EPA-R2-73-155
MAY 1973              Environmental Protection Technology Series
Environmental  Applications of
Advanced Instrumental Analyses
Assistance Projects,  FY 69-71
                       ^ PRO^°
                               Office of Research and Monitoring
                               U.S. Environmental Protection Agency
                               Washington, D.C. 20460

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            RESEARCH REPORTING SERIES
Research reports of the  Office  of  Research  and
Monitoring,  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
PROTECTION   TECHNOLOGY   series.    This   series
describes   research   performed  to  develop  and
demonstrate   instrumentation,    equipment    and
methodology  to  repair  or  prevent environmental
degradation from point and  non-point  sources  of
pollution.  This work provides the new or improved
technology  required for the control and treatment
of pollution sources to meet environmental quality
standards.

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                                             EPA-R2-73-155
                                             May,  1973
ENVIRONMENTAL APPLICATIONS  OF ADVANCED INSTRUMENTAL
       ANALYSES:   ASSISTANCE PROJECTS,  FY 69-71
                           by
       Lawrence  H.  Keith and  Shirley H.  Hercules
      Southeast  Environmental Research Laboratory
                  College Station Road
                 Athens, Georgia  30601
                    Project  16020-GHZ
                 Program Element 1B1027
        NATIONAL ENVIRONMENTAL RESEARCH  CENTER
           OFFICE  OF RESEARCH  AND MONITORING
         U.  S. ENVIRONMENTAL PROTECTION  AGENCY
               CORVALLIS, OREGON  97330
    For sale by the Superintendent of Documents, U.S. Government Printing Office, Washington, B.C. 20402
                 Price $1.25 domestic postpaid or $1 GPO Bookstore

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                      ABSTRACT
A multitude of analyses involving the identification
and measurements of organic pollutants in water are
discussed under eleven project categories.  In most
cases these analyses have helped to solve, or at least
understand more clearly, the related pollution incident
and in some cases provided evidence for enforcement of
regulatory legislation.
                            11

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                         CONTENTS

                                                   Page

Abstract                                           ii

List of Figures                                    v

List of Tables                                     viii

Acknowledgments                                    x

Sections

I      Conclusions                                 1

II     Recommendations                             3

III    Introduction                                5

IV     Projects

       1   Tennessee River Contaminant  (bis 12-     7
           ethyIhexyl]phthaiate)

       2   Fish Kill Contaminant Characteriza-     10
           tions  (pesticides)

       3   Confirmation of PCB's in Environmental  20
           Samples

       4   Escambia Bay, Florida, Industrial       23
           Pollution

       5   Kansas City Landfill Contaminants       28

       6   Identification of the Cause of Earthy-  29
           Musty Taste and Odor Problems in Ohio
           Water Supplies

       7   Western Louisiana Industrial Waste      35
           Survey

       8   Color Body Characterization and         50
           Related Studies from a Kraft Paper
           Mill Effluent

       9   Characterization of Foam from Kraft     59
           Pulp Mills
                            111

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                                                   Page
        10   Analysis of Oil Spills by NMR         63
             Spectroscopy

        11   Analysis of Oil Spills by Fluores-    72
             cence Spectroscopy

V       References                                 81
                             IV

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                           FIGURES

 No.                                                 Page

 1   Infrared Spectrum of the Tennessee River
     Sample                                           8

 2   Mass Spectrum (top)  and NMR Spectrum (bottom)
     of Tennessee River Sample                        9

 3   Comparison of the Infrared Spectrum of the
     Tennessee River Sample with that of
     bis(2-ethylhexyl)phthalate                      11

 4   Comparison of the NMR Spectrum of the
     Tennessee River Sample with that of
     bis (2-ethylhexyl)phthalate                      12

 5   Comparison of the Mass Spectrum of the
     Tennessee River Sample with that of
     bis(2-ethylhexyl)phthalate                      13

 6   Comparison of the Mass Spectrum of Boone Lake
     Extract with that of Phenylmercuric Chloride    14

 7   Comparison of the Mass Spectrum of Tombigbee
     River Extract with that of Diazinon             17

 8   Comparison of the Mass Spectra of the Locust
     Fork Branch and Black Warrior River Extracts
     with those of Malathion Obtained by GC-MS
     and by Direct Probe Insertion                   18

 9   Comparison of the FID Gas Chromatogram of
     Escambia Bay Sediment Extract with that of
     Aroclor 1254                                    21

10   Comparison of Select Chlorine Isotope Clusters
     from Escambia Bay Sediment Extract with that
     from Corresponding PCB's in Aroclor 1254        22

11   Comparison of the FID Gas Chromatogram of
     Great Miami River, Ohio Extract with that
     of Aroclor 1248                                 24

12   Comparison of Select Chlorine Isotope Clusters
     from Great Miami River, Ohio Extract with that
     from Corresponding PCB's in Aroclor 1248        25
                              v

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No.                                                page

13    The NMR Spectrum of  "Celinol"                   31

14    Comparison  of  the Mass  Spectrum of  "Celinol"
      with  that of Isoborneol                        32

15    Comparison  of  the Infrared  Spectrum of
      "Celinol" with that  of  2-Methylisoborneol       33
*
16    Comparison  of  the Mass  Spectrum of  Indian
      Lake  Extract with that  of Geosmin               34

17    Chemically  Characterized Gas Chromatogram
      of Petrochemical Company "A"                    37

18    Chemically  Characterized Gas Chromatogram
      of Petrochemical Company "B"                    41

19    Chemically  Characterized Gas Chromatogram
      of Petrorefinery "A"                            43

20    Chemically  Characterized Gas Chromatogram
      of Petrorefinery Company "B"                    46

21    Chemically  Characterized Gas Chromatogram
      of Synthetic Rubber  Company                    48

22    Chemically  Characterized Gas Chromatogram
      of Chemical Company  "A"                         51

23    Chemically  Characterized Gas Chromatogram
      of Chemical Company  "B"                         52

24    Comparison  of  Kraft  Pulp Mill Effluent  and
      Marsh Water Color Intensity as  a Function of
      pH                                              54

25    Chemically  Characterized Gas Chromatogram of
      Foam  from a Kraft Papermill Bleach  Stage
      Caustic Clarifier                              62

26    Comparison  of  NMR Spectra of Oil from the
      Suspect Barge  and from  a Florida Beach          64

27    Comparison  of  NMR Spectra of Oil from
      Duluth Harbor  and the Primary Suspect Source    66
                            VI

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No.
28   Comparison of NMR Spectra of Oil from
     Duluth Harbor and the Secondary Suspect
     Source                                         67
                            VII

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                        TABLES

No.                                                Page

 1   Compounds Identified in the Escambia Bay,
     Florida Industrial Waste Survey                26

 2   Compounds Identified in the Kansas City,
     Kansas Landfill Analysis                       27

 3   Chemical Characterization and Gross Pollution
     Measurements of Petrochemical Company A        38

 4   Gross Pollution Measurements and Chemical
     Characterization of Petrochemical Company B    42

 5   Gross Pollution Measurements and Chemical
     Characterization of Petrorefinery Company
     "A"                                            44

 6   Gross Pollution Measurements and Chemical
     Characterizations of Petrorefinery Company
     "B"                                            47

 7   Gross Pollution Measurements and Chemical
     Characterization of Synthetic Rubber Company   49

 8   Chemical Characterization of Chemical Company
     A                                              53

 9   Chemical Characterization of Chemical Company
     B                                              53

10   Comparison of Kraft Pulp Mill Effluent and
     Marsh Water Fluorescent Spectra                55

11   Color Leached from Lagoon Mud as a Function
     of Calcium Hydroxide Concentration             56

12   Comparison of the Amounts of Acid Insoluble
     Organics in the Lime Treated and the Untreated
     Effluent Samples                               57

13   Compounds Tentatively Identified in the
     Neutral Volatiles Fraction of the Riceboro
     Pulp Mill Effluent                             58
                           Vlll

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No.                                                Paqe
14   Compounds Tentatively Identified in the
     Phenols and Acids Fraction of the Riceboro
     Pulp Mill Effluent                             58

15   Compounds Identified in Perdido Bay Kraft
     Papermill Wastewater, Foam, and Scum Samples   60

16   Normalized Peak Areas of Proton Signals in
     Samples from the St. Mark/ Florida, Oil
     Spill                                          65

17   NMR Comparison of Oil from West Florida
     Beaches with Suspect Crude Oil and Bunker C
     Oil                                            70

18   Fluorescence Spectra of Duluth Harbor Samples  73

19   Fluorescence Spectra of Savannah River
     Samples                                        74

20   Fluorescence Spectra of the Duluth Superior
     Harbor Samples                                 76

21   Fluorescence Spectra of the Dundee Canal
     Samples                                        77

22   Fluorescence Spectra of St. John's River
     Samples                                        79

23   Other Analyses of St. John's River Samples     80
                            IX

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                      ACKNOWLEDGMENTS
The following NWCCRP staff were the principal investi-
gators of the projects listed in Section IV:

     Project

       1.     L. H. Keith and A. W. Garrison
       2.     A. W. Garrison and A. D. Thruston, Jr.
       3.     A. W. Garrison
       4.     A. W. Garrison
       5.     R. G. Webb
       6.     A. W. Garrison and L. H. Keith
       7.     L. H. Keith
       8.     C. G. Gustafson
       9.     L. H. Keith
      10.     L. H. Keith
      11.     A. D. Thruston, Jr.

The assistance of A. L. Alford, T. L. Floyd, F. R.
Allen, A. C. McCall, and M. M. Walker in preparing the
samples for analysis, providing spectral data, inter-
pretation of spectra, and confirmation with standards
is gratefully acknowledged.
                           x

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                      SECTION I
                     CONCLUSIONS
The identification and quantification of specific
chemical compounds responsible for a wide variety of
pollution incidents have helped to solve, or at least
to understand more clearly, the problems associated
with the incidents.  In some cases evidence was provided
for enforcement of regulatory legislation.  The projects
have demonstrated the applicability of newly developed
laboratory techniques to environmental pollution
problems.

Because of the complexity and small size of the samples
available, gas chromatography-mass spectrometry has
been the most valuable technique for analysis.  Nuclear
magnetic resonance and infrared spectroscopy were
helpful when a single compound was isolated in sufficient
quantity.

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                     SECTION II
                   RECOMMENDATIONS
Analyses of the type described will be necessary and
should be used for establishing and enforcing water
quality effluent standards for the industrial permit
program.  The data provided will also be needed to
update these standards with respect to specific
organic compounds discharged into receiving waters.
Many more laboratories, including those at the state
level, should be equipped to conduct the type of
analyses described herein.

Newly developed techniques should be investigated for
their applicability to pollution analysis.

An annual report of unpublished contaminants charac-
terization work is recommended.

