A STUDY OF THE DISTRIBUTION AND FATE OF
               POLYCHLORINATED BIPHENYLS
            AND BENZENES AFTER SPILL OF
                  TRANSFORMER FLUID
        \
         (S
       CONTRACT NO! 68-01-3232
DIVISION OF OIL AND SPECIAL MATERIALS CONTROL
    OFFICE OF WATER PROGRAM OPERATIONS
   U.S. ENVIRONMENTAL PROTECTION AGENCY
          WASHINGTON, D.C. 20460

   U.S. ENVIRONMENTAL PROTECTION AGENCY
      REGION IV- 1421  PEACHTREE ST., N.E.
         ATLANTA,GEORGIA 30309
JANUARY 1976

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            FOLLOW-UP STUDY OF THE
           DISTRIBUTION AND FATE OF
          POLYCHLORINATED BIPHENYLS
           AND BENZENES IN SOIL AND
        GROUND WATER SAMPLES AFTER AN .
     ACCIDENTAL SPILL OF TRANSFORMER FLUID
          CONTRACT NO.:   68-01-3232
DIVISION OF OIL AND SPECIAL MATERIALS CONTROL
     OFFICE OF WATER PROGRAM OPERATIONS
    U. S. ENVIRONMENTAL PROTECTION AGENCY
          WASHINGTON, D. C. .20460

                     and
     THE ENVIRONMENTAL PROTECTION AGENCY
    REGION IV - 1421 PEACHTREE STREET, NE
           ATLANTA, GEORGIA  30309
              PROJECT OFFICER,
               GEORGE J. MOEIN
       ENVIRONMENTAL PROTECTION AGENCY
              ATLANTA, GEORGIA
                JANUARY, 1976

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                           FOREWORD








The Environmental Protection Agency's interest-in the spills of




Polychlorinated Biphenyls (PCB) and related substances has been




demonstrated through a series of response oriented actions, and




joint EPA industry efforts to mitigate the damages caused by such




spills.  In recent years some significant quantities of PCB have




spilled, mostly from transportation related media in remote and




populated areas of the United States.  In the Southeastern United




States several such spills have occurred during the past three years




causing alarm to the public and large clean up expenses to the




industry.








This technically oriented study was designed to derive a PCB con-




centration profile in a spill area two years after the occurrence




of the spill.  This study is somewhat unique, in that many months of




field work and laboratory analysis were spent to examine numerous




environmental factors and parameters to determine the fate of PCB




and Benzenes in the "natural environment".  The findings and con-




clusions of this study should have significant value to EPA response




personnel who often have to determine and recommend a "safe level"




of clean up and removal operation; to the industry who frequently




pays for such operations; and to the general public who is ultimately




affected by the menace of spills.  The tabular data contained here-




in are arranged as such that little interpretation is needed for under-




standing by laymen.  Many diagrams, maps, and chromatograms have been

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included as supporting documentation and as reference for any

future work.



Many individuals have contributed to the success of this project

and should be acknowledged. 'Notably among them are Mr. Al J. Smith,

Chief, Environmental Emergency Branch of EPA, Region IV, who directed

the clean up and removal operation during the initial phases of the

spill and coordinated the Federal response activities; Mr. Kenneth

E. Biglane, Director, Oil and Special Materials-Control Division, EPA,

Washington, who conceived the idea for the follow-up study and pro-
                  1              .    .          v                ^
vided the necessary funding; Messrs. Bill Loy and Tom Bennett of

Surveillance and Analysis Division, EPA,.Region IV, who assisted in

the sample analysis and the quality control phase.



It is reasonable to expect that subsequent studies of a more detailed

nature will be made in this area to answer many remaining questions.
                               George J. Mbein
                               Chief, Hazardous Materials Section
                               Environmental Protection Agency
                               Region IV, Atlanta, Georgia'
                               Hans Crump-Weisner
                               Co. Project Officer
                               Environmental Protection Agency
                               Washington, D. C.
                                ii

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                             TABLE OF CONTENTS



                                                                      Page
                                                                       No.

  I.  SUMMARY	• .  .  •	   1


 II.  CONCLUSIONS	   2


III.  RECOMMENDATIONS .'...'	,	   3


 IV.  INTRODUCTION	 .  .	.  .  .  .	   4

      A.  History of Spill	   4

      B.  Brief Review of PCB Literature	   5

          1.  Distribution of PCBs in the Environment	   5

        .2.  Fate of PCBs in the Environment	   6

          3.  Literature References  Cited		•   8
                                              *•
      C.  Purpose of This Study	   *


  V.  TECHNICAL APPROACH	 . .   U


      A.  Introduction	   *•*• •

      B.  Sampling Protocol and Collections . .	   ^

          1.  Criteria for Selection of Soil Sampling Sites .....   1*

          2.  Initial Field Preparations  	 .....   **

          3.  Drilling and Soil Sampling	   15

          4.  Field Procedures Relating to Soil  Sampling	\ ^  20

          5.  Site Selections for Sediment and Water	   22

          6.  Sampling Procedures for Sediments, Water,
              and Controls	   23
                                     iii

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                            TABLE OF CONTENTS
                               (Continued)
                                                                     Page
                                                                      No.
     C.  Preliminary Laboratory Preparations and Splits       , ' '
         of Soil Samples	   24

     D.  Microbiological Studies	 .	   25

         1.  Procedures for Microorganism Investigations 	   25

         2.  Determinations of Soil pH and Moisture	   26

     E.  Analytical Methodology	   27

         1..  Analysis of Water, Sediment, arid Soil
             for Polychlorinated Biphenyls (PCBs)  .........   28

         2.  Analysis of Water, Sediment, and Soil '
             for Polychlorobenzenes	   35

     F.  Isomer Verification	   36


VI.  EXPERIMENTAL RESULTS  . . . .  .	 .  . . .  .  . .  .  .   41


     A.  Field Data	   41

         1.  Cores	 . .	   41

         2.  Geology	 . .	   41

         3.  Environmental Samples .	   46

         4.  Wells .	   46

         5.  Climatology	...........   49

         6.  Soil Temperature  .........	  .   52

         7.  Drilling Equipment and Procedures . . .  . . .  .  ...  .   56

     B.  Microbiological Data	\ .  .  .   56

     C.  Analytical Data from Electron-Capture Gas
         Chromatography (EC/GC)	• •  •   78

     D. .Special Soil Extraction Experiment  .... 	   78
                                     iv

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                             TABLE OF CONTENTS
                                (Continued)
                                                                      Page
                                                                       No.
      E.  Quality Assurance Data
      F.  Analytical Data for Gas Chromatography/
          Mass Spectrometry (GC/MS)  .  . . . ............    91


VII.  FATE OF POLYCHLORINATED BIPHENYLS (PCBs) AND
      POLYCHLOROBENZENES AFTER A TWO-YEAR EXPOSURE
      IN A NATURAL ENVIRONMENT ......... ........  .  .    92


      A.  Fate of Aroclor 1254 ..................  ,    92

          1.  Distribution of Aroclor 1254 In and
              Around the Spill Site in 1975  .  ...........    92

          2.  Degradation of Aroclor 1254  ..... .....  '.  .  .    108

      B.  Fate of Askarel Solvent  .................    113

      C.  Over-all Assessment of the Environmental
          Impact of the Spill  ...... ........  .....
          1.  Condition of the Area Immediately
              After the Spill  ..............  .....    117

          2.  Condition of Spill Area—August 1975  .......  .  .    U9

  APPENDICES ....... ......... ............    I21

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                            LIST OF TABLES

Table No.                            l

    1      Gas ChrOmatographic Operational Parameters—
           PCBs	;.,....	 .       31

    2      Method Sensitivity—PCBs  .  .	       32

    3      Gas Chromatographic Operational                  /
           Parameters—Askarel Solvent .............       37

    4      Method Sensitivity—Askarel Solvent	       38

    5      Description of Soil Types Characterizing
          - Core Samples Taken in August 1975	       45.

    6      Climatological Data Showing Monthly
           Averages for the Study Area	       51

    7      Rainfall Data Prior to and During
           Well Sampling	       53

    8      Collection and Microbiological Data from
           Soil Samples Taken in August 1975	       58

    9      Microbial Populations Relative to
           Soil Types	       65

   10      Microbial Populations Relative to
           pH of Soil Samples	'	       66

   11      Microbial Populations Relative to
           Moisture Content of Soils 	       68

   12      Microbial Populations Relative to
           Core Sample Depths  .	      ,70

   13      Microbial Populations Relative to
           PCB Concentration in Core Samples .........       77

   14      Collection and Analytical Data from
           Three Test Core Sites Sampled Over
           Four-Inch Intervals in August 197,5  ........       79
                                   vi

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                            LIST OF TABLES
                             (Continued)

Table No.                                                          Page
   15      Collection and Analytical Data from
           Core Sites Sampled Over Four-Inch
           Intervals in August 1975	       82

   16      Collection and Analytical Data from
           Core Sites Sampled Over Sixteen-Inch
           Intervals in August 1975	       83

   17      Aroclor 1254 and Askarel Solvent
           Analyses for Water, Sediment, and
           Control Soil Samples	       86

   18      Results from Special Soil Extraction
           Experiment—Aroclor 1254	  .       88

   19      In-House Quality Control Data—
           Aroclor 1254	'......       89

   20      Percent Coefficient of Variation
           Evaluation for In-House Quality Control
           Data—Aroclor 1254	 .  .. .  .       90

   21      Comparison of Analytical Data from
           Core Samples Collected in 1973 and 1975	      106

   22      Typical Homolog Composition of
           Aroclor 1254  .	      Ill
                                   vii

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                           LIST OF EXHIBITS

Exhibit No.

    I        Location of Spill Site in Relation
             to General Topography . ,	      17

   II        Excavation Area and Location of
             Core Sites with Contour Intervals ........      42
        »
  III        A Cross Section Profile of a
             Northeast Intersect through
             the Study Area	      43

   IV        A Cross Section Profile of a
             North-Northwest Intersect
             through a Steep Surface Gradient
             Section of the Study Area	 .      44
                                   viii

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                           LIST OF FIGURES

Figure No.                                                        Page

    1        Representative Chromatogram for  ,
             Aroclor ,1254	     33

    2        Representative Chromatogram for
             Askarel Involved in 1973 Spill	     34

    3        Representative Chromatogram for
             Solvent Portion of Askarel  .	     39

    4        Representative Chromatogram for
             Standard Chlorobenzene Mix  . . .1	     40
    5        Location of Environmental        ',
             Sampling Sites	...'...	     47

    6        Locations of Well Stations Relative
             to the Spill Site	     48

    7        Location of the Control Well  .,	     50

    8        Temperatures of Various Soil Types
             Over Four-Inch Intervals as Compared
             with Ambient Conditions 	     55

    9        Total Ion Current Chromatograms ... 	     93

   10        Specific Ion Search for the 292* Ion	     94

   11        Specific Ion Search for the 326* Ion	     95

   12        Specific Ion Search for the 360+ Ion	     96

   13        Specific Ion Search for the 394+ Ion	     97

   14        Spectrum Typical of Four Chlorine
             Biphenyl  ......... 	     98

   15        Spectrum Typical of Six Chlorine
             Biphenyl	     99

   16        Spectrum Typical of Seven
             Chlorine Biphenyl .	 .  .	     100
                                    ix

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                           LIST OF FIGURES
                             (Continued)
Figure No.                                                         Page

   17        Concentration Distribution of Aroclor
             1254 in Core Samples, 1975  ............     102

   18        Concentration-Depth Profiles of
             Core Site UC, 1973-1975	. . . . .     103

   19        Concentration-Depth Profiles of
             Core Site RL, 1973-1975	     104

   20        Concentration-Depth Profiles of
             Core Site JF, 1973-1975 . .  .	     105

   21        Comparative Chromatograms for
             Aroclor 1254 in a Typical "Aged
             Environmental Sample" (1975)                       ,
             and Aroclor 1254 in the Askarel
             Spilled in 1973   .... ....'.........     112

   22        Solvent Concentration Profile—
             Well Located Closest to Spill
             Site (Station 7)  . .	     116

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I.  SUMMARY




    The area of an askarel spill, which had been cleaned up two years




    prior to this study, was investigated for migration and/or degrada-




    tion of residual PCB and lingering intrusion of the solvent into




    ground waters.  The'PCB was found unchanged while the solvent had




    continued to leach into the underground water.

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II.  CONCLUSIONS

     No significant reduction in the concentration of Aroclor 1254 in the

     'soil has occurred as the result of migration or degradation.  There

     is no way to clearly assess the effect of the original insult on the

     soil microorganism population of the spill site since no microbiological

     studies were conducted in 1973.  There was, however, no evidence of
                                 «     X
     permanent environmental damage detected in the spill area in 1975.



     The more water-soluble components of the askarel solvent invaded the

     ground water supply almost immediately after the occurrence of the

     spill.  Leaching was the migration mechanism responsible for the

     intrusion of the lower chlorinated benzenes into the ground water.

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III.  RECOMMENDATIONS

      Because of the many diverse and interrelated effects imposed by a

      particular spill environment, no threshold concentration of PCBs
                                                           \ ,
      can be recommended which would be equally applicable to all land

      spill occurrences.  Although the data from this study can serve as

      a guideline, each PCB spill will require an individual evaluation

      and assessment since no appreciable migration or degradation was

      detected in the specific environment Investigated.  It is recommended

      that more research be conducted with varying soil types and a more

      favorable medium for sustaining microorganisms in greater abundance.

      More knowledge of the toxicity of askarel on various microbla would

      be required as an integral part of these degradation studies.

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IV.  INTRODUCTION


     A.   History £f Spill.


         On March 5, 1973,  an accidental spill of approximately 1500


         gallons of askarel occurred in a rural area near Kingston,


         Tennessee.  The spill resulted in the.environmental.contamina-
                     '                                        'I
         tion of two watersheds because of its location on the crest of

         a hill.  Through the influence of rainfall, geology,  and

         characteristics of the overlying stratum of soil, the chemical


         was subsequently dispersed through the soil both horizontally

         and vertically.



         The particular askarel spilled was composed of a commercial ,


         mixture of polychlorinated biphenyls (Aroclor 1254) and a pro-

         prietary solvent mixture of polychlorinated benzenes.  Both

         components of  the askarel involved were chlorinated hydrocarbons

                                                        \
         —a class of compounds noted for their persistence in the environ-

         ment.   Consequently, an extensive "clean-up" operation in the
                                          I
         affected area  was started March 14, 1973, by the responsible

         parties.  To assure the effective removal and proper disposal of

         the hazardous  substances, the contaminated soil was packed and

         sealed in metal drums before it was taken from the spill site.

         The removal procedure resulted in extensive excavations in three

         general areas.  After ll,531 drums of contaminated soil were re-

         moved, 'the excavated areas were sealed, backfilled, and packed

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    with uncontaminated soil.  The entire affected area was subse-




    quently covered with top soil, seeded with grass, and landscaped.






    On March 8, 1973, the Regional Office (Region IV) of the




    Environmental Protection Agency (EPA) in Atlanta commenced a




    sampling program of the affected areas to determine the concen-




    tration level of contaminants in the soil and water table.




    Stewart Laboratories, Inc. (SLI), a private laboratory located




    in Knoxville, TN, joined EPA in the sampling program on March 21,




    1973.  This sampling program, for monitoring and detection pur-




    poses, continued jointly by EPA and SLI for several additional




    weeks.  In addition to the EPA sampling program, Stewart




    Laboratories, provided sampling and analytical assistance during



    the excavation.






    A monitoring program, approved jointly by EPA and TWQC (Tenn.




    Water Quality .Control Board) was conducted for a 12-month period




    following the cleanup operation.








B.  Brief Review of_ PCS Literature.




    1.  Distribution £f_ PCBs jln the Environment.  According to the




        Interdepartmental Task Force on PCBs (1), the history of




        PCBs started in 1929 when industry introduced them for use




        as non-flammable oils in electrical transformers, condensers,




        and in paint.  During the next forty years,- industrial uses




        of PCBs grew steadily.  They have been widely employed as




        plasticizers, as sealers in waterproofing compounds and




        putty, in printing inks, in waxes, in synthetic adhesives,

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    in cutting oils, as dielectrics, as hydraulic fluids, as



    high-pressure lubricants, and as a heat-transfer medium (2).



    Sales of PCBs in the United States came to about 34,000



    tons in 1970, with cumulative production over the years



    amounting to an estimated 4 x 10^ tons (3).






    Although PCBs were never intended for direct release into




    the environment, they were first identified by Jensen as




    a potential food contaminant in 1966 (4).  Since that time,



    it has been demonstrated that .they are ubiquitous environ-



    mental pollutants.  Numerous studies (5-18) have confirmed




    their presence in animals and the aquatic environments as



    well as in humans.








2.  Fate of PCBs jin the Environment.  The sparsity of knowledge



    about the fate of PCBs in the environment is Illustrated by



    the recommendation of. the Interdepartmental Task Force on



    PCBs (1) that "more scientific information about PCBs is
                            /


    needed" relative to their occurrence, transfer, and cycling



    in the environment.  Only general statements can be made



    about how PCBs reach the environment and how they reach



    target organisms.






    The biologically important characteristics of PCBs are their



    insolubility in water, high solubility in fats, toxicity to



    metabolic processes, and extreme stability.  The combination



    of persistence and accumulation in fat (during transition

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through a food chain) can result in considerable increases in




concentration of the compounds at higher levels in the food




chain.  Like DDT, PCBs are.reported to degrade very slowly




under natural conditions (19).  The highly chlorinated PCBs




(Aroclor 1254 and 1260) seem to persist in the environment




longer and are less toxic than the more rapidly degradable




low chlorine forms (20).






The metabolism of several PCB isomers in fish, rats, and




pigeons has been studied by Hutzinger and co-workers (21).




Mono-, di-, and tetrachlorobiphenyl isomers were converted




into their corresponding mono-hydroxy derivatives in rats



and pigeons.  However, no hydroxylation of any of the isomers




was observed with fish.  It was also observed that 2,2',4,4',




5,5'-hexachlorobiphenyl was not oxidized by any of the




animals in the study.






Laboratory photolysis studies on pure chlorobiphenyl isomers,




as well as Aroclor 1254, have been reported by a number of




investigators (22-26).  Using hexane as the solvent, the de-




chlorination reaction was predominant.  However, in the




presence of air and water, a number of polar products were




observed.  Examination of the polar products.from the Aroclor




1254 irradiation indicated formation of both hydroxylated




and hydrated products.  Although these results suggested




pathways in which degradation of environmental PCB might

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                                                                 8
    occur, the actual observation of degradation products in

    "natural environmentally aged samples" has not been reported

    to date.



3.  Literature References Cited.

    1.  .Interdepartmental Task "Force\on.PCBs, U. S. Department
        of Commerce Pub. COM-72-10419, 1972.

    2.  Bonelli, E. J., Am. Laboratory, February 1971.

   . 3.  Hammond, A. L., Science, 175.  155 (1972).

