/88 1«16:99 + 32:39
SAMPLE: \tf»<*3
CONDS.:
ENHANCED (S 15B 2N 8T)
R8283STDB #1939
CALI: R8283STDB 12
188.8-1
BASE M/Z: 225
RIC: 84736.
225
Diphenyl phthalate (DPP)
GJ
00
58.8-
77
M/Z
39 51
-58
65
.iJ.69
184
89..
153
115
15
I .
127
169 183
287
r 35392.
158
| • -• . .... |
288-
-------
Class List
02/O4/88 1:16:OO
Sample:
!#
*
#<»3
Diphenyl phthalate (DPP)
Data: RO203STDB «1959
32:39 Cali: R02O3STDB * 2
Base PI/I: 225
RIG: 84736.
Conds. :
Enhanced (S 15B 2N OT)
37
228
Mass
37?
38?
39?
40
41
49
50
51
92
53
55
62
63
64
65
66
69
70
73 .
74
75
76
77
78
79
87
88
89
91
92
93
94
101
1O2
103
1O4
1O5
106
113
114
115
116
119
121
126
127
128
0. OO
X RA
0. 13
0. 59
5. 17
0. 30
0. O6
0. OS
4. 45
5. 89
0. 31
O. 27
0. 22
0. 23
1. O7
0. 6O
4. 45
0. 46
O. O9
0. 05
0. 10
0. 80
1. 65
13. 02
38. 88
2. 57
0.24
0. 04
0.06
0.28
0. 24
0.28
0. 82
0.47
0. O7
0. 14
O. O6
8. 54
2. 16
0. 18
0.26
O. 19
3. 35
0. 35
0. 11
O. 65
0. O6
O. 18
O. 02
O.
Inten.
47.
210.
1830.
1O7.
22.
19.
1574.
2084.
111.
97.
78.
82.
380.
212.
1574.
162.
31.
16.
36.
282.
583.
46O8.
1376O.
911.
85.
13.
23.
99.
85.
98.
289.
165.
25.
5O.
22.
3024.
764.
64.
91.
67.
1186.
125.
39.
23O.
23.
63.
8.
Minima
Maxima
Mass
140
141
142
149
ISO
151
152
153
154
155
168
169
170
179
181
182
183
184
196
197
198
207
222
223
225
226
227
228
Min Inten:
« O
X RA
O. 33
2.47
O. 32
O. 60
0. 15
O. 68
2. 6O
6. O6
0. 79
0. O6
0.71
O. 82
0. 12
O. 19
O. 46
O. 07
0.64
0. 10
O. 28
1. 55
0. 23
O. 14
0. O4
O. 4O
1OO. OO
15. Ol
1.59
0. 16
C-39
O.
Inten.
118.
875.
112.
214.
54.
239.
921.
2144.
279.
23.
253.
29O.
44.
69.
162.
24.
227.
34.
99.
549.
83.
51.
14.
14O.
35392.
5312.
564.
55.
-------
132
133
139
0. 68
0. 14
1. 21
242.
51.
428.
C-40
-------
iee.e-
58.8-
MASS SPECTRUM
02/84/88 I:16i98 + 34:17
SAMPLEi \W<»3
CONDS.i
ENHANCED (S 15B 2N 9T)
DATAi R8283STDB «2857
CALIt R0283STOB «2
225
BASE H/Zi 225
RICt 68488.
Diphenyl isophthalate (DPIP)
76
1(
T t ?
,1 . L J,,,1.84 93
14
1
'I5
L 1 132 ,
1
169
, 153 1 179 197
318
r 29376.
M/H
58
158
)8 258
-------
Mass List
02/O4/88
Sanple:
1: 16: OO + 34: 17
Data: RO203STDB O2O97
Call: R02O3STDB * 2
Base A/Z: 225
RIC: 68480.
Diphenyl isophthalate (DPIP)
Conds. :
Enhanced (S 15B 2N OT)
37
32O
Mass
37?
38?
39?