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                     SECTION III


                    INTRODUCTION
The National Water Contaminants Characterization
Research Program (NWCCRP) develops methods for the
identification and quantification of chemical pollu-
tants and identifies specific compounds associated with
various sources of pollution.  Information thus
obtained may be used in the development of regulatory
legislation.  In assessing the applicability of new
techniques to environmental samples and developing
information that the NWCCRP has a unique capability
for providing, a number of samples are analyzed that
are directly related to a wide range of environmental
problems.

Results of the aforementioned analyses are reported in
individual memoranda to the person requesting the
analysis and therefore receive limited distribution.
To acquaint other researchers and administrators with
the type of information that can be provided and to
help analytical chemists recognize the broad applica-
bility of the techniques used, a series of annual
reports is planned.  Brief summaries will be presented
of the problems under study by the NWCCRP.  This first
report, summarizing the problems studied during the
fiscal years 1968 through 1971, deals mostly with the
application of gas chromatography-mass spectrometry
since the NWCCRP was the only group in the Federal
Water Quality Administration with a GC-MS capability
during this period.  Future reports will discuss
applications for a wider variety of techniques.

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                     SECTION IV



                      PROJECTS
           1.  Tennessee River Contaminant
             (bis [2-ethylhexyl] phthalate)
When the Ohio River is at flood stage, the Tennessee
River dam above Calvert City, Kentucky, is closed,
releasing little or no flow down the Tennessee River.
During the spring of 1968 the dam was closed and the
industries on the lower Tennessee River complained
about the pooling of industrial wastes under the
stagnant river condition.

Samples of the contaminated water were taken by
personnel at the Robert A. Taft Sanitary Engineering
Center, Cincinnati, Ohio.  Analysis by flame ionization
gas chromatography revealed the presence of one
compound whose concentration far exceeded that of all
others.  IR and GC of chloroform extracts suggested
"the presence of a phthalate very similar to dinonyl
phthalate" (1).  An extract was submitted to the NWCCRP
for molecular structural identification.

Both thin layer and gas chromatography indicated that
the extract contained only one major component in about
90% purity.  The NMR, infrared, and mass spectra were
therefore recorded with no further purification.  The
infrared spectrum (Figure 1) shows peaks characteristic
of a carbonyl, an ester, and an ortho substituted
phenyl  (2).  NMR and mass spectra (Figure 2) confirm
the presence of a phthalate and indicate it to be a
dioctyl phthalate ester.

Fifteen possible isomeric structures exist for dioctyl
phthalate, assuming the diester to be symmetrical.
Further analysis of the NMR peaks in conjunction with
the electronic integration yielded the structure
bis[2-ethylhexyl] phthalate  (I).

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00
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                       TENNESSEE  RIVER SAMPLE
                 4000
                          3500
                                  3000
                                          2500
                                                   2000   1800    1600  •  1400   1200    1000    800
                                                        FREQUENCY CfTl
                                                                    .-I
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                      Figure  1.   Infrared  Spectrum of  the  Tennessee  River Sample

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10O

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                                       49
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Figure  2.   Mass  Spectrum (top)  and NMR Spectrum (bottom)  of Tennessee River
              Sample

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Comparison of the NMR, IR, and mass spectra of the
sample with those of a standard confirmed the above
structure (Figures 3-5).

The results of the structure determination were
submitted to the Taft Center.
      2.  Fish Kill Contaminant Characterizations
                       (Pesticides)

Boone Lake

During the summer of 1968 a massive kill of approxi-
mately one million fish occurred in the Boone Lake
reservoir, Tennessee.  After eliminating all of the
more common causes of fish kills, the contents of some
nearly empty 55-gallon drums found floating on the
reservoir were investigated.  The metal drums were of
the kind commonly used to float docks and boathouses.

The manufacturer listed on one of the drum labels,
Buckman Laboratories, Inc., indicated that the drum may
have contained a mixture of phenylmercurie acetate and
2,4,6-trichlorophenol, a microorganism control product.
Subsequent analysis of the river water and drum residue
revealed the presence of the trichlorophenol and
diphenyl mercury, a decomposition product of the phenyl-
mercuric acetate.  A residue sample analyzed by mass
spectrometry using the direct probe inlet was shown to
contain phenylmercurie chloride (Figure 6, top).  How-
ever, recent work in the National Environmental Research
Center, Analytical Quality Control Laboratory of
Cincinnati, Ohio, indicates that the acetate is readily
converted to the chloride.  Confirmation of the
structure was obtained by comparison of the mass

                             10

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                                        WAVELENGTH
                                             rH U
                                             6 ~   7
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Figure 3.   Comparison of  the  Infrared Spectrum of  the Tennessee River  Sample
             with that of bis  (2-ethylhexyl)phthalate

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      Figure 4.   Comparison of the NMR Spectrum of the Tennessee  River Sample with
                 that of bis(2-ethylhexyl)phthalate

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       Figure 5.  Comparison of the Mass Spectrum of the Tennessee River Sample with
                  that of bis(2-ethylhexyl)phthalate

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EXTRACT  OF SAMPLE FROM BOONE LAKE, TENN.
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Figure  6.   Comparison of the Mass  Spectrum of  Boone Lake  Extract with  that of
              Phenylmercuric Chloride

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spectrum with that of a standard  (Figure 6, bottom).

Bioassay conducted by TVA personnel confirmed the
toxicity of the phenylmercury compound to  fish.
Analysis of water samples indicated the presence of
diphenylmercury at greater than acutely toxic levels,
establishing it as the primary cause of the fish kill.

Beginning October 7, 1968, all drums containing and
suspected of containing toxic materials were to be
removed from the area of the kill by personnel of the
Division of Reservoir Properties.  The year 1972 was
set by the Tennessee Valley Authority as a deadline
for the complete removal from the reservoir of all
drums used as flotation devices.  A subsequent fish
kill in 1969 prompted the suggestion that  attempts be
made to remove the drums before the suggested deadline.
Tombigbee River, Alabama

In 1968, a fish kill involving about 30 thousand fish
occurred on the Tombigbee River in Alabama.  Geigy
Chemical Corporation, a manufacturer of pesticides
among other products, was a suspected source of the
contamination that killed the fish.

Gas chromatography indicated the presence of the
organophosphorus pesticide, diazinon (II), in the
water samples taken from the area of the fish kill, but
more substantial evidence was required for,legal
prosecution.

                                  CH,
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Accordingly, the sample was chromatographed by TLC and
the adsorbent containing the spot with an Rf corres-
ponding to that of diazinon was  scraped  from the plate,
                         15

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A similar area of adsorbant was scraped from a blank
plate.  The residues were introduced into the MS by
direct probe; subtraction of the peaks present in the
spectrum of the blank from the peaks in the river
sample spectrum left a fragmentation pattern (Figure 7,
top) identical to that of a diazinon standard (Figure 1,
bottom) in the region above m/e 100.

A federal court  awarded the state damages in the
amount of the value of the fish killed.
Black Warrior River and Locust Fork Branch Fish Kills

Fish kills in the Black Warrior River occurred at the
Locust Fork Branch, near Birmingport, Alabama in
October, 1969, and again near Demopolis, Alabama in
September, 1970.  The 1969 kill involved 750 thousand
fish and the 1970 incident killed 8 thousand fish.
Both kills were suspected to have been caused by the
spraying of malathion (III) in conjunction with a U. S.
Corps of Engineers mosquito control program in this
area.
                    s
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extracts of both the Locust Fork Branch (Figure 8-A)
and the Black Warrior River area near Demopolis
(Figure 8-B).  The molecular ion (m/e 330) was not
observed in spectra of either of the samples or of the
standard when it was introduced from the gas chromato-
graphic column (Figure 8-C).  Instead, the fragment
(m/e 173) resulting from cleavage of (0130)2PS2*
was the largest significant ion.  A small parent ion
was found in a sample introduced by the direct probe
(Figure 8-D).  The fragmentation pattern of 8-D is
                             16

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Figure 8.
Comparison of the Mass Spectra of  the  Locust
Fork Branch and Black Warrior River  Extracts
with those of Malathion Obtained by  GC-MS
and by Direct Probe Insertion
                          18

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significantly different from 8-C  (primarily due to
different sample pressures in the ion source),
illustrating the necessity of comparing mass  spectra
of samples with standards under identical conditions.
The major fragmentations of malathion are indicated
in Figure 8-D.

The results of these analyses were used in a  brief
prepared by the Alabama Department of Conservation and
submitted to the Alabama State Attorney.
Yalobusha River—Mississippi

A kill of an estimated 15 tons of fish occurred in
April, 1971, in the Yalobusha River, Grenada, Mississ-
ippi.  The organic chemical responsible for the kill
had been tentatively identified by gas chromatography
as pentachlorophenol  (IV) .  Confirmation of the
structure was requested  for legal action.
                           OH
                           IV
The samples and  a  standard  of pentachlorophenol were
each methylated  with  diazomethane  and  analyzed by GC-MS.
Both retention time and mass spectral  analysis confirmed
the unknown to be  pentachlorophenol.   The mass spectrum
indicated  also the presence of  tetrachlorophenol isomer
in some of the samples.

The results of the structure determination were
submitted  to  the Office of  the  Attorney  General in
Mississippi for  prosecution.
                         19

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        3.   Confirmation of PCB's in Environmental Samples

PCB's (polychlorobiphenyls), manufactured by the chlorina-
tion of biphenyl, are distillation fractions containing
20 or more isomers.  They have been widely used in a
large variety of industrial and consumer products since
1929.  Manufactured throughout the world, PCB's are sold
in the U. S. solely by Monsanto Company under the trade
name Aroclors(R).  Because of their resistance to
chemical breakdown, PCB's have become ubiquitous pollu-
ters of the environment  (4).

Only recently has the cumulative chronic toxicity of
the chlorinated biphenyls to animal life been observed.
It has therefore become important to develop methods
for their detection and identification in environmental
samples.

Mass spectrometry has been used to confirm the presence
of PCB's, tentatively identified by electron capture
GC, in water and mud samples.  The flame detector
chromatograms of the extracts are invariably compli-
cated, but the specificity of the mass spectrometer
allows unequivocable identification of submicrogram
amounts of PCB's.
Escambia Bay, Florida

In one case an Escambia Bay, Florida, sediment extract
cleaned on a Florisil column was analyzed for Aroclor
1254.  The GC flame detector pattern is shown in
Figure 9.  Eleven of the peaks were shown by MS to be
due to PCB's.  Their retention times and chlorine
numbers (number of chlorine atoms per PCB molecule, as
determined by MS) correspond with those of an Aroclor
1254 standard, also shown in Figure 9.