    4.  Jensen, Soren, PCB Conference, National Swedish
        Environment Protection Board,  Stockholm, 7 (Sept., 1970).

    5.  Risebough,  R. W., Rieche, P., Peakall, D. B., Herman,
        S. G., and Kirven, M. N., Nature, 220. 1098 (1968).

    6.  Veith, G. D.,  and Lee, G. F.,  Water Research, 4., 265
        (1970).

    7.  Martell, J. M., Rickert, D. A., Siegel, F. R., Environ.
        Sci. Technol., 2» 872 (1975).

    8.  Maugh, T. H.,  II, Science, 178, 388 (1972).

    9.  Veith, G. D.,  and Lee, G. F.,  Water Research, _5,
        1107 (1971).

   10.  Bailey, S.,,and Bunyan, P. J., Nature, 236, 34 (1972).

   11.  Edwards, R., Chem. and Ind., 1340 (1971)

   12.  Price, H. A.,  and Welch, R. L., Env. Health Perspectives,
        it 73 (1972).                      ,

   13.  Burns, J. E.,  Pesticides Mon.  J., _7, 122 (1974).

   14.  Gruger, E. H., Jr., Karrick, N. L., Davidson, A. I.,
        and Hruby, T., Envion. Sci. Technol., JJ» 121\(1975).

   15.  Koeman, J. H., et al., Nature, 221, 1126 (1969).

   16.  Bache, C. A.,  et al., Science, 177, 1191 (1972).

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        17.  Holden, A. V., Nature, 228. 1221 (1970).

        18.  Crump-Wiesner, H. J., Feltz, H. R., Yates, M. L., Jour.
             Research U. S. Geol. Survey, _!, 603 (1973).

        19.  Waldbott, G. L., Health Effects of Environmental
             Pollutants (C. V. Mosby Co., St. Louis, 1973), p. 225-9.

        20.  Monsanto Company, Presentation to the Interdepartmental
             Task Force on PCBs, Washington, D. C., May 15, 1972.

        21.  Hutzinger, 0., et al., Science, 178, 312  (1972).

        22.  Stalling, D. L., 163rd National Meeting, ACS, New York,
             N. Y., Symposium on PCB, No. 22, August 1972.

        23.  Hutzinger, 0., Jamieson, W. D., and Zitko, V.,
             Nature, 225. 664 (1970).

        24.  Safe, S., and Hutzinger, 0., Nature, 232. 641 (1971).

        25.  Ruzo, L. R., Zabik, M. J., and Schuetz, R. D., Bull.
             Environ. Contam. Toxicol., j}, 217 (1972).

        26.  Hustert, K., and Korte, F., Chemosphere, 1, 7 (1972).
C.  Purpose of This Study.
           «
    The primary purpose of this project is to study the biodegradation

    effects of a natural environment on the chemical components of an

    askarel spill after a two-year time lapse.  Extensive experimental

    background data were obtained during and after the 1973 cleanup

    procedures employed at the study site.  Correlation of 'the 1973

    data with the results of this follow-up study will provide a means

    of assessing the distribution and fate of PCBs and polychlorinated

    benzenes in natural environmentally aged samples.


    The findings of this project may form the basis for deriving a safe

    concentration level for PCBs in soil,  The practical application

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                                                                 10
of such a threshold level will assist EPA spill response personnel



to determine the degree of soil removal necessary in a land spill



situation.  Sampling of water supplies adjacent to the spill area



will provide information on the rate of intrusion of lower chlori-



nated benzenes into ground water.  Such information will be of




paramount importance in increasing the efficiency and effective-



ness of EPA's spill cleanup efforts.

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                                                                         11
V.  TECHNICAL APPROACH




    A.  Introduction.




        Potential mechanisms for the loss of askarel from the spill




        site include volatilization, leaching, and metabolic and/or




        nonmetabolic degradation.  The experimental approach to be




        employed in this study has been designed in a manner which will




        detect and assess the magnitude of each potential route for.the




        removal of residual askarel from the spill site.






        Data obtained by Stewart Laboratories, Inc., during the 1973




        site cleanup included:  (1) core samples in the bottom of



        excavation areas immediately prior to fill; (2) core samples




        at elevations geologically below the spill; (3) water and sedi-




        ment samples from a spring well in a watershed below the site;




        (4) water samples from domestic wells adjacent and peripheral




        to the spill area; (5) water samples from sources in the general




        vicinity of the spill area; and (6) water samples from a domestic




        control well.  The initial phase of this project involved a •




        re-sampling of the area in order to determine migration and/or




        degradation of the PCBs in soil and askarel concentration changes




        in water.






        The second phase of the study provided the analytical data




        necessary to evaluate any changes which might have occurred over




        the past two years.  Identical analysis procedures, concentration




        units, and detection limits were employed for the determination

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                                                                          12
         of PCB content in both the present study and the 1973 project

         so that direct comparison of the two sets of data would be

         facilitated.   Careful choices were made as to which samples

         should be confirmed and identified by GC/MS analysis.  The.

         requested isomer identifications were done by the Surveillance  .

         and Analysis  Division, Region IV, EPA, in Athens, Georgia.  The

         techniques employed in the field and laboratory positively identi-

         fied soil profile correlations which allowed for the interpre-

         tation of chemical and biological data.


         Core samples  of soil were examined by established procedures

         to determine  the relative numbers and major types of microbial

         flora.  Among these were the true bacteria, actinomycetes, and

         a wide variety of microscopic fungi, all of which were enumerated

         and identified.


         Algae and protozooans are also found in soil.  However, they

         are far less  numerous than the other microorganisms (27) and

         are more difficult to evaluate because of somewhat unique

         cultural requirements and were, therefore, not investigated.


         Since available literature indicates that very little is
(27)  Microbiology.   3rd Ed., 1972.  M. J. Pelczar and R. D. Reid.  McGraw
     Hill Pub.,  New York, New York.

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                                                                     13
    known concerning the degradation of PCBs in the natural environ-




    ment, potential biodegradative microorganisms are not identifi-




    able.  The two-year lapse since the spill incident, however,




    should have allowed for some valid degradation.  The over-all




    approach in determining microorganism abundance was to isolate




    certain groups which were probable major participants in




    chemical alterations.  It was realized, however, that other




    environmental conditions such as pH, moisture, type of soil,




    and sample depth influence the relative numbers and varieties




    of soil flora.








B.  Sampling Protocol and Collections.




    During the field sampling phase of the project, the contractor




    collected 120 soil samples and 40 water samples in and around




    the spill area.  These samples were to constitute the basic




    materials for the study of the distribution and fate of




    residual PCBs in the natural environment.  Probable causes




    of measured and documented changes in the chemical status




    of the spill components could then be addressed.






    An initial over-all assessment was made of the requirements




    for proper choice of primary sampling sites, procedures for




    collection of samples and attendant field data, as well as




    essential equipment and supplies.  A final protocol was estab-




    lished which required only minor modifications once the field




    work began.

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                                                                 14
1.  Criteria for Selection of Soil Sampling gites.  All analytical


    data obtained should have reasonable reference to analyses


    performed in 1973.  The core sites chosen for comparison


    were to represent a broad concentration range.  The areas


    from which these cores were taken needed to exhibit a full


    spectrum of topographic variables which would characterize


    the study area as to soil type, probable hydrology, microbial


    environments, and potential migration of PCBs.  Attempts


    needed to be made to sample areas of undisturbed terrain


    as well as those which had been excavated or top dressed


    in 1973.  Distribution patterns of the residual contamina-


    tion in the 1973 cores were studied in order to replicate,


    as closely as possible, the same contamination variability


    for the 1975 sampling.  The depth of sample takes would


    be dependent on the assessment of all other criteria thus


    mentioned.  It would, therefore, be essential that an


    appropriate number of available sites be chosen so that

                                       .t
    the interdependency of all these requirements could be up-


    held; and unforeseen field situations would not preclude


    an adequate sampling.
                                  t


2.  Initial Field Preparations.  The spill area had received
      '.

    little maintenance since it had been backfilled and seeded


    in 1973.  Successive cuttings with rotary mowers and hand


    raking were required to remove grass, weeds, and limbs from


    the study area.  Photographs, drawings, and field data were

-------
                                                                15
    used to approximate the location of the excavation perimeter



    and the 1973 core sites.  Surveyors'  flags and stakes were



    used to mark these assigned areas.   Photographs and rough



    sketches were recorded as guides for final and more exact



    designation.








3.  Drilling and Soil Sampling.  The extreme variability in depth



    of required cores and the soil matrix (clay with fractured



   .quartz) indigenous to the site area required the use of a



    truck-mounted drill rig.  The drill unit was thoroughly



    washed and cleaned prior to arrival on site.  It remained



    in the study area throughout the sampling period.






    A methodical drilling program was begun to determine the



    excavation limits in all areas peripheral to probable



    sampling sites.  Continuous cores were pulled using a split



    spoon sampler for the determination of soil profiles.  Since



    the floor of all excavated areas had been sealed with "Vis-



    queen" and a layer of sand (2-4 inches) prior to backfilling,



    the position of the perimeter of the disturbed area was



    readily discernible.  Appropriate adjustments on all drawings



    and records were made as documentation.  All field markers



    were securely installed at their verified locations.  During



    this preliminary drilling period, extreme precautions were



    taken against contamination of the surface of the study




    areas.  The positioning of the drill truck and the disposition

-------
                                                             16
of the test cores were under constant surveillance.  Once

logging of core data was completed, the cores were tamped

back in their original holes.


All original 1973 core sites,external to the excavation had

been backfilled, packed, and covered with top soil.  In

order to identify the core holes and differentiate fill

top soil from natural overburden, each core hole was found ' -

and drilled once with the split spoon.  By visual observa-

tion and actual measurement of the soil strata, the depth

of unnatural cover material was ascertained.  The sampler

was pressed gently and rotated in order to avoid compact-

ing.  Impact techniques were restricted to instances where
                                 i
chert refused penetration.


Available core sampling equipment included a split spoon

sampler, a Shelby Tube Sampler, and a California Sampler.

The order of desirability of the three core samplers were:



a.  California sampler—consists of four sequential brass

    units two inches in diameter and four inches long allow-

    ing for ease of sample removal and differential choice

    of sample take.  The cost per tube is moderate and

    they are readily available.  Two limitations were the

    questionability of penetration of quartz layers and

    the sample size.

-------
                        ROAD CLASSIFICATION

         Heavy-duty	______ Poor motor road ...  -,- = = ,-
         Medium-duty	^-_=__^ Wagon and jeep track	
         Light-duly	== Foot trail		
                          -EXHIBIT I   Location of  Spill Site in
                                         Relation to  General  Topography
                In developed areas, on

                                  BACON  GAP, TENN
                                      N3545-W8430/7.5








                                                                       t> ^ if,-2/A;sl .V'c-fn^.^^-r^--
                                                                         SCALE  1:24000
                                                                               o
                                                1000
                                                               1000
                                                                       2000
                                                                              3000
                                                                                             5000
                                                                                                               l fECI
utM GRID AND 1968 MAGNETIC NORTH
 DECLINATION AT CENTER OF SHEET
                           . CONTOUR INTERVAL 20 FEET
                    .DASHED LINES REPRESENT HALF-INTERVAL CONTOURS
                               DATUM IS MEAN SEA LEVEL

                 THIS MAP COMPLIES WITH NATIONAL MAP ACCURACY STANDARDS

            FOR SALE BY U.S. GEOLOGICAL SURVEY. WASHINGTON. D.C. 20242.
              TENNESSEE DIVISION OF GEOLOGY, NASHVILLE. TENN. 372.19.
U.S. TENNESSEE VALLEY AUTHORITY. CHATTANOOGA, TENN. 37401 OR KNOXVILLE. TENN. 3/90?
          A FOLDER DESCRIBING TOPOGRAPHIC MAPS AND SYMBOLS IS AVAILABLE ON REQUEST

-------
                                                             18
b.  Split spoon sampler—an open-chambered tube  two inches



    in diameter and ^24 inches long which provides a



    relatively rapid sample take.  Two disadvantages of



    this sampler are possibility of disturbed core and poten-



    tial cross contamination of samples.







c.  Shelby tube—a sturdy impact corer of three  inches in



    diameter and thirty-six inches in length and capable



    of delivering substantially undisturbed samples.  Three



    disadvantages are that each sample has to be extruded



    with a press, the tubes are very expensive, 'and the



    units are marketed with a baked-on plastic liner to



    prevent rust.





After careful consideration of the sampler types and the



inherent geology involved, the California Sampler was
                                        f


initially chosen for trial.  Because of extreme difficulty



encountered in removing clay and chert fragments from the



tubes in the field, the split spoon sampler was used for



all consequent core ^samplings.





Since the topography of the study area rolls at varying



angles around the spill site, a fairly comprehensive choice



of sampling spots was made.  The Knox Dolomite (undivided)



sub-surface versus the overburden influx on some of the sites



external to the excavation broadened the coverage of the

-------
                                                            19
study region from the standpoint of degradation and/or move-



ment of PCBs.  A topographic representation of the general



spill site is shown in Exhibit I.





Sample core drilling began on August 4, 1975.  The first



three core sites chosen for replicate testing were drilled



in a four-cornered quadrant pattern.  Each of the four cores



was taken approximately one foot from the center of the 1973



core location.  All samples were taken over four-inch inter-



vals to allow for differentiation of gradient effects and



reproducibility studies.





In order to representatively sample the site variations as



specified in the protocol, all subsequent collections were



designed to take soil from one core hole adjacent to an



identified 1973 core hole.  This phase of collections began



with core taken over four-inch intervals; however, sampler
                                            *


refusal and tendency toward loss of sample integrity be-



cause of inherent quartz in the clay matrix precluded



sampling over such a short interval.  A decision was made



to complete the sampling with cores composited over sixteen-



inch intervals.

                                           ^-              . •



Upon completion of all collections peripheral to the



excavation area, core samples were taken at a level approxi-



mating some of the original sample points at the bottom of

-------
                                                                 20
    the backfilled areas.  The location of these samples was

    confined to an area within three feet of the 1973 sampling

    point.  Few sites were available for this phase of the study

    since nearly all excavation areas received additional clean-

    ups after final data had been obtained in 1973.  Additionally,

    the floor and walls of one deeply excavated area had been

    impregnated with a material to inhibit movement of any

    residual askarel.  There was no ready access to the basin

    sampling areas without breaking the seal to the environment.



4.  Field Procedures Relating £o Soil Sampling.  The techniques

    employed in the actual sampling and handling of core samples
                     \ •      '
    and the collection of supplemental field data followed the

    protocol with no modifications.



    a. . Core samples—Each split spoon sample was carefully re-

        moved onto a piece of aluminum foil supported on a ply-

        wood base.  It was measured, cut into four sections of

        four-inch length, logged, characterized as to general

        soil type, and sealed in labeled foil for transport to

        the laboratory.                   \
                                           \


    b.  Cleaning and contamination precautions—Prior to each
                                             \              ' '
        use, all drilling equipment, core cutting blades, etc.,

-------
                                                             21
    were washed with water, and a brush, scrubbed in a bath

    of technical grade hexane, and finally rinsed in

    pesticide grade hexane.  This operation was not performed

    near sampling sites.  Disposable gloves were used while

    handling each sample take, and all throw-away materials

    were kept carefully sealed.  Foil strips were removed

    from the roll and appropriately labeled in an area remote

    to the core operations.


c.  Temperature measurements—A general profile was made

    of the temperature of the soil.at several points within

    the study area.  A four-inch temperature-sensing probe

    with a strip chart recorder readout was inserted in the

    test cores.  Cross checks on the accuracy of these read-

    ings were made by inserting the probe in soil at very-

    ing depths in advance of core takes.  General confirma-"

    tion between these two procedures was obtained.  The

    probe was cleaned in the manner described in "b" above.

                                                      »
d.  Mapping—A grid survey .on two-foot intervals was made

    of the entire study area.  The data from these measure-

    ments were translated into a contour map depicting the

    exact location of sampling sites and excavation limits

    on a grid basis.  Substantiating photographs were shot

    over the entire region with identifying landmarks in

    plain view.

-------
                                                                 22
    e.  Restoration—A concerted effort was made to return the
                    1                   /                 .       -
        area back to its original state.   All core holes were

        plugged; stakes, flags, and twine were removed; and all

        equipment, tools, and supplies were returned to, the

        laboratory facility.



5.  Site Selections for Sediment and Water.  A protected spring

    well located in the watershed below the spill site had been

    monitored during the 1973 cleanup and subsequently for about

    a year.  Since some positive data had been obtained from

   
-------
                                                                23
6.  Sampling Procedures for Sediments, Water, and Controls.



    Once the sites external to the spill were chosen and collec-



    tion permission obtained from the well owners, grab samples



    were taken after relatively dry weather (August 29, 1975)



    and again after heavy rains (September 26, 1975).






    a.   Techniques—^Sediments were taken with a metal, roll-


        sided scoop since the depth of water was quite shallow.



        Each sample was placed in a pre-labeled glass jar whose


        cap was lined with aluminum foil.




        All water samples were taken directly into a pre-


        labeled gallon glass jug.   Well samples were obtained



        at the tap from which they had previously been sampled


        in 1973.   Each tap line was allowed to flush for 2-3



        minutes so, that samples could be drawn directly from
                                                \

        the well system itself.




        Two control soil samples were taken from nearby areas



        not influenced by the spill.  The collections were made



        at 3-6 inch depths below the surface.  One of these was



        from an exposed clay and chert matrix while the other



        was primarily loam.






    b.   Cleaning and contamination precautions—The sediment


        scoop was cleaned according to the procedure previously



        given for field equipment.  All glass containers were

-------
                                                                     24

            washed with distilled water, rinsed with technical grade

            hexane, rerinsed with pesticide grade hexane, capped with

            foil liners, and labeled at the laboratory.  Special care

            was taken to insure that caps and glass container mouths

            did not come into contact with potential contaminants

            in the field.



C.  Preliminary Laboratory Preparations and Splits of Soil Samples.

    Sediments were passed through a quarter-inch screen in order to

    remove inorganic and organic debris.  These samples were allowed

    to air dry in a metal tray.  The dried material was then passed

    through a U. S. Standard No. 18 sieve to insure uniformity of

    particle size.  The sediments were then submitted to the

    laboratory for PCB analysis.


    Core samples were mixed and spread for air drying on their

    individual aluminum foil sheets.  As the samples were being

    spread, an approximate 40-gram sample for moisture, pH, and

    microbiological determinations was sealed in a pre-labeled

    container and submitted for analysis.  Forty representative

    soil samples of approximately 100 grams each were sealed in

    containers for potential subsequent anaerobic study.

   v                                            '. •                 i
    The dried core aliquots for PCB determinations were passed through

    a U. S. Standard No. 18 sieve after obvious rocks and debris had

    been removed.  The fines were then passed through a splitter in

-------
                                                                         25
        order to obtain an aliquot of an appropriate size for analysis.

