EPA-660/4-75-005
JUNE 1975
Environmental Monitoring Series
Analysis of Organic Compounds in
Two Kraft Mill Wastewaters
National Environmental Research Center
Office of Research and Development
U.S. Environmental Protection Agency
Corvallis, Oregon 97330
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development,
U.S. Environmental Protection Agency, have been grouped into
five series. These five broad categories were established to
facilitate further development and application of environmental
technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in
related fields. The five series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
This report has been assigned to the ENVIRONMENTAL MONITORING STUDIES
series. This series describes research conducted to develop new
or improved methods and instrumentation for the identification and
quantification of environmental pollutants at the lowest conceivably
significant concentrations. It also includes studies to determine
the ambient concentrations of pollutants in the environment and/or
the variance of pollutants as a function of time or meteorological
factors.
EPA REVIEW NOTICE
This report has been reviewed by the National Environmental
Research Center--Con/allis, and approved for publication. Mention
of trade names or commercial products does not constitute endorsement
or recommendation for use.
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EPA-660A-75-005
JUNE 1975
ANALYSIS OF ORGANIC
COMPOUNDS IN TWO KRAFT MILL WASTEWATERS
By
Lawrence H. Keith
Southeast Environmental Research Laboratory
National Environmental Research Center
Athens, Georgia 30601
Program Element 1BA027
ROAP/Task No. 07ABL-02, 03
NATIONAL ENVIRONMENTAL RESEARCH CENTER
OFFICE OF RESEARCH AND DEVELOPMENT
U. S. ENVIRONMENTAL PROTECTION AGENCY
CORVALLIS, OREGON 97330
For Sale by the National Technical Information Service,
U.S. Department of Commerce, Springfield, VA 22151
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ABSTRACT
Wastewaters from two Jcraft paper mills in Georgia were
sampled at various points in the waste treatment systems.
Gas chromatography of the organic extracts and
identification of many of the specific chemical components
by gas chromatography-mass spectrometry provided a "chemical
profile" of the effluents. The mills, in different
geographical locations, have very similar raw wastewater
compositions but different wastewater treatments. In spite
of these differences, the treated effluents are
qualitatively similar in composition although the quantities
of the various components differ. After two years the raw
and treated effluents of both mills were re-sampled.
Analyses showed that although concentrations of the organics
varied, the same compounds are still present. This report
was submitted in fulfillment of ROAP 07ABL, Tasks 02 and 03
by SERL, Athens, Georgia. Work was completed as of April
1974.
IJL
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CONTENTS
Sections
I conclusions 1
II Recommendations 4
III Introduction 6
IV Analytical Procedures 12
V Identification of Acids and Phenols 26
VI Identification of Terpenes 73
VII References 92
VIII Appendices 9 7
111
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FIGURES
Page
1 EPA Regional Districts 8
2 Waste treatment diagram of Mill "A" 9
3 Waste treatment diagram of 11
Interstate Paper Corporation,
Riceboro, Georgia
H Kuderna-Danish concentrator 13
5 Gas chromatograms of the extracts 14
of Mill "A" aerated lagoon effluent
from (A) carbon chloroform extract
(CCE) and (B) liquid-liquid chloro-
form extract
6 Apparatus for diazomethane methyla- 17
tion
7 Apparatus for dimethyl sulfate 19
methylation
8 Chromatograms of the methyl 20
derivatives from a kraft paper
mill wastewater sample using (A)
dimethyl sulfate, (B) diazomethane,
(C) Methyl-8, and (D) MethElute
as methylating reagents
9 Chromatograms of control samples 21
treated with (A) dimethyl sulfate,
(B) diazomethane, (C) Methyl-8,
and (D) MethElute as methylating
reagents
10 Computer reconstructed (A) gas 30
chromatogram, (B) LMRGC using
m/e 149 and (C) LMRGC using
m/e 74 and 87 superimposed on
the same axis
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FIGURES (continued)
No.. Page
11 Mass spectra of (A) methyl stearate 33
without impurity and background
subtraction and (B) methyl
stearate after computer subtrac-
tion of mass fragments from inter-
fering compounds on either side of it
12 Mass spectra of (A) methyl palmitate 37
and (B) tentatively identified methyl
10-methyltetradecanoate
13 Mass spectra of ethyl palmitate 38
found in (A) the methylated
lagoon effluent extract of Mill
A and (B) the neutral raw effluent
extract of Mill "A"
14 LMRGC of the methylated extract 40
from Mill "A" lagoon effluent
15 Mass spectra of (A) methyl 2- 41
methylhexadecanoate and (B) ethyl
palmitate (ethyl hexadecanoate)
16 (A) Gas chromatogram and (B) RGC 44
of fatty acid methyl esters from
chlorella algae
17 Mass spectrum of (A) methyl abie- 48
tate and (B) methyl dehydroabietate
18 Mass spectrum of (A) methyl pimarate 49
and (B) methyl isopimarate
19 Mass spectrum of (A) methyl sandaraco- 50
pimarate and (B) methyl neoabietate
20 Mass spectrum of (A) methyl 13- 51
abieten-18-oate and (B) methyl
6,8,11,13-abietatetraen-18-oate
21 Chemical profile of acids and 55
phenols from Mill "A"; sample points 1-4
22 Chemical profile of acids and phenols from 55
the Interstate mill at Riceboro, Georgia;
sample points 1-4
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FIGURES (continued)
No. Page
23 Print-out of the computer program used 61
to analyze the raw effluent extract
from the Interstate mill at Riceboro
24 Computer print-out of the normalized 62
peak areas from the chromatogram of
the raw effluent extract from the
Interstate mill at Riceboro
25 Computer analysis of the chromatogram 63
of Interstate's raw effluent acid and
phenol extract
26 Gas chromatogram of the acid and 64
phenol extract of Interstate's
(A) raw effluent and (B) treated effluent
27 Computer analysis of the chromatogram 65
of Interstate's treated effluent acid and
phenol extract
28 Computer analysis of the chromatogram 66'
of Mill "A's" raw effluent acid and
phenol extract (A) run no. 1 and (B)
duplicate run
29 Computer analysis of the chromatogram 67
of Mill "A's" outfall acid and phenol
extract
30 Gas chromatogram of the acid and phenol 68
extract of Mill "A's" (A) raw effluent
run no. 1 and (B) treated effluent
31 Chemical profile of neutral volatiles 75
from Mill "A"; sample points 1-4
32 Chemical profile of neutral volatiles from 76
the Interstate mill at Riceboro; sample
points 1-4
33 Gas chromatogram of neutral volatile extract 77
in Mill "A's" raw effluent (A) and (B)
treated effluent
VI
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FIGURES (continued)
No.
34 Gas chromatoqram of neutral volatile extract 78
from Interstate's raw effluent (A) and (B)
treated effluent
35 Gas chromatograms of Mill "A" neutral 80
volatile extracts analyzed by GC-IR; sample
points 3 and 4
36 GC-IR spectra of peaks 3-7; sample point 3 82
with inset of the corresponding portion of
the gas chromatogram
37 GC-IR spectra of peaks 8-11; sample point 3 83
with inset of the corresponding portion of
the gas chromatogram
38 GC-IR spectra of peaks 11-14; sample point 3 84
with inset of the corresponding portion of
the gas chromatogram
39 GC-IR spectra of peaks 16-19; sample point 3 85
with inset of the corresponding portion of
the gas chromatogram
40 GC-IR spectra of peaks 20-24; sample point 3 86
with inset of the corresponding portion of
the gas chromatogram
41 GC-IR spectra of peaks a-e.; sample point 4 87
with inset of the corresponding portion of
the gas chromatogram
42 GC-IR spectrum of camphor and corresponding 88
GC-IR spectrum from sample peak
43 GC-IR spectrum of borneol and corresponding 89
GC-IR spectrum from sample peak
44 GC-IR spectrum of 2-acetylthiophene and 90
corresponding GC-IR spectrum from sample
peak
45 GC-IR spectrum of 2-propionylthiophene and 91
corresponding GC-IR spectrum from sample
peak
vii
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TABLES
No.
Percent methylation and recovery 22
of model phenols and acids using
dimethyl sulfate
Acids and phenols identified in 27
both kraft paper mill effluents
with approximate concentrations
Fatty acids in the extract of 43
Chlorella algae culture
Percent removal of phenols 47
versus their structural
complexity
Collective-pollution-parameter 70
measurements and total concentrations of
the volatile components in the acid/
phenol extracts
Volatile acidic components as 71
percentages of TOC
Neutral volatiles identified in 74
both kraft paper mill effluents
with appropriate concentrations
Vlll
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ACKNOWLEDGMENTS
All sample preparations and gas chromatographic separations
were done by Terry Floyd. Mass spectral data were provided
by Ann Alford and Mike Carter. Infrared data were provided
by Leo Azarraga and Ann McCall.
Charles Davis, Lloyd Chapman, and William J. Verross
provided valuable assistance and wastewater samples from the
Interstate Paper Corporation at Riceboro, Georgia.
Officials at Mill "A", which prefers to remain anonymous,
were also very helpful in providing both samples and
information. Without the cooperation of the staff from
these two mills this study would not have been possible.
Duane F. Zinkel (U.S.D.A. Forest Products Laboratory,
Madison, Wisconsin) kindly supplied us with reference
standards of the resin acids.
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SECTION I
CONCLUSIONS
Computer-assisted GC-MS is the best method of identifying
unknown volatile organics in wastewaters.
It is now possible to obtain routinely by GC analysis much
pertinent data concerning the quantities of specific
chemicals being discharged into environmental waters from
kraft paper mills.
The efficiency of various types and combinations of existing
treatment facilities can now be measured and compared as to
their removal of undesirable compounds from kraft paper mill
wastewaters.
In mill laboratories, computer-assisted GC analysis can save
much time and expense over manual GC.
The GC-MS techniques and extraction/concentration methods
employed permitted identification of some compounds in less
than 1 part per billion in the wastewaters.
A number of general conclusions can be drawn from this
study:
Raw effluents from kraft pulp mills producing paper
from the same raw materials contain essentially the
same volatile organics.
The volatile organic composition of kraft
wastewaters remains relatively constant over the
years as long as the raw materials remain the same
and the process is not changed.
When peaks in the GC of a pulp mill wastewater have
been identified, routine GC analyses on a mill
laboratory basis are feasible.
The relative quantities of the volatile organics in
pulp mill wastewaters can fluctuate rapidly.
Because of rapid fluctuations in the quantities of
organics in the wastewaters, composite samples are
needed for representative quantitative results.
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The wastewaters of the two mills were similar in several
respects:
The raw effluents were very similar qualitatively
with regard to volatile organic chemical content.
This was expected since both mills produce a kraft
linerboard by the same process and from the same raw
materials.
Significant BOD reductions occurred in both
treatment systems.
Total chromatographable organics were significantly
reduced by both treatment systems.
Both treated effluents showed large reductions in
resin acid, phenolic, and neutral volatiles content.
Only one point of difference was noted. Total fatty acid
content was decreased in Interstate*s treated effluent but
not in that of Mill "A". Fatty acids in the oxidation of
Mill "A" may be biological metabolites from aquatic life in
these ponds.
Conclusions can also be reached from a comparison of the
effects of biological treatment (Mill "A's" trickling filter
and aerated lagoon) versus chemical/biological treatment
(Interstate1s lime flocculation/stabilization lagoon) on the
volatile organic compounds in these wastewaters. All values
used in these comparisons came only from the January 1974
composite samples.
BOD reductions were in the range of 85-90X with both
treatments.
TOC reductions were in the range of 80-85% with
chemical/ biological treatment and 60-65% with only
biological treatment.
Total volatile acids and phenols were reduced by 75-
80% in both treatments.
Total resin acids were reduced in concentration by
87% with biological treatment but were still present
at about 1.4 mg/liter in the treated effluent.
Total resin acids were reduced in concentration by
74% with chemical/biological treatment but were
still present at about 6.4 mg/liter in the treated
effluent.
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Total phenols were reduced in concentration by 73%
with biological treatment and were present at 0.6
mg/liter in the treated effluent. They were reduced
by 94% with chemical/ biological treatment and were
present at 0.2 mg/liter in the treated effluent.
Total fatty acids increased 17% in concentration
with biological treatment and were present at 1.6
mg/liter in the treated effluent. They decreased by
86% with chemical/biological treatment and were
present at 0.2 mg/liter in the treated effluent.
Biologically treated wastewaters from both mills
contained a greater number of fatty acids than did
the raw wastewaters. The new fatty acids were
branched and oddnumbered carbon compounds.
Total volatile materials ranged from 1-10% of the
TOC.
Terpenes were reduced by 90% or more by both
treatments.
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SECTION XI
RECOMMENDATIONS
We recommend that further studies by Federal and university
laboratories, and especially laboratories associated with
the paper mills or the National Council for Air and Stream
Improvement, be conducted to build up a reliable body of
knowledge on the amounts of specific volatile organics being
discharged from pulp mill wastewaters and to determine the
waste treatment systems that most effectively remove them.
In particular, the following questions must be answered:
Does the chemical composition of kraft paper mill
wastewaters remain fairly consistent across large
geographic areas or only within smaller geographic
areas such as within the Southeast?
How does the composition of these wastewaters vary
with changes in wood and changes in the pulping
process?
Does the chemical composition of these wastewaters
vary with season?
What types of compounds are likely to build up in
concentration if the wastewater is recycled?
Can the in-plant source of the more refractory com-
pounds be determined so that one or more small
volume wastewater streams can be selected for
special intensive treatment before they are combined
with other in-plant wastewaters?
We recommend that other wastewater treatment systems be
evaluated using the techniques developed for this study. In
particular, the effectiveness of activated sludge and of
activated carbon filters for reducing the concentrations of
these organics would be useful information.
We also recommend that the information on specific organic
pollutants contained in this report be used for developing
programs to monitor for evidence of kraft paper mill
pollution in receiving waters. Furthermore, some of the
more abundant and/or resistant compounds (such as
dehydroabietic acid, pimeric acid, isopimeric acid,
sandaracopimeric acid, 13-abieten-18-oic acid, guaiacol,
vanillin, stearic acid, oleic acid, margaric acid, alpha-
terpineol, and camphor) should be considered as additions to
measured parameters in future EPA effluent guidelines.
-------
Often chronic and acute toxicity data are lacking for most
of these compounds. We recommend that toxicologists examine
these compounds for possible detrimental effects. We also
recommend that taste and odor thresholds be determined,
especially for the phenolics, where this information is
lacking.
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SECTION III
INTRODUCTION
A substantial body of knowledge exists concerning the
identities of specific chemical compounds in various types
and physical sections of trees and, to a lesser degree, in
the wastewaters of paper mills. But little is known about
what happens to these chemicals as they pass through
wastewater treatment systems and enter into receiving
waters. To our knowledge this study represents the first
attempt to chemically characterize a wastewater, trace the
dissolved volatile organics through a treatment system, and
correlate this information with the traditional collective
pollution parameter measurements (BOD, TOC). The results
detailed in this report were gathered over a six-year period
and portions of them have been presented or published
previously.l~5
By tracing the chemicals through the treatment system one
can identify which compounds are being effectively removed
and which are resistant to the treatment in use. Any new
chemicals produced during treatment are readily apparent.
Once identifications are made, the approximate concentration
of each compound can be calculated at each stage of the
treatment. By comparing two or more different types of
treatment, their effects on the individual compounds, or on
the various classes of compounds in the wastewaters, can be
ascertained. This knowledge will be particularly useful for
advanced wastewater treatment and control studies,
especially those involving wastewater recycling. A build-up
in concentration of compounds resistant to the proposed
treatment could be detrimental to closed-loop systems. If
the segregated wastewater streams are analyzed before they
are combined for treatment, the main sources of compounds
resistant to treatment can be identified and singled out for
more economical, specialized treatment, if desired or
needed.
Knowledge of the specific chemical composition of treated
wastewaters is also basic to the evaluation of the
environmental impact of these wastewaters and to the problem
of analyzing and controlling their discharge. Once these
"refractory" compounds are identified, studies involving
their fate and ecological effects can commence. Acute and,
possibly more significant, chronic effects of these
chemicals on aquatic life can be determined.
Ultimately, when more statistical data have been gained on
the occurrence of specific "refractory" compounds in kraft
-------
paper mill wastewaters, and their environmental effects have
been evaluated, recommendations relative to some of these com-
pounds may be incorporated into the effluent guidelines for
paper mill wastewater discharges. Information gained from
these studies concerning specific organic contaminants has
already been used by Region IV Surveillance and Analysis
Division CSAD) in enforcement conferences^'7 and is currently
being used by Region VIII SAD8 to determine if the source of
some pollution problems lies with paper mill contaminants
CFig. 1J. In the summer of 1972, we provided the Organic
Analysis Unit of the Lower Mississippi River Field Facility
with evidence that contaminants from paper mills in that area
contributed measurably to the organic pollution of the
Mississippi River.* Identification of specific resin acids
and certain phenols in polluted waters provide strong evidence
for contamination by paper mills.