Mass spectra showing several chlorine isotope clusters
for some of the PCB's from the Escambia Bay sediment
are compared with corresponding spectra from an Aroclor
1254 standard in Figure 10.  The parent ion was
observed in all the PCB's.  The major fragmentation
path is loss of successive chlorine atoms from the
parent molecule.  Each such loss gives a fairly
definitive cluster of isotope peaks whose intensity
                             20

-------
                PCBs- SEDIMENT EXTRACT
                         CHLORINE
                         NUMBERS
                                            1
                                                •50
                                                40
                                                •30
                                                20
                                    • in *°
                                    IU m

                                       8
                                       TO
                                       O
                                       Om
                                       70
                  AROCLOR 1254

                            5
                       E  K
                 665
                                       I
                                     30
                                                 20
                                                 10
         14    12
       10
 8     6
- TIME, min.
4
Figure  9.
Comparison of the FID  Gas  Chromatogram  of
EsGambia  Bay Sediment  Extract with that of
Aroclor 1254.
                           21

-------
                           PCB's-SEDIMENT EXTRACT

100


80
£
I"
> 40
<
S
20

M*4C1
2CI












3CI
(M*-CI)




I





100


80
£
j"
— 40
5
DC
20

4CI


2CI 3CI








200 22O 24O Z6O 290 30O 20O 2
















100


M*.SCI go







20 240 260 28O XO 32O
§
|60
^40
i

20

34O 2-
M*6CI
I
I


4CI
SCI
SCI




1O 260 2UU 3OO 320 34O 360
to
to
PCB's-AROCLOR 1254

KM


80
i
« "
;
« *
i
20

M*,4CI

2CI
(M*-2CI>










SCI
(M*-CI)







|
100


80
|
5 60
g
5 *"
te
2C

4CI





2CI





SCI







|
100


M*SCI _ ^






1
|
»-
z
» 60
S
5 40
i
20
i
M*,6CI

!


4CI !
SCI ,
r
; L
,, ! i!!<
i s t>' pi
j 1 , 1 s i
1
)

Figure 10.   Comparison  of Select Chlorine Isotope clusters from Escambia Bay
            Sediment  Extract with that from Corresponding PCB's in Aroclor 1254

-------
ratios depend upon the number of chlorine atoms left
on the fragment.  A comparison of the sample and
Aroclor standard spectra  (Figure 10) shows similar
isotope ratios and fragmentation patterns.
Gre at Mi ami Rive r, Ohi o

Figure 11 shows the GC flame detector chromatogram of
a water extract from the Great Miami River in Ohio.
Mass spectra of the components eluting from the GC
showed 15 of them to be PCB's.  They appear to come
from a mixture of two Aroclors.  The retention time
and chlorine numbers of the first eleven of these
correspond to an Aroclor 1248 standard, also shown in
Figure 11.  However, the last four PCB peaks with
chlorine numbers of 5, 6, and 7 appear to be from
Aroclor 1254  (cf. Figure 9).  These four components
are in relatively large concentrations, as in the 1254
standard, whereas the standard Aroclor 1248 contains
only trace amounts, if any, of 6-chlorine and 7-chlorine
PCB's.

Mass spectra showing the clearly definable chlorine
isotope clusters of some of the PCB's from the river
sample are compared with corresponding spectra from
an Aroclor 1248 standard in Figure 12.  Only 6 yg of
Aroclors were present in the total sample according to
electron capture GC analysis.  Approximately 0.1 yg
of an individual PCB gave a good spectrum.
   4.  Esoambia Bay3 Florida  Industrial Pollution

The polluted state of  the Escambia  River and Escambia
Bay near Pensacola, Florida,  was  suspected to be the
major cause of numerous  fish  kills  during 1969.  The
governor of Florida requested technical assistance in
evaluating pollution from sources on  the river.  The
NWCCRP was asked to characterize  three effluent
samples by GC-MS.

The nine compounds identified and confirmed by GC-MS
are listed in Table 1  along with  five that were
tentatively identified.  In addition, diphenyl ether,
which may be implicated  in odor problems and acrylo-
nitrile, were indicated  in the Monsanto effluent.
                         23

-------
         PCB's-OHIO RIVER WATER
                                               •70
                          CHLORINE
                          NUMBERS
                             TIME,min.
Figure 11.
Comparison  of  the FID Gas Chromatogram of
Great Miami River, Ohio Extract with that
of Aroclor  1248
                           24

-------
                               RGB's-OHIO RIVER WATER
            £40
                1(1
                       M«3CI
                    2CI
                        It-
                                 £60
                                 Z


                                 1»
                                 s
                                      *
                                   TOD	5St	JMT
                                                             3£l
                       |60
                                4CI
                                                                aaa  300 — aid 3*0
Ul
PCB's-AROCLOR 1248
1C)










M*,3tl '°°









a




w
t
* 60
i
£ 40
. S
s
**rt

JCI










no
M*,4CI




3CI


J 	 _
80
£
i«o
5
i"
s
20
L
M*

JCI


1 •)

' 1 4CI !
' ' l'
"
•- -trim 	 4tm— — imn '-' wf~ 'vet^
      Figure  12.   Comparison of  Select Chlorine Isotope  Clusters from Great Miami

                   River, Ohio Extract with that from  Corresponding PCB's  in Aroclor

                   1248

-------
                           Table 1

      Compounds Identified in the Escambia Bay, Florida
                   Industrial Waste Survey
     Compound
                             Structure
Identi f ication
  confirmed
a-Terpineol


Naphthalene


Diphenyl ether


Oibenzofuran


o-Nitrotoluene


p_-Nitrotoluene


Acrylonitrile


n-Hexadecane


n-Octadecane


n-Heptadecane


m-Nitrotoluene
2,4'-Dimethyldi-
  phenylsulfone
Dibromopropene
2,6-Di-t-butylcresol
                                                     Yes
                                                     Yes


                                                     Yes
                             CH =CH-C=N
                             C16H34
                             C18H38
                             C17H36
                          (Isomer not determined)
                          (Isomer not determined)
                                                     Yes


                                                      No
                                                      No
      No
      No
                              26

-------
                        Table 2


      Compounds Identified in the Kansas City. Kansas
                    Landfill Analysis
        Compound
Phenol




£-Cresol




g-Cresol




Methylnaphthalene




DimethyInaphthalene




l-Methyl-4-ethylbenzene




3-Methylbiphenyl




1-Indanone




Diethyl phthalate




Di-n-butyl phthalate




Di-2-ethyl-n-butyl phthalate




A variety of n-alkanes
        Structure
                                      ,OH
                     f"3- f«l
                     .  "1




               (6lJW^
               ^xS<0^Tsx'
                    O      '*«—.
CH3{CH2)nCM3
                          27

-------
             5.   Kansas City Landfill Contaminants

Many complaints were registered in June, 1969, by
citizens of Kansas City, Missouri, concerning a medi-
cinal or "iodine-like" taste in their city water.  An
investigation was undertaken to determine the source
of the contamination.  The Kansas City Water Supply
Department, monitoring organics at their intake by gas
chromatography, showed a heavy organic load in the
water.

A suspected source of contamination was a landfill for
industrial wastes, recently reopened upstream of the
city.  Colorimetric tests of extracts of earth and
fiberglass samples from the operation proved positive
for phenols (compounds that, when chlorinated during
water treatment, produce tastes similar to those
reported.  Results indicated, therefore, that leaching
of soil and other materials being bulldozed into the
river was the source of the problem.

For corrective action, more positive identification of
the contaminants was required.  Accordingly, samples
of fiberglass from the dump were leached for 30 minutes
in distilled water to simulate being pushed into the
river.  An ether extract of the resulting solution was
examined by GC-MS.  Positive identification was made
of phenol, p-cresol, and benzothiazole.

The Johnson County, Kansas, Water Treatment Plant
(Kansas City, Kansas) requested that a similar study
for potential pollutants be performed on yet another
landfill operation whose drainage could reach their water
intake.  Should a similar problem occur in their city,
such information would permit immediate action.

Among the industries served by the disposal service
were a petroleum products plant, a paint company, and
several home appliance companies.  Examination by GC-MS
of extracts of earth, water, and fiberglass indicated
the presence of the organic compounds in Table 2 in
addition to elemental sulfur.

The most interesting compounds from a taste and odor
standpoint are phenol, the cresols, and dimethylnaph-
thalene, which has an odor of burnt rubber and has been
                              28

-------
found in the effluent from a refinery's activated
sludge plant.  Phenol and o- and p-cresol were
confirmed by comparison of mass spectra of standards
and the sample.
 6.  Identification of the Cause of Earthy., Musty
    Taste and Odor Problems in Ohio Water Supplies

In March, 1968, the Advanced Waste Treatment Research
Laboratory in Cincinnati, Ohio, was requested to
conduct studies of an intermittent musty, earthy odor
in the Wabash River in Indiana.  An investigation
showed that the odor was present constantly in Grand
Lake, Ohio, the principle headwater of the Wabash
River.

In 1929 an earthy odor in water was attributed to
actinomycetes  (6) and recently a compound with a
similar odor, termed "geosmin", was isolated from
numerous actinomycetes cultures.  Its structure was
identified as trans-1ylO^-dimethyl-trans-g-decalol  (V) ,
(7,8).  However, GC retention indices showed the river
pollutant to be different from "geosmin".
                           V
Rosen, Mashni, and Safferman of the Advanced Waste
Treatment Research Laboratory of the NERC, Cincinnati,
Ohio, isolated from a culture of streptomyces lavendulae
a compound that matched the odor and gas chromatography
retention indexes of the odorous compound from the lake
and river samples.  It was shown to be different from
"geosmin" and was termed "celinol" after the city of
Celina, Ohio, which uses Grand Lake as a water supply.
                         29

-------
Infrared spectra indicated that the compound was an
aliphatic alcohol; high resolution mass spectra and
an element map showed a parent ion at mass 168 with
an indicated empirical formula of C-Q^QO.  The sample,
believed to have a structure similar to "geosmin",
was submitted to NWCCRP for further structural
elucidation.

The NMR spectrum (Figure 13) considered in light of
the molecular weight and empirical formula, indicated
an aliphatic bicyclic structure.  The mass spectrum of
"celinol" was compared with those of compounds
considered possible on the basis of the above evidence
—either a borane or a pinane.  The fragmentation
pattern was similar to that of isoborneol, but the
major fragments were 14 mass units greater (Figure 14)
indicative of an added methyl group, which is consistent
with the NMR data.  The extra methyl group, from NMR
evidence, was geminal to the hydroxyl, yielding the
structure 2-methylisoborneol (2-exo-hydroxy-2~methyl-
bornane).

2-Methylisoborneol had not been reported previously as
a naturally occurring compound and no commercial
standard was available.  We therefore prepared the
compound by two different methods:  1) reacting camphor
(VI)  with methyl magnesium bromide according to Zelinsky
(9) and 2) reacting camphor (VI) with methyl lithium
reagent.
                     I) CH3MgBr


                °    or 2) CHoLi
                             O
          VI                             VII
Spectral comparisons (NMR, IR, and MS) confirmed the
structure of "celinol" to be identical to 2-methyl-
isoborneol (VII).   Figure 15 shows the infrared compar-
ison.
                             30

-------
Figure 13.  The NMR Spectrum of "Celinol"

-------
100
90-

80
r 70~
I 60-
- 50-
> 40-
.•
-H 30-
^20-

10-






"CELINOL" GC INLET


















C3H5*
41

H20
1]
i '
'



0 ' 20







1 1
"T 1 i








,

,,l!
60




79 C^HQ4.





1
{
f T
93




|| [

T

ii
,
C8H11*
107




M-H20,CH3.
135
M-H20
C9H13* 150
121


1 i
0 ' 100 ' 120 140 160 '
                           m/e
100-
90
80
£70
M
g 60-
c
- 50-
|40J
D
^30-
at
20
10-

ISOBORNEOL











i
1
0 20 4


_^*
c
!-—•
MW:






1

, ll
•"_i — ' f"*1
0


A/
C
\
154









60


OH
YH





79




l .



l.