        This aliquot was submitted to the laboratory for analysis of

        PCBs.


        All sample preparations were performed under clean room condi-
                                                          i
      1  tions.   Disposable gloves were worn and all equipment and con-

        tainers were pre-cleaned by washing and subsequent hexane rinses.
               . •   .                                       i
        Samples were kept distinct from each other, and each step of  -

        the preparations was done in a separately ventilated zone in

      .  the laboratory.                                   .


    D.   Microbiological Studies.

        One hundred forty-two soil samples were examined to determine the

        populations of heterotrophic aerobic bacteria, fungi, and actino-

        mycetes, which constitute the majority of microorganisms in soil.

        Laboratory data concerned with microbial flora enumeration and

        identification were researched in correlation with soil type, pH,

        moisture, and sample depth.


        1.   Procedures for Microorganism Investigations.  The various

            techniques and culture media described by Parkinson, et al.

            (28) were used for the isolation of the soil microorganisms.


            Ten grams of each soil sample were suspended in 95 ml of

            sterile distilled water to give a 1:10 dilution.  From this,
(28)  Methods for Study the Ecology of Soil Microorganisms.  1971.
     D.  Parkinson, T.  R.  Gray, and S. T.  Williams.  International Biological
     Programme Bladswell Scientific Publications, Oxford, England.

-------
                                                                 26
    serial 10-fold dilutions were prepared.  Dilutions of 1:100

    through 1:1,000,000 were employed for determining plate

    counts to enumerate the three microbial groups.  The numbers

    of heterotrophic aerobic bacteria were obtained by plating

    the various dilutions of the soil samples in Plate Count

    Agar (Difco Laboratories, Detroit, Mich.) and incubating

    at 25°C for 72 hours.  Colony counts were then determined

    with the use of a Quebec Colony Counter (American Optical

    Company, Buffalo, N. Y.).  Determination of fungal counts

    were conducted by plating dilutions of the samples in Sabour-

    aud's Dextrose Agar (Baltimore Biological Laboratories,

    Cockeysville, Md.) with incubation at 25°C for 96-120 hours.

    Enumeration of the organisms was performed as described for

    the heterotrophic bacteria.  Actinomycete counts were made

    by plating soil dilutions in Starch Casein Agar, containing

    0.0002 percent actidione for inhibition of fungi (Kuster and

    Williams, 1964X, followed by incubation at 25°C for 96-120

    hours.   Again, colony counts were made after incubation as

    described above.  If no growth was observed on the fungal

    or actinomycete plates after 96 hours, incubation was con-
                                                     \
    tinued and the plates read at 120 hours in order to detect

    any slower growing organisms.  The heterotrophic aerobic

    bacteria grew readily and never required more than 72 hours

    for maximum growth.




2.  Determinations of Soil pH and Moisture.

    The pH of each sample was obtained by preparing a slurry

    with three grams of soil and three ml of .distilled water

-------
                                                                    27
        in a 10 ml beaker and making the determination with a

        Labomatic Model 165 pH meter.



        Moisture content of a representative five-gram aliquot of
                                                             I
        each sample was obtained by drying to constant weight at
E.  Analytical Methodology.                                   j

    The method of choice for the analysis of PCBs in all known
                          »
    monitoring and regulatory applications is GC/EC (gas-liquid
                                                                  \
    chromatography utilizing electron capture detection) .  This

    analysis mode is, likewise, most frequently employed in the

    assessment of the environmental impact and health effects of

    PCBs.  This overwhelming utilization is by no means intended

    to imply that the analytical method is the ideal mode of analy-

    sis for PCBs.  Gas chromatography is not ah inherently definitive

    analytical technique.  It is subject to serious complications

    when other electron-capturing components are present in the

    samples in addition to the PCBs.  The shortcomings of the method

    can be successfully overcome when the analyst involved is fully

    cognizant of the ramifications of the situation.  Most analytical

    techniques incorporate a liquid chromatography cleanup of liquid-

    liquid extracts prior to GC/MS (gas chromatography/mass spectro-

    metry) analysis.  This procedure is % 90 percent effective in

    separating PCBs from organochlorine pesticides.

-------
                                                                 28
The utilization of two or more unlike columns in the gas


chromatpgraphie analysis is another means for establishing the


identity of gas chromatographic patterns.  The application of


GC/MS is considered by most experts to be the desired technique


for confirming qualitative identification of gas chromatographic


patterns.  Microcoulometry and thin layer chromatography are


also useful tools for positive identifications.  It is appro-


priate to conclude, however, that GC/EC is a most effective
                                 *

analysis mode for the detection and measurement of PCBs in


environmental samples when interfering substances are either


totally absent from the samples under study or when they have


been effectively removed prior to analysis.




1.   Analysis of Water, Sediment, and Soil for Polychlorinated
    Biphenyls (PCBs).


    a.  Background information—Since this project is a follow-


        up study and many conclusions will be based on correla-


        tion of current analytical data with that obtained at


        the time of the spill (1973), a consentaneous decision

        was made by all responsible parties that the GC/EC


        analytical methodology should not change from that em-


        ployed in 1973.



        The method employed in 1973 for the analysis of water,


        sediments, and soils for PCBs is basically the method


        from the R & D Laboratories of Monsanto Company (Method


        69-13) which is contained in the publication Manual

-------
                                                             29
    of Analytical Methods prepared by the Perrine Primate


    Research Laboratories of EPA (current designation:


    Pesticides and Toxic Substances Effects Laboratory, NERC,


    Research Triangle Park, N. C.).  The contractor modi-


    fied the Monsanto method to incorporate the Perrine


    column selection and instrumental recommendations.



    In principle, the contractor's method is unchanged


    from the Monsanto method.  The PCBs in water, sediment,


    and soil are extracted into hexane.  Interfering com-


    ponents, if present, are then removed from the extracts


    by chemical treatment and column adsorption chromato-


    graphy.  The amount of PCB present is determined by


    electron capture gas chromatography.



      •   •            '                \ '
b.  Analytical method—The complete Monsanto method consti-


    tutes the Appendix of this report, and only those pertinent


    modifications employed by the contractor will be discussed


    at this time.  The chromatographic column used is the most


    significant change.  The column packing is 1.5% SP


    2250/1.95% SP 2401 on 80/100 mesh Supelcon AW-DMCS


    (Cat. #01-1947, Supelco, Inc.).  This is a custom packing


    prepared to the Perrine Research Laboratories' specifica-


    tions and is especially suitable for the separation of


    chlorinated pesticides and other chlorinated hydrocarbons.


    The gas chromatographic conditions employed for the

-------
                                                             30


                                                        t

    analysis of Aroclor 1254 and method sensitivity data are


    contained in Tables 1 and 2, respectively.  Typical


    chrbmatograms of Aroclor 1254 and the askarel involved


    in the 1973 spill are shown in Figures 1 and 2.
        i




c.  .Operational narrative—


    Concentration and chemical clean-up.   After the initial


    liquid-liquid or liquid-solid extractions, exploratory


    chromatograms were run to determine whether the extracts


    would require further adjustment by dilution or concen-


    tration to bring the peaks into a quantifiable range


    consistent with the linear range of the detector.  When


    these initial chromatograms indicated the presence of


    interfering materials, extensive clean-up procedures


    were employed.  In most instances, the hexane extracts


    required no additional clean-up.





    Contamination.  To insure against the possibility of


    undetected contamination, blanks were routinely carried


    through all steps of the procedure.





    Measurement.  Initially, both the individual and total


    peak height methods were employed to determine the


    amount of Aroclor 1254 present.  It was soon found,


    however, that a calibration plot of the major isomer


    peak height could be used for the quantitation of the Aroclor.

-------
                                                                         31
      Table 1.  Gas Chromatographic Operational Parameters - PCB's

                      \
Instrument:  Beckman GC-45

DETECTOR:  Electron Capture (polarized helium plasma)
           Source Current: 7ma          Polarizing Voltage:  610 volts
           Scavenger:  He, C02, Rate 80, 1.2 ml/min.
GAS:  Helium                            Carrier Flow:  60 ml/min.
COLUMN:  Glass                          Length:  6 feet     Diameter:  1/4"
         Coating:  SP 2250/SP 2401               Cone.:  1.5Z/1.95Z
         Support:  Supelcon AW-DMCS              Mesh:  80/100
TEMPERATURE:
         Column:  195°C                          Injection Port:  220°C
         Detector:  250°C                        Detector Line:  240°C
SENSITIVITY:  x 8 K                              Recorder Range:  1 mv
CHART SPEED:  1/2 inch/minute

-------
                                                               32
       Table 2.  Method Sensitivity - PCBs









   Water:  0.5 parts per billion sensitivity




           Absolute sensitivity = 0.1 x 10"' grams




           Volume injected = 6 pi




           Final volume extract = 10 ml




           Sample size = 1000 ml



                     • (




Sediment:  0.05 parts per million sensitivity




           Absolute sensitivity = 0.1 x 10"' grams




           Volume injected = 6 pi




           Volume extract =» 200 ml




           Sample size = 100 grams

-------
Figure 1.   Representative Chromatogram for Aroclor 1254

-------
Figure 2.   Representative  Chromatogram for Askarel Involved in 1973 Spill
                                                      4--80-U-I-
                                                           Aroclor  1254

-------
                                                                35



        Chlorinated Pesticide Interferences.   Since surface


        water drainage of the area of interest was known to


        involve some agricultural land,  a chromatogram showing


        the elution pattern of 13 of the more common chlorinated


        pesticides was run using the instrumental conditions of


        the Aroclor method.  Pesticides  in the mixture included


        a - BHC, 6 - BHC, Lindane, Heptachlor, Aldrin, Hepta-


        chlor Epoxide, p,p'-DDE, Dieldrin, Endrin, o,p'-DDD,


        p,p'-DDD, o-p'-DDT, and p,p'-DDT.  The major isomer


        peak for Aroclor 1254 was free from interference from


        any of these pesticides under the analysis conditions


        employed.
                                                              \




2.  Analysis of_ Water, Sediment, and Soil for Polychlorobenzenes.


    a.   Background information—Since the solvent for Aroclor 1254


        in askarel is a mixture of chlorobenzenes, the fate of


        the solvent in the environment of the spill area needed


        to be determined.  Because of chemical similarities


        between PCBs and the solvent components, the analytical


        method of choice was again electron capture gas chroma-


        tography.





    b.   Analytical method—The solubility of the solvent compo-


        nents in hexane made it possible for a solvent analysis


        to be performed on the same extract prepared for the

-------
                                                                     36
            PCB1 analysis.  Likewise, the versatility of the chroma-

            tographic column selected for the PCB analysis allowed

            for its use in the solvent analysis.  Instrumental

            parameters for the solvent analysis are found in Table 3.

            Method sensitivity data are contained in Table 4.  A
                                                                •  /  •
            chromatogram of the solvent portion of the askarel is

            shown in Figure 3.  For reference, a chromatogram of

            a mixture of various chlorobenzenes is also included .

            as Figure 4.


F.  Isomer Verification.

    The Surveillance and Analysis Division, Region IV, Environmental

    Protection Agency, Athens, Georgia, provided the identification

    and verification of PCB isomers in the natural environmentally

    aged samples using GC/MS.                      ^  •

-------
                                                                         37
Table 3.  Gas Chromatographic Operational Parameters - Askarel Solvent
                                                  •


Instrument:  Beckman GC-45
•

DETECTOR:  Electron Capture (polarized helium plasma)
           Source Current:  7 ma    Polarizing Voltage:  610 volts
           Scavenger:  He, C02, Rate 80, 1.2 ml/min.
GAS:  Helium                        Carrier Flow:  60 ml/min.
COLUMN:  Glass                      Length:  6 feet     Diameter:  1/4"
         Coating:  SP 2250/SP 2401           Cone.:   1.5Z/1.95Z
         Support:  Supelcon AW-DMCS          Mesh:  80/100
TEMPERATURE:
         Column:  125°C                      Injection Port:  220°C
         Detector:  250°C                    Detector Line:  240°C
SENSITIVITY:  x 8 K                          Recorder Range:  1 mv
CHART SPEED:  1/2 inch/minute

-------
                                                                38
   Table 4.  Method Sensitivity - Askarel Solvent








   Water:  0.006 parts per billion sensitivity




           Absolute sensitivity « 0.003 x 10"9 grams



           Volume injected - 6 yl



           Final volume extract » 10 ml



           Sample .size - 1000 ml








Sediment:  0.010 parts per million sensitivity



           Absolute sensitivity - 0.003 x 10"' grams



           Volume injected - 6 yl



           Volume extract - 200 ml



           Sample size - 100 grams

-------
   Figure 3.  Representative Chromatogram

              Solvent  Portion of Askarel
H-H-i  -;••:•;-
I'!:  H.J i!;'

-------
                               riTTr•'  1111:
                              hi!  !   !  M
Figure 4.  Representative Chromatogram for Standard

-------
                                                                          41
VI.  EXPERIMENTAL RESULTS




     .JL  Field Data.




         The sampling phase of this project involved only minor adjustments in




         the initial protocol.  A total of 145 core, 2 soil control, 19 water,




         and 3 sediment samples were ultimately submitted to the laboratory for




         analysis.  Field log information and analysis data for these samples




         are presented in a later section of this report.






         1.  Cores.  Forty-five core sites were sampled over a 10-day period.




             Exhibit II shows the relative position of each of these sites with




             reference to the 1973 excavation area, the local topography over a




             2-foot interval, and all other core locations.  Cross section profiles



             at two intersects are shown in Exhibits III and IV as an illustration




             of the gradient in the immediate area of the spill site.






         2.  Geology.  The geological formations of the site area strike 40° north-




             easterly on the average, and normal dips are 25-35° to the southeast.




             The rocks underlying the total spill area are part of the Knox dolomite




             group.  A shaly limestone series known as the Chickamauga formation lies




             to the southeast and upon the Knox group.  The Knox of this area is




             the usual sequence of thin to massive-bedded dolomite (high MgC03 as




             compared to limestone with its high CaCC^) and is well fractured.






             The overburden above the Knox group is thick—in the 50 to 150 'foot




             range.  The primary overburden is made up of clays mixed heavily with




             chert fragments (see Table 5).  On top of the clay is a zone of top




             soil ranging from 0 to 4 feet in thickness.  The top soil allows for




             ease of perculation while the clays are penetrable primarily through




             the fracture crevices of the chert.

-------
                Exhibit II    _

Excavation Area and Location of Core Sites
          with Contour Intervals
                                                                   Sccle:   1 inch "* 20 :"-ec

                                                                   Datun is Mean Sea Level
                                                                                                          N2

-------
                                                 Exhibit III.  A Cross Section Profile of a Northeast Intersect
                                                               through the Study Area
                                                                                                                        Tcpsoil
Sc.ile:  1 inch'20 feet
                                                                                                                                                       U>

-------
                                                                         44
                   Exhibit IV.   A Cross  Section Profile  of  a  North Northwest
                                Intersect  through  a  Steep 'Surface  Gradient
                                Section  of the  Study Area,
                                                            Topsoil
                          Cross Section Profile
                               Intersect B
                N
Scale:  1 inch^O feet

-------
                                                                       45
       Table 5.  Description of Soil Types Characterizing
                 Core Samples Taken in August 1975
Code No.                      General Soil Characterizations

    1                    Reddish-brown clay with chert fragments

    2                    Yellowish-brown clay with chert fragments

    3                    Greyish-brown shale to reddish-brown clay

    4                    Reddish-brown clay

    5                    Greyish-brown silty clay with chert fragments

    6                    Greyish-brown to reddish-brown silty clay
                         with numerous chert fragments

    7                    Greyish-brown to reddish-brown clay with
                         chert fragments

    8                    Reddish-brown silty clay with chert fragments

    9                    Yellowish-brown silty clay with chert
                         fragments

   10                    Greyish-brown clay with chert fragments

   11                    Loam (Control)

-------
                                                                  46
3.  Environmental Samples.  The spill occurred at a,point where


    most of the drainage is geologically controlled by a north-


    east trending hollow near the site.  This valley is aligned


    along the strike of the formations, and there are no visible


    outcrops of bedrock to indicate that sub-surface liquids


    would come to the surface after they entered the clay over-


    burden.



    A spring well located in the hollow seemingly provided an


    excellent sampling site for detecting movement of suspected


    spill materials in the ground water.  In order to test this


    thesis, the spring and the mouth of the creek draining the


    hollow were sampled for water and sediment contamination.



    Two soil control samples were taken for analytical and


    microbiological analyses.  One site was characterized by


    clay and chert within the first three, inches of the over-


    burden while the clay at the other site was covered with


    two feet of loam.                 ' . x   •



    Locations of all of these environmental sites are given in


  ^ Figure 5.                              .





4.  Wells.  Six wells within close proximity of the spill were
      \          '                   i

    chosen for analysis.  These wells were included in the


    study because they were all located along probable geological

               -                             /
    strike formations, and 1973 data on these wells were extensive.


    The relative locations of the study wells are given in Figure 6.

-------
        Heavy-duty ..
        Medium-duly
        Light-duly ...
 ROAD CLASSIFICATION
.. _<•___> Poor motor road .... sa,,*,;
.. L   xi.     Wagon and jeep track	
      '     Foot trail		
               In developed treat, only through road! are clasiilied
FIGURE  5.   Location  of Environmental
              Sampling  Sites
                                                                              'SS^.Cj.'mTS;
                KENTUCKY
         MOYT- •"•""•"••" y^-
         7/~^     -•-•'
          7TENNESSE.E: •J^/
          L  '.      ' _ .. r .. •^f
          BACON GAP, TENN
               N3545-W8430/7.5
                                           1968

                                   AMS 4055 I SE-SERIES VS4I
                                                                         SCALE 1:24000
                                                                               o

                                                1000
                                                               1000
                                                                      2000
                                                                              3000
                                                                                      4000
                                                                                             5COC
                                                                                                            70*,
JTH GRID *NQ 19M MACNCUC NORTH
 OCCUNATION AT CENTER OF SHCCT
                                            CONTOUR INTERVAL 20 FEET
                                    DASHED LINES REPRESENT HALF-INTERVAL CONTOURS
                                               DATUM IS MEAN SEA LEVEL

                                 THIS MAP COMPLIES WITH NATIONAL MAP ACCURACY STANDARDS

                            FOR SALE BY U.S. GEOLOGICAL SURVEY. WASHINGTON. D.C. 20242.
                              TENNESSEE DIVISION OF GEOLOGY, NASHVILLE. TENN. 37219.   ~  -.^
                U.S. TENNESSEE VALLEY AUTHORITY. CHATTANOOGA. TENN. 37401 OR KNOXVILLE. TENN. 3/902
                          A FOLDER DESCRIBING TOPOGRAPHIC MAPS AND SYMBOLS IS AVAILABLE ON REQUEST     "T-

-------
         FIGURE 6.  Locations of Well Stations Relative to the Spill Site
6  Well Location

     and Depth



   Scale:_ 1 inch s o.l mile
STA4O  o
 165'

-------
    During the 1973 excavation and cleanup of the spill, a
    control well was sampled on numerous occasions.  This well
    was included in the present study for comparative purposes
    (see Figure 7).

