United States
Environmental Protection
Agency
Environmental Monitoring and
Support Laboratory
Cincinnati OH 45268
Research and Development
EPA/600/S4-85/068 Dec. 1985
Project Summary
Determination of Phenols in
Industrial and Municipal
Wastewaters
J. R. Florance, J. R. Hall, M. Khare, S. M. Maggio, J. C. Mitchell, R. A. Solomon,
J. R. SoloRio, D. L. Strother, and M. N. Wass
This report describes the research
conducted to develop an analytical test
procedure for the analysis of specific
organic toxic substances in effluent
wastewaters. The test procedure is
applicable to the analysis of 11 of the
114 priority pollutants identified by the
EPA as Category 8—Phenols. The test
procedure developed was originally
published in the December 3, 1979
issue of the Federal Register as EPA
Method 604 and subsequently included
as one of the EPA Test Methods for
Organic Chemical Analysis of Municipal
and Industrial Waste water.12
The procedure consists of several
steps, including extraction, concentra-
tion, and quantification by gas chroma-
tography with flame-ionization detec-
tion. An optional derivatization step is
presented, using pentafluorobenzyl
bromide followed by column chroma-
tography cleanup and quantification of
the derivatives by gas chromatography
with electron capture detection.
The final report describes the research
leading to the selection of the proce-
dural steps and includes the results of a
literature search, sample preservation
procedures, elution of the phenols on
various gas chromatographic columns,
several solvent extraction efficiencies
versus pH, stability of the compounds in
water-soluble solvents, sample extract
cleanup procedures, and application of
the procedures and effluent waste-
waters.
This Project Summary was developed
by EPA's Environmental Monitoring and
Support Laboratory, Cincinnati. OH, to
announce key findings of the research
project that Is fully documented in a
separate report of the same title (see
Project Report ordering information at
back).
Introduction
The Federal Water Pollution Control
Act Amendments of 1972 (PL 92-500)
and the Amendments of 1977 (PS 95-
217) required that the Administrator of
theU.S. Environmental Protection Agency
promulgate guidelines establishing test
procedures for the analysis of the priority
pollutants, which were separated into 12
categories. The final report fully describes
the research required for the method
developed for 11 of the priority pollutants
in one of those categories: Category 8—
Phenols.
The 11 compounds in Category 8—
Phenols are phenol, 2,4-dimethylphenol,
2-chlorophenol, 2,4-dichlorophenol, 2,4,
6-trichlorophenol, pentachlorophenol, 4-
chloro-3-methylphenol, 2-nitrophenol,
4-nitrophenol, 2,4-dinitrophenol, and
4,6-dinitro-2-methylphenol.
The research program included a litera-
ture search from 1960 through 1978,
sample preservation studies, evaluation
of solvents for liquid-liquid extraction,
stability studies of the compounds in
water-miscible solvents, and evaluation
of sample and extract cleanup procedures.
The use of XAD-26 resin as a possible
extraction and concentration step for all
the priority pollutant phenolics in waste-
water was evaluated and compared to
liquid-liquid extraction.
The gas chromatographic characteris-
tics data of the category compounds are
presented and include information on
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retention times with various gas chroma-
tography columns at different tempera-
tures, responses to both electron-capture
and flame ionization detectors, linearity
curves, and chemical data for all com-
pounds, and minimum detection limits.
To enhance the electron-capture re-
sponse of the phenols, two derivatization
techniques were investigated with the
results described in the full report. Penta-
fluorobenzyl bromide was selected as the
derivatizing reagent for the final method.
Based on the information gathered
during the research program, methods
were proposed for the analyses of the
Category 8 compounds in water. These
proposed methods were then used to
develop data on overall compound re-
coveries in spiked distilled water and
wastewater effluent samples.
Conclusions and
Recommendations
The results of the preservation study
experiments indicate the samples are
best preserved when buffered to a pH of 7
to 10, dechlorinated with 35 mg of sodium
thiosulfate per mg/L of chlorine per liter
of water, exposed to a minimum of UV
light, and stored and shipped at a temper-
ature of 4°C. When so handled, minimal
change or loss of phenol concentrations
are observed up to 7 days. The maximum
loss seen during the studies was 50% for
4-nitrophenol while the minimum loss of
8% was observed for 2,4-dinitrophenol.