This study should raise many questions and stimulate further
work that will result in an accumulation of data from many
other mills with other types of wastewater treatment. The
report should be used as a tool by other laboratories that
desire to study the composition of kraft pulp mill wastewaters
and the effects of waste treatment on them. Although we
relied heavily on gas chromatography-mass spectrometry (GC-MS)
for compound identification, once the identifications are made
and GC retention times are established, GC-MS is not absolutely
necessary. A great deal of useful work can be accomplished
with GC and standards of the compounds identified in this
report. Most paper mill laboratories and college forestry or
ecology laboratories have gas chromatographs available and the
column packings used Ccarbowax 20M/TPA and SE-30) are very
common ones.
PAPER MILL TREATMENT FACILITIES
Two mills, having similar processes but different waste treat-
ments, were used in this study. The first, Paper Mill "A",
daily produced about 1400 tons of containerboard in March 1972
when the samples were taken. Approximately 13 million gallons
of water passed through the treatment system daily. Treatment
consists of a primary clarifier, trickling biofilter, secondary
clarifier, and 5 aerated lagoons with a total retention time of
2-2 1/2 days (Fig. 2). The trickling biofilter is 30 feet high,
and 80 feet in diameter, and is packed with a vinyl core
material having 97% void space. It handles an average daily
load of 56,000 Ib BOD and provides 50% BOD removal. The
total BOD reduction through the whole system is reported to be
in excess of 70%. In 1972, grab samples for chemical
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00
IX
San Francisco\
Figure 1 EPA Regional Districts
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Kraft Paper Mill "A"
Sample point No. 1
(Grab sample)
e
Primary clarifier
(Retention time about 5 hours)
Sample point No. 2 / \
(Grab sample 5 hours after Sample No. 1)
Sample point No. 3
(Grab sample 3 hours after Sample No. 2)
Secondary Clarifier
(Retention time about 3 hours)
Trickling Biofitter
(Retention time
about 2-4 min)
1 M/
Aerated Lagoon
Sample point No. 4
(Grab sample 2 days
after Sample No. 3)
(Retention time about 2 days)
Figure 2 Waste treatment diagram of Mill "A"
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characterization were taken at various stages of the
treatment system and time delays were programmed so that a
"slug" of the effluent would be followed through the
treatment facilities. However, quantitative analysis
indicated that the slug was missed at the outfall. A second
sampling period, in January 1974, used 3-day composites.
Daily composites of the raw effluent from January 16-18
were combined. The quantitative results of these "average
slugs" were much more satisfactory.
The second mill. Interstate Paper Corp. at Riceboro, Ga.,
daily produced about 540 tons of containerboard in March
1972 when the samples were taken. Approximately 5.5 million
gallons of water passed through the treatment system daily.
Treatment consists of lime addition at 0.1%, a 40-minute
flocculation period followed by gravity clarification and a
650-acre (3-6 month retention) stabilization lagoon (Fig.
3) . The highly alkaline effluent (pH 12) first undergoes
partial neutralization to pH 10 by surface absorption of
atmospheric carbon dioxide, accompanied by precipitation of
nearly all the remaining calcium in the inlet section of the
stabilization basin. Lime treatment removes about 905S of
the color from the effluent. Overall BOD reduction is
reported to be 93X, with a concentration of about 6 mg/1 in
the lagoon effluent.10'11 The 1972 sampling consisted
of 24-hour composites or grab samples at various stages of
the treatment system as indicated in Figure 3.
During a second sampling period, in January 1974, 3-day
composites were collected. Daily composites of the raw
effluent and the outfall were collected from January 16-18.
The raw effluent samples were combined, the outfall samples
were combined, and the composites of each were analyzed.
10
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Interstate Paper Corp.
Riceboro, Ga.
Sample point No. 1
(24-hr composite)
Sample point No. 3
(24-hr composite)
\
e
I
Lime Flocculation Tank
(Retention time about 40 min.)
Lime
Addition
Clarifier
(Retention time about 6-8 hours)
Sample point No. 2
(Grab sample)
vy
i i i i /i\
Quiescent Storage Basin
I
r\
Sample point j)
No. 4 jf
(Grab sample) (r C
(Retention time 3-6 months)
I
Figure 3 Waste treatment diagram of Interstate Paper Corp., Riceboro, Georgia
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SECTION IV
ANALYTICAL PROCEDURES
SOLVENT EXTRACTION EFFICIENCIES
To determine the best of several common solvents used for
liquid-liquid extractions, 1-liter portions of the Mill "A"
aerated lagoon effluent were extracted with UOO-ml portions
of low boiling petroleum ether, diethyl ether, chloroform,
and ethylacetate.
Each of the extracts was concentrated in a Kuderna-Danish
evaporator (Fig. 4) and chromatographed under identical
conditions. Chloroform was the best of the four solvents
for these extractions.
CHLOROFORM VS CARBON CHLOROFORM EXTRACTS
Adsorption of organics on granulated carbon followed by
extraction with chloroform provides a large amount of sample
with which to work.
Accordingly, 1800 gal. of the Mill "A" aerated lagoon
effluent was passed through a bank of 8 carbon filters in
parallel. The carbon was dried and extracted with
chloroform. The resulting extract was concentrated in a
Kuderna-Danish apparatus, and chromatographed under
conditions identical to those used for the 1-liter
chloroform extract. Figure 5 shows a comparison of the two
chromatograms. Most of the same peaks are present in both
chromatograms. Although the relative intensity differs
somewhat, the chromatograms are similar enough that, from a
qualitative aspect, the two methods of sample concentration
were essentially equivalent.
However, the carbon adsorption method has numerous
disadvantages:
Extent of adsorption varies with size of carbon
particles, contact time with carbon (flow rate), and
turbidity of the water.
Carbons from various sources differ markedly in
their adsorptive ability.
« A carbon column can go septic, causing biological
degradation of the adsorbed organics.
12
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J
Figure 4 Kuderna-Danish concentrator
13
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Lagoon Chloroform Extract
B
CCE
Figure 5 Gas chromatograms of the extracts of Mill "A" aerated lagoon effluent
from (A) carbon chloroform extract (CCE) and (B) liquid-liquid chloroform extract.
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Even with the greatest practical contact times, not
all organic matter in water will be adsorbed; highly
polar compounds are not removed by carbon columns.
it is more time-consuming to set up the pumps and
equipment than it is to simply "grab" a sample for
solvent extraction.
Some compounds may be only partially desorbed from
the carbon.
A greater possibility of chemical change (iso-
merization, hydrolysis, etc.) exists when carbon
with its large, active surface area is used.
We therefore elected to obtain the rest of our samples by
the simpler technique of extraction with chloroform followed
by Kuderna-Danish concentration.
Papermill wastewater extracts contain two types of con-
pounds: neutrals (predominately terpenes and their
derivatives) and acidic compounds that are converted to
their methyl derivatives to facilitate gas chromatographic
(GC) separation.
COMPARISON OF METHYLATION TECHNIQUES
After the pH is raised to 11 and the neutrals are extracted,
the aqueous solution can be methylated directly with
dimethyl sulfate and sodium hydroxide, followed by
extraction of the methyl derivatives with chloroform. This
method has the advantage of being an in situ procedure;
however, it is complex and time consuming. Alternatively,
after extraction of the neutrals at a high pH, the solution
can be made strongly acidic with concentrated hydrochloric
acid, and the free acids and phenols extracted with
chloroform. Methyl derivatives of the acids and phenols can
then be made in a separate step using diazomethane, on-
column GC methylation techniques, or several other common
methylation procedures. An evaluation of each method was
made using representative standard compounds and also using
samples of kraft mill wastewater.
15
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Methylation of Standards with Diazgmethane
Ten milligrams each of guaiacol (I), vanillin (II), palmitic
acid (III) and dehydroabietic acid (IV) (2 phenols, a
saturated fatty acid, and a resin acid, respectively) were
dissolved in 600 ml of water made to pH 11 with sodium
hydroxide. The solution was placed in a separatory funnel,
made acidic with concentrated hydrochloric acid, and
extracted with three 133-ml portions of chloroform. After
the combined extract was dried by passing it through a
column of anhydrous sodium sulfate (previously baked at
600°C. for 2 hours to remove phthalate ester impurities) , it
was evaporated to near dryness in a Kuderna-Danish
concentrator. Two ml of 10% methanol in ether was added and
the sample was methylated by the standard procedure with
diazomethane (Fig. 6). After methylation, chloroform was
added to bring the volume to 10.0 ml, providing a
theoretical 1,000 ppm solution of each of the four methyl
derivatives based on 100X extraction and methylation.
Duplicate injections into a gas chromatograph were made and
the areas of the peaks (calculated by multiplying the peak
height times the peak width at half height) were compared to
the peak areas from standards of veratrole (V),
veratraldehyde (VI), methyl palmitate (VII) and methyl
dehydroabietate (VIII) (10 mg each) in 10 ml of chloroform
(1,000 ppm). The calculated percent conversions were as
follows: I-»V, 18%; II-+VT, U7%; III>-VII, 70X; IV*
VIII, 96%.
.OCH3
.OCHj
CHO
CH3
(CH,)14
COOR
H3C
H3C COOR
I: R = H II: R = H III: R = H
V:R = CH3 VI:R = CH3 VII:R=CH3
IV: R = H
III:R = CH3
Methy.lation of Standards with Dimethyl Sulfate
The reaction with dimethyl sulfate is dependent on tem-
perature control, addition time, and reaction time, and on
16
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REDUCTION VALVE
METERING
VALVE
'11'
NITROGEN TANK
GLASS OR
STAINLESS STEEL-
0.7mm I D
RUBBER STOPPER
0.7cm 0 D
TUBE I
(I6x ISOmm)
RUBBER STOPPER
0.7cm OD
O.I cm 0 D
TUBE 2
(15 x 85mm)
GENERATOR
RUBBER STOPPER
0.7cm OD
O.lcm OD
T.UBE 3
(15* 85mm)
TRAP
0.1 cm 0 D
TUBE 4
(I5x 85 mm) or
KD CONCENTRATOR TUBE
SAMPLE
Figure 6 Apparatus for diazomethane methylation
-------
how vigorously the reaction mixture is stirred. The
procedure we use1* is a variation of that described by Bicho
et al.17> is Figure 7 shows a diagram of the apparatus. To
determine the average yield and recovery of methylated model
compounds from this reaction, four mixtures of I through IV
were subjected to the same reaction and work-up procedures.
Three mixtures containing 10 mg of each compound and a
fourth mixture containing 25 mg of each compound were
compared. After methylation, the products were extracted
with chloroform, dried over sodium sulfate, and concentrated
to less than 10 ml in a Kuderna-Danish apparatus. The
volume was adjusted to 10.0 ml with chloroform. Comparision
of GC peak areas with peak areas of standards V through VTII
allowed calculation of the percent methylation and recovery.
The results are summarized in Table I.
tion of Wastewater Samples with Dimethyl Sulfate
A 500-ml portion of the Mill "A" wastewater from sample
point 2 (primary clarifier effluent) was made alkaline to pH
1 1 with sodium hydroxide and extracted with chloroform to
remove the neutral compounds. Methylation of the aqueous
layer with dimethyl sulfate was followed by re-extraction
with chloroform to remove the methylated organics. After
concentration in a Kuderna-Danish apparatus to 0.5 ml, 1.2
yl of the extract was chromatographed on a 50 ft support
coated open tubular (S.C.O.T.) column coated with Carbowax
20 M/TPA and programmed from 100 to 200°C. at 4°/minute with
an initial 2 minute hold at 100°. The chromatogram is shown
in Figure 8-A. A control sample was prepared exactly the
same way using 500 ml of distilled water; its chromatogram
is shown in Figure 9-A.
Methylation of Wastewater Samgles with Diazgmethane
A 1-liter portion of the same wastewater was extracted with
chloroform at pH 11 to remove neutral compounds and then
made acidic to pH< 2 with cone, hydrochloric acid. The
aqueous layer was re-extracted with four 200-ml portions of
chloroform to remove the acids and phenols. The extracts
were combined and divided into two equal portions, each
representing 500 ml of the wastewater extract.
One portion was concentrated to near dryness in a Kuderna-
Danish apparatus and methylated with diazomethane as
previously described; the final volume was adjusted to 0.5
ml. The chromatogram of 1.2 pi of this sample under
conditions identical to those of the previous sample is
shown in Figure 8-B. A control sample was prepared from 100
18
-------
Nitrogen Out
Heater-Stirrer
Figure 7 Apparatus for dimethyl sulfate methylation
19
-------
Figure 8 Chromatograms of the methyl derivatives from
a kraft paper mill wastewater sample using
(A) dimethyl sulfate, (B) diazomethane, (C)
Methyl-8, and (D) MethElute as methylating
reagents
20
-------
B
DIAZOMITHANI
ILANK
OIMITHTL
IIHIATI
Figure 9 Chromatograms of control samples treated
with (A) dimethyl sulfate, (B) diazomethane,
(C) Methyl-8, and (D) MethElute as methylating
reagents
21
-------
Table 1
PERCENT METHYLATION AND RECOVERY OF MODEL PHENOLS AND ACIDS
USING DIMETHYL SULFATE
Reaction
No.
I
> V
II
-> VI
III
* VII
IV -
» VIII
#1 (10 mg each) 74%
#2 (10 mg each) 91%
#3 (10 mg each 100%
#4 (25 mg each) 96%
60%
76%
81%
69%
47%
64%
66%
53%
39%
57%
84%
60%
AVERAGE
90%
72%
58%
60%
22
-------
ml of chloroform subjected to the same prodecure; the
chromatogram of 1.2 yl of the control is shown in Figure 9-
B.
Methvlation of Wastewater Samples with On-Column Reagents
The second portion of the chloroform extract was
concentrated in a Kuderna-Danish apparatus to near dryness
and MethElute (trimethylanilinum hydroxide in methanol;
Pierce Chemical Co.) was added to bring the volume to 0.5
ml. The chromatogram of 1.2 yl of this sample under
conditions identical to those of the previous two samples is
shown in Figure 8-D. MethElute provides on-column
methylation of the sample. A control sample was prepared
from 100 ml of chloroform subjected to the same procedures.
The chromatogram of 1.2 yl of the control is shown in Figure
9-D.
Another 500-ml portion of the wastewater, after extraction
of neutral compounds at pH 11, was made acidic and extracted
with four 100-ml portions of chloroform. The extracts were
combined, dried, and concentrated to near dryness as before
in a Kuderna-Danish apparatus. Enough Methyl-8 (DMF
dimethyl acetal in pyridene; Pierce Chemical Co.) was added
to bring the volume to 0.5 ml and the solution was heated at
60°C for 15 minutes in a reacti-vial. The chromatogram of
1.2 //I of this sample, under conditions identical to those
of the other three, is shown in Figure 8-C. A control
sample was prepared from 400 ml of chloroform subjected to
the same procedure. The chromatogram of 1.2 yl of the
control is shown in Figure 9-C.
Methvlation Evaluation
Comparison of the chromatograms in Figure 8 shows that
dimethyl sulfate and MethElute are better reagents for
methylating these samples than diazomethane or Methyl-8.
Phenols, especially guaiacol, were not methylated well with
diazomethane or Methyl-8. Although extraction of guaiacol
and other phenols with chloroform is not 100%, incomplete
extraction can be eliminated as a major contributor to the
smallness of the guaiacol peak in the diazomethane sample
because the samples for diazomethane and MethElute treatment
came from the same extract, which was divided into two
portions, and the MethElute sample shows a large veratrole
peak.
The dimethyl sulfate appears to be equivalent to MethElute
with respect to methylation of the resin and fatty acids.
23
-------
However, the larger phenolic peaks in the dimethyl sulfate
sample and absence of the MethElute reagent peak favored use
of dimethyl sulfate for all remaining methylations involved
in analysis of these wastewaters.
INSTRUMENTATION
Gas Chromatograph
Most of the chromatograms shown in this report were obtained
with a Varian 1400 gas Chromatograph (GC) equipped with a
flame ionization detector (FID). Because helium is usually
used as a carrier gas with gas chromatographs interfaced
with mass spectrometers (GC-MS), we used helium as a carrier
gas routinely in developing our chromatographic conditions.
Generally, a commercially prepared (Perkin-Elmer) 50-ft
support coated open tubular (S.C.O.T.) capillary column
coated with carbowax 20M/terephthalic acid (K 20 M/TPA) was
used for separation. The optimum carrier gas flow for our
GC-MS systems operating under a vacuum and using a Gohlke
jet separator is 16 to 18 ml/minute. If a helium flow of
about half this is used for optimizing conditions with an
auxiliary GC (operating under atmospheric pressure), the
other chromatographic variables (temperature program rate,
initial temperature, initial hold) will hold true when the
same column is transferred to the GC-MS system.