    The time of sampling of these water supplies was predetermined
    in the protocol—after dry weather and after heavy rainfall.
    This aspect of the study will be discussed under "climatology."


5.  Climatology.  Since the protocol for water sampling was
    established on the basis of single grab samples to be taken
    immediately after dry weather and again after heavy rains,
    climatological data were obtained for a 2-1/2 year period
    in order to substantiate monthly, as well as, seasonal patterns
    of precipitation for the study area (see Table 6).
                                                                \
    Eastern Tennessee receives its greatest rainfall during the
    winter and early spring.  This is due to the more frequent
    passage of large-scale storms over and near the state during
    these months.  A secondary maximum of precipitation occurs
    .in mid-summer in response to shower and thunderstorm activity.
    This activity is especially pronounced in Eastern Tennessee
    where July rainfall frequently exceeds the precipitation of
    any other month.  Normally, the lightest precipitation is
    observed in the fall and is brought on by the maximum occur-
    rence of slow-moving, rain suppressing high pressure areas.
    Although all parts of the state are generally well supplied

-------
     FIGURE 7. Location of the Control Well
 '^-/fLGKTDO
            UNITED STATES
        DEPARTMENT OF THE INTERIOR
          GEOLOGICAL SURVEY
STATE OF TENNESSEE
            Scale 1:500,000
           1 inch equals approximately 8 miles
O Well Location
  and Depth

-------
                                                                        51
        Table 6.   Climatological Data Showing Monthly Averages
                  for the Study Area (D  (2)
                                    Total Precipitation in Inches

        Month

      January

      February

      March

      April

      May

      June

      July

      August

      September

      October

      November

      December



(1)  Climatological Data,  U.  S.  Dept. Commerce, Annual Summary 1973, Vol. 78,
    No.  13, p. 2.

    Ibid., Annual Summary 1974, Vol. 79, No. 13, p. 2.

(2)  Climatological Data,  U.  S.  Dept. Commerce, 1975, Vol. 80, Nos. 1-9.
1973
4.51
3.30
12.44
4.55
9.82
7.33
5.81
3.58
4.32
3.12
9.74
8.38
1974
10.00
5.41
6.97
3.54
7.36
2.72
1.64
5.43
3.10
1.59
4.26
7.04
1975
5.93
5.90
13.19
2.45
6.43
4.47
3.63
2.00
5.17
5.23
3.87
4.65

-------
                                                                52
    with precipitation; there occurs, on the' average, one or more

    prolonged dry spells each year during the summer and fall.


    Based on the foregoing information, a decision was made to

    obtain local daily rainfall information beginning with the

    inception of the project.  It was apparent that over-all dry

    weather had prevailed from June 21 through August 27, 1975

    (see Table 7).  There had been minimal>measurable rain
                                 \
    (total =5.5 in.) with three days of trace precipitation

    over the 68-day period; therefore, water samples were

    collected on August 28 and 29.  During the next 29-day period,

    from August 29 through September 25, appreciable rain fell

    at the study site (total = 6.42 in.) with traces on two

    other days.  In order to take advantage of the wet weather,

    the water sites were resampled on September 26.



6.  Soil Temperatures.  Seasonal temperature variations in the

    shallow layers of earth overburden are affected primarily

    by incoming solar radiation and outgoing terrestrial radia-

    tion.  Normally, in the top two feet of surface materials

    there is a definite diurnal gradient which varies among

    various types of soils.  However, daily fluctuations in

    soil temperature during the summer months lags considerably

    behind affective atmospheric temperature variations.


    Differences in texture, structure, and organic matter tend

    to determine the moisture capacity of soils and also in-

    fluence their ability to absorb and transmit heat.  Rain,

-------
                                                                  53
Table 7.  Rainfall Data Prior to and During Well Sampling
         Date
        (1975)

          June 21
               26
               27
               28

          July  3
                6
                9
               10
               13
               16
               20
               21
               25
               26
               30
               31

        August  3
                5
                6
               10
               11
               16
               19
               24
               29

      September  7
               13
               17
               18
               21
               22
               23
               24
Precipitation
  (inches)

    0.02
    0.19 I
    0.45
    0.01

    0.05
    0.24
    0.41
    Trace
    0.05
    Trace
    0.04
    0.78
    0.02
    0.10
    0.78
    0.78

    0.10
    0.04
    0.56
    Trace
    0.37
    0.20
    0.12
    0.20
    0.46

    0.30
    0.09
    Trace
    1.93
    0.28
    Trace
    0.98
    2.38

-------
                                                             54
although usually at a lower temperature than the soil,




generally leads to an increase in the temperature of shallow




layers; however,, if it falls in sufficient quantity, it can




serve to increase the rate of conduction of heat from the .




deeper layers to the colder surface.  This phenomenon can




more than compensate for the initial fall of temperature




due to rain.






Variations of soil types in the study area, as observed by




core drillings, were minimal (see Table 5).  Secondly, the




over-all effect of rainfall on the area was judged to be




insignificant, since climatolbgical data indicated little




rainfall prior to soil temperature measurements.  On




August 6, 1975, three soil types were measured for tempera-




ture gradients over four-inch intervals to a depth of fifty-




six inches (see Figure 8).  The ambient temperature for a




24-hour period preceding the measurements ranged from a




high of 86.8°F to a low of 70°F.  During the sampling period,




the ambient temperature moderated to a high of 82°F with a




low of 73°F.






All determinations indicated that the soil temperature was




highest within the first four inches and followed a general




downward trend over the remaining fifty-two inches.  However,




slight gradients were observed at varying depths in the




three types of soil sampled.  These deviations were probably




due to moisture differentials caused by the non-uniform




clay-chert matrix within a given sampling site.

-------
86
                                        Figure 8.   Temperatures of Various  Soil  Types

                                                    Over Four-Inch Intervals as Compared

                                                    with Ambient Conditions  (see  Table

                                                    5 for Soil Type Codes)
   84
   82
   80
   78
   76
En
O
   74
                         Soil Type  2
                                                                                     w
                                                                                 u w
                                                                                 e a,
                                                                                 E)
                                                                                 H o
                                                                                     w -;
W
   72
                                                                Y
   70
   68
   66
              00
 i
oo
                          O
                          N

                          (£>
                                  0>J

                                  O
00
N
 I
CO
 I
00
(O
n
 i
N
n
                                 DEPTH  (inches)
o

(0
en
                    I
                    o
                                            00
                                            «f
                                            I
10
 I
00
                                                                         to
                                   CM

-------
                                                                        56
        7.  Drilling Equipment and Procedures.  The drilling equipment



            used in the field was quite adequate and versatile in that



            it provided for the use of an auger and three types of core



            samplers,   the weight of the truck became critical at one



            stage after a rain when the sites which had not been cored



            were on a steep slope.  A smaller drill truck was brought in,



            and provided relief from the problem.






            The split spoon sampler was an excellent compromise in



            that it was efficient, produced a core sample of sound in-



            tegrity, worked well in the clay and chert matrix, and
                             i


            cleaned with minimal difficulty.








    B.   Microbiological Data.



        The soil samples, which were various types of clay and chert,



        were found to have relatively low bacterial, fungal, and actino-



        mycete populations.  The data for each sample is shown in



        Table 8.  It may be noted that the bacterial population far



        exceeded populations within the other two groups.  Plate counts



        for these organisms determined in fertile agricultural soil have



        been shown to be much higher.  Average bacterial counts may



        exceed 15 million per gram of soil, while smaller fungal and



        actinomycete populations may average 400 thousand and 700



        thousand, respectively (29).  Only one of the 142 samples was



        shown to have a bacterial count as high as three million per
(29)  Surges,  A.   1958.   Microorganisms in the Soil.  Hutchinson and Co.

     Publishers, London.

-------
                                                                 57
gram, and only four others exceeded one million per gram of


sample.  Fungal and actinomycete counts in the samples examined


were correspondingly lower than those usually found in fertile


soils.  Additionally, the two control soil samples, taken from
                                   i

areas remote to the spill, were surprisingly low for all three


types of organisms.           v  .  .



Eleven different types of clay and chert were identified, with


the majority of microorganisms occurring in types designated


as 1 and 2 (see Table 9).



It may be seen that the average bacterial, fungal, and actino-


mycete counts for 81 samples in soil type 1 were lower than the


average for all 142 samples, while populations of the three


groups in soil type 2 were higher than the over-all average.


Bacterial and fungal counts for soil type 2 were approximately


two times as great, and actinomycete counts were greater than


four times the mean for all samples.  The remaining 34 samples


were distributed among 8 soil types and constituted too small a


group in each instance to provide statistically valid data.



Analysis of microbial populations relative to pH of the soil


samples is summarized in Table 10.  From these data it is


obvious that conventional distribution of the three groups of


organisms was observed.  Bacterial and actinomycete counts were


highest in samples having a pH of 5:0 to 5.9 and lowest in a


pH range of 4.0-4.5.  Conversely, the number of fungi were


highest in the latter range and lowest at a pH of 5.0 or above.

-------
                      TABLE a   COLLECTIOB AND MICROBIOLOGICAL DATA FROM SOIL SAMPLES TAKES  iN AUGUST, 1975.
                                                                                               Microorganism Counts/Gram of Soil
                                               er                  .    X Holatur*
Hucber            (Inches) W         of Soil
Field Sample         Sazple Death          Character
  GF 7316
  OF 7520
  CF 7524
  GF 7S28

  CF 7517              23-27
  CF 7521
  CF 7525
  CF 7529

  C? 7518              27-31
  CF 7522
  CF 7526
  CF 7530

  CF /519              31-35
  CF 7523
  CF 7527
  GF 7531

  GF 7618.              17-33

  CF 7532              21-25
  CF 7536              25-29
  CF 7540              29-33
  CF 7544              33-37
JES-
4.6
4.4
4.7
4.4
4.5
4.5
4.4
4.5
4.0
4.4
4.4
4.6
4.1
4.8
4.7
4.6
4.7
4.6
Z Mol«tur«
by Weight
7.30
6.99.
4.65
5.47
4.28
5.65
2.54
3.35
10.0
4.38
10.3
5.58
15.6
10.8
10.6
8.65
9.07
9.00
Bacteria
2.310,000
34,500
< 10,000
- ' 25,000
< 10,000
•'^ < 10,000
< 10,000
25,000
< 10,000
< 10.000
50.000
< 10,000
< 10.000
78.000
570.000
306.000
191.000
354.000
Actlnomycetes
2.500
1.500
< 1.000
< 1.000
500 .,
< 1.000
3.000
5,000
< 1.000
< 1,000
1,000
< 1,000
< 1.000
500
700
600
350
< 100
Fungi
25,000
3,500
50,000
400
< 1.000
5,000
7.200
32.GCO.OOO
< 1.000
5,000
< 1,000
< 1,000
< l.COO
li9CO
6.500
1,550
700
5.800
                                                                                                                                                             in
                                                                                                                                                             00

-------
                   TABLE 8.  COLLECTION* AND MICROBIOLOGICAL DATA FROM SOIL SAMPLES TAKEN IN AUGUST, 1975 (con't)
Field Sai.?le
  Number

  CF 7i.»3
  CF 7537
  CF 7541
  CF 7545

  GF 7534
  CF 7538
  CF 7542
  GF 7546

  CF 7535
  CF 7539
  CF 7543
  GP 7547

  CF 7548
  CF 7556
  CF 7564
  CF 7572
  C? 7552
  CF 7560
  GF 7568
  CF 7576

  CF 7549
  CF 7557
  CF 7565
  CF 7573
  CF 7553
  CF 7561
  CF 7569
  CF 7577
Sample Depth
(inches) U)

  25-29
  29-33
  33-37
  19-23
  23-27
 Character
of Soil <2>
  31-JS
                    4.4
                    4.8
                    5.2
                    4.5

                    4.6
                    4.4
                    4.5
                    4.2

                    5.0
                    4.4
                    4.3
                    4.2

                    4.4
                    4.4
                    4.2
                    4.3
                    4.3
                    4.0
                    4.1
                    4.8

                    4.4
                    4.9
                    4.4
                    4.1
                    4.7
                    4.3
                    4.3
                    4.5
Z Moisture
by Weight

   4.45
   3^30
   9.29
   6.60

   6.85
   6.21
   3.92
   7.20

   6.29
  12.7
  10.5
   2.84

  15.0
  18.2
  14.1
  12.8
  22.8
  19.3
  21.5
  20.8

  19.5
  19.1
  22.7
  17.8
  20.9
  19.6
  22.1
  22.7
                                                                                                   Microorganism Counts/Cram of Soil
Bacteria
20,500
2,000,000
1,270,000
800,000
6C.OCO
945,000
195,000
650,000
645,000
< 10.000
< 10,000
< 10,000
37,500
83,000
79,000
T., 000
8.000
3i.OOO
2.000
< 1.000
43.500
38.000
51.000
37.000
1,000
14.500
< 1,000
11.000
Actinomvcetes
6,000
43.800
230,000
5,500
< 1,000
7,500
2,500
3.000
l.OCO
1,000
< 1,000
< 1.000
600
300
. 850
1,400
100
650
300
< 100
< 1.000
450
450
300
350
< 100
< 100
400
Fungi
< l.COO
8,000
12,000
15,000
< l.OCO
3,500
< l.OCO
11.000
< 1,000
< 1,000
< 1.003
3,000
600
120
10. COO
3.500
250
200
1.000
1.050
1.000
50
100.000
/ 1,000
< 100
100
< 100
1.250
                                                                                                                                                             Ul
                                                                                                                                                             VO

-------
                   TABLE 8.  COLLECTION AND MICROBIOLOGICAL DATA FROM SOIL SAMPLES  TAKEN  IN AUGUST, 1975  (con't)
Field Staple
  Xuaber

  CF 7550
 •CF 7553
  CF 7566
  CF 7574
  GF 7554
  C- 7562
  CF 7570
  CF 7578

  CF 7551
  CF 7559
  CF 7567
  CF 7575
  GF 7555
  CF 7563
  CF 7571
  CF 7579

  CF 7580
  CF 7581
  CF 7582
  CF 7533

  CF 75S8
  CF 7589
  CF 75SO
  CF 7591
  CF 7592
  CF 7593
  CF 7594
  CF 7595
Saople Depth
(inches) U)

  35-39
  39-43
  43-47
  47-51
  15-19
  19-23
  23-27
  27-31

   9-13
  13-17
  17_-i
  23-kS
  25-29
  29-33
  33-37
  37-41
 Character
of Soil »)
                     pH

                    4.2
                    4.1
                    4.4
                    4.3
                    4.3
                    4.6
                    4.6
                    4.8

                    4.3
                    4.3
                    4.0
                    4.6
                    4.7
                    4.6
                    4.6
                    4.7

                    4.5
                    4.6
                    4.3
                    4.7

                    4.4
                    4.8
                    5.1
                    4.8
                    4.7
                    4.3
                    4.2
                    4.3
Z Moisture
by Weight

  20.4
  19.3
  20.9
  18.2
  20.4
  21.7
  22.8
  20.5

  19.7
  19.7
  20.8
  21.0
  20.8
  21.7
  24.2
  22.8

   7.14
  14.3
  16.0
  15.0

  17.2
  17.2
  18.7
  19.3
  13.3
  15.0
  18.3
  19.2
                                                                                                 Microorganism Counts/Cram of Soil
Bacteria
28,000
26,000
18,000
20,500
28,000
500
< 1.000
2,500
23,500
< 1.000
3.500
8.500
< 1.000
500
< 1,000
f 1,000
139,500
89.000
24.500
19,000
97.000
197,500
309,500
192.500
161,000
276,500
43.000
70.000
Actinoayceces
800
< 100
< 100
300
500
50 -_
200
< 100
900
< 100
< 100
< 100
150
< 130
< ioc
< 100
850
5.000
600
< 100
600
Ii550
450
250
ISO
200
< 100
100
Fung:
SCO
200
< 100
1.000
5, SCO
500
300
600
200
< 100
5, COO
650
< 100
< 100
< 100
< 100
2,5iO
180
1,900
105
2,300
2,550
3,150
2,600
10,000
. 10.000
1.200
1,200
                                                                                                                                                               ON
                                                                                                                                                               O

-------
TABU 8.  COLLECTIOB AND MICROBIOLOGICAL DATA FROM SOIL SAMPLES TAKES IN AUGUST, 1975 (con't)
Field Saaple
Xuaber
CF 7596
CF 7597
GF 7593
GF 7599
CF 7600
CF 7601
GF 7602
GF 7603
CF 7604
CF 76C5
CF 7606
CF 7637
.Gr 7608
CF 7539
CF 7610
CF 7611
C? 7612
CF 7613
GF 7614
GF 7615
CF 7616
CF 7617
C? 7619
CF 7620
Simple Depth
(Inches) U>
9-13
33-17
x/-21
21-25
20-24
24-28
28-32
32-36
33-37
37-41
41-45
45-49 .
49-53
53-57
57-61
61-65
9-25
25-41
9-25
21-:,
12-28
9-25
9-25
9-25
Character
of Soil (2)
5
5
1
1
6
6
6
6
7
7
7
1
1
1
1
1
1
1
1
1
1
8
4 '
10
                                            pH

                                           4.6
                                           4.6
                                           4.7
                                           4.8

                                           4.5
                                           4.4
                                           4.8
                                           4.6

                                           4.7
                                           4.9
                                           4.9
                                           4.8
                                           4.8
                                           4.8
                                           5.0
                                           4.9

                                           4.9
                                           4.6

                                           4.7

                                           4.2

                                           4.9

                                           4.6

                                           5.2

                                           5.2
Z Moisture
by Height

  6.89
  6.71
 14.6
 16.5

 17.9'
 19.4
 19.7
 21.9

 13.6
 14.5
 13.7
 14.7  .
 17.1
 18.4
 17.2
 17.9

 19.0
 19.3

 18.8

 22.8

 14.1

 11.2

 14.6

 13.9
                                                                                Microorganism Counts/Cram of Soil
Bacteria
112,000
113,000
29,500
35,500
80,500
81,000
40,500
7,500
40,000
25,500
29,000
2,500
1,500
< 1,000
159.000
35,000
320,500
12,500
4,000
6,000
126,500
219,000
262,500
219,500
Actlnoaycetes
550
40C
2,500
< 100
250
150
< 100
< 100
100
100
200
< 100
< 100
50
900
400
200
100
< 100
< 100 .
250
< 100
3CO
50
Tumi
7,403
4,850
950
750
. 2.950
1,700
2,250
350
1,900
2,250
900
100
600
250
3,700
500
7,700
500
5,400
350
4.030
9,000
1,400
600