' 80















93


121




107
















M-HoO
136

139
., ..
100 120 140 160
m/e
Figure 14.
Comparison of the Mass Spectrum of  "Celinol"
with that of Isoborneol
                         32

-------
OJ
U)
                                     "CELINOL"
2- METHYLISOBORNEOL
              4OOO
                     350O
                            3OOO
                                          20OO
                                                WOO   1«00   1400
                                                   WAVENUMBER (CM"1)
                                                                "So"
                                                                       1OOO
                                                                                        400
       Figure 15.   Comparison  of the  Infrared Spectrum of "Celinol" with that of
                     2-Methylisoborneol

-------
      K>OT
      80
                                            149
                                                 M+182
         Indian  Lak* Samp)
                      60   80   100   120  140   160   1
                          80   100   120  140   160   180
                                                 M+182
Figure 16.
Comparison of  the Mass  Spectrum of Indian
Lake  Extract with that  of Geosmin
                            34

-------
A simultaneous, but independent, investigation of a
compound from three other species of actinomycetes
by Medsker, Jenkins, and Thomas  (10) led to the same
structure as "celinol".  Further samples from Grand
Lake were found to contain both "geosmin" and 2-methyl-
isoborneol.  Samples from Indian Lake nearby contained
only "geosmin."  Figure 16 illustrates the matching of
a known sample of "geosmin" with that of an extract
from Indian Lake.

The concentration of 2-methylisoborneol and geosmin
were estimated by Rosen to be 0.1 ppb and 0.03 ppb,
respectively in Grand Lake.  The presence of 2-methyl-
isoborneol in a body of natural water had not previously
been reported.

Recently Piet, Zoeteman and Kraayeveld, Government
Institute for Drinking Water Supply, The Hague, Nether-
lands, detected "geosmin" and 2-methylisoborneol in
rivers of the Netherlands  (11).

Having once been identified and associated with a
particular problem, a pollutant becomes much more
readily identified in future cases.  Such an example
illustrates an important function of organic contami-
nants characterization.
     7.  Western Louisiana Industrial Waste Survey

In April, 1971, Dr. T. 0. Meiggs, EPA Division of Field
Investigations, Denver, Colorado, requested NWCCRP to
characterize chemically a number of industrial effluents
from an industrialized area in western Louisiana.  The
information was for an enforcement conference.  We
identified a total of 50 compounds in 7 different
industrial effluents; forty-nine of the identifications
were confirmed by comparison of GC-MS data and/or GC
retention times of standards with those of the
tentatively identified compounds.  Some compounds were
common to two or more of the effluents.

Values for total organic carbon  (TOG), chemical oxygen
demand (COD), specific conductivity, and concentrations
of total and suspended solids were supplied to the EPA
Division of Field Investigations from 24-hour composite
samples.
                         35

-------
One liter of each of the effluent samples was extrac-
ted with chloroform and concentrated.  A 50-ft.
support coated open tubular (S.C.O.T.) column with
carbowax 20M-TPA provided adequate separation of all
but one of the mixtures for which a 50 ft. S.C.O.T.
column coated with SE-30 was used.

Chemical characterization of these seven industrial
effluents

     •  revealed the discharge of unexpected compounds
        (not indicated from lists of products and raw
        materials supplied by the manufacturers) along
        with other data for enforcement of permit
        regulations.

     •  furnished knowledge of treatment effectiveness
        beyond that provided by gross pollution
        measurements.

     •  gave another means of determining the signifi-
        cance of an efiluent—whether the compounds
        present are likely to cause oxygen depletion,
        to contribute to taste and odor problems or to
        cause chronic or acute toxic effects in biota.
                  Petrochemical Company A

Petrochemical Company A, producing olefins and oxygena-
ted hydrocarbons, discharges an estimated 4 million
gallons per day.  Treatment consists of three ponds
connected in series, and covering a total of 5 acres
with a 5-6 day retention time.  The first and largest
of the ponds contains aerators.  The third has an over-
flow structure at the entrance of a bayou into which
the effluent is discharged; samples were gathered at
this point.

The effluent from Petrochemical Company A contained the
most complex mixture of organic compounds (Figure 17).
Because no standard was available for comparison, the
identification of 3-methylindene rests solely on mass
spectrometric data interpretation.  All other compounds
were confirmed by comparison with standards.  Phenol
and 2 ,6-dimethylnaphthalene were found to have the same
retention time under the gas chromatographic conditions
                              36

-------
         50' S.C.O.T CARBOWAX 20M-TPA
         70° ISO. / 2 MIN.; PROGRAM -> 200° AT 8V MINI.
                            i
                        8    10   12
                          MINUTES
                        14
16
18
20
Figure 17.
Chemically Characterized Gas Chromatogram
of Petrochemical Company "A"
                          37

-------
                                                              Table 3

                                           Chemical  Characterization and Gross  Pollution
                                              Measurements  of Petrochemical  Company A
                                                   GROSS  POLLUTION  MEASUREMENTS
                                                 Total  Organic  Carbon
                                                 Chemical Oxygen Demand
                                                 Total  Solids
                                                 Suspended Solids
                                                 Specific Conductivity
  180 mg/i
  612 mg/i
  868 mg/l
   78 mg/J,
1,100 v mhos/cm
CO
CHEMICAL CHARACTERIZATION

Products

Propylene
Ethylene
Butadiene
Butane
Octane
Ethylene glycol
Ethylene oxide
Polyglycols
Ammonia






Raw Materials

Raw gas
Ethane
Refinery gases
Refinery C-^
stream
Refinery C3
stream
Propane
Butadiene
Nitrogen
Hydroformer gas
Platformer gas



Compounds Identified

m-Xylene
p_-Xylene
o-Xylene
Styrene
o-Methylstyrene
Indane
Indene
Methylindene
3 -Me thy 1 indene
Naphthalene
2-Methylnaphthalene
1-Methylnaphthalene
2 ,6-Dimethylnaphthalene
Phenol
Approximate
Concentr at ion
(mg/Jl)
0.008
0.002
0.006
0.031
0.001
0.007
0.026
0.002
0.003
0.053
0.030
0.025
0.015
0.060
Approximate
Discharge
(Ibs/day)
0.3
0.1
0.2
1.1
0.1
0.3
0.9
0.1
0.1
1.9
1.1
0.9
0.5
2.1

-------
employed, but the mass spectrometric  fragmentation
patterns of each were readily discernable even though
they were present as a mixture.  A mixture of two
standards confirmed the spectral interpretation.

Sulfur and several polycyclic aromatics  such as
indenes and naphthalenes have been cited as the source
of odor in oil products  (12) .  They may  therefore be
responsible for odor problems in drinking water
supplies derived from sources into which petroleum
by-products are discharged  as waste.

Table 3 presents a summary  of the gross  pollution
parameters and a list of products and raw materials
supplied by the company, along with the  identified
compounds, their approximate concentrations, and their
rates of discharge.  A comparison of  the list of
products and raw materials  with that  of  the identified
compounds illustrates an important point—the lists
supplied by a company cannot be used  exclusively as
a key for the identification of organic  pollutants in
industrial discharge.
                Petrochemical Company B

The  second petrochemical  plant,  producing  alcohols and
paraffins ,  discharges  about 314  million  gallons per
day.  Wastes  from a paraffin unit and an alcohol unit
enter a  ditch at two points and  pass through  a  "flow
structure"  before emptying into  a bayou.   The effluent
from the paraffin unit passes through an American
Petroleum  Institute (API)  separator before entering
the  ditch;  the effluent from the alcohol unit,  compris-
ing  about  80% of the total effluent, is  not treated.
The  sample from the ditch was taken after  the effluent
passed through the "flow  structure."
                                     Flow Structure
                         39

-------
Only normal short chain alcohols with even carbon
numbers were found to any appreciable extent.
n-Hexanol was present in the largest concentration
along with lesser amounts of n-butanol and n-octanol
and a trace of n-decanol (Figure 18).  The absence of
any traces of paraffinic hydrocarbons in the effluent
is conspicuous.  Either the API separator did an
efficient job of removing them or their large molecu-
lar weight prevented their elution from the chromato-
graphic column.  Dilution by the effluent from the
alcohol unit would also help mask their presence if
they were being discharged in small amounts.

The gross pollution measurements and chemical charac-
terization data are summarized in Table 4.
                Petro re finery Company A

The first of the two refineries studied discharges an
estimated 3 million gallons per day and produces about
230,000 barrels per day.  Treatment consists of a
simple 8-hour retention-time lagoon that discharges
through an overflow structure into a tidal pond of a
river.  To avoid tidal influences, sampling was done at
the lagoon overflow structure.