GC-MS-Computer System
From 1968 to 1971, a Perkin-Elmer/Hitachi RMU-7 double
focusing mass spectrometer connected to a Perkin-Elmer 900
GC through a Watson-Bieman separator was used to analyze the
sample extracts from kraft papermi11 wastewaters. Spectra
acquisition, data reduction, and spectral interpretation had
to be done manually. Despite these limitations, much of the
early work reported in this project1-*»6*7 was accomplished
with this system.
Now, a semi-automatic GC-MS-computer system is used for
pollutant identification. The GC is a modified Varian 1400
Chromatograph with a temperature-controlled oven that can be
programmed from 50° to 500° C. It has no independent
detector and serves only as a specialized inlet to the mass
spectrometer. An all-glass, single-stage Gohlke jet
separator enriches organic samples by utilizing differences
in diffusion rates of sample and carrier gases in a
turbulent jet.
24
-------
The Finnigan 1015 quadrupole mass spectrometer with three
mass ranges extending to m/e 750 is capable of unit
resolution throughout the range. At a scan speed of 120
amu/sec, sensitivity is adequate to give identifiable
spectra for 20 ng of material introduced into the GC inlet.
The liquid inlet is used for introduction of calibration
compounds, the direct probe for solid materials.
A System Industries interface (analog-to-digital converter,
and the digital-to-analog converter) permits the Digital
Equipment Corporation (DEC) PDP8/e computer to control the
mass spectrometer during calibration and data acquisition;
to accept data from the mass spectrometer; and to control a
Houston plotter during data reduction.
The DEC PDP8/e computer has a 4096 word core and an ASR33
teletypewriter. Programs, raw data, and reduced data are
stored on either two DECtape units or a Diablo disc. Output
of the reduced data is achieved under computer control via
the plotter, the teletypewriter, or an acousticoupler. The
coupling device connects the PDP8/e to the CDC 6UOO computer
at Battelle Laboratories, Columbus, Ohio, and permits semi-
automatic spectrum identification by a matching program,
fully described elsewhere.19 /2<>
Using this system, data reduction times are much less than
for the manual reduction methods formerly used. About 2
hours of operator time is required to output reduced data
for an 80-peak chromatogram. Data output time ranges from
slightly more than four hours for the disc system to more
than eight hours for the tape system. Manual data reduction
and spectral plotting of the same 80 chromatographic peaks
would require several weeks.
25
-------
SECTION V
IDENTIFICATION OF ACIDS AND PHENOLS
The acids and phenols in each wastewater were analyzed as a
group for both the 1972 samples and the 1974 samples.
Results were then compared. The neutral extractable
organics (mostly terpenes) were analyzed separately and are
discussed in the next section.
GC-MS ANALYSES
Each of the 8 samples (4 from each mill at various sampling
points) was methylated with dimethyl sulfate as previously
described. Following this, each of the 8 extracts was
analyzed by GC-MS using the Finnigan 1015 GC-MS computer
system. After a sample is injected into the GC, the mass
spectrometer automatically scans its pre-set mass range
every five seconds (or other pre-set interval). As it does
so, the plotter draws a trace that is equivalent to a GC
signal. Since our samples were pre-analyzed on an auxiliary
GC, the auxiliary chromatograms were used to determine when
to terminate the GC-MS runs. When each run is complete, the
computer plots a reconstructed gas chromatogram (RGC). The
RGC peaks are all normalized to the amplitude of the largest
peak, arbitrarily plotted at 100. Each point on the
spectrum number scale under the RCG represents a complete
mass spectrum. Figure 10-A shows the RGC of the methylated
acids and phenols from the lagoon wastewater of Mill "A".
Any of the spectra can be displayed individually; therefore,
by choosing the appropriate spectrum numbers, the analyst
can see the composition of each peak in the chromatogram.
Specialized techniques of MS or data reduction can be used
to detect a specific material or class of materials in a
mixture. The most common technique is the generation of the
limited mass reconstructed gas chromatogram (LMRGC) . For
example, in Figure 10-A, the RGC shows as peaks those
spectra that contain significant numbers of any ion
fragments of m/e 33 to 450. Below the RGC is the LMRGC in
which the computer was instructed to respond only to those
spectra that contain the m/e 149 fragment (Figure 10-B) .
This fragment is usually due to protonated phthalic
anhydride ( IX), found in the spectra of phthalate esters.
The LMRGC, therefore, indicates that two of the sample
peaks, 44 and 61, are phthalates. The numbers 44 and 61
refer to the compound designations listed in Table 2.
26
-------
Table 2. ACIDS AND PHENOLS IDENTIFIED IN BOTH KRAFT PAPER MILL EFFLUENTS WITH APPROXIMATE CONCENTRATIONS
to
Approximate
Cone, in mg/1
Peak
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
1 9
20
21
22
23
24
25
26
Compound Identified as Methyl Confirmed
Derivative (Parent Compound) By
Furfuryl methyl ether
(furfuryl)
Anisole (Phenol)
1 , 3-Dimethoxy-2-propanol
Methyl trisulfide
Unidentified apparent MW=103
Benzaldehyde
Dime thy Isulf oxide
Ethyl carbamate
Borneol
a-Terpineol
Veratrole (Guaiacol)
o-Nitrotoluene
Methyl o-hydroxybenzoate
(o-hydroxybenzoic acid)
Methyl mandelate (mandelic acid)
Dime thylsulf one
Unidentified aromatic MW=166
Methyl isomyristate
(Ci4 fatty acid)
p-Methoxybenzaldehyde
(£-hydroxybenzaldehyde)
Eugenol
Methyl myristate (CIA fatty acid)
Unidentified aromatic, MW=196
Unidentified nonaromatic, MW=196
Methyl anteisopentadecanoate
(C15 fatty acid)
Methyl 10-methyltetradecanoate
(C15 fatty acid)
p-Methoxyacetophenone
(£-hydroxy acetophenone )
Methyl pentadecanoate
(Cj5 fatty acid)
GC
GC
GC
GC
MS,GC
MS, GC
MS,GC
MS,GC
GC
GC
MS,GC
MS,GC
GC
MS,GC
0.
0.
-
0.
0.
-
0.
-
0.
2.
-
0.
-
0.
0.
-
0.
0.
_
-
-
-
0.
-
1
010
008
-
035
002
-
010
-
020
700
-
010
-
240
130
-
055
025
_
-
-
-
020
-
Mill "A"
Sample Points
2 3
0.005
0.005
0.035
0.015
0.002
2.400
0.005
0.400
0.090
0.050
0.025
0.025
0.040
0.055
0.025
0.001
0.360
0.400
0.050
0.020
0.025
0.045
0.010
0.030
4
0.035
0.020
0.170
0.015
0.025
0.055
0.005
0.003
0.025
0.020
0.035
0.020
0.030
0.020
1
0.010
0.005
0.025
0.005
0.010
0.010
0.005
2.200
0.240
0.050
0.090
::
0.170
0.060
Approximate
Cone, in mg/1
Sample Points
2 3
0.005
0.010
0.055
0.080
0.080
0.020
0.020
3.200
0.540
0.030
0.240
0.175
0.130
0.020
0.020
0.020
0.060
0.115
3.700
0.400
0.160
0.190
0.245
0.080
4
~
0.015
0.020
0.010
0.002
0.035
0.130
0.001
0.005
0.010
0.065
0.005
0.002
0.010
0.010
-------
Table 2 (continued) . ACIDS AND PHENOLS IDENTIFIED IN BOTH KRAFT PAPER MILL EFFLUENTS WITH APPROXIMATE CONCENTRATIONS
00
Approximate
Cone, in mg/1
Peak
No.
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
Compound Identified as Methyl Confirmed
Derivative (Parent Compound) By
Methyl isopalmitate (C, , fatty
acid) lb
Methyl palmitate (C,, fatty acid)
Methyl palmitelaidate (Cifi trans-
9-unsaturated fatty acid)
Ethyl palmitate (C^g fatty acid)
Methyl anteisomargarate (C^7
fatty acid)
Methyl 3,4-dimethoxyphenylacetate
(Homovanillic acid)
Veratraldehyde (Vanillin)
2-Methylthiobenzothiazole
(2-Mercaptobenzothiazole)
Methyl vanillate (Vanillic acid)
Acetoveratrone (Acetovanillone)
Methyl stearate (Cla fatty acid)
Methyl oleate (C^8 cis-9-
unsaturated fatty acid)
3 , 4 , 5-Trimethoxybenzaldehyde
( Sy r ingaldehyde )
Methyl linoleate (C18 cis,cis-9,
12-diunsaturated fatty acid)
3 , 4-Dimethoxypropiophenone
( 3-Methoxy-4 -hydroxypropiophenone )
3 , 4-5-Trimethoxyaeetophenone
(Acetosyringone)
Methyl Arachidate
a Dihexylph thai ate
Unidentified resin acid "A" methyl
ester
Unidentified resin acid "B" methyl
ester
MS,GC
MS,GC
MS,GC
MS,GC
MS,GC
MS,GC
GC
GC
GC
MS,GC
MS,GC
MS.GC
MS,GC
GC
GC
1
0.070
0.005
0.020
0.050
1.500
0.035
0.420
0.025
0.470
0.070
0.350
0.060
0.055
0.030
0.030
Mill "A"
Sample Points
2 3
0
0
0
0
1
0
0
0
0
0
0
0
0
0
0
.190
.020
.025
.075
.700
.025
.490
.065
.600
.070
.470
.080
.060
.035
.055
0.015
0.180
0.025
0.040
0.090
0.450
0.030
0.005
0.450
0.045
0.430
0.070
0.230
0.025
0.050
0.030
0.095
4
0.030
0.430
0.125
0.030
0.050
0.080
0.410
0.025
0.005
0.370
0.110
0.400
0.040
0.100
0.050
0.025
0.100
1
0.140
0.030
0.090
2.100
0.820
0.100
0.570
0.070
0.450
0.090
0.035
0.070
Approximate
Cone, in mg/1
Interstate
Sample Points
2 3
0.160
0.120
0.365
4.400
1.600
0.100
0.510
0.100
0.920
0.200
0.025
0.075
0.035
0.040
0.150
2.600
0.860
0.020
0.120
0.070
0.160
0.130
0.130
4
0.003
0.017
0.010
0.001
0.003
0.070
0.120
0.080
0.020
0.005
0.020
-------
Table 2 (continued). ACIDS AND PHENOLS IDENTIFIED IN BOTH KRAFT PAPER MILL EFFLUENTS WITH APPROXIMATE CONCENTRATIONS
Appropriate
Cone, in mg/1
Peak
No.
47
48
49
50
51
to 52
10
53
54
55
56
57
58
59
60
61
Total
Compound Identified as Methyl
Derivative (Parent Compound)
Unidentified resin acid "C"
methyl ester
Unidentified resin acid "D"
methyl ester
Unidentified unsaturated fatty
acid methyl ester
Methyl pimerate (resin acid)
Methyl sandaracopimerate (resin
acid)
Unidentified resin acid "E"
methyl ester
Methyl-13-Abieten-18-oate
(resin acid)
Unidentified unsaturated fatty
acid methyl ester similar to
araconidate
Methyl isopimerate (resin acid)
Methyl abietate (resin acid)
Methyl dehydroabietate (resin
acid)
Methyl 6,8,11,13-Abietatetraen-
18-oate (resin acid)
Methyl neoabietate (resin acid)
Methyl lignocerate (C24 fatty
acid)
a Dioctylphthalate
Confirmed
By
MS,GC
MS,GC
MS,GC
MS,GC
GC
MS,GC
MS,GC
MS,GC
MS,GC
1
0.100
0.245
0.050
0.025
0.035
0.430
0.370
1.500
0.065
0.105
9.380
Mill "A"
Sample Points
2 3
0
0
0
0
0
0
1
0
0
10
.055
.475
.060
.430
.660
.430
.300
.070
.125
.692
0.010
0.005
0.570
0.125
0.800
0.770
0.420
4.000
0.170
0.055
10.246
4
0.035
0.800
0.045
1.400
0.780
0.050
1.000
0.095
0.030
7.123
1
1.270
0.340
0.100
0.075
4.400
3.300
3.300
0.160
1.300
21.690
Approximate
Cone, in mg/1
Interstate
Sample Points
2 3
0.825
0.245
0.035
1.700
1.500
2.700
0.115
1.200
21.480
0.610
0.275
0.050
1.200
1.900
3.600
0.280
0.450
17.700
4
0.145
0.500
0.110
1.050
0.800
3.900
0.180
7.243
-------
B
100
1M
20
0
100-
LMRGC m/e 149
C !-
I«
20
LMRGC m/e 74 & 87
23
CI?
""
<
60
100
200
300 400
SPECTRUM NUMBER
600
TOO
Figure 10 Computer reconstructed (A) gas chromatogram,
(B) LMRGC using m/e 149 and (C) LMRGC using
m/e 74 and 87 superimposed on the same axis
-------
Similar characteristic fragments exist for other classes of
compounds. The most useful ones in this study were m/e 74
and 87, which come from fragments x and XI respectively
and are characteristic of long-chain saturated fatty acid
methyl esters.21
OH
-C =
IX
0
h
CH30C (
XI
CH
The LMRGC corresponding to these fragments for the lagoon
wastewater from Mill "A" is shown in Figure 10-C. The
numbers above the shaded peaks correspond to the
identifications in Table 2. The 3 shaded peaks probably
correspond to methyl margarate (C-17), methyl arachidate (C-
20), and methyl behenate (C-22), but, because these were all
minor components of unresolved peaks, the mass spectra were
not definite enough to confirm their identification.
Margarate and arachidate were later confirmed in a 1974
sample of these wastewaters. Arachidate is assigned peak
number 43 in Table 2.
We have found that using the m/e 74 and 87 characteristic
fragments together results in a more reliable indication of
long-chain saturated fatty acid methyl esters than either
one by itself. The two LMRGC's are superimposed and
different colored pens are used for each trace. The peaks
at spectrum number 6 (m/e 87 search) and at spectrum number
51 (m/e 74 search) contain only one of the characteristic
fragment ions and are therefore not long-chain fatty acid
methyl esters.
From the RGC and/or LMRGC traces, spectra of interest are
chosen for output. The mass spectrum can be either plotted
or tabulated. This spectrum will be that of the sample
compound plus various fragments from GC column bleed, traces
of air, oil, and moisture, and possibly residual material
from a previously eluted compound. In the worst cases the
GC peaks may be incompletely resolved so that the mass
spectrum contains not only the fragments of interest but
also fragments from other compounds comprising the peak
envelope.
31
-------
An example of this situation is shown in Figure 11-A. Some
of the background fragments, from acetoveratrone and methyl
oleate, mask fragment peaks of the fatty acid methyl ester
mass spectrum. Computer subtraction of the appropriate
quantities of these impurities, indicated by an examination
of the RGC (0.6 of the mass spectrum of acetoveratrone and
0.2 of the mass spectrum of methyl oleate), produced the
mass spectrum shown in Figure 11-B. This spectrum is now
clearly recognizable as that of a saturated fatty acid
methyl ester, since the base peak is at m/e 74 and the
second most intense fragment occurs at m/e 87.
The mass spectrum is a chemical fingerprint that is
characteristic of a compound and can be interpreted to give
the structure of an unknown compound. Another way to
identify an unknown compound is to match its mass spectrum
with that of a known compound from a library of mass
spectra. We employ both methods; but to minimize the time
spent identifying the hundreds of spectra resulting from the
GC-MS data output from complex mixtures such as these
extracts, we first execute computer matching of the spectra.
This is followed by a manual comparison of the computer
matched spectra with the unknown mass spectra.
Widespread use of GC/MS/spectra-matching in pollutant
identification requires easy access to a central spectra
library,19 rapid matching, and an indication of the
similarity of the unknown spectrum to the reference spectrum
for each match. An EPA program that provides such
information was developed using the algorithm of a matching
program described in the literature.22 This rapid program
was developed jointly by Battelle and the Southeast
Environmental Research Laboratory, and utilizes a CDC 6400
time-shared computer.23
An application of the matching program is shown in Figure
11-B. The computer match output, inset on the spectrum,
shows that there were a total of 62 hits. The five best
matches are printed with the best match (correctly
identified as methyl stearate) listed first and the others
in descending order. The "similarity index" (SI) gives the
user an immediate indication of the quality of the matches.
The SI will show whether it is a poor match (<0.2 if the
data base does not contain any closely related compounds),
one of several fair matches (0.2-0.35 if the correct
compound is not in the data base but related ones are), or a
good match (>0.35 if the SI of the second best hit is
significantly lower).