-------
TABLE 8.  COLLECTION ASD MICROBIOLOGICAL DATA FROM SOIL SAMPLES TAKEN IN AUGUST. 1*75 (con't)
                                                                                Microorganism Counts/Gran of Soil
Field Sample
Number
GF 7621
CF 7622
CF 7623
CF 7624
CF 7625
CF 7626
GF 7627
CF 7630
CF 7631
CF 7632
CF 7633 .
CF 7634
CF 7635
CF 7656
GF 7637
C? 76id
GF 7639
CF 7641
CF 7640
Sample Depth
(Inches) I"
9-25
'25
25-41
41-57
57-73
19-35
3-19
9-25
25-41
: 9-25
25-41
3-19
4-20
9-25
25-41
3-19
9-25
25-41
3-19
Character
of Soil <2)
5
5
5
5
8
5
1
5
i
i
i
i
i
i
i
i
i
i
_ES_
5.1
5.6
5.5
5.4
4.9
5.5
5.2
5.3
5.0
5.0
5.2
4.8
4.6
5.0
4.8
_ ; 5.2
5.2
5.3
5.2
Z Moisture
by Ueigbt
13.8
16.0
13.9
14.8
23.4
15.3
24.7
9.00
7.52
9:00.
8.40
25.0
23.5
12.6
22.1
21.2 -
22.1
23.6
22.8
Bacteria
215.000
334.000
136.000
46.000
2.500
367.000
/ 4.500
112.500
157.500
218.500
155.000
100.000
113.500
82.500
29.500
160,000
60,000
74.500
48.000
Actinomycete*
250
850
250
200
< 100
400
< 100
750
SO
650
500
< 100
2CO
450
250
100
250
200
< 100
Fungi
1.800
350
1.400
350
SO
13.000
100
2.650
600
1.900
1.400
200
1.05C
950
1.100
750
1.500
1,100
5.000
                                                                                                                                           N>

-------
TABLE 8.  COLLEGIUM AND H1CEOBIOLOCICAL DATA FROM SOIL SAMPLES TAKEN IS AUGUST. 1975  (eon't)
                                                                               Microorganism Countg/Craa of Soil
Field Sample
Kuaber
CF 7642
CF 7643
CF 7644
. CF 7645
CF 7646
GF 7647
. CF 7648
CF 7649 '
CF 7650
CF 7651
GF 7652
CF 7653
CF 7654
CF 7655
CF 7656
CF 7657
CF 7658
CF 7659
GF 7660
Sample Depth
(Inches) W
9-25
25-41
2-9 .
9-25
9-25
25-41
9-25
25-41
. 9-25
0-16
29-45
21-37
25-37
33-45
9-2-
3-19
10-26
26-42
42-58
Character
of Soil <2)
1
1
1
• 1
1
1
1
2
1
2
2
1
1
1
1
1
1
1
-ESL
5.0
5.1
5.3
5.3
5.1
5.8*
5.7
5.8
5.9
4.6
5.5
5.8
4.9
5.0
4.6
4.7
4.8
4.5
4.2
X Moisture
by Weight
19.8
18.8
21.4
18.5
20.0
23.3
16.6
15. 5
13.3
23.1
7.95
12.9
10.6
11.3
21.0
12.2
11.5
13.1
20.4
Bacteria
315.000
96.500
204.000
56,500
57,500
11.000
1,480,000
170.000
665,000
503
. 65.000
4.503
17.500
138.500
4.500
291.003
3.030.000
165.500
1.400
Actinonycetes
400
1.050
200
250
700
100
500
350
20.000
< 100
150
< 100
200
400
< 100
250
5.500
100
< 100
Fungi
700
4.000
1.350
1.200
1.650
800
2.700
900
3,300
100
1.400
200
620
850
< 100
700
800
250
50

-------
                 TABU 8.   COLLECTION AND MICROBIOLOGICAL DATA FROM SOIL SAMPLES  TAKEN IN AUGUST,  1975 (con't)
                                                                                                 Microorganism Counts/Crag of Soil
Field Sample
Number
CF 7661
CF 7662
CF 7663
CF 7664
GF 7665
CF 7666
GF 7683
(control A)
GF 7684
(control B)

Sample Depth
(inches) U)
9-25
25-41
9-25
25-41
3-19
9-25
3-6
3-6

Character
of Soil (2)
10
10
5
5
5
5
1
11
Overall

_Efi_
5.2
5.0
5.0
5.0
5.0
5.3
5.0
6.2
Averages * '

Z Moisture
by Weight
15.6
18.9
14.1
12.8
18.9
16.5
8.40
3.60
Hif;h
Low
Bacteria
325,000
26,500
620,000
' 59,500
164,000
1.540,000
186,000
340,000
181.654'
180,823
Actinonycetes
800
100
4,000
100
200
450
4,000
11,000
2.860.
2,760
Fungi
2,600
< 100
1,700
350
550
14.500
1,500
1,500
3,343
3,259
Pleasured from the original 1973 surface prior to  backfill.

 See Table 5 for soil code descriptions.               _

 The high averages are calculated with less  than values  taken  to be positive  (<.l  taken as  .1).   The  low averages are
 calculated with the less than value taken to  be zero.   The actual mean of  the  sample oust  lie between the high average
 and the low average.                                            .                        '

-------
                                                                             65
             Table 9.  Microbial Populations Relative to Soil Types
                                  Average Microbiological Counts
Soil Type

     1


     2


     3


     4


     5


     6


     7


     8


     9


    10


    11
         (1)
Criterion (2)
for
Average
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
(Per
Bacteria
' 136628
136530
364796
360722
84333
84333.
138667
138667
207182
207182
52375
52375
31500
31500
110750
110750
78000
78000
190333
190333
340000
340000
Gram of Soil)
Actinomycete
469
427 .
12557
12183
2150
2150
158
125
723
723
. 150
100
133
133
100
0
500
500
317
317
11000
11000

Fungi
2707
2696
6093
5685
1543
1543
3984
3984
3127
3127
1813
1813
1683
1683
4525
4525
1900
1900
1100
1067
1500
1500
 Number of
Samples/Sell
    Type

     81


     27


      3


      6


     11


      4


      3


      2


      1


      3


      1
(1)  See Table 5 for soil codes.

(2)  The high averages are calculated with less than values taken to be positive
    (<.l taken as .1).   The low averages are calculated with the less than
    values taken to be zero.  The actual mean of the samples must lie between
    the high average and the low average.

-------
                                                                            66
                  TABLE 10.  Microbial Populations Relative
                             to pH of Soil Samples
                                 Average Microbiological Counts
                 Criterion^1)          (Per Gram of Soil)                 Number
   pH               for             •.	.	         of
 Ranges           Average      Bacteria	Actinomycete	Fungi       Samples
 4.0-4.5           High       .   90324           1043   ,      4444          49
                   Low           88447            861         4275

 4.6-5.0           High         215644           1299         2901          66
                   Low          215250           1225         2845

 5.0 Up            High         264315    .       9970         2426          27
                   Low          264315           9959         2426
(1)  The high averages are calculated with less than values taken to be
    positive (<.l taken as .1).   The low averages are calculated with
    the less than values taken to be zero.   The actual mean of the samples
    must lie between the high average and the low average.

-------
                                                                67




It is well recognized that fungi thrive best in a relatively



acid environment while bacteria and actinomycetes, in general,



prefer ~a more neutral pH range for survival and growth.





Microbial counts were compared in relation to moisture content



of the soil samples.  Determinations were made in the ranges



of 0-5, 5-10, 10-15, 15-20, 20-25, and 25-30 percent moisture



by weight.  From Table 11 it can be observed that the highest



bacterial and actinomycete concentrations were present in



samples having 5-10 percent moisture.  Fungal counts were the



greatest in samples having the least moisture.  A single sample
        ,\                                      |


having a 25-30 percent moisture was insufficient to provide



valid data at that level.  In fertile soils, rich in nutrients,



it might be expected that extremely high numbers of organisms



would be detected when moisture levels were high since water



is required to solubilize the nutrients, making them more readily



available to the soil populations.  However, in clay and chert



where nutrients are extremely deficient, and where the pH range



is predominately between 4.0 and 5.0, soluble acid minerals
                /

contributing to the low pH may function bacteriostatically or



bactericidally to maintain relatively low levels of organisms.



Very significant decreases in bacterial counts were observed



when moisture content of the samples exceeded 20 percent, and



for actinomycetes when moisture was above 10 percent.  Fungal



populations decreased steadily as moisture content increased



to 20 percent.  Above this level a slight increase in  fungi



was noted between 20-25 percent moisture.

-------
                                                                             68
                   Table 11.  Microbial Populations Relative
                              to Moisture Content of Soils
Moisture
(% by Weight)
0%- 5%
5%-10%
10%-15%
15%-20%
20%-25%
25%-30%
»
Criterion'1' Average Microbiological Counts
for (Per Gram of Soil)
Average
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
Bacteria
289500
289500
385630
385630
236156
236156
135940
135940
27240
27240
100000
100000
Actinomycete
7756
7756
11476
11476
1503
1503
407
407
200
200
100
100
Fungi
8633
8633
4813
4813
2202
2202
1976
1976
3791
3791
200
200
Number
of
Samples
9
23
32
42
35
1
(1)  The high averages are calculated with less than values taken to be positive
  .  (<.l taken as .1).   The low averages are calculated with the less than
    values taken to be zero.  The actual mean of the samples must lie between
    the high average and the low average.               ' .'

-------
                                                                        69


        It has been reported (30, 31, 32) that microbial counts in

        fertile agricultural soil are the highest in the first 6-8 inches

        below the surface, decreasing rapidly below that point to only a

        few hundred per gram at depths of 6 feet or more.  Although

        these data were obtained from samples deficient in nutrients at

        any depth, a similar trend could be observed.  The major differ-

        ence was revealed in Table 12 by bacterial counts which averaged

        157,330 per gram between 0 and 8 inches and increased to 271,700

        per gram between 8 and 29 inches.  Below 29 inches the bacterial

        population was reduced progressively to a total of 2500 per gram

        in a single sample taken at a depth below 66 Inches.  Actino-

        mycete counts were highest between 0 and 2 inches and at a depth

        of 24-29 inches, subsequently decreasing to 100-700 per gram at

        all depths below 29 inches.  Fungal counts were highest between

        8 and 27 inches, and 'these also decreased progressively below

        that level except for a very slight increase in numbers in 13

        samples taken at a depth between 56 and 61 inches.


        The fact that bacterial counts near the surface of the soil were

        slightly lower than at depths of 8-29 inches may possibly be

        attributed to the seasonal effect'of high summer temperatures

        which would tend to kill some of the organisms by baking of the

        soil to shallow depths.


(30)  Waksman, S. A., and Starkey, R. L.  1931.  Soil and the Microbe.
     Wiley and Sons, Inc., Publishers, New York.

(31)  Kuster, E.  and Williams S. T.  1964.  Selections of Media for Isolation
     of Streptomycetes.  Nature, London, 202:928-929.

(32)  Frobisher,  M.  1962.  Fundamentals of Microbiology (p. 652),  W. B.
     Saunders Co. Publishers, Philadelphia, Pennsylvania.

-------
TABLE 12.  Microbial Populations Relative
           to Core Sample Depths
                                                         70
              Average Microbiological Counts
Depth below
Surface
(inches)
0
(
i
2
3
4
5
6
7
8
9
10
11
12
13
Criterion u;
for
Average
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
(Per Gram of Soil)
Bacteria
175500
175500
175500
175500
182625
182625
149800
149800
146500
146500
146500
146500
146500
. 146500
146500
146500
146500
146500
211554
211554-
287932
287932
289357
289357
284833
284833
256058
256058
Actinomycete
5033
5000
5033
5000
3825
3800
1615
1575
1486
1450
1486
1450
1486
1450
1486
1450
1486
1450
1392
1373
1535
1516
1194
1174
1168
1149
1079
1040
Fungi
1033
1033
1033
1033
1113
1113
1175
1175
1164
1164
1164
1164
1164
1164
1164
1164
1164
1164
2538
2536
2524
2521
2582
2579
2621
2619 v
4973
4970
Number of
Samples/ Dept
Category ,
3
3
4 ;
10
11
11
11
11
11 .
37
37
35
36
43

-------
TABLE 12.  Microbial Populations Relative
           to Core Sample Depths (Continued)
                                                          71
Depth below
Surface
(inches)
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
Criterion*1)
for
Average
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
**V W*. b»^^_ **rib1
(Per
Bacteria
287944
287944
283932
283932
283932
283932
251307
251307
278405
278405
319549
319549
350714
350714
297307
297284
340730.
340703
309183
308671
264718
264179
266586
266207
285828
285172
176157
174700
191766
190797
Iv^VhS^V^VE^A^tl.*^. W
Gram of Soil)
Actinomycete
1165
1118
1157
1111
1157
1111
1081
1038
1168
1143
1283
1261
1461
1444
1216
1189
1328
1304
1274
1204
1113
1038
5743
5691
9380
9308
' 8564
8439
9273
9195
>«*• w^
Fungi
4997
4994
4931
4928
4931
4928
4349
4344
1943
1938
2770
2765
3048
3042
2800
2793
3137
3132
4131
4102
3651
3621
3301
3262
3985
3919
4001
3912
2780
2715
                                                      N'-r-.ber of
                                                     Samples/Depth
                                                      Category

                                                          36
                                                          37


                                                          37


                                                          44


                                                          37


                                                          41


                                                          35


                                                          44


                                                          37


                                                          41


                                                          39


                                                          58


                                                          32


                                                          35


                                                          32

-------
TABLE 12.   Microbial Populations Relative
           to Core Sample Depths (Continued)
                                                         72
              Average Microbiological Counts
Depth below
Surface
(inches)
29
30
- 31
32
33
34
35
36
37 .
38
39
40
41
Criterion ^
for
Average
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
(Per Gram of Soil)
Bacteria
201563
200763
116774
115774
105714
103971
114328
113359
116919
115477
87303
85455
87303
85455
86929
85821
76734
75672
69816
69658
69816
69658
69816
69658
66738
66595
Actinomycete
7795
7703
798
682
821
633
742 \
598
685
529
414
247
414
247
330
241
303
219
224
208
224
208
224
208
221
207
Fungi
2586
2478
2044
1938
2039
1859
1826
1660
1826
1633
1754
1569
1754
1569
1317
1206
1224
1115
916
889
916
889
916
889
888
864
Number of
Samples/Dept*
.Category
40
31
35
32
43
33
33
28
32
19
19
19
21

-------
TABLE 12.   Microbial Populations Relative
           to Core Sample Depths (Continued)
                                                         73
              Average Microbiological Counts
Depth below
Surface
(inches)
.42
A3
44
45
46
47
48
49
50
51
52
53
54
55
Criterion^1'
for
Average
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
(Pe
Bacteria
87914
87914
74983
74983
74983
74983
64629
64629
54975
54975
54975
" 54975
54975
54975
44280
44280
16300
16300
16300.
16300
16300
16300
12475
12225
16133
15800
16133
15800
r Gram of Soil)
Actinomycete
214
200
233
217
233
217
214
186
188
138
188
138
188
138
170
110
133
67
133
67
133
67
113
63
117
83
117
83

Fungi
671
671
742
742
742
742
650
650
350
350
350
350
350
350
400
400
333
333
333
333
333
333
313
313
217
217
• 217
217
Number of
Samples/Dept
Category
7 . .
6 j
6
7
4
4
4
5
3
3
3
4
3
3

-------
TABLE 12.   Microbial Populations Relative
           to Core Sample Depths (Continued)
                                                         74
              Average Microbiological Counts
Depth below
Surface .
(inches)
56
57
58
59
60
61
62
63
64
65
66
67
68
Criterion^
for
Average
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
High
Low
l) (Per Gram of Soil)
Bacteria
16133
15800
41980
41780
54300
54300
80750
80750
80750
80750
65500
65500
18750
18750
18750
18750
18750
18750
18750 .
18750
2500
2500
2500
2500
2500
2500
Actinomycete
117
83
270
230
367
. 300
500
450
500
450
467
433
250 .
200
250
200
250
200
250
200
100
0
100
0
100
0
Fungi
217
217
880
880
1267
1267
1875
1875
1875
1875
1417
1417
275
275
275
275
275
275
275
275
50
50
50
50
v 50
50
Number of
Samples/Deptl
Category
3
5
3
2
2
3
2
2
2
2
1
1
1

-------
                    TABLE 12.   Microbial Populations Relative
                               to Core Sample Depths (Continued)
                                                                             75
Depth below
  Surface
 (inches)

     69


     70


     71


     72


     73
Criterion^1)
   for
 Average

  High
  Low

  High
  Low

  High.
  Low

  High
  Low

  High
  Low
Average Microbiological Counts
      (Per Gram of Soil)
  Number of
Samples/Depth
Bacteria
2500
2500
2500
2500
2500
, 2500
2500
2500
2500
2500
Actinomycete
100
0
100
0
100
0
100
0
100
0
Fungi
50
50
50
50
50
50
50
50
50
50
Category
1
1
1
1
1
(1)  The high averages are calculated with less than values taken to be
    positive (<.1 taken as .1).   The low averages are calculated with the
    less than values taken to be zero.   The actual mean of the samples must
    lie between the high average and the low average.

-------
                                                                 76
 Analysis of microbial populations exposed to polychlorinated


 biphenyl (PCB)  in the soil suggested a possible stimulating

 effect of this  compound in concentrations between 0^05 and 5.0


 mg/kg on both the bacteria and actinomycetes,  as shown'in Table
».                              •        .
 13.   The lowest bacterial counts were obtained in samples where


 PCB  levels were less than 0.05 mg/kg.   Although only three samples


 were contaminated with PCB in excess of 10 mg/kg, the bacterial


 counts were greater than in relatively uncontaminated soil having


 less than 0.05  mg/kg.  Fungal counts were essentially unaffected


 in all except the one sample having the highest concentration of

 PCB.



 In summary, the relatively low microbial counts are compatible


 with the clay and chert soils which are largely devoid of nu-

 trients.   The soil acidity is also reflected by the low bacterial

 counts.  Excessive moisture did not appear to influence an in-


 crease in the microbial populations of the soil; and, in fact,

 the  lowest bacterial and actinomycete counts were observed in

 samples having  a moisture content of 20-25 percent while the

 highest numbers were found in samples having 0.5-10.0 percent

 moisture.



 The  most interesting observation was the correlation between


 microorganism counts and the soil samples containing
                                 \.          .
 0.05-5.0 mg/kg  of the polychlorinated biphenyl compounds.


 Bacterial counts in 55 samples were shown to be 50 percent

 higher than the mean for all samples,  and actinomycete counts

 were similarly  100 percent greater.