4O
41
SO
51
52
53
55
57
62
63
64
65
66
69
70
73
74
75
76
77
78
84
88
89
91
92
93
94
98
1O2
1O3
1O4
1CS
1O6
113
114
115
116
119
121
127
132
133
139
0. OO
X RA
0. 11
O. 61
4. 97
0. 31
0. 08
4. 91
2. 11
0. 15
0. 23
0. 18
0. 03
0. 25
1. 06
0. 69
4. 30
0. 58
0. 12
0. 14
0. O7
0. 86
3. 04
26. 82
4. 38
O. 29
0.08
0. 07
0. 19
O. 35
0.25
O. 58
0. 51
0. 2O
0. 14
0. 90
22. 58
2. 48
0. 16
O. 61
O. 17
4. 52
0. 45
O. 13
0. 15
0. 08
O. 37
0. 11
1. 04
O.
Inten.
31.
178.
146O.
92.
24.
1442.
621.
45.
67.
52.
8.
72.
311.
202.
1262.
170.
35.
40.
20.
253.
892.
7880.
1286.
85.
23.
22.
55.
1O4.
74.
169.
150.
59.
41.
263.
6632.
728.
48.
178.
51.
1328.
132.
37.
44.
24.
110.
32.
3O6.
Minima
Maxima
Mass
143
149
151
152
153
154
167
168
169
17O
171
172
179
195
196
197
198
223
225
226
227
228
317
318
319
320
Min Inten:
« 0
X RA
0. 10
0.09
0.29
0. 62
0. 79
0. 11
0. 04
1. 13
3. O2
O. 38
0. BO
O. 09
0. 13
O. 10
0. 22
0. 84
0. 13
0. 34
1OO. OO
14. 92
1. 58
0. 15
0. O3
2. 54
0. 54
0.03
C-42
O.
Inten.
28.
27.
86.
181.
233.
33.
13.
331.
687.
112.
234.
27.
37.
28.
64.
246.
39.
99.
29376.
4384.
463.
44.
9.
747.
159.
1O.
-------
140 0. 37 109.
141 9. 9O 29O8.
142 1. 18 348.
C-43
-------
MASS SPECTRUM
02/04/88 0:23:00 + 34(21
SAMPLE: PHTHALATE STD A 100NG/UL
CONDS.: FINN4500
ENHANCED (S 15B 2N 0T)
100.0-1
DATA: R0203STOA 12961
CALI: R0203STDA #2
225
BASE M/Zi 225
RIC: 53376.
Diphenyl terephthalate (DPTP)
o
I
50.0-
104
76
39 50
M/2
132
'j41
169
197
1
50
' I ' ' ' '
150
318
I*
r 21088.
250
1 I '
300
-------
Mass List Data:
02/04/88 0:23:00 + 34:21 Call:
Sample: PHTHALATE STD A 1OONG/UL
Conds.: FINN45OO
Enhanced (S 1SB 2N OT)
RO203STDA «2O61
R0203STDA « 2
Base »/z:
RIC:
225
33376.
37
320
Mass
37?
38?
39?
40
41
SO
51
32
53
33
62
63
64
65
66
7O
73
74
73
76
77
78
84
89
91
92
93
94
96
98
1O2
1O3
1O4
1O3
1O6
113
114
115
116
121
132
133
134
139
140
141
142
149
132
168
0. OO
% RA
0. 10
0. 69
5. 32
0. 34
0. O6
3. 43
1. 99
0. 14
0. 29
0. 17
0. 20
0. 98
0. 58
4. 69
0. 52
0. 11
0. 07
0. 94
2. 53
20. 22
2. 98
0. 20
0. 05
0. 16
0. 20
0. 17
O. 64
0. 49
0. O7
0. 08
0. 12
O. 46
43.93
3.37
0.22
O. 33
O. 11
2.35
0.24
0.21
20. 14
1. 78
0. 14
0. 72
0.21
2. 10
O. 23
O. 31
O. 18
O. 43
O.
Inten.
22.
145.
1122.
71.
13.
1146.
419.
29.
61.
35.
43.
207.
122.
989.
110.
24.
14.
199.
533.
4264.
629.
42.
1O.
34.
43.
36.
136.
103.
15.
16.
26.
97.
9264.
711.
47.
69.
23.
495.
SO.
43.
4248.
375.
3O.
131.
44.
443.
S3.
65.
38.
9O.
Minima
Maxima
Mass
169
170
195
196
197
198
225
226
227
228
318
319
320
Min Xnten:
* 0
X RA
O. 98
0. 13
O. 11
0. 41
3. 28
O. 47
100. 00
14. 81
1. 56
O. 14
2. 51
0. 52
O. 04
0.