Additional information provided with the match shows (in
sequence after the chemical name) the source of the mass
32
-------
u>
U)
8
o o
n HI
CD g cr e 01
H- 0) *< h(
rt en M H- 01
tr 01 rt>O
CD 01 *< CD
H hh rt o
hi CD p ft
01 pi pi p hi
H-lQ hi Q, pi
CD CD rt D* O
O CD pi hh
O rt O
hh 01 pi ?f .->
hhiQ >
H- hh rt hj >-
rt hi CD O .
SFCCIHK MHER 3Z7
OSM CJ-2-72)
bl|k
110 120 139 110 ISO IGO no 180 ISO 200 210 220 730 ZW 2S9 ZBO Z78 290 Z99 300 310 320
sa HITS
FETHVL STEMMTE (SEWD-ITUO!- 2S8 C19 H38 08 KK 6
FILE KEV> S773
SI-e 533
PCTHVL fTVRISTATC CSEUL>-13U01~ 242 CIS KM 02
FILE KEV« 8771
51-0 5O6
fCTHVL ISOSTEAftATE (SERL)-1V»1«J01- 29* CIS KM 0
FILE KEY- 8937
si-e saa
tCTHVL PALHITATE tSEUL>~15U01> 37* C17.K34 OS I
FILE KEV- 8772
SI-8 481
rCTHVL tCPTADECMIOATE ~lflU01~ 2*4 CU K38
SI-0 475
FILE KEV 8773
rCTHVL STEMATE ISEUL )*17U01- SOS CIS M38 03 »SR
M.'E INT n/E IHT IVE 1HT ft/E INT
^1 30 « 48 55 3e 57 El
fi
20 ao » so en TO at
MX E
III ll Jl l.ll, III ,1 I.I I
100 110 120 130 110 ISO I6B 1TO 195 13) 2BO 210 220 330 210 Z5D 260 270 280 230
-------
spectral data (SERL indicates the Southeast Environmental
Research Laboratory), the Wiswesser Line Notation, the
molecular weight, the empirical chemical formula, the
Battelle spectrum number, and the file key. A computer
print-out of the spectral data used for the match can be
obtained with the file key number. The spectral data from
file key 8773 is also inset on the spectrum in Figure 12-B.
The match of these data (the 2 most intense fragments every
1U mass units) with the plotted spectrum is good. The other
matches, in decreasing order of SI number and in groups of
5, would also be printed out if desired.
ANALYSIS OF THE 1972 SAMPLES
All compounds identified in the acidic fractions of both
mills' wastewater samples in 1972 are listed in Table 2.
The peak numbers assigned are used throughout this report.
The approximate concentrations for each compound are
calculated in milligrams per liter (mg/1) based on the
sample peak area relative to the peak area and flame
response of four representative compounds: veratrole, 3,4-
dimethoxybenzaldehyde, methyl palmitate, and methyl
dehydroabietate. Corrections were not made for inefficiency
in methylation and recovery (70-90X with phenols; 60X with
fatty and resin acids) in the calculated concentrations.
Consequently, these values are lower than they may actually
be.
Fattv Acjds
A total of 17 different fatty acids were found in the waste-
waters of both mills; 15 were identified and two unsaturated
acids remain unidentified.
The fatty acid content of both raw wastewaters (sample point
1) were qualitatively similar. Seven of the eight fatty
acids present in the raw wastewaters were found at both
mills (palmitic, anteisomargaric, stearic, oleic, linoleic,
arachidic, and unidentified unsaturated acid f54).
More dissimilarity existed in the fatty acid content of the
two lagoon effluents (sample point 4). Ten acids were found
in both effluentsisomyristic, myristic,
anteisopentadecanoic, 10-methyltetradecanoic, pentadecanoic,
isopalmitic, palmitic, palmitelaidic, anteisomargaric, and
oleic. Six others were found in the lagoon effluent of one
mill but not the otherethyl palmitate (as the ester),
stearic, linoleic, lignoceric, arachidic, and unidentified
unsaturated acid f5U. In both mill wastewaters a greater
34
-------
number of fatty acids was found in the lagoon effluent than
in the influents. The majority of these new compounds are
saturated low molecular weight (C-1U, C-15, C-16) branched
and straight-chain fatty acids. They are probably
metabolites of aquatic life in the lagoons.
Significance of Fatty Acids
Fatty acids are the main components of the parenchyma cell
resin of both hardwoods and softwoods and occur mainly as
glyceride esters.2* Although those with even numbers of
carbons are present in larger amounts than those with odd
numbers of carbons, all the fatty acids from C-12 to C-21
have been found. Recent work identified 30 fatty acids in
tall oil from pine and birch including odd-numbered carbon
fatty acids and one branched fatty acid.25 Fatty acids are
therefore expected to be in abundance in the raw effluents
from kraft mills.
Fatty acids are not generally considered to be very toxic
although the California "Water Quality Criteria" lists the
minimum lethal dose (MLD) for several of them at 5.0 mg/1
for fish.26 Recent work by Leach and Thakore27 has
indicated that the sodium salts of unsaturated fatty acids
are more toxic to fish than those of saturated fatty acids.
Furthermore, among the C^g unsaturated fatty acids, the
toxicity of the sodium salts increased with increasing
unsaturation in the order oleic
-------
The mass spectrum of the peak identified as methyl palmitate
is a good example of this fragmentation (Figure 12-A).
CH30-C-(CH2)n
0
XII
Introduction of a methyl group in the carbon chain causes
changes in the fragmentation pattern because of easy
cleavage alpha to the tertiary carbon atom.28 This results
in a gap of 28 mass units when the CH3-CH- moiety is
cleaved. Figure 12-B shows the mass spectrum of the
compound tentatively identified as methyl 10-
methyltetradecanoate. A gap of 28 mass units occurs between
m/e 171 and 199. This corresponds to cleavage on each side
of the number 10 carbon. The mass spectrum of methyl 10-
methyltetradecanoate is not in the mass spectral data base;
however, all of the first five matches were fatty acid
methyl est ers.
Ethyl Palmitate
The computer unexpectedly matched two mass spectra with
ethyl palmitate (ethyl hexadecanoate). To compound the
confusion, one spectrum was from a peak that only occured in
the methylated acid fraction of the Mill "A" lagoon effluent
(Fig. 13-A) and the other spectrum occured in the neutral
fraction only of the Mill "A" raw effluent (Fig. 13-B).
The base peak (m/e 88) and second largest fragment (m/e 101)
are shifted 14 mass units higher than their counterparts in
the mass spectra of saturated fatty acid methyl esters.
These fragments would correspond toxil and xiv respectively.
'25
I
OH
XIII
C0HcOCCHo CH0
2 -> n t. L
0
XIV
36
-------
SKCIIW HfflBR 270-J^
WR-1 C3-2-TZ1 fCTH
co
aa ao w so as ra ao ao IOB 110 IZB 130 iw iso teo ne IBB IM aoo 210 220 zao z« zso ago zro ZBO zso aw 310
',» iz i« iz IB \n i» » ao aci M b>"Jn SB
Figure 12 Mass spectra of (A) methyl palmitate and (B) tentatively
identified methyl 10-methyltetradecanoate
-------
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However, they could also be rationalized as coming from
structures XV and XVI . corresponding to the compound
methyl 2-methylhexadecanoate. The mass spectrum of the 2-
methylhexadecanoate was not in the computer data bank.
CH 0 C=CH
3 I I
O-H
CRn
CH00CCHCH0
3 II | 2
0 CE,
XV XVI
An LMRGC generated for each sample using m/e 88 and m/e 101,
indicated that no more compounds of this type were present
in the extracts. The LMRGC of the methylated extract from
Mill "A" lagoon wastewater is shown in Figure 1t. Only one
peak (shaded) contains both fragments.
The mass spectrum of methyl 2-methylhexadecanoate, (Fig. 15-
A) closely resembles those of the sample spectra (Figs. 13-
A and 13-B) with only some small variations. However, the
methyl 2-methylhexadecanoate eluted from the GC much earlier
than the unidentified compound in the samples.
The mass spectrum of a standard of ethyl palmitate (Fig. 15-
B) also closely resembles the mass spectra of the sample
peaks (Figs. 13-A and 13-B). However, methyl 2-
methylhexadecanoate (Fig. 15-A) shows a fragment at m/e 253
(loss of OCH ) and ethyl palmitate (Fig. 15-B) shows a
fragment at m/e 239 (loss of OC2H5). Figure 13-B contains
the m/e 239 fragment and no m/e 253 fragment. Figure 13-A
does not show a m/e 239 fragment, but it shows no fragment
at m/e 253 either. The m/e 239 peak was probably deleted
during background subtraction since it is quite small.
The GC retention time of ethyl palmitate was identical to
the peak in question. Spiking the samples with ethyl
palmitate increased the size of this peak in the sample
extracts, confirming the identity.
Unsaturated Fatty Acids
Six unsaturated fatty acids were detected in the acid/phenol
fraction of which three were identified (oleic, linoleic,
and palmitelaidic acids). Most of the unsaturated fatty
39
-------
100-
30
I»1
LMRGC m/e 88 & 101
20-
\hk.
100
200
300
400
500
600
Figure 14 LMRGC of the methylated extract from
Mill "A" lagoon effluent
-------
H
SPECTIU1M1CER 91-93
ieiHH.-2-ICTHlL (ORDEDMHTE
me-
nu
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Ihi
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3>ECnU1 MJCEP 191 - 130
OWL nuironE
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Figure 15
90 IBB 110 120 l» 110 ISO 160 170 160 190 200 210 Z20 230 210 2S) 2GO 2TO a» 290 300 310 32
Mass spectra of (A) methyl 2-methylhexadecanoate
and (B) ethyl palmitate (ethyl hexadecanoate)
-------
acid concentration is contributed by oleic and linoleic
acids.
Two additional unsaturated fatty acid methyl esters were
found in the neutral extracts. These have been tentatively
identified as a monounsaturated and a diunsaturated
derivative of nonadecanoic acid (C-19). Their isomeric
identities were not determined.
Fatty Acid Content of Chlorella Algae
The increase in numbers of branched and odd-numbered carbon
fatty acids in the lagoon effluents of both mills led us to
consider that they could be metabolites of the aquatic life
in the lagoons. Algae appeared to be a likely source of
these compounds.
Accordingly, 1 liter of a culture of Chlorella algae common
to the mill area was made basic with sodium hydroxide and
extracted with chloroform to remove the neutrals. The
alkaline solution was then made strongly acidic with
hydrochloric acid and re-extracted to remove the acids. The
acid extracts were evaporated almost to dryness in a
Kuderna-Danish concentrator. A 0.10 ml aliquot of the
concentrated extract was mixed with 0.03 ml MethElute and
injected into a gas chromatograph for on-column methylation
and analysis.
The gas chromatogram and the RGC are shown in Figures 16-A
and 16-B respectively. The peak designated "R" in Figure
16-A is from the MethElute reagent. The peak designated
with an asterisk in Figures 16-A and 16-B is di-n-
butylphthalate, a contaminant. There was no evidence of
branched fatty acids in this species of algae, and
pentadecanoic acid was the only odd-numbered carbon fatty
acid present. The acids identified and their relative
abundances are listed in Table 3. This evidence does not
invalidate the hypothesis that the odd-numbered and branched
fatty acids originate as metabolites from aquatic life in
the lagoons. Different species with different food sources
can produce different compounds.
Resin Acids
A total of 13 different resin acids were found in the waste-
waters of both mills in the 1972 samples; 8 were identified
and confirmed and 5 were not identified. The resin acid
content of the finished wastewaters were qualitatively
similar for the two mills. Seven resin acids were common to
42
-------
Table 3
FATTY ACIDS IN THE EXTRACT OF A CHLORELLA ALGAE CULTURE
CO
Fatty
Acid
12:0
14:0
15:0
16:0
16:2
16:3
18:0
18:1
18:2
Relative
0.
1.
0.
19.
17.
23.
4.
6.
13.
4
6
7
0
0
2
9
2
8
Found in Paper
Mill Wastewater
No
Yes
Yes
Yes
No
No
Yes
Yes
Yes
18:3
13.2
Sometimes
Compound Name & Comments
Laurie acid
Myristic acid
Pentadecanoic acid
Palmitic acid
cis or trans not determined
cis or trans not determined
Stearic acid
Oleic or elaidic acid
Linoleic or linolelaidic
acid
Linolenic or gamma-
linolenic acid
-------
I6»
B
Ss.
k
1KO
«*»
)MO
I ,
0 13 M 30 « SO 60 70 BO 30 iOOII01ZOI30l«lS016fll7O180l3020e2tOZ20230Z«2SOZ60?T02a023D3
3*£-TftH MJttP
15:3
IKO
1KO
» a /*
" »< lit M
Figure 16 (A) Gas chromatogram and (B) RGC of fatty
acid methyl esters from chlorella algae
44
-------
the raw wastewater extracts of both mills and six of these
were found in both finished wastewater extracts.
Differences were primarily found among the small,
unidentified resin acid GC peaks.
Significance of Resin Acids
Resin acids have long been known to be toxic to various
degrees to aquatic life. In addition, resin acid salts or
"soaps" are responsible for much of the foaming in kraft
mill effluents. The foam causes additional expense in the
treatment of these effluents because defoamers must be
added. If the foam is discharged to the receiving waters it
can float long distances before it is dispersed or
degraded.3
Dr. I. H. Rogers29 has recently reported that bioassays
using sockeye salmon have shown the lethal effects of a
mixture of resin acids isolated from Douglas fir oleoresin
to occur at concentrations as low as 2.0 mg/1. Others have
found resin acid soaps to be toxic to minnows at 1.0 mg/130
or to the water flea, Daphnia pulex,, at 2.0 mg/131. Resin
acids have been reported toxic to various fish at
concentrations of 1 mg/132 to 5 mg/1.33
Leach and Thakore34 recently reported that 80% of the
toxicity of unbleached white water (the major waste stream
from the pulping operation of a kraft mill) to juvenile coho
salmon was caused by the sodium salts of three resin acid
soaps: sodium isopimarate (55%), sodium abietate (22%), and
sodium dehydroabietate (5%). The acute toxicity of free
resin acids was much less than that of the sodium salts.
The pimaric-type acids were also found to be more toxic than
the abietic-type resin acids. The remaining toxicity (18%)
was contributed by sodium salts of the unsaturated fatty
acids: sodium palmitolate, sodium oleate, sodium linoleate,
and sodium linolenate.
Identification of Resin Acids
In addition to the isopimaric, abietic, dehydroabietic and
sandaracopimaric acids mentioned by Leach and Thakore3*, we
found pimaric, 6,8,11,13-abietatraen-18-oic, neoabietic, 13-
abieten-18-oic and several unidentified resin acids. To our
knowledge neoabietic, 13-abieten-18-oic, and 6,8,11,13-
abietatraen-18-oic acids have not been reported before in
kraft pulp mill wastewaters. All of these resin acids (as
their methyl esters) were initially identified by comparison
of their mass spectra with published mass spectra.33 They
45
-------
were all confirmed by comparison of gas chromatographic
retention times and GC-mass spectra from our instrument with
standards obtained from Dr. Duane F. Zinkel, D.S.D.A. Forest
Products Laboratory, Madison, Wisconsin.
The mass spectra and fragmentation patterns of these resin
acid methyl esters are covered in the literature36 and will
not be discussed here. The plotted spectra, however are
shown in Figures 17-20 because some of them have ion
abundances different from those reported in literature
references.35 These differences are primarily the
incorporation of masses between m/e 41 and 70 in our plots,
and may be due to the GC mode of introduction (versus direct
insertion probe35 3* ) with our samples and to our using a
quadrupole mass spectrometer instead of a magnetic
instrument.
The unidentified resin acid methyl esters may be
decomposition products or rearranged molecular structures
from a parent resin acid. The mass spectra of some may also
be contaminated with mass spectra of other unresolved
compounds or may be resin acids not previously identified
and studied.
Phenols
Eleven different phenols were identified in the kraft mill
wastewaters (Table 2). Guaiacol (identified as its methyl
ether, veratrole) varied in concentration from 0.5 to 2.7
mg/1 in the raw effluents and was very effectively reduced
in concentration by both treatments. Vanillin (identified
as its methyl ether, veratraldehyde) varied in concentration
from 0.8 to 2.1 mg/1 in the raw effluents but was not as
effectively reduced in concentration as guaiacol. The other
nine phenols occured in lesser concentrations and varied in
simplicity from phenol to acetosyringone (3,5-dimethoxy-4-
hydroxyacetophenone).
Based upon data obtained in 1974, some correlation can be
made between the percent removal of the phenols in these
waste treatment systems and the complexity or degree of
substitution of the phenol molecules. Table 4 lists the
removal data for these phenols and shows their structures.