-------
                                                                                77
                      TABLE 13.  Microbial Populations Relative to
                                 PCB Concentration in Core Samples
PCB Concentration
  (mg/kg soil)

       <.05
     .05-5.0


     5.0-10.0


    10.0-30.0


    30.0-66.6
Criterion*1)
   for
 Average

   High
   Low

   High
   Low

   High
   Low

   High
   Low

   High
   Low
                                     Average Microbiological Counts
                                           (Per Grain of Soil)
Bacteria
  82825
  81475

 273535
 273353

 920750
 920750

 140750
 140750

 160000
 160000
Actinomycete
Fungi
     1066
      936

     5708
     5643

     1500
     1475

      375
      375

      100
      100
 3378
 3242

 3269
 3249

 4638
 4638

 2700
 2700

 750
 750
Number
 of
Samples

   80
   55


    4


    2


    1
   (1). The high averages are calculated with less than values  taken  to be positive
      (<.l taken as  .1).  The low averages are calculated with  the  less than
      values taken to be zero.  The actual mean of the samples  must lie between
      the high average and the low average.

-------
                                                                        78


    C.  Analytical Data From Electron-Capture Gas Chromatography (EC/GC).

        During Phase Two of this project, a total of 145 core, three sedi-
                       •  /                        ,
        ment, and two control soil samples were analyzed quantitatively

        for Aroclor 1254.  Analytical and collection data for soils are

        contained in three tables.  Data for the three test core sites

        are in Table 14,  data for the five core sites sampled at four-

       ,inch intervals are in Table 15, and the remainder of the core

        samples are in Table 16.  The nineteen water.samples collected

        for this project were analyzed for both Aroclor 1254 and the

        askarel solvent.   Results for these analyses plus those for

        sediment and control soils are given in Table 17.  No analysis

        difficulties were encountered during the course of this project.



    D.  Special Soil Extraction Experiment.

        In 1973, data relating to the concentration of PCBs in soil and

        sediment were obtained using hexane as the solvent for the ex-

        traction.  Two other extraction systems were recommended for

        consideration by the project officer for Contract No. 68-01-3232.

        The first employs a dual hexane/acetone system (33), and the

        second extracts with 15% methylene chloride in hexane (V/V) (34).

        Eight of the larger samples were selected for comparative analysis

        using the three extraction systems.  In two instances, there was


(33) Crump-Wiesner,  H. J., Feltz, H. R., and Yates, M. D., 1973, A Study
     of the distribution of polychlorinated biphenyls in the aquatic
     environment:  Jour.  Research U. S. Geol. Survey, v. 1, no. 5, p. 603-607.

(34) National Pollutant Discharge Elimination System, Appendix A, Fed. Reg.,
     38,  No. 75j Pti II (11-28-73).

-------
                                                                      79
  TABLE 14.  Collection and Analytical Data From Three Test Core
             Sites Sampled Over Four-Inch Intervals in August 1975
                  Field    Sample
   Core Site      Sample   Depth2   Character3   PCB Concentration
Identification1   Number  (Inches)   of Soil     	(mg/kg)

     RL-1         GF7516   19-23         2              1.51     .
     RL-2         GF7520                                0.95
     RL-3         GF7524                '                1.52
     RL-4         GF7528                               14.5
     RL-1         GF7517   23-27         2             <0.05
     RL-2         GF7521                               <0.05
     RL-3         GF7525                               <0.05
     RL-4         GF7529                               10.9
     RL-1         GF7518   27-31         2             <0.05
     RL-2         GF7522                               <0.05
     RL-3         GF7526                               <0.05
     RL-4         GF7530                                0.18
     RL-1         GF7519   31-35         2             <0.05
     RL-2         GF7523                               <0.05
     RL-3         GF7527                               <0.05
     RL-4      ,   GF7531                               <0.05
     RL-5         GF7618   17-33         2              0.16
1See Exhibit II for general location
2Measured from the original 1973 surface prior to backfill
3See Table 5  for soil code descriptions

-------
                                                                      80
TABLE 14.  Collection and Analytical Data From Three Test Core
           Sites Sampled Over Four-Inch Intervals in August 1975
                             (Continued)
   Core Site
Identification1

     UC-1
     UC-2
     UC-3
     UC-4
     UC-1
     UC-2
     UC-3
     UC-4
Field    Sample
Sample   Depth2
Number  (Inches)
GF7532
GF7536
GF7540
GF7544

GF7533
GF7537
GF7541
GF7545
         Character3
          of Soil
21-25
25-29
PCB Concentration
	(mg/kg)

       0.09
      <0.05
      <0.05
       0.16

      <0.05
       0.70
       0.16
       2.24
UC-1
UC-2
UC-3
UC-4
UC-1
UC-2
UC-3
UC-4
GF7534
GF7538
GF7542
GF7546
GF7535
GF7539
GF7543
GF7547
                           29-33
                           33-37
                                     <0.05
                                      0.52
                                     <0.05
                                     <0.05

                                     <0.05
                                     <0.05
                                     <0.05
                                     <0.05
1 See Exhibit n for general location
2 Measured from the original 1973 surface prior to backfill
 See Table  5 for soil code descriptions

-------
                                                                      81
  TABLE 14.  Collection and Analytical Data From Three Test Core
             Sites Sampled Over Four-Inch Intervals in August 1975
                                 (Continued)
   Core Site
Identification1

     JF-1
     JF-2
     JE-3
     JF-4
     JF-5  ,
     JF-6
     JF-7
     JF-8
     JF-1
     JF-2
     JF-3
     JF-4
     JF-5
     JF-6
     JF-7
     JF-8
     JF-1
     JF-2
     JF-3
     JF-4
     JF-5
     JF-6
     JF-7
     JF-8
     JF-1
     JF-2
     JF-3
     JF-4
     JF-5
     JF-6
     JF-7
     JF-8
Field
Sample
Number
GF7548
GF7556
GF7564
GF7572
GF7552
GF7560
GF7568
GF7576
GF7549
GF7557
GF7565
GF7573
GF7553
GF7561
GF7569
GF7577
GF7550
GF7558
GF7566
GF7574
GF7554
GF7562
GF7570
GF7578
GF7551
GF7559
GF7567
GF7575
GF7555
GF7563
GF7571
GF7579
Sample
. Depth2 Character3
(Inches) of Soil
19-23 1



23-27 1

V

27-31 1



31-35 1

,
•
35-39 1



39-43 1



43-47 1



47-51 1


i
PCS Concentration
     (mg/kg)	

       0.36
      <0.05
       0.08
       0.17
    '  <0.05
      <0.05
      <0.05
      <0.05
       0.27
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
      <0.05
       0.18
      <0.05
      <0.05
       0.25
       0.24
      <0.05
      <0.05
      <0.05
      <0.05
 See Exhibit II  for general location
2Measured from the original 1973 surface prior to backfill
3See Table  5  for soil code descriptions

-------
                                                                      82
    TABLE 15.  Collection and Analytical Data From Core Sites
               Sampled Over Four-Inch Intervals in August 1975
   Core Site
Identification1

     EK-1
     EK-2
     EK-3
     EK-4
     SR-1
     SR-2
     SR-3
     SR-4
     SR-5
     SR-6
     SR-7
     SR-8
     CC-1
     CC-2
     CC-3
     CC-4
     SM-1
     SM-2
     SM-3
     SM-4
     FH-1
     FM-2
     FM-3
     FM-4
     FM-5
     FM-6
     FM-7
     FM-8
Field
Sample
Number
GW580
GF7581
GF7582
GF7583
GF7888
GF7589
GF7590
GF7591
GF7592
GF7593
GF7594
GF7595
GF7596
GF7597
GF7598
GF7599
GF7600
GF7601
GF7602
GF7603
GF7604
GF7605
GF7606
GF7607
GF7608
GF7609
GF7610
GF7611
Sample
Depth2
(Inches)
15-19
19-23
23-27
27-31
9-13
13-17
17-21
21-25
25-29
29-33
33-37
37-41
9-13
13-17
17-21
21-25
20-24
24-28
28-32
32-36
33-37
37-41
41-45
45-49
49-53
53-57
57-61
61-65

Character3
of Soil
3
3
3
4
1
1
1
1
4
4
4
4
5
5
1
1
6
6
6
6
7
7
7
1
1
1
1
1
                                                 PCS Concentration
                                                      (rag/kg)

                                                        0.13
                                                       <0.05
                                                        0.27
                                                        0.13
                                                        0.08
                                                        0.24
                                                        0.89
                                                       <0.05
                                                       <0.05
                                                       <0.05
                                                       <0.05
                                                       <0.05
                                                        0.17
                                                        0.40
                                                       <0.05
                                                        0.14
                                                       <0.05
                                                        0.08
                                                        1.10
                                                        0.11
                                                        0.22
                                                       *0.05
                                                        0.14
                                                        0.09
                                                       
-------
  TABLE 16.   Collection and Analytical Data from Core Sites
             Sampled Over Sixteen-Inch Intervals in August 1975
                                                                      83
                  Field-    Sample
   Core Site      Sample   Depth2   Character3   PCB Concentration
Identification1   Number  (Inches)   of Soil
LJ-1-4
LJ-5-8
MB-1-4
KH-1-4
SW-1-4
JR-1-4
GW-1-4
TE-1-4
JD-1-4
MM-1-4
MM- 5-8
MM- 9-12
MM-13-16
GE-1-4
CT-1-4
MT-1-4
MT-5-8
BT-1-4
BT-5-8
LP-1-4
JS-1-4
FA-1-4
FA-5-8
GF7612
GF7613
GF7614
GF7615
GF7616
GF7617
GF7619
GF7620
GF7621
GF7622
GF7623
GF7624
GF7625
GF7626
GF7627
GF7630
GF7631
GF7632
GF7633
GF7634
GF7635
GF7636
GF7637
9-25
25-41
9-25
21-37
12-28
9-25
9-25
9-25
9-25
9-25
25-41
41-57
57-73
19-35
3-19
9-25
25-41
9-25
25-41
3-19
4-20
9-25
25-41
1
1
1
1
1
8
4
10
5
5
5
5
8
5
1
5
1
1
1
1
1
1
1
7.36
<0.05
3.69
<0.05
23.8
5.00
3.23
<0.05
X 0.17
0.27
<0.05
<0.05
<0.05
0.32
0.16
4.35
,0.89
1.34
10.1
3.41
5.62
<0.05
<0.05
     Exhibit E  for general location
2Measured from the original 1973 surface prior to backfill
3See Table 5  for soil code descriptions

-------
                                                                      84
  TABLE  16., Collection and Analytical Data from
             Sampled Over Sixteen-Inch Intervals
                                  (Continued)
   Core Site
Identification1
Field    Sample
Sample   Depth2   Character3
Number  (Inches)   of Soil
                               Core Sites
                               in August 1975
PCB Concentration
     (mg/kg)
PT-1-4
FO-1-4
FO-5-8
AA-1-4
xEL-l-4
EL-5-8
PS-1-4
W-l-4
SS-1-4
SS-5-8
HH-1-4
HH-5-8
AM-1-4
VT-1-4
JE-1-4
BA-1-4
TR-1-4
CV-1-4
PQ-1-4
MD-1-4
BS-1-4
BS-5-8
BS-9-12
GF7638
GF7639
GF7641
GF7640
GF7642
GF7643
GF7644
GF7645
GF7646
GF7647
GF7648
GF7649
GF7650
\
GF7651
GF7652
GF7653
GF7654
GF7655
GF7656
GF7657
GF7658
GF7659
GF7660
3-19
9-25
25-41
3-19
9-25
25-41
2-9
9-25
9-25
25-41
9-25
25-41
9-25
0-16
29-45
21-37
25-37
33-45
9-25
3-19
10-26
26-42
42-58
, 1
1

1
1
1
1
1
1
1
2
1
2
2
1
1
1
1
1
1
66.6
<0.05
<0.05
0.34
<0.05
<0.05
2.04
<0.05
<0.05
<0.05
0.67
0.12
<0.05
0.17
<0.05
<0.05
<0.05
<0.05
0.21
0.15
7.45
0.26
0.13
'See Exhibit H  for general location
2Measured from the original 1973 surface prior to backfill
3See Table 5  for soil code descriptions

-------
                                                                      85
  TABLE  16.  Collection and Analytical Data from Core Sites
             Sampled Over Sixteen-Inch Intervals in August 1975
                                   (Continued)
   Core Site
Identification1

     HT-1-4
     HT-5-8
     ZZ-1-4
     ZZ-5-8

     BR-1-4

     SL-1-4
Field    Sample
Sample   Depth2   Character3
Number  (Inches)   of Soil
GF7661    9-25
GF7662   25-41

GF7663    9-25
GF7664   25-41
GF7665

GF7666
3-19

9-25
10
10

 5
 5

 5

 5
PCB Concentration
	(mg/kg)

       0.16
    ,  <6.05

       0.29
       0.50

       0.91

       2.05
1See Exhibit H for general location
2Measured from the original 1973 surface prior to backfill
3See Table 5  for soil code descriptions

-------
Table 17.  Aroclor 1254 and Askarel Solvent Analyses for
         .  Water, Sediment, and Control Soil Samples

                         WATER  .
      Concentrations are expressed as yg/liter (ppb)
Date
Collected
8-28-75
8-29-75
8-29-75
8-29-75
8-29-75
8-29-75
8-29-75
8-29-75
8-29-75
8-29-75
9-26-75
9-26-75
9-26-75
9-26-75 '
9-26-75
9-26-75
9-26-75
9-26-75
9-26-75

Field
Location
Sta. 7
Sta. IB
Sta. 22
Sta. 43
Sta. 23
Sta. 28
Sta. 42
Sta. 40
Sta. 70
Sta. 89
Sta. 43
Sta. 23
Sta. 22
Sta. 40
Sta. 42
Sta. 28
Sta. 70
Sta. 7
Sta. 89
Sta. IB
Sample
Number
PP 2257
PP 2258
PP 2260
PP 2261
PP 2263
PP 2264
PP 2265
PP 2266
PP 2267
PP 2268
PP 2270
PP 2271
PP 2272
PP 2273
PP 2274
~ PP 2275
PP 2276
PP 2277
PP 2278
1,2,4- 1,2,3- 1,2,4,5- 1,2,3,4- penta-
Aroclor Total trichloro- . trichloro- tetrachloro- tetrachloro- chloro-
1254
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
. <0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
<0.5
Solvent benzene benzene benzene benzene benzene
1.23 - 0.18 0.47 0.33 0.25
<0.006 - - -
<0.006 - - - -
<0.006 - - -
<0.006 - - - - -
<0.006 - - - • . - - -
<0.006 - - - - -
<0.006 - -
<0.006 - - -.-.-. -
<0.006 - - - -
<0.006 - - - -
<0.006 - - - - -
<0.006 - - - - -
<0.006 - -
<0.006 - - - . -
.. <0.006 - - - -
<0.006, - ' • '- - -
0.92 0.006 0.097 0.417 0.152 0.245
<0.006
Collection Permission Refused
               SEDIMENT AND CONTROL SOILS
   Concentrations are expressed as mg/kg (dry basis)
8-29-75
8-29-75
9-26-75
8-14-75
8-14-75
Sta. IB
Sta. 43
Sta. 43
Control A
Control B
PP 2259
PP 2262
PP 2269
GF 7683
GF 7684
<0.05
<0.05
<0.05
<0.05
<0.05
0.016
<0.010
<0.010


                                                                     0.016
                                                                                         00

-------
                                                                     87
    insufficient sample available for triplicate determinations.




    Comparative data are found in Table 18.  Based on this experi-




    ment, all three solvent systems are equally effective in the




    extraction of PCBs from the clay soil present at the spill site.




    Round-robin studies in 1973, including the contractor and the control




    agencies involved with the askarel spill, showed similar




    correlation between the use of hexane only as the extractant




    when compared to a dual hexane/acetone solvent system.








E.  Quality Assurance Data.




    The quality assurance for this project was provided by an in-




    house quality control program.  Duplicate blind sample splits




    for use as quality control checks were prepared as soon as the




    air-dried samples were screened; therefore, the quality control




    data are representative of both method and sample reproduci-




    blllty.  Results of these analyses are tabulated in Table 19.






    The eight samples involved in the special solvent extraction




    experiment were also included in the in-house quality control




    program.  In order to more accurately assess the precision




    of the data, the results from these two sources were combined;




    and an average percent coefficient of variation was determined for




    the samples involved.  Tabulation of the data is found in Table




    20.  The average coefficient of variation based on these results




    is ± 6.6%.

-------
                                                                      88
     Table 18.  Results from Special Soil Extraction Experiment
                —Aroclor 1254
                                         *t

      Concentrations are expressed as rag/kg (dry-weight basis)


                                      Solvent System
Sample
Identification
GF 7616
GF 7617
GP 7635
GP 7640
GF 7644
GP 7648
. GF 7654
GF 7675
Hexane
20.7
7.24
5.36
0.31
2.08
0.82
<0.05
4.35
' . * •.*.,> -. .-•
Hexane /Ace tone
20.7
5.46
6.12
0.35
2.22
'F
0.74
<0.05
3.50
Hexane-15*
Methylene Chloride
22.0
5.94
5.19
0.43
*
0.73
<0.05
*
•Insufficient sample for experiment

-------
                                                                      89
            TABLE 19.  In-house Quality Control Data—Aroclor 1254




        Concentrations are expressed as mg/kg (dry basis)









Sample Code          Original Analysis          Duplicate Analysis
GF 7545
GF 7589
GF 7608
GF 7609
GF 7610
GF 7611
GF 7616
GF 7617
GF 7627
GF 7635
GF 7640
GF 7644
GF 7648
GF 7654
GF 7675
2.24
0.24
<0.05
<0.05
1.27
1.21
23.8
5.00
0.16
5.62
0,34
2.04
0.67
<0.05
4.35
. 1.64
0.34
* <0.05
<0.05
1.22
1.21
18.0
5-65
0.06
5.39
0.31
2.08
0.82
<0.05
5.44

-------
Table 20.  Percent Coefficient of Variation Evaluation for In-house
           Quality Control Data—Aroclor 1254
                                            \

         Concentrations are expressed as mg/kg dry basis
                                                                      90
                                                          Hexane     Average
Sample
Code
GF 7616
Z C.O.V.
GF 7617
Z C.O.V.
GF 7635
Z C.O.V.
GF 7640
Z C.O.V.
GF 7644
Z C.O.V.
GF 7648
Z C.O.V.
GF 7654
Z C.O.V.
GF 7675
Z C.O.V.
Average
Value
21.0
5.86
5.54
0.35
2.11
0.76
<0.05
4.41
Original
Analysis
23.8
±13.
5.00
±15.
5.62
±1.4
0.34
±2.9
2.04
±3.3
0.67
±12.
<0.05
±0.
4.35
±1.4
QC Blind
Split
18.0
±14.
5.65
±3.6
5.39
±2.7
0.31
±11.
2.08
±1.4
0.82
±7.9
<0.05
±0.
5.44
±23.
Hexane
Solvent
20.7
±1.4
7.24
±24.
5.36
±3.2
0.31
±11.
2.08
±1.4
0.82
±7.9
<0.05
±0.
4.35
±1.4
Hexane/
Acetone
20.7
±1.4
5.46
±6.8
6.12
±10.
0.35
±0.
2.22
±5.2
0.74
±2.6
<0.05
±0.
3.50
±21.
Methylehe
Chloride
22.0
±4.8
5.94
.±1.4
5.19
±6.3
0.43
±23.
-
0.73
±3.9
<0.05
±0...
-
C.O.V.
(Z)
±6.9
±10.2
±4.7
±9.6
±2.8
±6.9
±0.
±11.7

-------
                                                                    91
F.  Analytical Data from Gaa Chromatography/Mass Spectrometry (GC/MS)..