The adjustment of the water samples to
pH 12, and extraction with dichloro-
methane was found to serve as a prelimi-
nary cleanup procedure. Most base/
neutral extractable interferences are
removed by this preliminary step, leaving
the phenols in the water.
The extraction of one-liter water
samples, with three 60-mL portions di-
chloromethane per liter, after adjustment
to pH 2 with 1N (-(280,4, was found to give
the best overall extraction efficiencies for
all the solvents studied. Optimum extrac-
tion efficiencies ranged from 42% to 100%
for the phenols. The dichloromethane
was concentrated in a Kuderna-Danish
evaporator with solvent exchange to 2-
propanol.
When the sample extracts require no
further cleanup, they are best analyzed by
flame ionization gas chromatography. The
evaluation of several chromatographic
columns led to the recommendation of
Supelcoport* coated with 1 % SP-1240DA.
Using this column, the response of the
phenols by GC/FID was found to be linear
throughout the low to high mg/L range.
Table 1 summarizes the gas chromato-
graphic operating conditions, retention
times, and MDL obtained under these
conditions. The separation of the 11
phenols achieved by this column is shown
in Figure 1.
An alternate extraction and concentra-
tion step, the use of XAD-26 resin, was
also evaluated. Results of this investiga-
tion indicate that the traditional solvent
extraction/concentration procedure
yields, average phenol recoveries that are
1.5 to 2 times the recoveries from the
resin.
In confirmation of the results by flame
ionization and as an aid in the elimination
of interferences in the analysis of phenols
in complex extracts, an optional deriva-
tization and column chromatographic
procedure was developed. Derivatization
of the phenols with chloracetic anhydride
was unsuccessful due to poor reaction
efficiencies and significant artifact for-
mation. However, the phenols, in genera I,
were successfully derivatized using pen-
tachromobenzyl bromide in the presence
of 18 Crown-6 catalyst. In order to
eliminate extraneous peaks in the deriva-
tized standard phenolic solution, a silica
gel cleanup procedure was adopted.
Evaluation of gas chromatographic
columns and conditions for the analyses
of the phenol derivatives led to the
recommended use of Chromosorb W-
AW-DMCS coated with 5% OV-17 in
conjunction with electron capture gas
chromatography. Table 2 summarizes the
recommended gas chromatographic con- '
ditions and operating conditions. The
table includes retention times and MDL
obtained under these conditions. An
example of the separation achieved by
this column is shown in Figure 2.
The procedures developed during this
study (Figure 3) were successfully applied
to three treated industrial effluent
samples and a municipal primary effluent
sample which contained several of the
phenols at concentration levels of 25 to
800 //g/L
The accuracy of the complete method,
based on spiked solutions of distilled
deionized water expressed as percent of
recovery, varied from 30% for 4-nitro-
phenol to 82% for 2-nitrophenol. The
same experiment using spiked municipal
wastewater gave an accuracy range from
50% for phenol to 76% for 4-chloro-3-
methylphenol. The single operator coef-
ficient of variation ranged from 10% for
2-nitrophenol to 54% for 2,6-dinitro-2-
methylphenol in a spiked municipal
wastewater sample.
References
1. Guidelines Establishing Test Pro-
cedures for Analysis of Pollutants,
Proposed Regulation. Federal
Register. 44(223):69484-69489,
Decembers, 1979.
2. Longbottom, J. E. and J. J. Lichten-
berg. Methods for Organic Chemical
Analysis of Municipal and Industrial
Wastewater. EPA/600/4-82/057,
USEPA, Environmental Monitoring
and Support Laboratory, Cincinnati.
1982.
Table 1. Chromatographic Conditions and Method Detection Limit
Parameter
2 - Chlorophenol
2-Nitrophenol
Phenol
2,4 -Dimethylphenol
2.4 -Dichlorophenol
2,4,6 -Trichlorophenol
4-Chloro-3-Methyfphenol
2,4-Dinitrophenol
2-Methyl-4,6-Dinitrophenol
Pentachlorophenol
4-Nitrophenol
Retention Time
(min.)