The analytical data for the calculations were obtained from
3-day composite samples, quantitated with the aid of a
Perkin-Elmer PEP-1 computer system. (The details of these
analyses are described in a later section.) The phenols are
generally more resistant to treatment as the complexity of
the molecule increases. This is especially true with Mill
"A", which has only biological treatment, with the single
-------
Table 4
PERCENT REMOVAL OF PHENOLS VERSUS THEIR STRUCTURAL COMPLEXITY
Parent Compound
Guaiacol
p_-Hydroxybenz aldehyde
Vanillin
Ace tovani Hone
Homovanillic Acid
Syringaldehyde
Identified As Structure
Veratrole j*
OH
p_-Methoxybenzaldehyde .,**.
0
CHO
OH
Ve r at r aldehyde (f\*\ 3
CHO
OH
Acetoveratrone fi^]" 3
COCH3
OH
Methyl Homovanillate [fil 3
CHjCOOH
OH
3 , 4 , 5-Trimethoxy- CH3° "Y/'VY^"3
benz aldehyde K j \
CHO
% Removed
Mill "A" Inter-
state
96% 98%
21% 75%
65% 93%
77% 90%
50% ?
62%
Acetosyringone
OH
3,4,5-Trimethoxy- CH3°
acetophenone
57% 67%
47
-------
SPECTHHIUBER 1
«mVL flBIETHIE SID. XS, XB.iaB-SVlS,
00
SPECTHLH HfBER 39
(ETHYL DEHVCFDfBIETfnE, X5,. «J-.fi. laj-BYlS
B
(SB.
XOOCHj
B. 11. 13,-AeffiTATRIEN-le^ATE
IDEHVDIHMfilETATE!
20 3B H
M^ E
...|,.l,|l,.l|,,l.|,l,l lll|llll|,,,l|lll,l llll|l,M|,lll
50 6B TB 8B 9B Iffi 111
110 120 130 1« 19 160 170 IBB 131
lll|.,,,|i lll,,.ll|llll|l nlllll, I
ie0 170 tag tat an 210
'i 'I i""i""i i i I'"
Figure 17 Mass spectrum of (A) methyl abietate and
(B) methyl dehydroabietate
-------
SPECTTU1 HJttB 1
a.
a_
a.
...I..., I!,
i1 i i'"
»-c s e c a1"
H/ E
is. a>
300 310 320 330 34i
SPECTFU1 K1CER 1
"COOCH,
o
°
.1,1.1 lill lluJJIli ^.rlllllnillL^iligiiH.,^!!,,,,.,!!! I Ill II, II
« se eg 7B ae SB IBB 110 28 iae i-w ise IBB ITB iaa 13B 2BB 2ia 22B 23)
20 30 « SB 80 Ttt
KS £
Figure 18 Mass spectrum of (A) methyl pimarate and
(B) methyl isopimarate
-------
3TORH fOGER 1
0?
_L L
38 v sa ee it et se MB
M^ E
ne lee ise
U1
O
SPECTHJ1 RKER 2B9
B
"COOCH,
4). 13(15)-ABIETADIEN-18-OATE
(NEOABIETATE)
m IB at 30 1» llfl 120 130 1« 1SB J8B 1TB 190 130 2BB Zlfl ZZB 230 Z1B 2S 280 2TB
Figure 19 Mass spectrum of (A) methyl sandaraco
pimarate and (B) methyl neoabietate
-------
srErm* MMEK lit - IBB
FCTHtL 6.e.ll.l3-feiETH1ETHEN-lB-ORIE
Figure 20 Mass spectrum of (A) methyl 13-abieten-18-
oate and (B) methyl 6,8,11,13-abietatetraen-
18-oate
-------
exception of E-hydroxybenzaldehyde, which may be produced in
the aerated lagoons by biological degradation of lignin or
vanillin. Most of these phenols are probably involved in
complex equilibria, and their concentrations depend upon
their initial concentrations in the wastewater, their rates
of degradation in the lagoons, and their rates of formation
from degradation of lignin and other molecules in the
lagoons.
Significance of Phenols
Phenols have long been associated with kraft paper mill
wastewaters and many studies of their effects have been
made. Chopin37 reported that phenolic compounds extracted
from various pulping processes are slightly toxic but much
less so than phenol or cresols. Concentrations of phenol
that have been reported as lethal or damaging to fish range
from 0.079 to 1900 mg/1 but the most reliable information
relating to pure phenol under carefully standardized
conditions indicates that the 24-, 48- and 96-hour TLm
concentrations are in the general range of 10-20 mg/1 at
20°C.38
Probably a greater problem caused by the phenolic
constituents of kraft mill wastewaters is the impairment of
the flavor of fish, shrimp, and other edible aquatic life.
A table in a recent report39 lists the highest
concentrations of various organics in water that will not
impair fish flavor. Values for phenolics range from 0.005
mg/1 for cresol to 5.6 mg/1 for phenol. The only compound
in the table that was found in the treated mill wastewaters
is guaiacol. The highest concentration of guaiacol that
would not impair the flavor of resident fish was listed as
0.1 mg/1.
An earlier investigation*0 showed that the flavor of cooked
coho salmon was impaired after exposure of the fish to
untreated kraft pulp mill effluent for 72-96 hours at
concentrations of 1-2% or more by volume. No flavor
impairment was noted when these fish were exposed to 2-9% by
volume of biologically-treated effluent. These results are
in agreement with our findings that, in general, the
phenolic constituents of kraft paper mill wastewaters are
reduced in concentration from 50 to 98% by biological
treatment.
In some kraft paper mills, the pulp is bleached with
chlorine. Analyses of several wastewater extracts from
bleached kraft mills in collaboration with Dr. I. H. Rogers,
Environment Canada, Pacific Environmental Institute, West
52
-------
Vancouver, B.C., have led to the identification of 3,4,5
trichloroveratrole XVII and tetrachloroveratrole XVIII .4
Since the samples were methylated prior to analysis, the
parent compounds were chlorinated guaiacols.
OCH,
OCH,
OCH,
XVII
XVIII
Phenols are known to be highly susceptable to chlorination
under the conditions employed in the bleaching operations of
most mills. Guaiacol was subjected to chlorination under
conditions approximating those used in the mill from which
the samples were taken and a series of chlorinated isomers
resulted. The only trichloroisomer found was 3,4,5-
trichloroguaiacol, identified and confirmed as its methyl
derivative (XV) by GO MS, NMR, and GC-IR. The only possible
tetrachloro isomer (XVI) was also found. The results of
these and other related analyses are fully described in a
separate report.41
Generally, chlorinated phenols are more toxic than the
parent compounds and the concentrations at which flavor in
fish is impaired or at which taste and odor can be detected
in drinking water is much lower for the chlorinated
compounds. For example, the highest concentration of 2,4-
dichlorophenol that does not impair fish flavor is 1/560,000
that of phenol for trout and 1/56,000 that of phenol for
bass.39 The upper limit for isomeric 2,3-dichlorophenol for
trout is 1/175 that of phenol for trout. A great deal of
research must be done with the phenols identified to
determine their chlorinated derivatives produced in mill
bleaching operations, and the effects of these chlorinated
phenolics on aquatic life and in drinking water.
CHEMICAL PROFILES OF ACIDS AND PHENOLS
A "chemical profile" is a vertical display of the gas
chromatograms of samples taken at consecutive points in a
53
-------
waste treatment system. The compounds present and their
relative concentrations are therefore shown as a function of
the treatment.
This display allows one to see at a glance the effectiveness
(or lack of it) of each step in the treatment process with
respect to an individual compound, to a class of compounds,
or to all chromatographable compounds in the wastewater,
whether they are identified or not.
This information in conjunction with the traditional
collective pollution parameters (e.g., BOD and TOC) provides
a much better understanding and evaluation of the effluent
composition and its possible environmental effects than the
chemical profiles or collective parameters alone.
Figure 21 shows the chemical profile of the acids and
phenols from the wastewater grab samples of Mill "A".
Figure 22 shows the chemical profile of the acids and
phenols from the grab samples of the Interstate mill at
Riceboro, Georgia. The peak numbers correspond to the
compounds listed in Table 2.
The compositions of the raw effluents from the two mills
were similar. The treated effluent samples from the two
mills exhibited more differences. In general, most
compounds decreased significantly in concentration as the
wastewater passed through the treatment system.
One exception appeared to be the resin acid content. Some
of the peaks appeared to increase in concentration. This
apparently was the result of large variances in the
concentration of the organics in the mill wastewaters with
time and our failure to have precisely sampled the "slug" of
effluent we tried to follow through the treatment systems.
Another factor could be that the biological parts of one or
both of these systems was not working at usual efficiency
when the samples were taken. In any event, the quantitation
of the compounds in 1972 samples was not accurate. The
major effort was directed at the identification of these
compounds and their verification with standards.
REPEAT ANALYSES OF THE ACIDS AND PHENOLS
Additional samples from both mills were obtained in January
1974almost 2 years after the previous samples were taken.
The analyses were repeated for two reasons:
54
-------
G-la
4a
Figure 21 Chemical profile of acids and phenols from
Mill "A"; sample points 1-4.
55
-------
xfl.22
'S
viu
i r
\
3a
f I
1 1 r 1 1 r
xO.33
Figure 22 Chemical profile of acids and phenols from
the Interstate mill at Riceboro, Georgia;
sample points 1-4
56
-------
to ascertain whether the same compounds were still
present and, if so, if they were present in
approximately the same concentrations.
to obtain reliable, representative, quantitative
data on the concentrations of these compounds.
The first objective is important if the information obtained
from these studies is to be of general use. A radical
change in the wastewater composition with time would require
each analysis to be a research projectan impractical
situation. However, if the composition of the wastewaters
is essentially the same over a two-year time span, then the
assumption that the wastewater composition will remain the
same is basically valid, unless major changes in raw
materials, production practices, or treatment processes
occur.
The second objective is important if the information
obtained from these studies is to be used to assess the
effectiveness of the treatments towards reducing the
concentration of a class of chemicals or even of a specific
compound.
Sampling
Samples from both mills were collected by automatic sampling
devices at the mills. All samples were collected during the
same week. Samples of the raw wastewaters and of the
outfalls, collected over 24-hour periods, were frozen each
day. A 3-day composite composed of equal volumes from the
Monday, Tuesday, and Wednesday raw wastewater samples of
Mill "A" was prepared. A second 3-day composite was
composed of equal volumes from the Wednesday, Thursday and
Friday treated wastewater samples of Mill "A". This
provided samples of a slug of wastewater, with a 3-day
averaged composition, both before and after it had passed
through the 2.5-day treatment system.
Three-day composites of equal volumes from the Monday,
Tuesday and Wednesday raw wastewater and of the treated
wastewater samples of the Interstate Mill at Riceboro were
prepared. Because the stabilization pond retention time is
at least 3 months, no attempt was made to obtain samples of
the same slug of this wastewater.
Samples were not taken at intermediate points in the
treatment system of either mill because automatic sampling
57
-------
devices were not available there and because analyses time
was limited.
Method of Analysis
A one-liter aliquot of each of the four composite samples
was taken and was prepared in essentially the same manner as
were the 1972 samples:
After the pH was raised to 10.5 with 50% sodium
hydroxide, the terpenes and neutral compounds were
extracted with chloroform.
The aqueous sample was divided into two 500-ml
portions and each portion was methylated using
dimethyl sulfate.
After extraction of the methylated portions with
chloroform, the extracts of each sample were
combined and concentrated to 1.0 ml. By utilizing
two separate methylation reactions for each sample,
some averaging of either a low- or a high-yield
methylation reaction is gained.
The samples were spiked with acenaphthene as an internal
standard and chromatographed on a 50-ft SCOT column packed
with carbowax 20M/TPA. Retention time and peak area data
were collected on a Perkin Elmer PEP-1 computer interfaced
with the gas chromatograph. Computer analysis of these data
allowed identification and quantitation of the compounds
based on information gained from the 1972 samples. PEP-1
computer identifications were verified by obtaining mass
spectra of the compounds in these samples and comparing them
with the mass spectral data of the identified compounds from
the 1972 samples.
COMPUTER ASSISTED GAS CHROMATOGRAPHIC ANALYSES
Based on the compound identifications from the 1972
analyses, a computer-assisted gas chromatographic analysis
was used to analyze the 1974 wastewater samples. GC-MS was
used to verify compound identifications and showed that,
once identified, computer-assisted GC analysis was
sufficient for additional analyses.
58
-------
Response Factors of Model Compounds
A standard solution consisting of 5.75 mg/1 veratrole, 1.12
mg/1 acenaphthene, 2.82 mg/1 methyl palmitate, and 8.96 mg/1
methyl dehydroabietate was prepared and chromatographed
using the PEP-1 computer system. From the known
concentrations and the measured peak areas, response factors
were calculated relative to that of acenaphthene, which, as
the internal standard, is assigned a value of 1.00. The
response factors were as follows:
Acenaphthene 1.00
Veratrole 1.49
Methyl palmitate 2.05
Methyl dehydroabietate 2.32
(internal standard)
(phenols)
(fatty acid methyl esters)
(resin acid methyl esters)
Assuming all compounds in the same chemical class have the
same response factor, the above factors were assigned to the
various phenols and fatty and resin acid methyl esters so
the computer could calculate the concentrations of these
compounds in the samples.
Analyses of the .1.924 Samples
A 300-^1 aliquot of each extract was spiked with 30 /
-------
the Single Reference Peak program because the time required
for each chromatogram approaches 50 minutes. Peaks
corresponding to veratrole, veratraldehyde and methyl
dehydroabietate were chosen as retention time reference
peaks.
An example of the computer program used to analyze the raw
effluent extract from the Interstate mill at Riceboro is
shown in Figure 23. The raw effluent reduced data, showing
peak areas normalized to the sum of all the peak areas in
the run, is shown in Figure 24. The product of the computer
analysis of the data in Figure 24 using the program in
Figure 23 is shown in Figure 25. The peak number
designations, the notation of the presence or absence of the
compound in the corresponding 1972 sample, and the total
from the sum of all the concentration values in Figure 25,
were manually added to the data printed out by the computer.
The gas chromatogram of the extract from the Interstate
mill's raw wastewater is shown in Figure 26-A. Peak number
designations match those in Figure 25 and are consistent
with identifications made with the 1972 samples. The letter
"A" designates the acenaphthene spike used for the internal
standard. The letter *M" designates methyl margarate, which
was not included in the list of identifications in the 1972
samples because the peak resolution was not as good and the
confirmation was doubtful.
The gas chromatogram and computer analysis of the extract
from Interstate's treated effluent (sample point #4 in
Figure 3] are shown in Figures 27 and 26-B respectively. A
comparison of either Figures 25 and 27 or of 26-A and 26-B
shows significant reductions in the concentrations of these
compounds after treatment.
Similar comparisons can be made from the computer analyses
of Mill "A" raw effluent and treated effluent extracts
(Figs. 28 and 29} or from the gas chromatograms of these
same extracts (Figs. 30-A and 30-B). Analyses of the raw
effluent were run in duplicate (Fig. 28) to determine the
range of agreement found using the PEP-1 system. Twenty of
the values were within a 1% to 10% range of agreement with
each other. Three of the values were greater than the 10%
range of agreement (35%, 38% and 53%).
60
-------
PSOt
«TO INTERSTATE RAW EFFLUENT-ACIDS AND PHEN0LSI
INST 1 , METH0D 50 , FILE 40 3l
STD C0NC l.OOOOl
TIMES 48.00, 9.00, 23.55, 26.55, 30.20, 33.75,
THRESHOLDS 16, 32,
IKK/AIR 1.0000, .00,
T0L .140, .010, 5.0,
REF
1.000, 9.50, 10.50, 20.00,
2.450, 24.00, 25.00, 30.00,
4.100, 40.50, 42.00, 48.00, I
STD NAME ACENAPHTHENEl
NAME
VERATR0LE:
DIMETHYLSULF0NEI
METH0XYBENZALDEHYDE»
I
I
I
AR0MATIC MV 182I
ACENAPHTHENEl
I
PALMITATEl
I
ANTEIS0MARGARATEt
ME H0M0VANILLATEI
MARGARATE:
VERATRALDEKYDEs
VERATR0NEI
STEARATE AND 0LEATE1
M0STLY LIN0LEATEI
3,4,5-TMAJ
!
ARACHIDATEt
ME RESIN ACIDi
ME RESIN ACIDi
ME RESIN ACID MV 314l
PIMERATEl
SANDARAC0PIMERATEI
13-ABIETEN-I8-0ATE!