    At the;conclusion of Phase II, Task A—Laboratory Analysis of


    Soil and Water Samples by EC/GC—a meeting was held at the EPA


    Southeastern Environmental Research Laboratory In Athens, Georgia,


    which involved Mr. Don Brown, Mr. Bill Loy, and Dr. Anna M.


    Yoakum.   After a careful review of the EC/GC data, the two samples


    containing the highest concentration of Aroclor 1254 were selected


    for further study utilizing gas chromatography/mass spectrometry


    (GC/MS).  One sample selected was from the original excavation


    area, and the other came from below the surface of an area un-


    disturbed in the original excavation.  The two samples selected


    for evaluation were correlated to an actual sample of the


    askarel involved in the 1973 spill.




    The GC/MS data outputs consist of total ion current chromato-


    grams and specific ion searches of the data (for ions charac-


    teristic of the Indicated polychlorinated biphenyls).  The


    chromatograms obtained (Figures 9-13) show that the same isomers
                  /

    are present in the same ratios in the environmentally aged


    sample extracts and in the askarel.  These data indicate that


    no selective degradation of the Aroclor 1254 isomers has


    occurred.  Mass spectra typical of four, six, and seven chlorine


    biphenyls are presented in Figures 14-16P

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                                                                            92
VII.  FATE OF POLYCHLORINATED BIPHENYLS (PCBs) AND POLYCHLOROBENZENES
   ..  AFTER A TWO-YEAR EXPOSURE IN A NATURAL ENVIRONMENT

      The experimental evidence presented in the preceding section of this

      report will now be evaluated in an effort to determine the fate of

      askarel after a two-year exposure in a natural environment.  This evalua-

      tion will be conducted in two parts.  The first part deals with PCBs and

      the second with the polychlorobenzene solvent.  Only one commercial mix-

      ture of PCBs, Aroclor 1254, was found associated with the spill.  Data

      evaluation for PCBs will, therefore, be restricted to this material.
 \  •     '                 .           -                  .        '

      Potential mechanisms for the loss of the askarel remaining in the

      spill area after the cleanup include volatilization, leaching,

      metabolic and nonmetabolic degradation.  Each of these mechanisms

      and any other loss pathway which appears appropriate will be con-

      sidered in conjunction with the experimental evidence collected.



      A.  Fate £f Aroclor 1254.

          1.  Distribution of Aroclor 1254 In and Around the Spill Site in

              1975.  Aroclor 1254 was detected in 68 of 145 core samples

              collected in and around the 1973 excavation areas.  Concentra-

              tions ranged from 0.05 to 67 mg/kg in the positive samples.  A

              concentration distribution of Aroclor 1254 in the core samples

              is shown in Figure 17.
              ^

              In order to effectively evaluate the 1975 distribution of

              Aroclor 1254, pertinent facts relative to the spill itself

              need to be considered.  The magnitude of the 1973 spill

              and the elevation contours of the semi-mountainous terrain

              of the spill site assured an initial mass flow transport
                                                       i
              process which resulted in complete saturation of the

-------
       Figure 9. Total Ion Current Chromatograms
                                           93
GF7638
Til
   GF7616
                                         Til
   PYRANOL
   Til
    60      80      100     120     140

-------
                                                                    94
    Figure 10. Specific Ion Search-for the 292  Ion




        Indicative of the Four Chlorine Biphenyl Isomers
      GF7616
292!
X'ed puk« are  the Mjor four chlorine blphenyl Isooers

-------
Figure 11.   Specific Ion  Search for the 326  Ion




      Indicative of the Five Chlorine Biphenyl Isomers
                                                                95
  PYRANOL
                     X
326!
   Z'cd peaks are~thc major five chlorine bipheuyl isoaera

-------
    Figure 12.  Specific Ion Search for the 360  Ion




         Indicative of the Six Chlorine Biphenyl Isomers
                                                          96
GF7638
      360
                                             x  x
                                         ww.
                                  •r—-*•
-r—••;—-r
     GF7616
            3601
     PYRANOL
            360:
     X'ed peaks are the major six chlorine biphenyl isomers

-------
      Figure 13.   Specific^Ioti Search for the 394  Ion



            Indicative of the Seven Chlorine Biphenyl Isomers
                                                                    97
GF7638
     GF7616
     PYRANOL
ft  394'
      X'ed peaks are the major seven chlorine biphenyl isomers

-------
                Figure' 14.   Spectrum typical of four chlorine Biphenyl
       SPECTRUM
  I

  8.
~
   *
i4
             S3 .-  SS

               ecu


                                                ll
                                                                                              X
                                                                                                                              &
                                                                                                                            0
     170  183  1S3  283  213  223  223  213  2S3  253 270 2e3  223  333  318  323  323  3*3 333  2S3  373,353  323  <1G3
              E
                                                                                                                          VO
                                                                                                                          oo

-------
       Figure 15. Spectrua typical of nix chlorine Biphenyl
      SPECTRUM MJK35R 103-107
 8.
-v»J_
 a.
 o

                                            M

                                                                                    T
T
                                                                                                                         J&
       **

    170  163  1S3  233  213  228  223  213  2S3  268  273  283 223  333 310  323  323  313  333  333  373  333  2£>3
             e  •
                                                                                                                           VO
                                                                                                                           VO

-------
                                  Figure 16.  Spectrum typical of  seven chlorine Biphenyl



                MtfBeR 131  -  132




       F-313232. CONC,MIX COL, 12J3Y10.C3,10-6-S
  8.

  8.


r-
fc5>
    .
go
                                                  lil
I|l11l|ll1l|lli<|;iltflll.«lll>pl»fllll|llf>llll<|l»>ill»|. I  I H|HII|lll'j...¥,JlllJ.fT«II ll|f..'f..Tj.I.,,.tl.|l	"('" I


250  270  283  233  333  313  323  333  3*3  323  332  373  £83

173  163   133

         e
                          218
233
                                                                                                                               o
                                                                                                                               o

-------
                                                            101




top soil with varying degrees of penetration into the clay




overburden.  This transport process covered a relatively




large area in both horizontal and vertical directions.




The spread of the askarel was also affected by the movement




of contaminated surface water resulting from massive rain-




fall in the weeks immediately following the spill.  Exca-




vation operations revealed that the distribution in the




clay was non-uniform; and numerous so-called "hot spots"




resulted from movement along the root systems of plants




and trees, as well as from movement in the fractured chert




frequently found in the clay matrix.






Based on 1973 data, three test core sites—RL, UC, and JF—



were selected for the initial phase of the study.  Concen-




tration-depth profiles relating the 1975 data to the




original 1973 data are shown in Figures 18-20.  As was




typical of the 1973 distribution, one so-called "hot spot"




was detected; but the magnitude of the Aroclor 1254 was




not excessive (<25 mg/kg).  A comparative tabulation of




1973 and 1975 core data are given in Table 21.






One stated purpose of the study was to determine if a




reduction in concentration of Aroclor 1254 in the soil had




occurred either as a result of migration from the spill




area or from degradation.  By direct comparison, the 1973




data is higher in 61% of the locations; but the order of

-------
   100-
    90-
                                        Figure  17_.




           Concentration Distribution o£_ Aroclor  1254  in  Core Samples, 1975
                                                      102
    80_
    70-
    60-
01
0)
i-t
a



I   50-

   30-
   20-
   10-
    0-
          (<0.05)
(0.05-0.99)
(1.0-9.9)
(10.-24.)
                      Concentration  Ranges,  Aroclor 1254 (mg/kg)

-------
 Figure 18.  Concentration-Depth  Profiles of Core  Site UC,
             1973-1975
                                                                       103
             Core  depth profiles given  in inches
    UC-/
25
29
33
37
    UC-3
21
25
29
33
37
                               1973 Core
                              21
                              23
                              29
                              33
37   \mz
                    AROCLOR  12S4  CONCENTRATION  mg/Kg
                   
-------
  Figure 19.   Concentration-Depth Profiles .of Core  Site RL,
               1973-1975
                                                 104
               Core depth  profiles given  in inches
     RL-I
 19
 23
 27
 31
 35
    RL-3
 19
23
27
31
33
                                           1975
                                       PL-Composite
1973 Core      "  M
                        21
                                       21
                        25
                                       25
                       29
                                       29
                       33
                                      33
                     AROCLOR  1254  CONCENTRATION   mg/Kg
                    <0.05       O.OS-.99     1.0-9.9       1.0-24.
                                         RL-2
                                                              19
                                                              23
                                                              27
                                                              31
                                                              35
                                         RL-4
                                         *f pfc.
                                     19   c-?T
                                                              23
                                                             27
                                                             31
                                                             35
                                          XX
                                          XX
                                          XX
                                          XX
                                          XX
                                          XX
                                          XX
                                          xx
                                          xx
                                          XX
                                          XX
                                         'XX
                                          XXx
                                          XX
                                          XX
                                          XX
                                          XX
                                          XXx

-------
                                                                         105
   Figure  20.   Concentration-Depth Profiles of  Core Site JF,
                1973-1975

                Core  depth profiles given in inches
   JF-I            JF-2           JF-3           JF-4        1973 Core
19
23
27
31
35
39
43
47
51
                19
                23
                27
                31
                35
                39
                43
                47
                51
                                19 S:::
                                23
                                27
                                31
                                35
                                39
                                43
                                47
                                51
                                                19
                                               23
                                               27
                                                31
                                                35
                                               39
                                               43
                                                47
                                              \
                                                51
                    AROCUOR  1254  CONCENTRATION   mg/Kg
                                                              .  19
                                                               23
                                                               27
                                                               31
                                                               35
                                                               39
                                                               43
                                                               47
                   
-------
                         Table 21.   Comparison of Analytical  Data from
                                    Core  Samples Collected  in 1973 and 1975
Core Site
 Collection Depth
Ranges (in Inches)
Identification Year 1973 Year 1975
Part I.
RL
UC
JF
Part II.
EK
SR
CC
SM
FM
Part III.
IJ
MB
KH
SU
JR
GW
TE
JD
4-Inch Test Cores
9-35
9-60
9-45
4-Inch Core Samples
9-35
9-45
9-36
9-36
9-66
16-Inch Core Samples
9-43
9-37
9-37
9-30
9-20
9-40
9-62
9-35

19-35
21-37
19-51

15-31
9-41
9-25
20-36
33-65

9-41
9-25
21-37
12-28
9-25
9-25
9-25
9-25
 Analytical Results—1973
PCB Concentration  No. of
	(mg/kg)       Samples
                                              1.25-5.87
                                              0.06-0.27
                                              2.04-7.67
                                              0.86-2.76
                                              0.27-0.43
                                              0.88-2.20
                                              0.44-0.85
                                              1.64-3.44
                                              0.55-1.45
                                              1.60-5.86
                                              4.10-6.56
                                              0.19-1.42
                                              0.52-2.40
                                              0.09-0.29
                                              0.07-0.21
                                              0.27-0.82
                                              3
                                              4
                                              3
                                              4
                                              3
                                              3
                                              3
                                              3
                                              3
                                              3
                                              3
                                              3
                                              2
                                              3
                                              4
                                              3
 Analytical Results—1975
PCB Concentration  No. of
     (mg/kg)	.  Samples
                                  <0.05-14.5
                                  <0.05-2.24
                                  <0.05-0.36
                                  <0.05-0.27
                                  <0.05-0.89
                                  <0.05-0.40
                                  <0.05-1.10
                                  <0.05-1.27
                                  <0.05-7.36
                                      3.69
                                     <0.05
                                     23.8
                                      5.00
                                      3.23 ~
                                     <0.05
                                      0.17
                      17
                      16
                      32
                       4
                       8
                       4
                       4
                       8
                       2
                       1
                       1
                       1
                       1
                       1
                       1
                       1

-------
                           Table 21.  Comparison of Analytical Data from
                                      Core Samples Collected in 1973 and 1975
                                                (continued)
  Core Site
Identification

      MM
      GE
      CT
      MT
      BT
      LP
      JS
      FA
      PT
      FO
      AA
      EL
      PS
      DV
      SS
      HH
      AM
      VT
      JE
      BA
      TR
      CV
      PQ
      MD
      BS
      HT
      ZZ
      BR
      SL
 Collection Depth
Ranges (in Inches)
Year 1973
9-66
9-35
3-6
9-40
9-36
3-6
3-6
9-42
. 3-6
9-36
6-9
9-50
0-18
9-34
9-36
9-66
9-38
3-6
39-45
34-37
33-41
33-52
9-22
0-18
9-48
9-55
9-40
0-18
9-61
Year 1975
9-73
19-35
3-19
9-41
9-41
3-19
4-20
9-41
3-19
9-41
3-19
9-41
2-9
9-25
9-41
9-41
9-25
0-16
29-45
21-37
25-37
33-45
9-25
3-19
10-58
9-41
9-41
3-19
9-25
                                             Analytical Results—1973
PCB Concentration
(mg/kg)
0.23-6.72
0.68-2.32
0.12
1.37-3.00
3.44-311.
3.26
3.85
1.85-8.50
0.50
0.05-0.21
0.60
0.11-1.00
0.09-0.69
0.21-0.58
0.46-5.70
<0. 05-1. 35
0.09-0.15
0.72
0.16-0.20
0.12
<0.05
<0. 05-1. 39
0.40-0.44
0.76-7.50
0.27-15.8
0.13-4.80
0.47-7.50
0.63-6.00
<0. 05-0. 20
No. of
Samples
4
3
1
3
3
1
1
3
1
3
1
3
3
3
3
3
3
1
2
1
2
4
2
3
4
4
3
3
3
Analytical Results—1975
PCB Concentration
(mg/kg)
<0. 05-0. 27
0.32
0.16
0.89-4.35
1.34-10.1
3.41
5.62
<0.05
66.6
<0.05
0.34
<0.05
2.04
<0.05
<0.05
0.12-0.67
<0.05
0.17
<0.05
<0.05
<0.05
<0.05
0.21
0.15
0.13-7.45
<0. 05-0. 16
0.29-0.50
0.91
2.05
No. of
Samples
4
1
1,,
2
2
1
1
2
1
2
1
2
1
1
2
2
1
1
1
1
1
1
1
1
3
2
2
1
1

-------
                                                                108
    magnitude for the Aroclor 1254 concentrations are comparable

    for both years; and no significant migration patterns are

    observed.




2.   Degradation of Aroclor 1254.  Two possible degradation modes

    exist—metabolic and nonmetabolic.  Each degradation mechanism

    will now be considered from the standpoint of applicability to

    the system under study.




    a.  Chemical Transformation—The non-biological alteration

        of a chemical introduced into any part of the environ-

        ment is dependent on the moisture, pH, and temperature

        of that environment; on the nature of reactive groups

        on the agent; and on the presence of catalytic sites.

        In addition, the nature and intensity of available

        illumination determines photochemical reactions.


        Irradiation (photolysis) of Aroclor 1254 under laboratory

        conditions has produced dechlorinated products; and, in

        the presence of air and water, hydroxylated and hydrated
               •\     ' .              •           • •
        products have been identified in the polar products of

        the irradiation.  Although these theoretical routes for

        chemical degradation exist, the extreme stability

        (chemical inertness) of Aroclor 1254 coupled with the

        absence of illumination at the potential reaction site

        make chemical transformation a highly improbable degrada-

        tion mechanism.

-------
                                                            109
b.  Biological Alteration—By analogy to the dechlorination of




    DDT to ODD by soil microorganisms, biodegradation of Aroclor




    1254 is considered to be a possible degradation mechanism




    in a natural environment.  The unique biochemical asset of




    certain aerobic microorganisms to catalyze early steps in




    degradation allows for the formation of metabolites which




    can enter the common pathways of metabolism.  The establishment




    of measureable biodegradation is dependent on the ease of




    physical or chemical sequestration of the PCB components due




    to the structure of the molecule as it relates to microbial




    enzymatic action.  The intrinsic toxicity of the askarel,




    environmental factors affecting microbial populations and




    their specificity, and available time for the maximum develop-




    ment of the degradation process are also significant considera-




    tions in the interpretation of biological alterations of PCBs.






c.  Aroclor 1254 Degradation Assessment—Aroclor 1254 is a mix-




    ture of chlorinated biphenyl homologs.  If no degradation




    in the environment occurred, or if all homologs degraded




    at the same rate, the ratio of homologs in "aged environmental




    samples" should be the same as that in the askarel introduced




    into the environment at the time of the spill.  On the other




    hand, if the homolog ratio in the "aged environmental samples"




    from the spill site differs from that of the askarel spilled,




    some process(es) must be operating in the environment to




    remove different homologs at different rates.  According to

-------
                                                                          110
                 data supplied by the Monsanto Company (35), the typical

                 percent (w/w) composition .of Aroclor 1254 is similar to

                 that given in Table 22.


                 All chromatograms from the EC/GC analyses of 1975 samples

                 were carefully examined for peak alterations and for the

                 appearance of new peaks with reference to the chromato-

                 gram of the askarel released into the environment at

                 the time of the spill.  Comparative chromatograms for

                 Aroclor 1254 in a typical "aged environmental sample"

                 and Aroclor 1254 in the askarel spilled in 1973 are

                 shown in Figure 21.  By comparing the two chromatograms,

                 it is readily apparent that the ratio of homologs in the

                 "aged environmental sample" is the same as that in the

                 askarel introduced into the environment at the time of

                 the spill.  These results indicate that either there has

                 been no degradation in the environment or that all

                 homologs have degraded at the same rate.


                 Using -gas chromatography-mass spectrometry (GC/MS), the

                 homolog distribution of selected 1975 samples was com-

                 pared to the distribution exhibited in a sample of

                 askarel from the 1973 spill.  These data also confirm

                 that the ratio of homologs in the "aged environmental

                 samples" is the same as that of the askarel spilled in

                 1973.
(35) Monsanto Company, "Presentation to the Interdepartmental Task Force
     on PCBs," Washington, D.  C.,  May 15, 1972.

-------
                                                                        Ill
     Table 22.  Typical Homolog Composition of Aroclor 1254  (35)
                 Homolog                                 Aroclor
        (No. of Chlorine/biphenyl)                        1254*
                    2                                      <0.5

                    3                                       1.

                    4                                      21.

                    5                                      48.

                    6                                      23.

                    7                                       6.

                    8                                      NDt
*Percent (w/w) by GC/Mass using area correlation factors by homolog
 response.

tND * None Detected, <0.01%.
(35) Monsanto Company, "Presentation to the Interdepartmental Task Force
     on PCBs," Washington, D. C., May 15, 1972.