1.70
2.00
3.01
4.03
4.30
6.05
7.50
10.00
10.24
12.42
24.25
Method Detection Lmiit
(fig/Li
0.31
0.45
0.14
0.32
0.39
0.64
0.36
13.0
16.0
7.4
2.8
•Mention of trade names or commercial products
does not constitute endorsement or recommenda-
tion for use.
Column conditions: Supelcoport (80V100 mesh) coated with 1 % SP-1240 DA in 1.8m long x 2 mm
ID glass column with nitrogen carrier gas at a flow rate of 30 mL/min flow rate. Column
temperature was 80°C at injection, programmed immediately at 8°C/min to 150°C final
temperature. Method detection limits were determined with a flame ionization detector.
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1
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i
1
;
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B
C
;
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Column: 1%SP-1240DA on Supelcoport
Program: 80°C. -0 Minutes 8° /Minute to 150°C
Director: Flame lonization
A. 2-Chlorophenol
B. 2-Nitrophenol
C. Phenol
D. 2,4-Dimethylphenol
E. 2,4-Dichlorophenol
F. 2.4.6-Trichlorophenol
E G. 4-Chloro-3-methylphenol
H. 2,4-Dinitrophenol
1. 4,6-Dinitro-3-methylphenol
J. Pentachlorophenol
K. 4-Nitrophenol
G \ 1
1
UL
1
H
(
- ^ ^vj
11
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1
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0 5 W 15 20 25
Retention Time-Minutes
Figure 1. Gas chromatogram of phenols.
Table 2. Electron Capture Gas Chromatography ofPFBB Derivatives
Electron Capture
Parent Compound
Retention Time
(min.)
Method Detection Limit
2-Chlorophenol
2-Nitrophenol
Phenol
2,4 -Dimethylphenol
2,4-Dichlorophenol
2,4,6-Trichlorophenol
4 -Chloro-2 -Methylphenol
Pentachlorophenol
4-Nitrophenol
3.3
9.1
1.8
2.9
5.8
7.0
4.8
28.8
14.0
0.58
0.77
2.2
0.63
0.68
0.58
1.8
0.59
0.70
Column conditions: Chromosorb W-A W-DMCS (80/100 mesh) coated with 5%OV-17 packed in a
1,8m long x 2.0 mm ID glass column with 5% methane/95% argon carrier gas at a flow rate of 30
1 mL/min. Column temperature isothermal at 200°C.
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Column: 5% 0 V-17 on Chromosorb W-A W
Temperature: 200°C
Detector: Electron Capture
E F
A. Phenol
B. 2,4-Dimethylphenol
C. 2-Chlorophenol
D. 4-Chloro-3-methylphenol
E. 2,4-Dichlorophenol
F. 2,4,6 -Trichlorophenol
G. 2-Nitrophenol
H. 4-Nitrophenol
I. Pentachlorophenol
10 15 20
Retention Time-Minutes
25
30
Figure 2. Gas chromatogram of PFBB derivatives of phenols.
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Sample
1 Liter -pH12
3x60 ml
MeClz
AQ Layer
pH2
3 x 60 ml
MeC/2
Exchange to IPA
K/D - 1 ml
(Optional)
FID Analysis
for
Phenols
1
Derivative to
PFBB's
Extract
W/10 mlHexane-
8 mlDIHsO
E C Analysis
' (Deriv. EH.)
Silica Gel
Clean-Up
Column Fractions
EC Analysis
Figure 3. Analysis of phenols.
J. R. Florence, J. R. Hall, M. Khare, S. M. Maggio, J. C. Mitchell, R. A. Solomon, J.
R. SoloRio, D. L Strother. andM. N. Wass are with IT Enviroscience, Knoxville.
TN 37923.
James J. Lichtenberg is the EPA Project Officer (see below).
The complete report, entitled "Determination of Phenols in Industrial and
Municipal Wastewaters," (Order No. PB86-119 120/AS; Cost: $16.95. subject
to change! 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 and Support Laboratory
U.S. Environmental Protection Agency
Cincinnati. OH 45268
&U. S. GOVERNMENT PRINTING OFFICE:1986/646-l 16/20723
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United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
Official Business
Penalty for Private Use $300
EPA/600/S4-85/068
OC00329 PS
U S ENVIR PROTECTION AGENCY
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