I
IS0PIMERATEI
I
AB- AND DEHYDR0AB-s
I
6,8,11,13-AB, NE0AB-I
LIGN0CERATEI
RRT
.000,
.350,
.650,
.760,
.770,
.817,
.850,
.890,
.980,
2.190,
2.260,
2.330,
2.350,
2.430,
2.450,
2.620,
2.650,
2.750,
2.870,
2.930,
2.990,
3.030,
3.080,
3.148,
3.240,
3.360,
3.400,
3.410,
3.560,
3.880,
4. 100,
4.170,
4.350,
4.600,
RF
1.4900,
2.0500,
1.4900,
1.0000,
1.0000,
1.0000,
1.4900,
1.0000,
1.0000,
2.0500,
1.0000*
2.0500,
1.4900,
2.0500,
1.4900,
1.4900*
2.0500,
2.0500,
1.4900,
1.0000,
2.0500,
2.3200*
2.3200,
2.3200,
2.3200,
2.3200*
2.3200,
1.0000,
2.3200*
1.0000*
2.3200*
1.0000*
2.3200*
2.0500*
C
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
1.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
.0000*
-P
C
0)
I
0)
A
-P
0)
N
fS
g o
(3 M
o
O .Q
CO
W
Cn-H
& 0)
p
b a
0) -P
4J M
3 H
ft
-------
N35I
RUN INTERSTATE RAW EFFLUENT
MST
, METH8D
ACIDS AND PHEN0LS
FILE 35 3t
TIME
6.48
7.35
8.40
8.84
9.50
10.10
11.56
11.89
12.47
12.79
13.13
13*37
13.65
14.27
14.63
15.38
15.60
16.24
16.66
17.39
17.85
18.36
18.72
19.07
19.77
20.05
20.60
20.82
20.99
21.33
21.59
22.05
22.49
22.80
23.30
23.68
23.97
24.37
24.66
25.67
26.41
26.71
27.57
28.49
28.88
29.37
29.61
29.96
30.47
31.01
31.73
32.78
33.60
33.98
34.54
36.23
37.00
38.77
41.98
42.80
44.62
45.92
47.13
AREA
.0631
.1066
.0188
.1778
.1666
8.3184
.0902
.4989
.0310
.0878
.0373
.0521
2.7822
.0687
.0437
.1452
.2228
.0688
.5923
.3927
.5308
1.6764
.6656
9.0883
.0555
.4136
.0518
.0247
.0230
.0737
.0378
.8961
.1292
.2664
.3619
.7918
.0738
.2071
7.7353
.0644
3.8972
2.2447 !
1.9567
.0262
.4292
.2510
.0847
.2554
.4545
.5225
1.1206
7.8243
2.1625
.9214
.6673
24.6361
.0222
.8480
50.9977
.7900
6.8304
.3696
.0180
RRT
.642,
.735,
.840,
.884,
.950,
.010,
.156,
.189,
.247,
.279,
.313,
.337,
.365,
.427,
.463,
.538,
.560,
.624,
.666,
.739,
.785,
.836,
.872,
.907,
.977,
2. DOS,
2.060,
2.082,
2.099,
2. 133,
2.159,
2.20S.
2.249,
2.280,
2.330,
2.368,
1.397,
2.437,
5.466,
2.567,
2.641,
2.671,
2.757,
2.849,
2.888,
2.937,
2.961,
2.996,
9.047,
J.101,
3.173,
3.278,
3.360,
i.398.
3.454,
3.623,
3.700,
1.877,
.198,
.280,
.462,
.592,
.713,
RF
.0000,
. 0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
. 0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
. 0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000.
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
. 0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
. 0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000,
.0000.
c
.0437,
0738,
.0130,
.1230,
.1153,
5.7582,
.0624,
.3453,
.0215,
.0608,
.0258,
.0361,
I. 9259,
.0475,
.0302,
.1005,
.1542,
.0476,
.4100,
.2718,
.3674,
1.1605,
.4607,
6.2912,
.0384,
.2863,
.0358,
.0171,
.0159,
. OS I 0,
. 0262,
.6203,
.0894,
.1844,
.2505,
.5481,
.0510,
.1433,
5.3546,
.0445,
2.6978,
1.5538,
1.3545,
.0181,
.2971,
.1737,
.0586,
.1768,
.3146,
.3617.
.7757,
5.4162,
1.4970,
.6378,
.4619,
17.0536,
.0153,
.5870,
35.3024,
.5468,
4.7282,
.2558,
.0125,
Figure 24 Computer print-out of the normalized peak
areas from the chromatogram of the raw
effluent extract from the Interstate mill
at Riceboro
62
-------
INTERSTATE RAW EFFLUENT-ACIDS AND PHEN0LS
U)
CD H
!x! 3
rS t '
rt rt
H (D
PI H
O DJ
ft rt
PI
ff
CO
^j
Pj
(D
Hi
Hi
£
ID
5.
P)
Q
l-i-
Pl
QJ
*T3
3*
CD
3
o
H-
c
(D
to
Ul
n
n
1
d-
fD
hj
PI
B
01
i '
01
H-
O
Hi
rt
(D
O
3-
hj
3
PI
rt
O
iQ
i-i
0)
§
O
Hi
INST 4
TIME
10.
11.
13.
15.
16.
17.
17.
18.
18.
19.
20.
22.
22.
23.
23.
24.
24.
26.
26.
27.
28.
29.
29.
30.
31.
31.
32.
33.
33.
34.
36.
38.
41.
42.
44.
45.
10
89
65
60
66
39
85
36
72
07
05
05
80
30
68
37
66
41
71
57
88
37
96
47
01
73
78
60
98
54
23
77
98
80
62
92
, METK0D
AREA
8.3184
.4989
2.7822
.2228
.5923
.3927
.5308
1.6764
.6656
9.0883
.4136
.8961
.2664
.3619
.7918
.2071
7.7353
3.8972
2.2447
1.9567
.4292
.2510
.2554
.4545
.5225
1.1206
7.8243
2.1625
.9214
.6673
24.6361
8480
50.9977
.7900
6.8304
.3696
50 ,
RRT
000,
177,
351,
544,
649,
721,
767,
817,
853,
.888,
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
3.
3.
3.
3.
3.
3.
3.
3.
3.
4.
4.
4.
4.
985,
186,
262,
312,
351,
420,
450,
626,
656,
743,
875,
925,
984,
031,
081,
148,
246,
322,
357,
409,
566,
802,
too.
176,
345,
465,
FILE 35
RF
1.4900,
1.
2.
1.
1.
1.
1.
1.
1.
1.
1.
2.
1.
2.
0000,
0500,
0000,
4900,
0000,
0000,
0000,
4900,
0000,
0000,
0500,
0000,
0500,
1.4900,
2.
0500,
1.4900,
1.
2.
2.
1.
1.
2.
2.
2.
2.
2.
2.
2.
1.
2.
1.
2.
1.
2.
2.
4900,
0500,
0500,
4900,
0000,
0500,
3200,
3200,
3200,
3200,
3200,
3200,
0000,
3200,
0000,
3200,
0000,
3200,
0500,
3i
1.
1.
C
3637,
0548,
6275,
0245,
0971,
0432,
0584,
1844,
1091,
0000,
0455,
2021,
0293,
0816,
1298,
0467,
1.2681,
.6389,
1.
6.
13.
5063,
4413,
0703,
0276,
0576,
1160,
1333,
2860,
9973,
5520,
2352,
0734,
2888,
0933,
0176,
0869,
1.7436,
0833,
NAME
VERATR0LEI
1
DIMETHYL SULF0NEI
I
M ETH0 XYBEN Z AL DEH YDE 1
I
l
1
AR0MATIC MV 182 l
ACENAPHTHENEt
l
PALMITATEl
l
ANTEI S0MARGARATE!
ME H0M0VANILLATEI
MARGARATEl
VERATRALDEHYDEl
VERATR0NES
STEARATE AND 0LEATEI
MB STL Y LIN0LEATEI
3,4,5-TMAl
l
ARACHIDATEl
ME RESIN ACIDt
ME RESIN ACIDl
ME RESIN ACID MV 3I4l
PIMERATEl
SAN DARAC0 PIM ERATE l
13-ABIETEN-18-0ATEI
l
IS0PIMERATEI
t
AB- AND DEHYDR0AB-S
t
6,8, 11,13-AB, NE0AB-I
L I GN0 CERATE 1
PEAK
11
15
18
31
32
M
33
36
37,38
40
42
43
45
50
51
53
55
56,57
58,59
60
IN «72
YES
YES
YES
N0
YES
YES
YES
N0
YES
YES
YES,YES
YES
YES
YES
N0
YES
N0
YES
YES
YES
YES
YES,YES
YES,YES
N0
TOTAL:
30.7714 MG/L
-------
20
B
IS
56,57
Figure 26 Gas chromatogram of the acid and phenol
extract of Interstate's (A) raw effluent
and (B) treated effluent
64
-------
RUN
INTERSTATE TREATED EFFL-ACIDS AND PHENOLS
DJST
METH0D
50
, FILE
41
3:
TIME
10.12
16.65
18.41
19.06
20.07
22.07
22.74
24.41
24.67
26.45
26.89
28.96
29.51
30.02
31.06
31.72
32.74
33.91
35.98
41.39
43.54
AREA
.1315
.1423
.1368
8.8748
.1581
.2320
.2390
.2475
.5476
.3995
.3413
. 1394
.2972
.2021
1.6762
1.3443
2.9344
3.6459
2.8126
11.7420
.2298
RRT
I. 000,
1.350,
1.645,
1.819,
1.883,
1.983,
2.186,
2.254,
2.423,
2.450,
2.630,
2.675,
2.720,
2.885,
2.941,
2.991,
3.093,
3. 157,
3.257,
3.371,
3.572,
4. 100,
4.309,
4.460,
RF
1.4900,
2.0500,
1.4900,
1.0000,
1.0000,
1.0000,
2.0500,
1.0000,
2.0500,
1.4900,
1.4900,
2.0500,
1.4900,
1.4900,
1.0000,
1.0000,
2.3200,
2.3200,
2.3200,
2.3200,
2.3200,
2.3200,
2.3200,
2.0500,
C
.0220,
.0239,
.0154,
1.0000,
.0178,
.0535,
.0269,
.0571,
.0919,
.0670,
.0788,
.0234,
.0334,
.0227,
.4381,
.3514,
.7670,
.9530,
.7352,
3.0694,
.0600,
NAME
VERATR0LEI
DIMETHYL SULF0NE:
METH0XYBENZALDEHYDEJ
I
ACENAPHTHENEt
I
PALMITATEt
:
MARGARATEs
VERATRALDEHYDE:
VERATR0NE!
STEARATE AND 0LEATEI
ME SYRINGALDEHYDE:
3,4, 5-TMAl
l
t
ME RESIN ACID;
ME RESIN ACID:
PIMERATEt
SAND-P AND J3-AB-18-:
IS0PIMERATE!
AB- AND DEHYDR0AB- :
6,8, 11, 13-AB, NE0AB-:
L I GN0 CERATE:
PEAK
11
15
18
A
28
M
33
36
37,38
39
42
45
48
50
51,53
55
56,57
58,59
60
IN '72
YES
YES
YES
--
YES
N0
YES
YES
N0,YES
YES
YES
YES
YES
YES
YES, YES
YES
N0,YES
YES,N0
N0
T0TAL:
6.9079 MG/L
Figure 27 Computer analysis of the chromatogram of Interstate's
treated effluent acid and phenol extract
-------
HIST
1 MILL A RAW EFFLUENT-ACIDS AND PHENILS
METHID SO , FILE 37 3|
TIME
10.14
13.66
16.70
18.48
19.10
10.08
28.10
£3.75
24.73
26.51
26.88
27.47
27.66
27.94
28.98
30.59
31.13
31.88
32.86
34.10
36.20
41.92
44.11
44.59
45.81
AREA
3.1384
1.7237
.2877
.4908
9.2566
.3537
1.3317 !
1
.4753 1
5.2457 1
3.0875 t
3.5064 1
.9110
.7588
.2840
.9289
.5729
.5133
.4960
4.0798
6.7065
3.8577
26.4614
.3482
.5035
.4653 4
RRT
.000*
.347.
646*
.822.
.883.
.980.
i. 184.
!.220.
8.350,
8.450,
!.629.
i.661.
.726.
.746.
.774.
879.
.037,
.087,
.152.
250.
.366.
.563.
.100,
.305,
350,
t.464.
RF
1.4900,
2.0500.
1.4900.
1.0000.
I. 0000,
1 . 0000.
2.0500,
2.0500.
1.4900.
1.4900.
1.4900.
2.0500.
1.4900.
2.0500.
1.4900.
1.4900.
1.0000.
2.3200.
2.3200.
2.3200.
2.3200.
2.3200,
2.3200.
8.3200,
2.3200,
2.0500.
C
.5051.
.3817.
.0463,
.0530,
1.0000.
.0382.
.2949,
.0765,
.8443,
.4969,
.7765,
.1466,
. 1680,
.0457,
. 1495,
. 06 1 8,
.1286,
. 1243,
1.0225,
1.6808,
.9668,
6.6320,
.0872,
.1262,
. 1030,
NAME
VERATMLCl
DIMETHYL SULFCNEl
H ETH« XYBEMIAL DEHYDEI
I
AC EM A PH THE* El
I
PALMlTATCl
PALM IT EL AI DATE I
HE HfHfVAMlLLATEl
VERATRAL DEHYDEl
VERATRCMEl
STEARATE AMD ILEATEl
ME SYRIMOALDEMYDEl
LIMSLEATEi
3,4-ENPl
3. 4,5-TM AI
HE RESIN AClDl
HE RESIN ACIDS B,C|
PlHCRATEt
SAMD-P AND I3-AB-18-I
IStPlHCRATEl
AB- AND DEHYDRCAB-I
6.8. II. I3-AB-I
MECABIETATEl
LI Mi CERATE I
II
19
18
29
38
33
36
37,38
39
40
41
46,47
50
51.93
59
56.97
98
9»
60
IN '71
YES
rts
res
TES
YES
YIS
YtS
YIS
YES.YIS
YES
YES
YES
YES
YES.YtS
YES
YES.M
YES
YES.YTS
YES
YES
TOTAL I
14.9834 MG/L
KM
WST 4
TIME
10.07
13.60
16.63
18.43
19.05
20.02
22.04
23.70
24.69
26.46
26.76
27.42
27.62
27.90
28.93
29.50
30.54
31.08
31.76
32.78
34.00
36.08
38.76
41.63
44.26
45.43
2 HILL
. HETHID
AREA
3.2236
1.8019
.3241
.4818
9.7027
.3204
1.3312
.4879
5.3897
3.1908
3.4873
.9637
.7164
.2862
.9194
.3288
.6329
.7586
.8566
4.4606
6.9305
3.9542
.3831
26.4755
.8086
.3379
A RAV
50 ,
RRT
.000,
.350,
.651.
.830,
.891,
.987,
2. 187,
2.220,
2.352,
2.450,
2.625.
2.654.
2.720,
2.739,
2.767,
2.869,
2.925,
3.027,
3.079.
3.145,
3.243.
.1.361.
3.563,
3.822,
4. 100,
4.354.
4.467,
EFFLUTMT-
riLE
RF
1.4900,
2.0500,
1.4900.
1.0000.
1.0000.
1.0000,
2.0500.
2.0500.
1.4900.
1.4900.
1.4900.
2.0500.
1.4900.
2.0500.
1.4900.
1.4900,
1.0000,
1.0000,
2.3200,
2.3200,
2.3200,
2.3200,
2.3200,
1.0000,
2.3200,
2.3200,
2.0500,
C
.4950,
.3807,
.0497,
.0496,
1.0000,
.0330,
.2812,
.0749.
.8276,
.4899,
.7368,
.1479,
.1513,
.0439,
.1411,
.0338,
.0652,
.1814,
.2048,
1.0665,
1.6571,
.9454,
.0394,
6.3304,
.1933,
.0714,
NAME
VCRATRfLEl
DIM ETHYL SULFCN El
HETHIXYBENEALDEHYDEl
t
ACENAPHTHENEl
I
PALM I TAT El
PALMITELAIDATEl
HE H»MiVAMILLATEl
VCRATRALDERYDEl
VERATRMEl
STEARATE AMD LEATKl
HE SYRIMaALDEMYDEl
LINfLEATEl
3. 4-DNPl
3,4. 5- THAI
ME RESIN ACIDl
HE RESIN ACIDS B.CI
PIHERATEt
SAMD-P AMD I3-AB-I8-I
1S4PIMERATEI
I
AB- AMD DEHYDRCAB-I
6.8. II, 1 3- A3. HUABI
LIONtCERATEl
||
15
lg
37,
3*
46,47
SO
51.53
55
_
56.97
98,9*
6O
IN -72
YtS
YES
YES
Y*S
YE5
*es
YIS
YtS
YtS. YtS
YtS
YES
YtS
YtS
YtS, YES
YES
YXS.N8
YtS
YtS, YtS
YtS, YtS
T»TALl
14.6913 HQ/L
Figure 28 Computer analysis of the chromatogram of Mill
"A's" raw effluent acid and phenol extract (A)
run no. 1 and (B) duplicate run
66
-------
RUN MILL A TREATED EFFLUENT-ACIDS AND PHENOLS
INST 4 , METH0D 50 , FILE 36 3i
c
b
[si
ID
!> O
3
ra_lo
" rt
O (D
rt
Hi fu
fa 3
H H
P> W
O H-
H- 01
Oi
O
p) (-h
3
DJ rt
f
T3 (D
!?