Inten.
206.
28.
24.
86.
691.
99.
21O88.
3124.
329.
29.
529.
no.
9.
Diphenyl terephthalate
C-45
-------
CT>
50.0-
MASS SPECTRUM
82/04/88 1:16:89 + 34j47
SAflPLEi \HMKt3
CONDS.t
ENHANCED (S 15B 2N 0T)
197
DATAs R0203STOB 12087
CALI: R8203STOB #2
BASE N/Z: 107
RICs 80128.
91
65
IL
51
1.
I
Dibenzyl phthalate (DBZP)
149
132
H/Z
165 J.ff 193 211
••^•^^—T™^T"*"*lp^^^^™r'w^T*^T^'^
238
150
r 22016.
250
-------
Mass List
02/04/88
Sample:
1:16:OO + 34:47
Data: R0203STDB #2087
Call: RO203STDB « 2
Base tn/z: 1O7
RIC: 80128.
..Dibenzyl phthalate (DBZP)
Conds. :
Enhanced (S 15B 2N OT)
38
256
Mass
38?
39?
40
41
44
SO
51
52
53
62
63
64
65
66
74
75
76
77
78
79
80
89
90
91
92
93
94
96
104
1O5
1O6
1O7
1O8
1O9
115
118
119
121
122
128
132
133
134
149
15O
151
0. 00
% RA
0. 38
3. 77
0. 32
1. 43
0. 10
2. 13
3. O5
0. 72
0. 3O
0. 25
1. 77
0. 80
15. 72
0. 90
O. 37
0. 55
3. 94
5. 96
0. 97
5. Ol
0. 31
2. 60
2. 18
80. 67
20. 93
3. 35
O. 19
O. O2
4.79
3.85
0. 11
100. OO
7. 53
0. 43
0. 12
O. O4
0. O7
2.34
O. 37
0. O6
2.80
1.3O
O. 15
70.28
6. 02
0. 77
O. 6O
0.
Inten.
84.
829.
71.
314.
22.
469.
672.
159.
67.
54.
389.
177.
3460.
199.
81.
122.
867.
1312.
214.
1102.
69.
573.
481.
1776O.
4608.
737.
42.
4.
1054.
847.
25.
22O16.
1658.
94.
26.
9.
15.
515.
82.
14.
617.
287.
33.
15472.
1326.
169.
132.
Minima
Hasina
Mass
166
167
178
179
180
181
193
194
197
198
211
238
239
240
255
256
Min Inten:
* 0
X RA
O. 35
O. 4O
O. 17
O. 22
0. 69
O. 4O
O. 24
O. 2O
O. 13
0. 06
0. 15
O. 57
0. 22
0. 04
O. 65
0. 11
O.
Inten.
77.
89.
37.
49.
153.
88.
53.
44.
28.
14.
33.
126.
49.
8.
143.
24.
C-47
-------
153
164
165
0 26
O. 06
1. 19
61.
14.
262.
C-48
-------
o
\o
188.8-
59.8-
M/Z
188.8-
38.8-
MASS SPECTRUM
82/04/88 8:23i88 + 35:27
SAMPLE! PHTHALATE STD A 188NG/UL
CONDS.I FINN4588
ENHANCED
-------
Mass List Data:
02/04/88 0:23:00 + 39:27 Call:
Sample: PHTHALATE STD A 100NG/UL
Conds.: FINN45OO
Enhanced (5 15B 2N OT)
R02O35TDA «2127
R0203STDA tt 2
Base «/z: 167
RIC: 124416.
Dioctyl isophthalate (DOIP)
36
391
Mass
39?
40
41
42
43
44
45
5O
31
52
33
54
55
36
57
58
65
66
67
68
69
7O
71
72
74
75
76
77
78
79
80
81
82
83
84
85
91
93
93
97
98
103
104
105
106
HO
111
112
113
114
0. OO
X RA
1. 87
0. 46
15. 47
6. 57
17. 19
0. 69
0. 11
0. 66
0. 33
0. 13
0. 77
1. 93
19. 36
15. 5O
19. 79
0. 87
5. 48
0. 66
2. 28
3. 39
20. 77
16. 91
15. 07
0. 82
0. 10
0. 79
4. 7O
1. 2O
0. 15
1.46
0. 15
0. 79
3. 12
11. 14
9. 32
O. 73
0. 54
O. 91
0.25
1.33
O. 13
0. 7O
7. O7
2. 31
0. 31
O. 30
O. 77
4.78
1. 32
0. 13
0.