                          Overflow
                        /Structure
                       £
                           Tidal
                           Pond
Refinery
                                                River
The chloroform extractables of the effluent consisted
primarily of phenol, p-cresol, and normal long chain
aliphatic hydrocarbons  (Figure 19).  Undecane  (Cii )
through nonadecane  (C^g) were identified and confirmed.
Small amounts of unspecified branched isomers appear
between the peaks of these normal hydrocarbons.
Although both 1-methyl naphthalene and 2-methylnaphtha-
lene were present, no naphthalene was detected in
measurable concentration.  The gross pollution measure-
ments and chemical characterizations are presented in
Table 5.
                              40

-------
                    50' S.C.O.T. CARBOWAX 20M-TPA

                    50°  ISO./2 MIN.; PROGRAM ->200° AT 8VMIN.
                                                o
                                                B
                                                a
                                                      o
                                                      e
                                                      a
                                                i
                                    10
12   14   16

  MINUTES
                                                   18
                                                       20
                                                           22
                                                              24
                                                                  26
                                                                      28
Figure  18.  Chemically Characterized Gas Chromatogram  of Petrochemical Company

             "B"

-------
                                          Table  4

              Gross Pollution Measurements and Chemical Characterization
                              of Petrochemical Company B
                             GROSS POLLUTION MEASUREMENTS
                      Total Organic Carbon      130 mg/i
                      Chemical Oxygen Demand    Not available
                      Total Solids              2650 mg/X.
                      Suspended Solids          34 mg/8.
                      Specific Conductivity     4,000 u mhos/cm
                                 CHEMICAL CHARACTERIZATION
                                                                Approximate    Approximate
     Products         Raw Materials     Compounds Identified   Concentration    Discharge
                                                                   (mg/i)         (Ibs/day)


Normal paraffin      Ethylene                 1-Butanol            16.0             90
Industrial alcohols  Aluminum                 1-Hexanol            65.0             375
Ethylene             Hydrogen                 1-Octanol            19.0             110
Methyl chloride      Raffinate                1-Decanol             2.5             15
Ethoxylates          Sulfuric acid
                     Ethylene oxide
                     Acetic acid
                     Caustic
                     Phosphoric acid
                     Kerosene
                     Ethane
                     Propane
                     Methanol
                     HC1

-------
              50' S.C.QT CARBOWAX 20M-TPA
              60° ISO. / 2 MIN.t PROGRAM -* 200° AT 8VMIN.
                                            oo
                          i
                                  i
                          10   12  14
                            MINUTES
                        16
18
                                             20
                                    22
24
Figure  19.
Chemically Characterized Gas  Chromatogram
of Petrorefinery  "A"
                            43

-------
                         Table  5

Gross Pollution Measurements and Chemical Characterization
                of Petrorefinery Company "A"
               GROSS POLLUTION MEASUREMENTS
         Total Organic Carbon
         Chemical Oxygen Demand
         Total Solids
         Suspended Solids
         Specific Conductivity
15 mg/i
Not available
9,220 mg/i
38 mg/i
13,700 vi mhos/cm
                CHEMICAL CHARACTERIZATION

Products

LPG propane
Propylene
Orthoxylene
Aromatics










Raw Materials Compounds Identified

Crude oil Undecane
Excess gas oil Dodecane
Extracts Tridecane
Tetradecane
Pentadecane
Hexadecane
Heptadecane
Octadecane
Nonadecane
2 -Me thy In aph tha lene
1-Methylnaphthalene
Phenol
o-Cresol
Approximate
Concentration
(mg/Jl)
0.027
0.031
0.042
0.039
0.030
0.026
0.022
0.017
0.013
0.013
0.005
0.200
0.120
Approximate
Discharge
(Ibs/day)
69
79
107
99
76
66
53
43
33
33
12
510
300

-------
               Petrorefinery Company B

The treatment facilities of refinery B, which discharges
about 1 million gallons per day  and produces about
70,000 barrels per day, are more extensive than those of
Company A.  The effluent from  the refinery flows through
an API separator and a corrugated plate interceptor
(CPI) into an activated sludge unit.  It then passes
through a clarifier into a 24-hour retention-time
aerated lagoon from which it discharges into a bayou.
The chloroform extractables  of  the  effluent produced a
chromatogram resembling  that of a crude oil and mass
spectra characteristic of  only  aliphatic hydrocarbons.
The normal chain hydrocarbons were  present in larger
amounts than the many branched  isomers and thus stood
out as spikes superimposed upon a base of complex,
poorly resolved lumps in the gas chromatograph  (Figure
20) .  Undecane  (C-, ^) through heineicosane  (C^i) were
confirmed.  No phenols or  naphthalenes were identified
in this sample.  The gross pollution measurements and
chemical characterizations are  summarized in Table 6.
               Synthetic  Rubber Company

A synthetic  rubber  company discharges about  6 million
gallons per  day with only primary treatment  followed
by discharge via  a  drainage ditch to a  bayou.

Styrene  (one of the 3 raw materials listed), furfural,
and 1-methylnaphthalene  were identified and  confirmed
in the effluent.  The compound producing the largest
peak in the  gas chromatogram (Figure 21)  and four
isomers of apparent molecular weight 156 could not be
identified from their mass spectra alone;  lack of time
and manpower precluded further investigation.  Table 7
summarizes the results of the analysis.
                         45

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              50' S.C.QT CARBOWAX 20M-TPA
              60° ISO./2 MIN.; PROGRAM -*200° AT 8VMIN.
                        10  12
14
16  18  20
 MINUTES
22  24  26  28  30  32  34  36
Figure 20.  Chemically Characterized Gas Chromatogram of Petrorefinery Company "B"

-------
                          Table 6

Gross Pollution Measurements and Chemical Characterizations
                of Petrorefinery Company "B"
               GROSS POLLUTION MEASUREMENTS
         Total Organic Carbon
         Chemical Oxygen Demand
         Total Solids
         Suspended Solids
         Specific Conductivity
210 rag/Jl
676 mg/i
2430 mg/«.
182 mg/X,
3900 y mhos/cm
CHEMICAL CHARACTERIZATIONS

Products

LPG
Propane
Butane
Gasoline
Kerosene
Diesel fuel
Heating oil
#6 Fuel oil
Coke



Raw Materials Compounds Identified

Crude oil Undecane
Isobutane Dodecane
PVC Tridecane
Tetradecane
Pentadecane
Hexadecane
Heptadecane
Octadecane
Nonadecane
Eicosane
Heine icosane
Approximate
Concentration
(mg/fc)
0.05
0.22
0.39
0.58
0.49
0.42
0.34
0.33
0.31
0.30
0.19
Approximate
Discharge
(Ibs/day)
0.4
2.2
3.8
5.6
4.8
4.0
3.3
3.2
3.0
2.9
1.8

-------
          50' S.C.O.T CARBOWAX 20M-TPA
          70°  ISO./2 MIN.; PROGRAM -»200°AT 6.5VMIN.
                       8   10    12
                         MINUTES
                        14
16
18
20
Figure 21.
Chemically Characterized Gas Chromatogram
of Synthetic  Rubber Company
                          48

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                                                         Table  7

                               Gross Pollution Measurements  and  Chemical  Characterization
                                               of Synthetic  Rubber Company
                                              GROSS POLLUTION MEASUREMENTS
                                        Total Organic Carbon       52 mg/l
                                        Chemical  Oxygen  Demand    168 mg/i
                                        Total Solids               3210 mg/t,
                                        Suspended Solids           76 mg/i
                                        Specific  Conductivity      5000 \i mhos/cm
vo
                                                CHEMICAL CHARACTERIZATION
Products
Synthetic rubber
Raw Materials
Butadiene
Styrene
Carbon black
Compounds Identified
Styrene
Furfural
1-Methylnaphthalene
Approximate
Concentration
(mg/i)
0.0026
0.0017
0.0017
Approximate
Discharge
(Ibs/day)
1.3
0.9
0.9

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                  Chemical Company A

Chemical Company A, a producer of chlorinated hydro-
carbons discharges about 4 million gallons of
effluent per day from their organics plant into a
ditch.  Its treatment, after mixing with the effluent
from a caustic and chlorine unit, consists only of
dilution with cooling water before discharge into a
river.

1,1,2-Trichloroethane and 1,1,2,2-tetrachloroethane
were identified as the two main contaminants (Figure
22).  The total discharge at the above mentioned rate
amounted to over 300 pounds of these chlorinated
hydrocarbons per day.  Table 8 summarizes the findings
                  Chemical Company B

The second chemical company, a polymer producer,
discharges an estimated 2 million gallons of effluent
per day.  The treatment consists of a series of six
lagoons, with a 4-5 day total retention, that discharge
into a river.  The only two compounds of significant
amount, as determined by gas chromatography, were
identified as decane and undecane (Figure 23) .   No
gross pollution measurements were available.  Table 9
summarizes the findings.
      8.  Color Body Characterization and Related
       Studies from a Kraft Paper Mill Effluent

The Interstate Paper Company at Riceboro, Georgia,
utilizes a recently developed lime treatment process
to remove "color bodies" from the effluent of Kraft
pulp mills.  The lime treatment removes 80-90% of the
color from the effluent; however, during passage through
the stabilization lagoon, the water becomes colored
again.  Because of the unusually long retention time of
the lagoon (3-6 months) the reappearance of the color
may result from one or both of two processes:  1) decom-
position of previously uncolored compounds that were
not removed by lime treatment and 2) leaching of natu-
rally occurring "color bodies" present in the soil at
the bottom of the lagoon.  The waste treatment system
is presented below.
                             50

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      50' S.C.O.T CARBOWAX 20M-TPA
      70° ISO./2MIR; PROGRAM-^200° AT 8VMIN.
           I    I   	I
                 I	I
      0   2    4    6    8    10   12
                 MINUTES
Figure 22.
Chemically Characterized Gas Chromatogram
of Chemical Company "A"
                        51

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         100' S.C.QI SE-30

         60° ISO./2MIN.; PROGRAM -*200° AT I2VMIN.
                c
                •


               "o
               UJ
                        I
                       6    8

                       MINUTES
                     10
12
14
Figure 23.
Chemically Characterized Gas  Chromatogram

of Chemical Company "B"
                         52

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                                                   Table   8

                                   Chemical Characterization of Chemical Company A
Products
Chlorine
Caustic soda
Raw Materials
Sodium chloride
Ethylene
Compounds Identified
1,1, 2-Trichloroe thane
1,1,2, 2-Tetrachloroethane
Approximate
Concentration
(mg/*)
5.4
2.2
Approximate
Discharge

-------
                                Key:  •	• Lime Treatment Effluent
                                     o	o Lagoon Effluent
                                     A	A Marsh Water
          0.090
          0.080
          0.070
       O  0.060
       c  0.050
          0.040
          0.030
                                   9

                                  PH
                                         10
                                                11
Figure 24.   Comparison of Kraft Pulp Mill Effluent and
              Marsh Water  Color  Intensity  as a  Function of
              pH
                              54

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                       Lime
                   Flocculat jon
                                         Stabilization
                                           Pond
                        R
                        I
                        V
                        E
                        R
                                                    I

 Some  physical characteristics of the lagoon effluent
 were  compared with those of nearby marsh water and of
 the effluent following lime treatment.  Plots of color
 intensity at 430 nm as a function of pH for the three
 samples  are shown in Figure 24.  The curve for the
 lagoon effluent parallels the curve for marsh water
 but differs greatly from that of the effluent following
 lime  treatment.   Fluorescence spectra were recorded for
 each  sample, both without any pH adjustment and after
 acidification to pH 2 with HCl.  The wavelengths of the
 fluorescence maxima and their intensities relative to
 that  of  the lagoon effluent are given in Table 10.


                       Table 10

    Comparison of Kraft Pulp Mill Effluent and
           Marsh Water Fluorescent Spectra
         Sample
Fluorescence
   Maxima
Intensity
  Ratio
Lime treatment  effluent
Lagoon effluent
Marsh water

  After Acidification

Lime treatment  effluent
Lagoon effluent
Marsh water
   427 nm
   450 nm
   450 nm
   390 nm
   450 nm
   450 nm
   3.8
   1.0
   1.8
   1.4
   1.0
   1.7
                         55

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The absorption and fluorescence data indicate that the
color characteristics of the lagoon sample closely
resemble those of the marsh water.

Two further experiments were performed to determine
the source of the color bodies.  One liter of lime-
treated effluent was sparged with air for 120 days to
test for the presence of compounds capable of decom-
posing to colored products.  During that time, however,
the color decreased probably because of a change in pH
caused by CO2 absorption.