(D O
O H
H O
(D QJ
X rt
rt 0
H IQ
(U hj
0 Q)
rt 3
O
Hi
3
H-
TIME
10.12
13.64
16.70
17.42
18.39
18.71
19.11
19.79
20.12
21.03
22.11
22.57
23.66
24.44
24.72
26.48
26.93
27.44
27.64
27.92
28.55
28.98
29.54
30.53
31.04
31.78
32.78
33.96
35.48
36.06
41.43
44.17
45.48
49.59
I
N0TEs
AREA
. 1291
.1732
.2452
. 1129
.1112
.2002
9.9590
. 1403
. 1804
.1711
1.8876
1. 1136
.2549
.2631
1.9480
.7781
2.2860
.3729
.1832
.1779
.3314
.4289
.2222
.6486
4631
.2201
.8428
1.4073
.2199
.7912
2.5728
.2192
.2818
.1314
RRT
.000,
.347,
.650,
.721,
.817,
.848,
.888,
.955,
.988,
2.079,
2.187,
2.234,
2.343,
2.421,
2.450,
2.626,
2.671,
2.723,
2.743,
2.771,
2.834,
2.877,
2.934,
3.031,
3.081,
3.154,
3.252,
3.368,
3.516,
3.573,
4. 100,
4.368,
4.496,
*******,
RF
1.4900,
2.0500,
1.4900,
2.0500,
1.4900,
2.0500,
1.0000,
2.0500,
1.0000,
2.0500,
2.0500,
2.0500,
1.4900,
2.0500,
1.4900,
1.4900,
2.0500,
1.4900,
2.0500,
1.4900,
1.0000,
1.4900,
1.0000,
2.0500,
1.4900,
2.3200,
2.3200,
2.3200,
2.0500,
2.3200,
2.3200,
2.3200,
2.0500,
1.0000,
TOTAL I
C
.0193,
.0356,
.0366,
.0232,
.0166,
.0412,
1.0000,
.0288,
.0181,
.0352,
.3885,
.2292,
.0381,
.0541,
.2914,
.1164,
.4705,
.0558,
.0377,
.0266,
.0332,
.0641,
.0223,
. 1335,
.0692,
.0512,
.1963,
.3278,
.0452,
.1843,
.5993,
.0510,
.0580,
.0131
3.8114
10-METHYLTETRADECAN0ATE UNDER ACENAPHTHENE
1
VERj
DIM
MET!
MYR
AR01
ANT
ACD
PEN
t
IS0
PALI
PALI
ME
MAR
VER
VER
STE
ME
LINI
PHT1
<
3,4
s
ARA
DIH
RES
PIM
SAN
UNS
1 50
AB-
NE0
LIGI
, 1
MG/L
IS E
NAME
ATR0LEI
DIMETHYLSULF0NE!
METH0XYBENZALDEHYDEI
MYRISTATEt
AR0MATIC M V I68l
ANTEIS0 C-15 I
ACENAPHTHENEt
PENTAOECAN0ATEI
t
IS0PALMITATEI
PALMITATEl
PALM1TELAIDATEt
ME H0M0VANILLATEI
MARGARATE:
VERATRALDEHYDEl
VERATR0NES
STEARATE AND 0LEATEI
ME SYRINGALDEHYDEl
LIN0LEATEI
PHTHALATEt
<
3,4,5-TMAi
s
ARACHI DATEt
DIHEXYLPHTHALATEl
RESIN AND FATTY ACIDl
PIMERATEt
SAND-P AND 13-AB-18-:
UNSAT FATTY ACIDl
IS0PIMERATEI
AB- AND DEHYDR0AB-I
PEAK
II
15
18
20
23
A
86
27
28
29
32
M
33
36
97,38
39
40
42
43
44
46,49
50
51,53
54
55
56,57
59
60
IN '72
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
N0
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
YES
N0
YES
YES
N0
YES
IS UN SATURATED.
-------
56,57
20
30
40
B
15
I
23
18
120
ir
27 29
33
37,36
\/
49
53
51/
26
55
56,57
I
10
30
Figure 30 Gas chromatogram of the acid and phenol
extract of Mill "A's" (A} raw effluent
run no. 1 and (B) treated effluent
68
-------
Comparisons of the 1974 and the 1972 Samples
Table 5 lists TOC, BOD and total volatile components in the
acid/phenol extracts from both mills in this study for both
sampling times.
Comparison of the collective pollution parameters indicates
that BOD removal is around 70-9OX in both mill wastewaters
after treatment. TOC reduction in the treated wastewaters
of both mills probably varies between 60-90%. Because of
large amounts of suspended solids in the raw effluent of
Mill "A", the 1972 TOC values may be erroneous.
Reduction in total volatile acidic materials is about 65-BOX
in both mills. The decrease of only 24% in the 1972
sampling of Mill "A" is probably not representative because
of faulty sampling, as explained earlier.
If treatment effectiveness is considered with respect to
classes of compounds, the phenols appear to be the most
susceptable to treatment. Reduction in the volatile
phenolic content of each mill was very consistent, ranging
in Mill "A" wastewaters from a 7356 to a 77% reduction.
Volatile phenols were reduced in the Interstate wastewaters
by 94% to 95%.
Resin acid reductions in the treated wastewaters ranged from
about 50% to 90%. The apparent increase in resin acid
content of Mill "A" in 1972 is due to a non-representative
grab sample in which the "slug" of effluent being sampled
was apparently missed.
Fatty acid content of the Interstate wastewaters was
decreased by about 85% to 90%, but there were increases in
overall fatty acid concentrations in both the 1972 and the
1974 samples of Mill "A" treated wastewaters. This is
probably due to the production of fatty acids by the
microbiota in the aerated lagoons. An increase in number of
branched and odd-carbon fatty acids was also noted.
To maintain the proper perspective with respect to a mass
balance, the total volatile acidic material as well as the
three major classes of compounds comprising it is presented
as a percentage of the TOC in Table 6. The total of all the
volatile acidic components in the wastewater extracts was
less than 10% of the total organic carbon content of the
wastewaters. The neutral components, discussed in the next
section, are usually less than 1% of the TOC. The
69
-------
Table 5
COLLECTIVE POLLUTION PARAMETER MEASUREMENTS AND TOTAL CONCENTRATIONS OF THE VOLATILE
COMPONENTS IN THE ACID-PHENOL EXTRACTS
CONCENTRATIONS (mg/1)
BOD5
TOC
Total GC Organics
Total Phenols
Total Fatty Acids
Total Resin Acids
Mill "A"
1972
Raw
Outfall
*
Change
1974
Raw
Outfall
*
Change
Interstate Paper at Riceboro,
1972
Raw Outfall
%
Change
Georgia
1974
Raw
Outfall
*
Change
323
240
9.38
4.96
1.01
2.04
88
230
7.12
1.15
1.46
4.27
-73
- 4
-24
-77
+45
+109
320
350
14.98
2.31
1.36
10.77
45
350
3.81
0.62
1.59
1.39
-86
0
-75
-73
+17
-87
438
470
21.69
5.53
1.40
14.24
70
200
7.24
0.27
0.14
6.71
-84
-57
-67
-95
-90
-53
440
490
30.77
3.57
1.34
24.37
52
85
6.91
0.23
0.19
6.37
-88
-83
-78
-94
-86
-74
*Related to concentrations of total gas chromatographable organic material.
-------
Table 6
VOLATILE ACIDIC COMPONENTS AS PERCENTAGES OF TOC
PERCENTAGE OF TOC
Total
Total
Total
Total
COMPONENT
Acidic Volatiles
Phenols
Fatty Acids
Resin Acids
Mill A -
Raw Effluent
4.
0.
0.
3.
28
66
39
08
1974
Outfall
1.09
0.18
0.45
0.40
Interstate
Raw Effluent
6.28
0.73
0.27
4.97
- 1975 J
Outfall
8.13
0.27
0.22
7.49
-------
sura of all the volatile components therefore is still less
than 10% of the TOC. Yet in this minority of the mass of
dissolved organic material probably lies the bulk of problem-
causing compoundstoxic compounds and those causing taste
and odor. The dark brown color of kraft pulp mill wastewaters
is believed to be due to partially degraded lignin molecules.
These high molecular weight non-volatile compounds form a
significant portion of the balance of the TOC. Other
contributors to the TOC are the carbohydrates and, to a
lesser extent, tannins and various other highly polar or
non-volatile compounds.
The increase in the proportion of the TOC represented by
total volatiles and resin acids in the Interstate
wastewaters is a reflection of the greater decrease in the
non-volatile portion of the organic content as compared to
the volatile and resin acid content. Lime flocculation
probably removed a large amount of the non-volatile organics.
The major difference in the volatile organic content of
wastewaters from the two mills was the fatty acid content.
Whereas the fatty acids decreased significantly during the
Interstate treatment, they increased during treatment in
Mill "A".
Reduction in BOD, TOC, total GC peak areas, and phenolic
and resin acid content were similar in the wastewaters from
the two mills.
72
-------
SECTION VI
IDENTIFICATION OF TERPENES
The neutral volatile fraction of the paper mill wastewater
extracts was not subjected to as complete a characterization
as was the acidic and phenolic fraction. There were several
reasons for this decision:
Dr. Bjorn Hrutfiord, University of Washington, has
been conducting a similar study on an EPA grant for
the past 2 years and has concentrated primarily on
the terpene fraction and on a sugar fraction.42
Our results indicated that the terpenes and other
neutral volatiles are about one-tenth the
concentration of the acids and phenols in the raw
wastewaters.
The terpenes appear to be generally susceptible to
biological treatment and are minor constituents in
the treated wastewaters being discharged.
GC-MS ANALYSIS
Chemical profiles of the neutral volatiles from the extracts
of both mills are shown in Figures 31 and 32. Table 7 lists
the individual compounds corresponding to the peak numbers
in Figures 31 and 32. Since these samples were taken in
1972, they may not be quantitatively representative of a
slug of the effluent. The concentrations of compounds in
each sample are probably slightly low because extraction
efficiencies are never 100%. Several compounds were
detected and identified even though their concentrations
were less than 1 part per billion.
Gas chromatograms from the 1974 sampling of the raw
effluents and the treated effluents of both mills are shown
in Figures 33 and 34. No attempt was made to identify these
compounds because of lack of time. However, most of the GC
peaks have similar relative retention time and areas as
those from the 1972 neutral volatile extracts.
GC-IR ANALYSIS
Two of the 1972 neutral volatile extracts from Mill A were
analyzed by a Digilab Fourier Transform infrared spectro-
73
-------
Table 7. NEUTRAL VOLATILES IDENTIFIED IN BOTH KRAFT PAPER MILL EFFLUENTS WITH APPROXIMATE CONCENTRATIONS
Approximate
Cone, in mg/1
Peak
No.
4
62
63
64
65
66
67
68
69
70
71
72
73
9
10
11
74
75
76
77
78
79
80
81
30
82
83
84
Total
Compound Identified
Methyl trisulfide
Fenchone
Hexachloroethane
Sabinene
Unidentified terpene ketone
Camphor
Unidentified terpene ketone
Unidentified terpene ketone
Fechyl alcohol
Unidentified terpene ketone
Terpene- 4 -ol
2-Formy Ithiophene
Methyl chavicol
Borneol
a-Terpineol
Veratrole
2 -Acety Ithiophene
Myrtenol
2 -Propiony Ithiophene
Anethole
Benzyl alcohol
Methyl eugenol
Unidentified terpene alcohol
Unidentified aromatic similar
to methyl isoeugenol (MW=178)
Ethyl palmitate
Unidentified monounsaturated
CIQ fatty acid methyl ester
Unidentified diunsaturated C19
fatty acid methyl ester
Unidentified phthalate diester
Confirmed
By
MS.GC
MS,GC,IR
GC
MS.GC
MS,GC
MS,GC,IR
MS,GC
MS,GC
GC,IR
GC,IR
GC
MS,GC
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1
1
.003
.007
.055
.045
.045
.020
.065
.025
.050
.010
.045
.275
.645
.020
.025
.010
.025
.007
.013
.002
.006
.006
.006
.038
.016
.464
Mill "A"
Sample Points
2 3
0.004
0.007
0.050
0.045
0.040
0.020
0.065
0.025
0.045
0.040
0.200
0.700
0.015
0.025
0.010
0.025
0.012
0.001
0.007
0.008
_.
1.344
0.008
0.015
0.055
0.060
0.050
0.025
0.035
0.020
0.040
0.030
0.155
0.625
0.015
0.030
0.008
0.025
0.008
0.010
0.009
___
1.223
4
0.001
0.015
0.045
0.090
0.045
0.020
0.010
0.004
0.010
0.090
0.008
0.025
0.010
._
0.711
1
0.015
0.040
0.090
0.045
0.020
0.105
0.015
0.030
0.030
0.470
0.490
0.015
0.012
0.008
0.020
0.025
0.030
0.009
__
1.469
Approximate
Cone, in mg/1
Interstate
Sample Points
2 3
0.002
0.010
0.015
0.006
0.003
0.040
0.005
0.010
0.010
0.200
0.215
0.004
0.002
0.005
0.004
0.015
0.006
__
0.552
0.001
0.015
0.020
0.020
0.008
0.060
0.009
0.015
0.020
0.260
0.280
0.008
0.004
0.010
0.007
0.015
0.008
__
0.760
4
<0.001
<0.001
0.003
0.004
0.035
0.008
0.002
<0.001
0.080
<0.001
<0.001
0.008
__
0.145
-------
G-ln
0 5 10 15 20 25 30 35 40 45 50 55 60 65
Figure 31 Chemical profile of neutral volatiles from
Mill "A"; sample points 1-4
75
-------
I-In
'1'1
I-2n
\
I-3n
I-4n
Figure 32 Chemical profile of neutral volatiles from
the Interstate mill at Riceboro; sample
points 1-4
76
-------
X 16
Figure 33 Gas chromatogram of neutral volatile extract
from Mill "A's" raw effluent (A) and (B)
treated effluent
77
-------
10
20
30
40
50
60
B
X 16
A.
A
10
20
30
40
50
60
Figure 34 Gas chromatogram of neutral volatile extract
from Interstate's raw effluent (A) and (B)
treated effluent
78
-------
photometer interfaced with a gas chromatograph (GC-IR).
Spectra of the eluting compounds are obtained "on the fly"
as they are with the GC-MS system.
The gas chromatograms of the two samples analyzed by GC-IR
are shown in Figure 35. GKR-3 and GKR-4 correspond to G-3n
and G-tn, respectively, in Figure 31. A different peak
notation system was used in Figures 35-45. The peak numbers
in these figures correspond to the identifications in Table
6 in the following way:
GC-IR
Designation
13
14
15
16
17
18
19
20
21
22
23
24
a
b
c
d
e
Table 6
Designation
67
68
69-71
73
9,10
74
75
76
78
65
66
67
68
Compound Name
unidentified ketone
unidentified ketone
3 compounds
methyl chavicol
borneol and a-terpineol
2-acetylth.iophene
myrtenol
2-propionylthiophene
benzyl alcohol
unidentified ketone
camphor
unidentified ketone
unidentified ketone
79
-------
Figure 35 Gas chromatograms of Mill "A" neutral volatile
extracts analyzed by GC-IR; sample points 3-4
80
-------
The GC-IR spectra of the peaks in Figure 35 are shown in
Figures 36-41.
Although the exact structures of peaks 11,13, and 14 (a,d,
and e in GKR-4) were not identified from the GC-IR spectra
(Figs. 38 and 41), they were all ketones. Computer matching
of the mass spectra of these same peaks had led to tentative
identifications of 1-cyclohexenyl methyl ketone, 4-nonyne,
and 3-cyclohexen-1-yl methyl ketone, respectively. The
presence of ketone carbonyl absorption in the infrared
spectra eliminates the possibility that one is 4-nonyne and
confirms the methyl alkyl ketone structures of all three
compounds.
Four of the GC-IR identifications were confirmed by
comparing the sample spectra with standards. The spectra of
camphor, borneol, 2-acetylthiophene, and 2-
propionylthiophene are shown in Figures 42-45 respectively
with the corresponding sample spectra. The two terpenes had
previously been confirmed by GC-MS but the two thiophene
isomers were confirmed only by GC-IR.