Inten.
523.
128.
4328.
1838.
48O8.
193.
31.
185.
93.
35.
215.
539.
5416.
4336.
5536.
243.
1534.
185.
637.
948.
58O8.
4728.
4216.
229.
29.
22O.
1314.
335.
43.
4O9.
41.
222.
872.
3116.
26O8.
204.
ISO.
235.
69.
373.
37.
196.
1976.
647.
86.
83.
216.
1338.
424.
36.
Minima
Maxima
Mass
115
117
119
12O
121
122
123
124
129
131
132
133
134
135
147
148
149
150
151
152
162
163
164
165
167
168
169
176
177
179
189
193
203
204
2O7
2O8
261
262
263
277
279
28O
281
282
291
39O
391
Min Inten:
* 0
X RA
0. 13
O. 12
O. 13
0. 16
8.25
2.86
1.47
0. 11
O. 20
0. 13
O. 39
5.36
0. 55
O. 27
O. 31
0. 76
2O. 62
2. 13
1. 34
0. 14
O. 55
0. 11
0. 11
O. 37
1OO. 00
8.81
1. 19
0. 36
0. 14
O. 12
0. 13
0. 13
0. 11
O. 3O
0. 15
O. 1O
13. 34
2.40
0.33
0.20
40.50
6.61
O. 9O
0. 12
0. 14
O. 11
O. 11
/- cn
29.
Inten.
36.
33.
36.
44.
2308.
799.
412.
31.
36.
35.
165.
1498.
154.
76.
88.
213.
5768.
595.
374.
38.
133.
31.
32.
103.
27968.
2464.
334.
1O1.
38.
33.
37.
33.
31.
83.
43.
29.
3788.
672.
93.
37.
11328.
185O.
231.
34.
39.
32.
30.
-------
APPENDIX D
RESULTS OF INORGANIC ANALYSIS
FOR THE ESTUARINE WATER, LEACHATE,
AND GROUNDWATER
D-l
-------
Table 1. Metals Results
Parameter
Aluminum
Antimony
Arsenic
Barium
Beryllium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Molybdenum
Nickel
Selenium
Silicon
Silver
Sodium
Thallium
Tin
Titanium
Vanadium
Zinc
i
EPA
Method
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
GROUNDWATER
SAMPLE Spike
ug/L
<50
<50
<100
79.1
<5.0
300
<5.0
113000
<10
<10
<10
31.4
<50
38000
45.7
<10
<20
<100
13100
<10
51900
<100
<50
<50
<10
<10
% Recov
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
Dup
Spike
RPD
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
Method
Method Detection
Blank Limit
ug/L
<50
<50
<100
<10
<5
<50
<5
<130
<10
<10
<10
<10
<50
<140
<10
<10
<20
<100
<50
<10
<250
<100
<50
<50
<10
<10
ug/L
50.0
50.0
100.0
10.0
5.0
50.0
5.0
130.0
10.0
10.0
10.0
10.0
50.0
140.0
10.0
10.0
20.0
100.0
50.0
10.0
250.0
100.0
50.0
50.0
10.0
10.0
NS - Not Spiked
D-3
-------
Table 2. General Chemical Results
GROUNDWATER
Parameter
Bromide
Chloride
B.C., umhos/cm
Flouride
Nitrate, as N
Nitrite, as N
pH, units of pH
Phosphate, as P
Sulfate
TDS, at 180 'C
TOC
FPa
Lif*\
Method
300.0 D
300.0 D
120.1 W
300.0 D
300.0 D
300.0 D
150.1 M
300.0 0
300.0 D
160.1 G
415
SAMPLE
mg/L
<0.05
79.7
910
<0.05
11.6
<0.1
7.4
<0.2
126
746
4.0
Spike
% Recov
104
114
N/A
108
106
125
N/A
92.7
97.7
N/A
NS
Dup
Spike
RPD
0.06
4.0
N/A
6.4
0.37
0.07
N/A
0.34
0.29
0.19
NS
Control
Sample
mg/L
105
99.0
N/A
NA
102
NA
N/A
108
104
N/A
NA
Method
Blank
mg/L
<0.05
<0.1
N/A
NA
<0.1
<0.1
N/A
<0.2
<0.3
N/A
N/A
D - Ion Chromatography
G - gravimetric
M - electrometric