In addition, mud from the lagoon bottom was dried,
powdered, and then leached with calcium hydroxide
solutions of two different concentrations.  The 5-gram
mud samples were shaken with 100 ml of solution for 10
days.  Absorption measurements at 420 nm were made
after 5 and 10 days.  The data summarized in Table 11
indicate that the leaching of color from the soil is
pH dependent.
                       Table 11

     Color Leached from Lagoon Mud as a Function
           of Calcium Hydroxide Concentration
Solution
H20
10" *M Ca(OH)2
10 3M Ca(OH)2
Absorbance at 420 nm
5 days
0.117
0.124
0.181
10 days
0.142
0.136
0.182
As a result of the study, the color increase in the
lagoon effluent was attributed to the leaching action
of the alkaline waste from the lime treatment.

An effort was made to identify the compounds responsi-
ble for the color in the effluent.  Freeze concentra-
tion followed by gel permeation chromatography of the
                             56

-------
lagoon effluent yielded molecular weight values for
the "color bodies" of 2,500 to 5,000.  Further work on
the molecular weight characteristics of these samples
is being conducted in detail by the Institute for
Paper Chemistry under FWQA Contract 1240-DKO.

The amount of acid-insoluble organic matter is relevant
to the color body determination since lignin and other
color-producing large molecules can be precipitated
from pulp mill effluents by acidification.  The
difference between the total organic carbon  (TOC)
contents of a sample before and after acidification
gives an indication of the concentration of acid-
insoluble organics.  Such measurements made on the
pulp mill effluent both before and after lime treat-
ment  (refer to Table 12} show that most of the acid-
insoluble components are removed by the treatment.
                      Table  12

   Comparison of the Amounts of Acid Insoluble
       Organics in the Lime  Treated and the
             Untreated Effluent Samples
             TOC Before      TOC After    Acidification
 Sample     Acidification  Acidification   Insolubles
                               CpH 1)
Lime
(Treated)     117 mg/1       111 mg/1          5%

Untreated     216 mg/1       112 mg/1         48%
Analyses of effluent samples from before and after lime
treatment and after lagooning were made to determine
the effect of the treatment on the organic wastes
discharged.   (The technique is described in references
13 and 14.)  Each sample was made basic to pH 11.5 and
extracted with chloroform to remove the neutral
volatiles.  The extracts were concentrated and analyzed
by gas chromatography-mass spectrometry.  A list of the
compounds identified is presented in Table 13.
                         57

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                       Table 13

       Compounds Tentatively Identified in the
           Neutral Volatiles Fraction of the
              Riceboro Pulp Mill Effluent
           2^-cymene
           limonene
           guaiacol
           a menthene isomer
           l-methoxy-4-(1-propenyl)-benzene
           l-methoxy-4-pentylbenzene
The aqueous residue was methylated with dimethyl sulfate
and re-extracted with chloroform.  GC-MS analysis of the
extract after concentration yielded the compounds listed
in Table 14.
                       Table 14

        Compounds Tentatively Identified in the
           Phenols and Acids Fraction of the
              Riceboro Pulp Mill Effluent
             dimethylsulfone
             2,5-diethylthiophene
             4-me thoxybenz aIdehyde
             3,4-dimethoxyethylbenzene
             ^-methoxypropiophenone
             3,4-dimethoxybenzaldehyde
             3,4-dimethoxyacetophenone
             3,4,5-trimethoxyacetophenone
             methyl 3,4-dimethoxybenzoate
             3,4-dimethoxypropiophenone
             methyl pimarate
             methyl isopimarate
             methyl abietate
             methyl dehydroabietate
                             58

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The results of this analysis indicated that

     •  most of the neutral volatiles were not
        affected by lime treatment.

     •  most of the resin acids are significantly
        diminished in concentration after lime
        treatment.
 9.  Characterization of Foam from Kraft Pulp Mills

Source of an Environmental Contamination

Brown foam, floating to shore in masses, has presented
a persistent problem in Perdido Bay in Florida's
western panhandle.  Tall oil in the discharge from a
kraft paper mill at the north of the bay was suspected
to be the cause of the foam.

Three samples were collected and analyzed.  The first,
a water sample, was taken from below the second of 2
sedimentation ponds in a stream composed almost
totally of the|plant's effluent.  A foam and scum
sample was collected in Perdido Bay about 17 miles
below the mill at a point where the foam was in
abundance.  The last sample was of foam only and was
taken from a point about 24 miles below the papermill.

To determine the presence of resin acids and fatty
acids, the samples were acidified, extracted with
chloroform, esterified with diazomethane, and analyzed
by flame ionization gas chromatography.  Structures
were confirmed by GC-MS.  The compounds listed in
Table 15 were identified  (the acids as the methyl esters)
for the three samples.

The identification of the same and similar compounds in
all three samples indicates that the paper mill is the
source of the foam contamination in the bay.  The mill
has begun installing secondary treatment.  In-plant
changes have reduced the organic content of the
effluent to one-third of its former load.
                         59

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                                 Table 15
            Compounds Identified in Perdido Bay Kraft Papermill
                    Wastewater, Foam, and Scum Samples
          Mill
       Wastewater
  Foam & Scum (17 miles
       below mill)
      Foam (24 miles
        below mill)
Pimaric acid

Isopimaric acid

Dehydroabietic acid

6,8,11,13-Abietatetraen-
  18-oate

Unidentified resin acid
Palmitic acid

Stearic acid



Lignoceric acid
Pimaric acid

Isopimaric acid

Dehydroabietic acid

6,8,11,13-Abietatetraen-
  18-oate

Unidentified resin acid

Unidentified resin acid



Stearic acid

Eicosanoic acid

Lignoceric acid
Pimaric acid

Isopimaric acid

Dehydroabietic acid

6,8,11,13-Abietatetrean-
  18-oate

Unidentified resin acid

Unidentified resin acid

Palmitic acid

Stearic acid

Eicosanoic acid

-------
Constituents from a Bleach Stage Caustic Clarifier

The International Paper Company at Springhill,
Louisiana, was involved in developing a "massive  lime
process" to remove color from the effluent of the
"bleach plant caustic stage."  After the lime was
added to the effluent and the mixture was passed to a
"caustic clarifier", however, excessive foaming
developed that caused flow stoppage and solids carry-
over.  Identification of the foam constituents was
desired in order to take action to reduce the foaming.

The analytical procedure was similar to previous
characterization of paper mill effluents (13,14).  The
foam was mixed with distilled water in a separatory
funnel and made strongly alkaline with sodium hydroxide.
The chloroform extracts of the basic solution contained
the neutral volatile constituents, which were only
minor components of the mixture and were not examined
further.

The acidic fraction in the aqueous phase, comprising
the majority of the organic componnents, was methylated
by two procedures to produce derivatives that could be
separated by gas chromatography.  Methylation of the
chloroform extract of the acidified aqueous phase by
diazomethane proved superior to direct methylation
with dimethyl sulfate; the amounts of methyl esters
were greater using the former method.

A GC-MS analysis showed no resin acid or phenolic
compounds in detectable amounts, in contrast to previous
studies involving total effluents.  Instead, the foam
consisted primarily of long-chain carboxylic ("fatty")
acids.  Palmitic and stearic acids were in predominance
(Figure 25).  Also present in significant amounts were
identified isomers of the di- and triunsaturated
analogs of octadecanoic acid.  The identifications of
the numbered peaks in Figure 25 are listed below:

     Peak Number            Methyl Ester of

          1       heptanoic acid  (heptylic acid)
          2       octanoic acid  (caprylic acid)
          3       nonanoic acid  (pelargonic acid)
          4       decanoic acid  (capric acid)
          5       undecanoic acid
          6       dodecanoic acid  (lauric acid)
                         61

-------
                            DIAZOMETHANE  METHYLATION
en
to
            <4  42  40  3a  36  34  32  30 28  26 24  22  20 |g  u  14  12  10
                                         FOAM
                                                                     if
                                                                     Ji,

6420
   till
      Figure 25.  Chemically Characterized Gas Chromatogram of Foam from a Kraft
                  Papermill Bleach Stage  Caustic Clarifier

-------
Peak Number          Methyl Ester  of

     7       tetradecanoic acid  (myristic acid)
     8       hexadecanoic acid  (palmitic acid)
     9       octadecanoic acid  (stearic acid)
    10       diunsaturated analog  of octadecanoic
               acid
    11       unidentified ester; molecular weight
               262
    12       unidentified ester; molecular weight
               280
A mechanical means of  foam control was tried; however,
the plant was closed down for lack of funds and no
chemical solution to the problem was attempted.
    10.  Analysis  of Oil Spills l>y NMR Speatrosoopy

Nuclear magnetic resonance  (NMR)  spectroscopy is a
useful method  for  helping to determine the source of
an oil spill.  NMR can  rapidly distinguish between
different types of oils and sometimes can identify
the type of oil in a spill without reference to a
standard.  Its greatest disadvantage is that it cannot
distinguish between different oils of the same general
type.
Shell Point Beach Oil  Spill

An oil spill occurred  in February,  1969, off the coast
of St. Mark, Florida,  on Shell Point Beach.  The U. S.
Coast Guard collected  samples from  a nearby punctured
barge and from  Shell Point Beach  3, 5, and 7 days
after the spill was noticed.  An  analysis was performed
in conjunction  with the Chemical  Services Branch to
determine whether the  barge was the source of the
spill.

A small amount  of each sample was dissolved in carbon
tetrachloride containing 2-3% tetramethylsilane as a
reference standard.  The NMR spectra were scanned and
electronic integrals of the areas under each signal
in the spectra  were made in triplicate  (Figure 26).
Since each sample was  run at least  three times, the
mean signal areas are  averages of at least nine
integrals.  The average areas under the signals were
normalized (Table 16).
                          63

-------
        St. Mar*, Fia. OiI Spi

        S-nple No. 5ST4

        Crude 0>I from Barqe
    r o
   T o
                                                   10
Figure 26.
Comparison of  NMR Spectra of  Oil from the
Suspect Barge  and from a Florida Beach
                            64

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                                          Table  16

                 Normalized Peak Areas  of Proton Signals  in Samples from the
                              St.  Mark, Florida, Oil Spill


             Sample            T9.05-9.60    T 8.00-9.05     T6.80-3.00    f 2.00-4.00


      Crude oil from barge        0.27            0.60           0.08           0.04
      (Figure 26, top)

      Sample from Shell           0.23            0.62           0.09           0.06
      Point Beach—
^     Weathered 3 days
Ul
      Sample from Shell           0.23            0.62           0.11           0.05
      Point Beach Weeds—
      Weathered 5 days
      (Figure 26, bottom)

      Sample from Shell           0.21            0.65           0.11           0.04
      Point Beach Weeds—
      Weathered 7 days

-------
         •'  I  •  I

         Harbor
      ',',  Source A
              1  •  i  •  1  •  i  -  1
Figure 27.
Comparison of NMR Spectra of Oil from
Duluth Harbor and the Primary Suspect Source
                          66

-------
         Harbor

         Source  B
Figure 28.  Comparison of NMR Spectra of Oil from Duluth
            Harbor and the Secondary Suspect Source
                         67

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The similarity between the chemical shifts and the
relative peak areas of each signal indicates that the
samples are all of the same oil type and that the barge
could have been the source of the spill.  A comparison
of infrared spectra and the sulfur and vanadium content
of pollution samples with those of oil from the suspect
barge also indicated that they were the same.