81
-------
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-------
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I I
Figure 41 GC-IR spectra of peaks a-e; sample point 4
with inset of the corresponding portion of
the gas chromatogram
87
-------
00
00
3500 3000 2500
2000
Cm -I
1500 1000
Figure 42 GC-IR spectrum of camphor and corresponding GC-IR spectrum from
sample peak
-------
00
vo
3000
2500
2000
-I
Figure 43 GC-IR spectrum of borneol and corresponding GC-IR spectrum from
sample peak
-------
2-ACETYLTHIOPHENE
_1_
GKR-3
PKN-19
3500 3000 2500 2000 1500 1000
Dm -'
Figure 44 GC-IR spectrum of 2-acetylthiophene and corresponding GC-IR
spectrum from sample peak
-------
'3500
3000
2500
2000
1500
1000
Figure 45
GC-IR spectrum of 2-propionylthlophene and corresponding GC-IR
spectrum from sample peak
-------
SECTION VII
REFERENCES
1. Keith, L. H. Identification of Organic Contaminants
Remaining in a Treated Kraft Paper Mill Effluent.
Southeast Environmental Research Laboratory.
)Presented at the 157th National Meeting of the
American Chemical Society, Division of Water, Air, and
Waste Chemistry. April 14-18r 1969.) 6 p.
2. Keith, L. H., A. W. Garrison, M. M. Walker, A. L.
Alford, and A. D. Thruston, Jr. The Role of Nuclear
Magnetic Resonance Spectroscopy and Mass Spectrometry
in Water Pollution Analysis. Southeast Environmental
Research Laboratory. (Presented at the 158th National
Meeting of the American Chemical Society, Division of
Water, Air, and Waste Chemistry. New York. September
8-12, 1969.) 4 p.
3. Garrison, A. W., L. H. Keith, and M. M. Walker. The
Use of Mass Spectrometry in the Identification of
Organic Contaminants in Water from the Kraft Paper Mill
Industry. Southeast Environmental Research Laboratory.
(Presented at the 18th Annual Conference on Mass
Spectrometry and Allied Topics. San Francisco. June
14-19, 1970.) 9 p.
4. Keith, L. H. , arid S. H. Hercules. Environmental
Applications of Advanced Instrumental Analyses:
Assistance Projects, FY 69-71. Environmental
Protection Agency. Washington, D.C. Publication
Number EPA-R2-73-155. May 1974. p. 50-63.
5. Alford, A. L. Environmental Applications of Advanced
Instrumental Analyses: Assistance Projects, FY 72.
Environmental Protection Agency. Washington, D.C.
Publication Number EPA-660/2-73-013. September 1973.
p. 27-31.
6. Adams, B. H., H. C. Vick, R. P- Lawless, T. B. Bennett,
Jr., W. Hanks, and J. Shailer. Supplement to Effects
of Pollution on Water Quality, Perdido River and Bay,
Alabama and Florida. Environmental Protection Agency,
Southeast Water Laboratory Technical Service Program,
Athens, Georgia. (1971). p. 14-17.
7. Hill, J. S., and B. H. Adams. Wastewater Survey St.
Regis Paper Company, Cantonment, Florida. Environmental
Protection Agency,
92
-------
Surveillance and Analysis Division, Athens, Georgia,
(1972). p. 29-30.
8. Private communication with Mr. Lawrence Wapensky, EPA,
Denver Federal Center, Denver, Colorado (1973).
9. Webb, R. G., A. W. Garrison, L. H. Keith, and J. M.
McGuire. Current Practice in GC-MS Analysis of
Organics in Water. Environmental Protection Agency.
Washington, D.C. Publication Number EPA-R2-73-277.
August 1973. p. 48-54.
10. Davis, C. L. Color Removal from Kraft Pulping Effluent
by Lime Addition. Environmental Protection Agency.
Washington, D.C. Publication Number 12040 ENC.
December 1971. p. iii.
11. Environmental Protection Agency Technology. Color
Removal from Kraft Pulping Effluent by Lime Addition.
Technology Transfer Capsule Report 2.
12. Dostal, K. A., R. C. Peerson, D. G. Hager, and G. G.
Robeck. Carbon Bed Design Criteria Study at Nitro, W.
Va. J. Am. Water Works Assoc. 57; 663-674, 1965.
13. Bishop, D. F., L. S. Marshall, T. P. O'Farrell, R. B.
Dean, B. ©Connor, R. A. Dobbs, S. H. Griggs, and R. V.
Villiers. Studies on Activated carbon Treatment. J.
Water Poll. Contr. Fed. 39: 188-203, 1967.
14. Booth, R. L., J. N. English, and G. N. McDermott.
Evaluation of Sampling conditions in the Carbon
Adsorption Method. J. Am. Water Works Assoc. 57:
215-220, 1965.
15. Webb, R. G., A. W. Garrison, L. H. Keith, and J. M.
McGuire. Current Practice in GC-MS Analysis of
Organics in Water. Environmental Protection Agency.
Washington, D.C. Publication Number EPA-R2-73-277.
August 1973. p. 88.
16. Webb, R. G., A. W. Garrison, L. H. Keith, and J. M.
McGuire. Current Practice in GC-MS Analysis of
Organics in Water. Environmental Protection Agency.
Washington, D.C. Publication Number EPA-R2-73-277.
August 1973. p. 90.
17. Bicho, J. G., E. Zavarin, and D. L. Brink. Oxidative
Degradation of Wood II. TAPPI. 49: 218-226, 1966.
93
-------
18. Brink, D. L., Y. T. Wu, H. P. Noveau, J. G. Bicho, and
M. M. Merriman. Oxidative Degradation of Wood IV.
TAPPI. 55: 719-721, 1972.
19. McGuire, J. M., A. L. Alford, and M. H. Carter.
Organic Pollutant Identification Utilizing Mass
Spectrometry- Environmental Protection Agency.
Washington, D.C. Publication Number EPA-R2-73-234.
July 1973. p. 10-13.
20. Hoyland, J. R., and M, B. Neher. Implementation of a
Computer-Based Information System for Mass Spectral
Identification. Environmental Protection Agency.
Washington, D.C. Publication Number EPA-660/2-74-048.
June 1974. p. 5-33.
21. Ryhag, R., and E. Stenhagen. Mass Spectrometry of
Long-Chain Esters. In: Mass Spectrometry of Organic
Ions. McLafferty, F. W. (ed.). New York, Academic
Press, 1963. Chapter 9, p. 399-443.
22. Hertz, H. S., R. A. Kites, and K. Biemann. Anal. Chem.
43: 681, 1971.
23. Hoyland, J. R., and M. B. Neher. Implementation of a
Computer-Based Information System for Mass Spectral
Identification. Environmental Protection Agency.
Washington, D.C. Publication Number EPA-660/2-74-048.
June 1974. p. 1.
24. Mutton, D. B. Wood Resin. In: Wood Extractives and
Their Significance to the Pulp and Paper Industries.
Hillis, W. E. (ed.) New York, Academic Press, 1962.
Chapter 10. p. 331-360.
25. Holmbom, B., and E. Avela. Studies on Tall oil From
Pine and Birch. Acta Academiae Aboensis. Ser. B. 31_;
(13): 1-14, 1971.
26. McKee, J. E., and H. W. Wolfe. Water Quality Criteria.
California State Water Resources Control Board.
Sacramento. Publication Number 3-A. Second Edition.
1963. p. 187.
27. Leach, J. M., and A. N. Thakore. Identification of the
constituents of Kraft Pulping Effluent That Are Toxic
to Juvenil Coho Salmon. J. Fish. Research Brd.
Canada. 30: 479, 1973.
94
-------
28. Odham, G. , and E. Stenhagen. Fatty Acids. In:
Biochemical Applications of Mass Spectrometry, Waller,
G. (ed.). New York, Wiley-Interscience, 1972. Chapter
8. p. 211-228.
29. Rogers, I. H. Secondary Treatment of Kraft Mill
Effluents: Isolation and Identification of Fish-Toxic
Compounds and Their Sublethal Effects. Pulp and Paper
Magazine of Canada. 74: T303-T308, 1973.
30. Van Horn, W. M., J. B. Anderson, and M. Katz. The
Effect of Kraft Paper Mill Waters on Fish Life. Tappi.
33: 209-212, 1950.
31. Maenpaa, R., P. Hynninen, and J. Tikka. On the
Occurrence of Abietic and Pimaric Acid Type Resin Acids
in the Effluents of Sulphite and Sulphate Pulp Mills.
Pap. ja pun. 50: 143-150, 1968.
32. Hagman, N. Resin Acids in Fish Mortality. Finnish
Paper and Timber J. .18: 32-34, 40-41, 1938.
33. Ebeling, G. Recent Results of the Chemical
Investigation of the Effect of Wastewaters from
Cellulose Plants on Fish. Vom Wasser. 5: 192-200,
1931. (C. A. 36: 2262).
34. Leach, J. M., and A. N. Thakore. Identification of the
Constituents of Kraft Pulping Effluent That Are Toxic
to Juvenile Coho Salmon (Oncorkynchus kisutch). J.
Fisheries Res. Brd. Canada. 30: 479-484, 1973.
35. Zinkel, D. F., L. C. Zank, and M. F. Wesolowski.
Diterpene Resin Acids. U. S. Department of
Agriculture. Forest Service. Madison, Wise. Forest
Products Laboratory. Washington, B.C. 1971. p. C1-
C32.
36. Chang, T., T. E. Mead, and D. F. Ziabel. Mass Spectra
of Diterpene Resin Acid Methyl Esters. J. Am. Oil
Chem. Soc. 48: 455-461, 1971.
37. Chopin, J. Phenolic Substances in Pulp Mill Effluents.
J. Bull. Ass. Tech. Ind. Pap., No. 3; 147-155, 1959.
38. McKee, J. E., and H. W. Wolf. Water Quality Criteria.
California State Water Resources Control Board.
Sacramento. Publication Number 3-A. Second Edition.
1963. p. 238.
95
-------
39. Shumway, D. L., and J. R. Palensky. Impairment of the
Flavor of Fish by Water Pollutants. Environmental
Protection Agency. Washington, B.C. Publication
Number EPA-R3-73-010. February 1973. p. 68-69.
40. Shumway, D. C.r and G. G. Chadwick. Influence of Kraft
Mill Effluent on the Flavor of Salmon Flesh. Wat. Res.
5: 997-1003, 1971.
41. Rogers, I. H., and L. H. Keith. Organochlorine
Compounds in Kraft Bleaching Wastes. Identification of
Two Chlorinated Guaiacols. Environment Canada,
Fisheries and Marine Service Technical Report Number
465. September 1974. p. 8-17.
42. Hrutfiord, B. J. Organic Compounds in Pulp Mill Lagoon
Discharge. Environmental Protection Agency
Publication. In Press.
96
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SECTION VIII
APPENDICES
£§36
A. Procedure for diazomethane methylation 98
B. Procedure for dimethyl sulfate methylation 99
97
-------
APPENDIX A
Procedure for diazomethane methylation
The apparatus is shown in Figure 6.
1. Evaporate the sample extract just to dryness with a
stream of nitrogen in a centrifuge tube, the bottom
tube of a Kuderna-Danish apparatus, or the sample
storage vial. A small amount of methylene chloride may
be retained, but the presence of chloroform may produce
artifacts. Dissolve the extract in 0.5-1.0 ml
distilled-in-glass ethyl ether.
2. Add about 5 ml of distilled-in-glass ether to the first
tube of the apparatus to saturate the nitrogen carrier
gas with ether. Add 0.7 ml of carbitol [2-(2-
ethoxyethoxy)ethanol], 1.0 ml of 31% aqueous KOH (not
over 2 days old), and 0.1-0.2 g of N-methyl-N-nitroso-
p-toluenesulfonamide ("Diazald," Aldrich Chemical Co.)
to the second tube. The base immediately begins to
release diazomethane from the sulfonamide.
3. Immediately position the second test tube and adjust
the nitrogen flow to about 10 ml per minute. Caution;
Diazomethane is an extremely toxic and explosive gas.
A good fume hood and safety glasses are mandatory. No
chipped glassware should be used, as rough glass
surfaces catalyze decomposition of diazomethane.
4. Position the third tube (a safety trap to prevent
reagent carry-over) and the sample tube to bubble the
nitrogen and diazomethane gas mixture through the
sample. Continue the reaction until the slight yellow
color of diazomethane persists in the sample solution
(from a few seconds to 30 minutes, depending upon the
sample concentration). In the case of dark colored
extracts in which the diazomethane is not visible, a
reaction time of 30 minutes is recommended.
5. Allow the esterified sample to stand unstoppered in the
hood for 15 to 30 minutes to allow excess diazomethane
to escape from the ether solution. Discard all waste
from the reaction with care and rinse the apparatus
with acetone. Evaporate the sample to the volume
necessary for gas chromatography.
98
-------
APPENDIX B
Procedure for dimethyl sulfate methylation
The apparatus is shown in Figure 7.
1. Bring the original sample (300 ml) to pH 11 with NaOH
and extract with chloroform to remove neutral and basic
compounds.
2. A 500-ml 3-neck (standard taper 24/40) round bottom
flask, equipped with a fourth neck for a thermometer,
is fitted with two pressure-equalizing addition
funnels, the probe of a single-probe pH meter, and a
magnetic stirrer.
3. Nitrogen is introduced into the top of the first
addition funnel and exits from the top of the second
one. Place forty ml of Eastman reagent grade dimethyl
sulfate into the first addition funnel and a SOX
solution of sodium hydroxide (80 ml) into the second.
4. Pour the sample into the flask and flush the system
with nitrogen.
5. After raising the temperature to 85° C, begin dropwise
addition of both the dimethylsulfate and the sodium
hydroxide solution. Maintain temperature between 80
and 90° C. (Cautionexothermic reaction. Have ice
available to add to water bath.) and the pH between
10.5 and 11. Since dimethylsulfate is not readily
soluble in water vigorous stirring must be used. The
addition time is about 1 hour.
6. After all the dimethylsulfate is added, maintain the
reaction vessel at 85-90° C for an additional 15-20
minutes and then cool to room temperature.
7. Add 5 ml concentrated ammonium hydroxide to destroy
excess dimethylsulfate and re-extract the reaction
mixture with chloroform to remove the methyl esters of
acids and the methyl ethers of phenols.
8. Dry the chloroform extract and evaporate to the
appropriate volume for GC analysis.
99
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TECHNICAL REPORT DATA .
(Please read Instructions on the reverse before completing)
REPORT NO.
EPA-660/4-75-005
3. RECIPIENT'S ACCESSION-NO.
4. TITLE AND SUBTITLE
Analysis of Organic Compounds in Two Kraft
Mill Wastewaters
B. REPORT DATE
April, 1975
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Lawrence H. Keith
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORG \NIZATION NAME AND ADDRESS
Analytical Chemistry Branch
Southeast Environmental Research Laboratory
Athens, GA 30601
10. PROGRAM ELEMENT NO.
1BA027
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCV NAME AND ADDRESS
Environmental Protection Agency
Southeast Environmental Research Laboratory
College Station Road
13. TYPE OF REPORT AND PERIOD COVERED
Final, FY67-74
14. SPONSORING AGENCY CODE
Athens, GA
30601
15. SUPPLEMENTARY NOTES
Prepared in fulfillment of ROAP 07ABL-Tasks 02, 03
16. ABSTRACT
Wastewaters from two kraft paper mills in Georgia were sampled at
various points in the waste treatment systems. Gas chromatography of
the organic extracts and identification of many of the specific chemical
components by gas chromatography-mass spectrometry provided a "chemical
profile" of the effluents. The mills, in different geographical
locations, have very similar raw wastewater compositions but different
wastewater treatments. In spite of these differences, the treated
effluents are qualitatively similar in composition although the
quantities of the various components differ. After two years the raw
and treated effluents of both mills were re-sampled. Analyses showed
that although concentrations of the organics varied, the same compounds
are still present. This report was submitted in ^fulfillment of ROAP
07ABL, Tasks 02 and 03 by SERL, Athens, Georgia. Work was completed as
of April 1974.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS
c. cos AT I Field/Group
Pollutant identification*
Mass spectrometry*
Gas chromatography*
Organic compounds*
Water pollution sources*
paper mill pollution
infrared spectroscop
fatty acids
resin acids
phenols
18. DISTRIBUTION STATEMENT
Release Unlimited
19. SECURITY CLASS (ThisReport)'
21. NO. OF PAGES
109
20. SECURITY CLASS (Thispage)
22. PRICE
EPA Form 2220-1 (9-73)
ft U. S. GOVERNMENT PRINTING OFFICE: 1975-699,183 /32 REGION 10
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