W - Wheatstone bridge conductivity meter
N/A - Not Applicable
NA - Not analyzed
NS - Not spiked
D-4
-------
Table 3. Metals Results
Parameter
Aluminum
Antimony
Arsenic
Barium
Beryllium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Molybdenum
Nickel
Selenium
Silicon
Silver
Sodium
Thallium
Tin
Titanium
Vanadium
Zinc
EPA
Method
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
ESTUARINE
WATER
ug/L
2100
<50
<100
<10
<5.0
3610
<5.0
331000
<10
<10
<10
1720
<50
1090000
35.5
38.8
<20
<200
9260
<10
8650000
<200
<50
50.3
17.8
<10
Spike
% Recov
94
94
NS
90
93
NS
102
97
104
97
92
96
104
104
98
NS
99
- NS
95
87
106
NS
NS
NS
96
101
Dup
Spike
ug/L
8.9
1.3
NS
9.4
12
NS
20
1.0
12
13
13
1.1
13
0.77
7.6
NS
7.9
NS
0.17
4.5
0.37
NS
NS
NS
11
11
Method
Method Detection
Blank Limit
ug/L
71.7
<50
<100
<10
<5.0
<20
<5.0
<100
<10
<10
<10
<20
<50
<100
<10
<10
<20
<200
87.6
<10
<200
<200
<50
<50
<10
<10
ug/L
50
50
100
10
5.0
20
5.0
100
10
10
10
20
50
100
10
10
20
200
50
10
200
200
50
50
10
10
NS - Not spiked
D-5
-------
Table 4. General Chemical Results
ESTUARINE
WATER LEACHATE
Parameter
Bromide
Chloride
Fluoride
Nitrate, as N
Nitrite, as N
pH, units of pH
Phosphate, as P
Sulfate
TDS, at 180°C
Uf ri
Method
300.
300.
340.
300.
300.
150.
300.
300.
160.
0
0
2
0
0
1
0
0
1
D
D
E
D
D
M
D
D
G
mg/L
31.4
23900
0.61
<0 . 1
<0 . 1
8.0
<0.2
146000
31100
mg/L
<0.1
14.5
<0.05
<0 . 1
<0 . 1
5.5
<0.2
<0.3
32900
Spike
% Recov
NA
94.3
98.9
116
NA
NA
103
81.3
NA
Dup
Spike
RPD
NA
0.72
0
NA
NA
NA
1.6
0.41
4.5
Control
Sample
% Recov
NA
117
NA
108
NA
NA
NA
105
NA
D - Ion Chromatography
E - ion selective electrode
G - gravimetric
M - electrometric
NA - Not analyzed
D-6
-------
Table 5. Metals Results
Parameter
Antimony
Arsenic
Barium
Beryllium
Cadmium
Chromium
Cobalt
Copper
Lead
Molybdenum
Nickel
Selenium
Silver
Thallium
Vanadium
Zinc
Aluminum
Boron
Calcium
Iron
Magnesium
Manganese
Silicon
Sodium
Tin
Titanium
EPA
Method
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
200.7
LEACHATE
ug/L
<50
<100
2040
<5.0
<5.0
201
76.4
41.1
<50
15.0
<20
<200
11.1
<200
153
81.9
68600
<20
23400
51700
13000
3030
24900
7900000
<50
2330
Spike
% Recov
93
100
121
84
90
108
85
*
125
90
89
116
82
86
90
*
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
Dup
Spike
RPD
0
0
1.1
0
11
1.6
5.3
1.1
2.0
7.1
6.9
0
1.2
0
4.7
1.9
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
Method
Method Detection
Blank Limit
ug/L
<50
<100
<10
<5.0
<5.0
<10
<10
<10
<50
<10
<20
<200
<10
<200
<10
<10
<50
<20
<100
<20
<100
<10
<50
<200
<50
<50
ug/L
50
100
10
5.0
5.0
10
10
10
0.2
10
20
200
10
200
10
10
50
20
100
20
100
10
50
200
50
50
* Sample greater than 50x added spike.
NS - Not spiked
D-7
------- |