Small but distinct differences appeared between the
barge sample and the spill samples and also among the
spill samples themselves.  The weathered samples showed
a decrease in the relative CH3 signal area and an
increase in the CH2 signal area as weathering time
increased.  A mixture of hydrocarbons containing both
volatiles and nonvolatiles would be expected to show
such relative change in signal areas.  The volatile
materials, having a larger ratio of 013 to CH2 groups,
evaporate faster, leaving the long chain hydrocarbons
and other larger molecules behind.  The result is a
decrease in the 013 to CH2 ratio.
Duluth Harbor Oil Pollution

In 1969, the harbormaster in Duluth, Minnesota,
submitted samples of oil from the persistently polluted
harbor to the National Water Quality Laboratory in
Duluth.  These samples and samples of a suspected
source (a railway company) were sent to NWCCRP for NMR
analyses.  Also sent later was a sample from a second
possible source, an oil company.

The NMR spectrum of the harbor sample was very different
from that of the railway company sample (Figure 27) .
It closely resembled, however, the spectrum of the oil
company sample (Figure 28) implicating the oil company
as the probable source of pollution.  Prosecution of
the oil company resulted in conviction and a fine.
Tampa Bay Oil Spill

The A.'iooa Trader was suspected to be the source of an
oil slick on Tampa Bay, Florida, in 1969.  However, NMR
spectra of samples from the slick and from the ship's
oil cargo displayed significant differences.  The
proportions of the various types of protons in the
                              68

-------
samples as calculated from the  integrated intensities
are presented below:

                      Methyl  Methylehe  Misc.  Aromatic

Tampa Bay Sample        0.20      0.55     0.16     0.10

Alooa Trader Sample     0.28      0.61     0.07     0.04


The differences in the  spectra  are not believed to be
due merely to weathering  effects on the slick samples
since the changes in some proton proportions do not
correspond to those predicted from weathering,  in
studies of the weathering on other oil samples  (crude
oil and Bunker C) three changes were noted:

     •  weathering lowers the proportion of methyl
        proton,

     •  it increases the  methylene proton proportion,
        and

     •  it has a small  or no effect on the aromatic
        proton percentage.

On the bases of these data, therefore, the oil from the
slick was judged to be  from a source other than the
Alcoa Trader or was highly contaminated with oil from
another source.
Florida West Coast Oil Contamination

In the spring of 1970, oil pollution was reported along
an 80-100 mile stretch of Florida's northwest coast.
Two possible sources were the Greek tanker, Delian
Apollon, wrecked off Tampa in February, 1970, carrying
Bunker C oil, and a large Chevron crude oil fire from
a well off the coast of Louisiana.  Because the two
sources represented two different types of oil, the
analysis was performed by NMR.

Spectra were scanned of samples from the two suspected
sources and from the polluted areas (Pensacola Beach,
Panama City .Beach, St. Andrews Park, Indian Pass,
Destin Beach, and submerged oil out from Destin Beach).
Data are summarized in Table 17 along with data from
previous weathering studies on Bunker C and crude oil.

                          69

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                          Table 17

NMR Comparison of Oil from West Florida Beaches with Suspect
                 Crude Oil and Bunker C Oil
Sample
Environmental samples
Delian Appollon
Chevron oil well and
oil slick
Weathered Bunker C
Weathered crude

Methyl
0.21-0.22
0.22
0.26-0.27
0.17-0.21
0.21-0.24
Normalized Proton
Methylene
0.68-0.71
0.61
0.65-0.66
0.56-0.59
0.69-0.71
Signal Areas
Misc.
0.05-0.07
0.10
0.05
0.10-0.15
0.04-0.06

Aromatic
0.02-0.04
0.07
0.02-0.03
0.10-0.13
0.02-0.03

-------
The spectra of the spill samples were all similar and
matched those of weathered crude oil but not those of
Bunker C or weathered Bunker C oil.

The Chevron oil well crude oil was therefore indicated
as the more probable source of the pollution.


Miscellaneous Oil Analyses

In the fall of 1970 an oil slick was discovered by the
U. S. Coast Guard around the Dutch Cargo carrier Maya.
Samples of the oil slick, of water being discharged from
the Maya and of the oil in the bilge of the Maya were
taken for analysis by EPA Region IV Surveillance and
Analysis Division at SEEL.  An NMR analysis was also
requested since prosecution was expected and confirma-
tory data were desired to show that the ship was the
source of the slick.

The NMR spectra of the samples, were all similar,
indicating a partially refined crude oil.  Infrared
analysis likewise confirmed that the samples were all
very similar and probably from the same source.  The
data were entered as evidence in court by the State of
Florida.  The case, however, has been postponed by a
Federal injunction charging that Florida's coastal
pollution laws are unconstitutional.

In the winter of 1970 Mr. Warren T. McFall, EPA Alaska
operations Office, requested an analysis of oil on the
feathers of a bird that had been killed by oil pollu-
tion and crude oil from a suspect source near Cook
Inlet.  The NMR spectra of the two samples were quite
different—more than would be expected from weathering
and contamination from the bird's natural oils.
Fluorescence data also indicated that the two were not
from the same source.  We concluded that the Cook
Inlet crude oil was not the source of the oil that
killed the bird.  No further work was done concerning
this problem.

Also in the winter of 1970, the Chemical Services Branch
received an oil sample taken by the National Marine
Fisheries Service during exploratory fishing at 330
fathoms northwest of the Dutch West Indies island of
Aruba.  From NMR analysis we concluded that the oil
was unlike any crude, Bunker C or refined oils that we
examined previously.  The source remains unknown.


                          71

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              11.   Analysis of Oil Spills
              by Fluorescence Spectroscopy

Fluorescence spectroscopy represents yet another method
of analysis that may be used to characterize an oil
spill and determine its source.  Whereas NMR can differ-
entiate only between oil types, different oils of the
same type have sufficiently distinctive fluorescence
characteristics to distinguish between them (15).

Several samples are cited comparing fluorescence data
from oil spills with data from other techniques.
Duluth Harbor Oil Pollution

In a Duluth Harbor oil spill mentioned previously the
NMR spectra indicated that the oil of the slick
matched that of the suspect oil company and did not
match any of three oil samples from the suspect railway
company.  Fluorescence analysis showed that the railway
samples were radically different from the harbor oil,
corroborating NMR conclusions.

NMR spectra of both oil slick samples and the samples
from the oil company, however, were identical.  Also,
the relative fluorescent intensities of the oil company
samples and the intensity ratios closely matched those
of the first of the two harbor samples (Table 18).

Harbor Sample #2 did not match the oil company samples
as well.  But a mixture of 1 part oil from the railway
company and 14 parts oil from the oil company produced
a fluorescence spectrum similar to that of the second
harbor sample.  Thus, although the oil company was the
main source of the pollution there was evidence of a
measurable contribution from the railway company.
Savannah River Spills

In April of 1971, two different oil spills occurred on
the Savannah River.  Fluorescence analyses were
performed on the spill samples and on samples from
suspect sources.  The data obtained were compared with
that from gas chromatographic analysis using flame
ionization and photometric detectors.
                              72

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                  Table 18
Fluorescence Spectra of Duluth Harbor Samples


Samples


Harbor
Harbor
Oil Co
Oil Co
Oil Co


Sample #1
Sample #2
. Sample #1
. Sample #2
. Sample #3
Fluorescence Spectra (Excitation at 340 mji)
Intensity ratio at 400 mja of
harbor sample/oil co.
Dilution
1:1000
0.97
1.98
1.14
1.00
1.00
Dilution
1:10,000
1.00
3.67
0.97
1.00
0.90
Intensity ratio, shoulder
at 440 mu/max at 400 mju
Dilution
1:1000
0.39
0.58
0.40
0.41
0.39
Dilution
1:10,000
0.30
0.48
0.31
0.30
0.37

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                                   Table 19

                 Fluorescence  Spectra  of Savannah River Samples
Spill
Spill A




Spill B

Sample
Fuel line
Drip pan
Bilge
River sample 11*
River sample #2*
Suspect
River sample
10,000 Dilution
Maximum
intensity
100
17
72
92
95
100
46
440 nm
368 nm
.40
.15
.35
.40
.40
.80
.55
50,000 Dilution
Maximum
intensity
100
21
75
93
94
100
42
440 nm
385 nm
.40
.22
.32
.40
.39
.66
.49
     *The 440nm/368nm ratios of the spill and suspect match very well.  The
intensity readings are slightly low for the two spill samples but this is not
uncommon for oils that have been weathered.

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                         Spill  "A"

Two samples from spill  "A" were compared by fluorescence
analysis with three oils taken  from the fuel line, a
drip pan, and the bilge of a nearby suspect ship.

Fluorescence data are presented in Table 19.  The
relative intensity measurements from the spill samples
matched those of the fuel line  from the ship, confirming
results of GC analysis.
                         Spill "B"

In the second spill, which appeared to be a Bunker fuel,
the suspected source was the fuel tank of a nearby
vessel.  The fluorescence data (Table 19), however,
indicated that the two oils were not similar, again
confirming GC data.
Duluth Superior Harbor Spill

In an oil spill on Duluth Superior Harbor in 1970 a
British and an American ship were suspects.  Spill
samples were collected from the bay and from the slip
in which both ships had been docked at different times.
Comparisons were made with oils obtained from the ships.
Each sample was diluted with oils obtained from the
ships.  Each sample was diluted wt/vol 1:7500, 1:10,000,
and 1:100,000 with cyclohexane.  The relative fluores-
cence intensity data for the spill and suspect samples
are presented in Table 20.

All parameters of the sample from the British freighter
match closely those of the spill sample, implicating
that ship as the source of the pollution.  These results
were turned over the Coast Guard for legal action.
Dundee Canal, Savannah River

An oil spill covering a two to three mile length of the
Dundee Canal occurred in February, 1971.  A tank trailer
carrying Bunker fuel was the suspect source.  Five spill
samples and a sample from the trailer were collected.
The broad range of intensity values  (Table 21) at
1:50,000 dilution indicates weathering of the spill
                          75

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                         Table 20



Fluorescence Spectra of the Duluth Superior Harbor Samples
Sample
British freighter
American freighter
Spill sample
Intensity @
Dilution
1:7,500
94
30
93
Dilution
1:10,000
87
20
81
Intensity ratio
386nm 440nm/386nm
Dilution
1:100,000
65
6 >-:.
52
Dilution Dilution
1:7,500 1:10,000
0.67 0.64
0.24 0.25
0.63 0.60
Dilution
1:100,000
0.57
0.28
0.55

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                     Table 21
Fluorescence Spectra of the Dundee Canal Samples
Samples

Suspect tank trailer #82
#1 — Puddle on ground
behind trailer
#2 — Ditch along Hwy 80
under highway
#3 — Ditch along Hwy 80
#4 — Dundee Canal
#5 — Dundee Canal at
Savannah River
1:10,000 Dilution
Ratio
Max. 440 nm
Intensity 386 nm
100 .66
91 .60
94 .64
97 .65
100 .66

77 .59
1:50,000 Dilution
Ratio
Max. 440 nm
Intensity 386 nm
100 .56
69 .53
73 .56
84 .56
94 .57

54 .50


Ni
(ppm)
44
28
32
33
43

25

V
(ppm)
365
198
285
308
348

210

Ni/V
.12
.14
.11
.11
.12

.12

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samples.  A decrease in fluorescence intensity is common
in weathered oil.

Fluorescence data confirmed the metal ratio  (Ni/V) analy-
ses and GC data indicating that the spilled oil was non-
homogeneous.  Spill sample #4  (Dundee Canal), however,
matched the suspect oil in all parameters.
St. Johns River, Jacksonville/ Florida

An oil spill occurring in the St. Johns River covered a
one square mile area.  Four samples  (numbered 1-4) were
taken from the slick and six suspect samples (numbered
5-10) were collected from oil depots, ships, and indus-
tries surrounding the spill area.  All samples were
cleaned, dried, and analyzed by the Region IV Chemical
Services Branch for four parameters:  1) percent
sulphur, 2) nickel-vanadium ratios, 3)  infrared ratios,
and 4) color comparison under UV light.  The NWCCRP was
requested to analyze the samples by the fluorescence
technique.

Fluorescence spectra were taken at various dilutions of
the oils with cyclohexane (1:10,000, 1:50,000 and
1:100,000).  The ratio of the maximum intensity (386 nm)
to that of a shoulder (440 nm)  was calculated for each
spectrum (15).   Table 22 summarizes the data for
fluorescence analyses and Table 23 summarizes the
results of the other analyses.   Both sets of data
indicate that the four spill samples came from the same
source.

Of the six suspect samples, only one matched the spill
sample in all fluorescence parameters.   The same
suspect sample matched the spill with all other
analytical techniques employed, confirming the
identification.
                               78

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10
                                                        Table 22

                                    Fluorescence Spectra of St. John's River Samples
Sample
no.

Maximum
Dilution
1:10,000
intensity
Dilution
1:50,000
at 386
nm
Dilution
1:100,000
shoulder
Dilution
1:10,000
Intensity ratio
at 440 nm/386 nm
Dilution
1:50,000
max.
Dilution
1:100,000
Parameters
that agree
                                                      Spill samples
   1
   2
   3
   4

Range of
 spill
samples
70
65
66
72
                               65-72
77
81
72
72
         72-81
0.67
0.70
0.71
0.67
0.60
0.61
0.63
0.61
0.54
0.55
0.55
0.53
72
90
76
75
         72-90      0.67-0.71 0.60-0.63 0.53-0.55

           Suspect samples
5
6
7
8
9
10
67
50
73
78
50a
100
81
60
68
76
73
100
90
64
65
71
99
100
0.71
0.62
0.57
0.58
0.82a
0.56
0.63
0.55
0.53
0.52
0.72
0.51
0.55
0.47
0.45
0.45
0.59
0.44
6 of 6
0 of 6
0 of 6
1 of 6
1 of 6
0 of 6
                   aMaximum is at 405 nm instead of 386 nm.

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00
o
                                                                               Table 23

                                                              Other Analyses of St. John's Diver Samples
Infrared ratios
Sample
no.
1
2
3
4
Range of
spill
s
6
7
8
9
10
%
Sulfur
a
2.59
2. 78
2.62

2.59-2.78
2.65
2.30
2.56
2.02
2.08
1.68

Ni/V
a
0.31
0.38
0.29

0.29-0.38
0.31
0.19
0.27
0.21
0.61
0.24
720 cm~l
1375 cm"1
0.28
0.29
0.34
0.31

0.28-0.34
0.29
0.19
0.33
0.26
0.35
0.16
3050 cm"1
2925 cm"1
0.023
0.020
0.034
0.026

0.020-0.034
0.023
0.023
0.023
0.017
0.041
0.034
810 on"1
1375 CUT1
0.24
0.27
0.36
0.26

0.24-0.36
0.27
0.25
0.20
0.21
0.47
0.26
810 cm"1
720 am'1
0.88
0.94
1.09
0.86

0.86-1.09
0.95
1.34
0.60
0.82
1.36
1.66
1600 cm"1
1375 cm"1
0.13
0.15
0.18
0.16

0.13-0.18
0.16
0.15
0.09
0.11
0.27
0.14
Parameters
uv Colors
yellow
yellow
yellow
yellow

yellow
same yellow
darker (brown)
lighter (yellow)
lighter (yellow)
darker (brown)
lighter (yellow)
that agree






8 of 8
3 of 8
2 of 8
0 of 8
0 of 8

                            •Sample  no.  1 did not contain enough oil for sulfur,  nickel,  or vanadium analysis.

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                         SECTION V

                         REFERENCES
 1.   Lichtenberg, J. J. , personal communication, July 16,
     1968.

 2.   Nakanishi, K. , "Infrared Absorption Spectroscopy,"
     Holden-Dayf Inc./ San Francisco, 1964, p. 182.

 3.   Beynon, J. H. , "Mass Spectrometry and Its Applica-
     tions to Organic Chemistry," Elsevier Publishing
     Co., New York, 1960, pp. 297-301 and 554-563.

 4.   Gustafson, C. G., "PCB's—Prevalent and Persistent,"
     Environ. Sci. Techno 1. , 4_, 814  (1970) .

 5.   "Effects of Pollution on Water Quality, Escambia
     River and Bay, Florida," U. S. Department of
     Interior, F.W.P.C.A., Southeast Water Laboratory,
     pp. 31-32  (January, 1970).

 6.   Adams, B. A., "The Cladothrix Dichotoms and Allied
     Organisms as a Cause of 'Intermediate1 Taste in
     Chlorinated Water," Water and Water Eng., 31, 327
     (1929).

 7.   Gerber, N. N. , and Lechevalier, H. A., "Geosmin, an
     Earthy-Smelling Substance Isolated from Actinomy-
     cetes," Appl. Microbiol., 13, 935 (1965).

 8.   Gerber, N. N. , "Geosmin, from Microorganisms in
     Trans-l,10-dimethyl-trans-9-decanol," Tetrahedron
     Lett. , 25_, 2971  (1968) .

 9.   Zelinsky, N., "Uber eine Synthese der Cyclischen
     Tertiaren Alkohole mit Hilfe von Magnesiumhalogen-
     alkylen," Ber. Dtsch. Chem. Ges., 34, 2877 (1901).

10.   Medsker, L. L. , Jenkins, D., and Thomas, J. F.,
     "Odorous Compounds in Natural Waters.  2-Exo-
     hydroxy-2-methylbornane, the Major Odorous Compound
     Produced by Several Actinomycetes," Environ. Sci.
     Technol., 3, 476  (1969).
                           81

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11.  Piet, G. J., Zoeteman, B. C. J. , and Kraayeveld,
     A. J. A., "Earthy-Smelling Substances  in  Surface
     Waters of the Netherlands/" Water Treatment  and
     Examination, 21  (4), 281  (1972).

12.  Zoeteman, B. C. J., and Kraayeveld, A. J. A.,  and
     Piet, G. J., "Oil Pollution and Drinking  Water
     Odor," H20, 4., 367  (1971).

13.  Keith, L. H., "Identification of Organic  Contami-
     nants Remaining in  a Treated Kraft Paper  Mill
     Effluent," American Chemical Society,  Division of
     Water, Air, and Waste Chemistry Preprints, 76  (1969),
     Minneapolis, Minnesota.

14.  Garrison, A. W., Keith, L. H., and Walker, M.  M.,
     "The Use of Mass Spectrometry in the Identification
     of Organic Contaminants in Water from  the Kraft
     Paper Mill Industry," American Society for Mass
     Spectrometry and Allied Topics Preprints, B205
     (1970), San Francisco, California.

15.  Thruston, Jr., A. D., and Knight, R. W.,  "Charac-
     terization of Crude and Residual-Type  Oils by
     Fluorescence Spectroscopy," Environ. Sci. Techno!.,
     5, 65 (1971).
                                U. S. GOVERNMENT PRINTING OFFICE : 1973—514-156/364
                         82

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 SELECTED WATER
 RESOURCES ABSTRACTS
 INPUT TRANSACTION FORM
                                     /. RefoftNo.
2.
                                                      w
 4. Title                                                  5," heport Date
   ENVIRONMENTAL APPLICATIONS  OP ADVANCED INSTRUMEN-
   TAL ANALYSES:  ASSISTANCE PROJECTS, FY 69-71          ,
                                                        8.: Fig/or/a * af Org* g» . -.atioa
    . ..  . .
 7. Authttr(s)
   Lawrence H.  Keith and Shirley  H.  Hercules
 9. Orgnaizatioa
   National Water Contaminants  Characterization
   Research Program, Southeast  Environmental
   Research Laboratory
                                                      10. PtojM ,Vo.

                                                        16020  GHZ
                                                                  ftro
n.
                     Environmenal Protection Agency
                                                       |-f. Type
                                                          Period Covered
 IS. Supplementary
    Environmental Protection  Agency report number EPA-R2-73-155 ,
    May,  1973
  IS. Abstract
    A multitude of analyses  using gas chromatography-mass spectrometry
    (GC-MS),  nuclear magnetic  resonance (NMR), infrared, and fluorescence
    spectroscopy to identify and measure organic pollutants in water are
    discussed under eleven project categories.  In most cases these
    analyses  have helped to  solve, or at least understand more clearly,
    the related pollution incident and in some cases provided for
    enforcement of regulatory  legislation.  Projects included identifi-
    cation of pesticides and PCB's from natural waters, organics from
    industrial wastewaters and landfill runoffs, organics from paper
    mill wastewaters and foam, and analyses of oils from oil spills
    and suspect sources.
  Ha. Descriptors   *Water pollution,  *Chemical wastes, *Chemical analyses,
    *Mass spectrometry,  *0il spills, *Pesticides
  i~b. identifiers   *Fishkill,  *Polychlorinated biphenyls, *0rganoleptic
    properties, *Nuclear  magnetic resonance, *Fluorometry, Taste,
    Odor, Oil pollution,  Paper mill pollution, Phthalates, Fatty acids
    Resin acids
  171. COWRRField & Group 05A
	 ,w- ' 	 m
18 Availability ' tf- St<.uri"sCfa8. *

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