-------
                         Tf.tF
                         1OO
                         ±r±
                          Askarel Spilled in 1973
               ;:
                                                                             l  :
      -Representative Aged Environmental Sample—1975 J-J--J -j --!-j-
f
Figure 21.   Comparative Chromatograms for Aroclor 1254 in a Typical

            "Aged Environmental Sample"  (1975)  and Aroclor 1254 in
            the Askarel Spilled in 1973
                         t-rt
                                       H-
                                                                           i  I '
                                                                          1 -
            11
                                                                        ill!'

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                                                                     113
            No evidence was found by GC/MS and/or EC/GC to indicate




            the presence of degradation products in any of the "aged




            environmental samples" collected in or around the spill




            site.  Primary biodegradation can be defined as minimum




            alteration of the chemical structure of the material in


              \

            question to an extent that characteristic properties of




            the original material are no longer evident.  Based on




            the experimental evidence collected, and considering the




            definition just presented, it may be concluded that no




            detectable reduction in the concentration of Aroclor




            1254 in the soil has occurred as the result of chemical




            transformation or biodegradation.  An evaluation of the




            occurrence of major soil microorganisms is consistent with




            this conclusion in that the three microbial groups studied




            were probably not present in sufficient numbers to support




            measureable degradation of the PCB.







B.  Fate of Askarel Solvent.




    The scope of this project did not include the analysis of soils




    for the askarel solvent.  Only water samples, collected adjacent
                                *



    to the study area, were included to provide information on the




    rate of intrusion of polychlorobenzenes into ground water.




    As a consequence, since biodegradation analyses were limited to




    soil samples, an assessment of solvent degradation is not




    possible.  This discussion of the so-called "fate" of askarel

-------
                                                                  114
solvent in the spill environment will be limited to the distribu-
tion of polychlorobenzenes in ground water adjacent to the spill
site.

The 1973 data showed only minimal movement of Aroclor 1254 in

the ground water.  Only two ground water sampling stations gave
positive data for Aroclor 1254.  These locations were a.spring

just below the spill area and a well less than one hundred feet
from the spill excavation area.  The highest Aroclor 1254 con-

centration detected in a water sample was 2.1 ppb.

Migration of the'askarel solvent, however, was completely differ-
ent from that observed for the PCS, Aroclor 1254.  Differential
migration of the askarel solvent in the spill environment was
observed in a well water sample-collected March 22, 1973.  A
subsequent screening of selected water sources—wells, creeks,
ponds, etc.—revealed an interesting phenomenon.  There were
two distinct patterns for the solvent concentration in the
                               \   •-
water samples.  One closely resembled the component distri-
bution of the askarel solvent spilled.  The other was almost
95% trichlorobenzenes.
                              *

Because of this observed phenomenon, a selected number of repre-
sentative core drillings at the spill site were analyzed for
askarel solvent.   These data indicated that tetra-and penta-
chlorobenzene were preferentially retained in the soil while
the other solvent components—especially the trichlorobenzenes—
                      /'
had selectively moved out of the area.  This study also showed

-------
                                                                    115
    that Aroclor 1254 was preferentially retained in the soil.  Data

                                              i
    from monitoring programs confirmed that water samples with sol-


    vent patterns resembling the .askarel solvent component distribu-


    tion indicated possible contamination by surface water run-off
                     V

    and seepage of surface water into a supply.   Water with the high


    trichlorobenzene concentrations indicated intrusion of these sol-


    vent components into the ground water supply.  A graphic repre-


    sentation of solvent concentration in the well located closest


    to the spill site is shown in Figure 22.  Samples from this


    station collected during the follow-up study still show solvent


    present in the water at the 1.0 ppb concentration level.



    These data clearly indicate that the more water-soluble compo-


    nents of the askarel solvent invaded the ground water supply


    almost immediately after the occurrence of the spill.  Leaching


    was the migration mechanism responsible for the intrusion of


    the lower chlorinated benzenes into the ground water supply.



    Based on the 1973 and 1975 data, it can be concluded that in-


    trusion of lower chlorinated benzenes into a ground water


    supply used for drinking water purposes occurred rapidly after


    the spill.  This same water supply contains minimal, but de-


    tectable, quantities of askarel solvent two years after the


    occurrence of the spill.





C.  Over-all Assessment of the Environmental Impact of the Sp-tl-1-.—	


    The contractor did not observe the spill area prior to


    the actual spill; however, observations began on March 21, 1973,

-------
   EB0
   330
^^
   100
a  M
tn  H0
§  30

o=  20
   10
    a
    5
    E
    H
    3
             FIGURE Z2.
   SOLVENT CONCENTRRTIDN PRDFILE
WELL LDCHTED CLD5E5T TD 5P1LL H1TE
            (5THTIDN 7)
                                                                                            *-^
~I'  I     T
 ii   i
   1373
    T    i
                                       OUL1BN DHTE
                                                            i    r

                 1H7H
 s
•1

-------
                                                                117
approximately two and one-half weeks after the spill occurred.




Invasion of the spill area by workers and heavy equipment involved




in the cleanup had already transpired by this time. " There had been




heavy rainfall during the two and one-half week period immediately




after the spill.  The area received 12.44 inches of precipitation




during the month of March which constituted a 7.23 inch departure




from the norm.  All assessments will utilize, as an environmental




basis, the condition of the area at the time the contractor arrived.






1.  Condition of the Area Immediately After the Spill.




    a.  Primary Spill Area—The primary spill area was located




        adjacent to the intersection of a main north-south




        highway and a secondary access road utilized mainly by




        residents living in sparsely developed areas along the




        lake (see Exhibit I). The property on which the spill




        occurred was a segment of land designated for pasture




        farming.







        The spill site is situated in a watershed approximately




        0.6 mile long which drains into the lake in a north-




        easterly direction and generally runs parallel to the




        secondary access road.  The distance from the spill site




        to the lake is approximately 0.5 mile.  The main water-




        shed also received drainage from several secondary




        watersheds adjacent to it.  The spring well, Station  IB,

-------
                                                        118
is located approximately 0.2 mile from the spill site



in this north-east trending hollow.  Most of the chemical



spilled migrated down the slope toward the north-east,



influenced primarily by the surface topography.






The primary spill area, which was later excavated,



covered an area approximately 250 feet long and 125 feet



wide.  This area was sparsely wooded with pines and hard-



woods including:  Shortleaf Pine (Pinus echinata),



Eastern White Pine (Pinus strobus), Virginia Pine  (Pinus



virginiana), White Oak (Quercus alba), Southern Red Oak



(Quercus falcata), Mockernut Hickory (Garya tomentosa),



and Flowering Dogwood (Cornus florida).  Natural under-

                       /         '

story plant associations were observed in the area at



this time.  Two Southern Red Oaks and a Shortleaf Pine



with base diameters of 12-18 inches were removed from



the excavation area in 1973.






Although the cleanup operation was not intended in any
                                                   N^


way to be a terrestrial ecology study, cursory observa-



tions, were made on the biota of the immediate spill area



and the surrounding area at various times during the



period of March through July, 1973. •> No mammals, reptiles



or amphibians and very few birds were observed in  the



vicinity at this time.  Crustacea (crayfish) were observed,



and watercress blanketed the area of the spring well



(Station IB).  There was some evidence of damage to trees



and understory plants in the area of heaviest contamination.

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                                                                119





    b.  Area Peripheral ^o Spill Site—It was suspected that the




        chemical may have also migrated to the south-east across




        the road and into another watershed.  South-east from the




        crest of the hill where the spill occurred the topography




        is gently rolling through 'fields and wooded areas down to




        the lake.  The distance from the spill site to the lake




        in this direction is approximately 0.4 mile.  Little wild-




        life was observed in this area.  Evidence of wildlife




        within pastured segments including dry weather ponds




        was absent.









2.  Condition of Spill Area - August 1975.




    In 1973, the cleanup procedure resulted in extensive exca-




    vation in the spill area.  Contaminated soil was removed;




    and the excavated areas were sealed, backfilled, and packed




    with, uncontaminated soil.  The entire affected area was




    covered with top soil, seeded with grass, and landscaped.




    this area had received little maintenance since 1973; and,




    as a result, grass and weeds were overgrown, hindering field




    operations.   A rotary mower was used to cut the .overgrowth,




    and the area was raked.






    According to the property owner, several damaged trees had




    been removed in the period between the 1973 cleanup opera-




    tion and the latest study.






    Again, the 1975 follow-up study was not intended to examine




    the terrestrial ecology of the area; however, over a period




    of approximately one month (July and August, 1975),

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                                                             120
observations made in the spill area .indicated no obvious

detrimental effects to any biota of the ecosystem.  In the

course of sampling, several specific observations were made.

Crayfish and minnows were found in abundance in the spring
                      l                                '
well waters, as were various aquatic insects and plants.

The immediate spill area was infested with insects including

ants, spiders, ticks, grasshoppers, crickets, flies, mos-

quitoes, bees, and yellow jackets.  Several garter snakes .

were seen in this area also.  Segmented worms were observed

in core samples taken within a few feet of the surface.

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                APPENDIX
   Method for Analysis of Water and




Sediment for Polychlorinated Biphenyls








           Monsanto Company




  Analytical Chemistry—Method 69-13




            February 1970

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               ANALYSIS OF WATER AND SEDIMENT FOR
                   POLYCHLORINATED BIPHENYLS
SCOPE

This methodology was developed for  the determination of  the  amount and
type of polychlorinated biphenyls (PCB)  in water and sediment  samples.
Absolute confirmation of PCB structures  is no.t obtained  with this method.
Structure proof can be obtained using additional techniques  such as mass
spectrometry to further identify the GC  fractions.
PRINCIPLE

The PCB(s) in water and sediment samples are extracted into an organic
solvent.  Interfering components are then removed from the extracts by
chemical treatment and column adsorption chromatography.  The amount
and type of PCB present is determined by electron capture gas chroma-
tography (EC/GC).                                        .
REAGENTS

Hexane


Acetonitrile


Sodium Sulfate
Alumina Adsorption
Nanograde, Mallinckrodt Chemical Works,
Catalog No. 4159.

Nanograde, Mallinckrodt Chemical Works,
Catalog No. 2442.

Anhydrous, granular:  AR grade,
Mallinckrodt Chemical Works, Catalog No.
8042.  Heat at 400°C for one hour prior
to use.

(for chromatographic analysis) 80/200
mesh, Fisher Scientific Co., Catalog No.
A540.  Heat at 400°C for a minimum period
of 4 hrs. and deactivate with 5% (w/w)
distilled water.

Alumina column preparation:  Fill a
chromatographic column with hexane up to
the point where the reservoir joins the
column and push a glass wool plug to the
bottom with a glass rod.  In a 50 ml beaker
measure 35 ml of deactivated alumina (i«30g).,
and pour this slowly into the column.  Tap
or vibrate the column to settle the alumina
and top the alumina with 2-3 cm of anhydrous
sodium sulfate.  Wash the column with 50-
100 ml of hexane prior to the addition of
the sample.

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 Distilled Water               Extracted with hexane to remove hexane
                               soluble electron capturing impurities.

 Sulfuric Acid                 Analytical Reagent Grade, SG = 1.84

 Potassium Hydroxide           Analytical Reagent Grade

 Ethanol     .    .              Formula 2B

 2.5% (w/v) Alcoholic          Dissolve ^12.5 grams of AR grade KOH in
 Potassium Hydroxide           500 ml of ethanol

 9/1 (v/v) Sulfuric            Carefully add 270 ml of AR-grade sulfuric
 Acid - Water                  acid to 30 ml of distilled water in a 500
                               ml iced beaker

 .PCS Standards                 Aroclor 1242, 1248, 1254, and 1260


 APPARATUS

 1.  Separatory funnels equipped with ground glass stoppers and Teflon
     stopcocks:  125, 250, 500, 1000 and 2000 ml capacities.

 2.  Kunderna-Danish Evaporative Concentrators, 500 ml capacity equipped
     with 3-ball Snyder columns and graduated 5 ml capacity vials:  Ace
     Glassware Company, Catalog No. 6707.

 3.  Chromatographic columns, glass, 10" x 20 mm (OD) with a 5" x 50 mm
     (OD) reservoir at the top, equipped with Teflon stopcocks.

 4.  Sintered glass filter funnels, 600 ml capacity, 90M.

 5.  Flat bottomed boiling flasks, 125 ml capacity:  Ace Glassware
     Company, Catalog No. 6896, Code - 04.

 6.  Liebig Condenser, 200 mm in length:  Ace Glassware Company,
     Catalog No. 5915, Code - 12.

 7.  Hot plates, Corning PC-100:  Fisher Scientific Company.

 8.  Water bath, Thelco, Precision Scientific, Model No. 84, Fisher
     Scientific Company.

 9.  Reciprocating variable speed shaker, Eberback Corporation, Fisher
     Scientific Company.

10.  10 yl Hamilton Syringes, Catalog No. 701N.

11.  32 oz. all glass mortars and pestles.

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12.  8 x 12 x 2"  (2-1/2 qt.) Pyrex baking dishes.

13.  U. S. Standard Sieve, No. 30, Fisher Scientific Company.

14.  Usual laboratory glassware.
SAMPLING

It is to be assumed that a rather wide variety of sampling techniques
were employed in collecting the samples submitted for analysis.  In
general the procedures used were probably selected for ease of adaption
to the local situation.  For this reason water and sediment samples
were usually treated as follows:
Water             ,

Where possible the entire water sample, including the container in which
it was collected, was extracted with hexane.  With larger samples, where
this was not physically possible, the containers were simply agitated
and a 250 ml portion used for analysis.
Sediment

Any excess water was decanted and the entire sediment transferred to a
glass baking dish to air dry at room temperature.  The dried material
was transferred from the dish into a mortar and pestle and ground.
The ground sediment was sieved, remixed, and a 250g portion taken for
analysis.
PROCEDURES          :

Extraction of Water Samples

1.  Extraction of water samples - after agitating, transfer the entire
    aqueous sample or a 1000 ml aliquot into a graduated glass cylinder.
    Record the volume of the sample and quantitatively transfer it to
    a separatory funnel with distilled water.

2.  Rins> the graduated cylinder with one 50 ml portion of hexane and
    add each to the separatory funnel.

3.  Stopper the separatory funnel and shake vigorously for at least 2
    minutes.  Allow the layers to separate and transfer the lower
    aqueous phase to a second separatory funnel.

4.  Extract the water sample a second' time with a 50 ml portion of
    hexane.  After the layers have separated, add the first hexane
    extract to the second separatory funnel and transfer the aqueous
    layer to the original separatory funnel.

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 5.  Repeat the extraction with a third 50 ml portion of hexane.  Discard
     the aqueous layer and combine the hexane extracts.

 6.  Filter the combined extracts through a 4" funnel plugged with glass
     wool which is covered with sodium sulfate.  Collect the filtrate
     in a Kunderna-Danish evaporative concentrator^, add a small boiling
     chip, put the Snyder column in place, and reduce the hexane volume
     to less than 5 ml by heating the apparatus in a 80-90°C water bath.
     (CAUTION:  SOLVENT VAPORS MUST BE VENTED TO A HOOD.)

 7.  After cooling, remove the 5 ml graduated, tube and transfer hexane
     extract to an alumina adsorption column washing it in with several
     5 ml portions of hexane.

 8.  Carefully add 100 ml of hexane to the column reservoir and collect
     the total eluent in either a 250 ml volumetric flask or a Kunderna-
     Danish evaporative concentrator.

 9.  If the column eluent is collected in a volumetric flask, dilute to
     volume with hexane and proceed with the gas chromatographic analysis.

10.  If the column eluent is collected in a Kunderna-Danish evaporative
     concentrator, reduce solvent volume, cool, dilute to volume and pro-
     ceed with the gas chromatographic analysis.
 Extraction of Sediment and Soil Samples

 1.  Decant off any excess water and transfer the entire sediment sample
     to a glass baking dish.  Air dry at ambient temperature (heat should
     not be applied).

 2.  When dry, transfer the soil/sediment to a mortar and pestle and grind.
     Sieve the ground material through a No. 30 mesh sieve and weigh 250g
     (to the nearest O.Olg) into a 16 oz. narrow neck screw cap (aluminum
     foil liner) glass bottle.

 3.  Moisten the soil with water C\/ 10 ml) and add 150 ml of acetonitrile.
   .  Cap the bottle tightly, and mechanically shake for a minimum period
     of 1 hour.

 4.  Quantitatively transfer the acetonitrile extract into a 600 ml
     sintered glass filter funnel containing a 1/4" layer of anhydrous
     sodium sulfate.  Collect the filtrate in a 600 ml beaker (vacuum
     filtration may be necessary).

 5.  After the acetonitrile has completely drained into the beaker, wash
     the bottle twice with 50 ml portions of acetonitrile, adding each
     wash to the funnel after the previous has completely percolated
     through the sediment.                      .

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                                  I                '           •
 6.  Quantitatively transfer the extract to a Kunderna-Danish
     evaporative concentrator, add a small boiling chip, put the Snyder
     column in p-lace, and reduce the solvent volume to less than 5 ml
     by heating the apparatus in a 80-90°C water bath.  (CAUTION:
     SOLVENT VAPORS MUST BE VENTED INTO A HOOD.)

 7.  After cooling remove the 5 ml graduated tube and transfer the con-
     centrate of extracts to a 125 ml extraction flask with the aid of
     several small portions of solvent.

 8.  Evaporate the extract just to dryness with a gentle stream of dry
     filtered nitrogen and add 25 ml of 2.5% alcoholic potassium
     hydroxide.

 9.  Add a boiling chip, put a water condenser in place, and allow the
     solution to reflux for 45 minutes.

10.  After cooling, transfer the solution to a 250 ml separatory funnel
     with the aid of 25 ml of distilled water.

11.  Rinse the extraction flask with 25 ml of hexane and add it to the
     separatory funnel.

12.  Stopper the separatory funnel and shake vigorously for at least
     1 minute.  Allow the layers to separate and transfer the lower
     aqueous phase to a second separatory funnel.

13.  Extract the saponification solution with a second 25 ml portion of
     hexane.  After the layers have separated add the first hexane
     extract to the second separatory funnel and transfer the aqueous
     alcohol layer to the original separatory funnel.

14.  Repeat the extraction with a third 25 ml portion of hexane.  Discard
     the saponification solution and combine the hexane extracts.

15.  Carefully add.25 ml of the sulfuric acid solution (9:1 concentrated
     sulfuric acid/water) to the hexane extracts.

1$.  Stopper the separatory funnel and shake vigorously for at least one
     minute.  Allow the layers to separate and discard the lower aqueous-
     acid layer.  Repeat this step until the acid layer is colorless.

17.  Wash the hexane with 25 ml portion of water.  Discard the water
     wash.                                                       .

18.  Filter the hexane extract through a 4" funnel plugged with glass
     wool which is covered with a layer of sodium sulfate into a
     Kunderna-Danish evaporative concentrator.    '    •

19.  Add a small boiling chip, put the Snyder column in place and
     reduce the hexane volume to less than 5 ml by heating the
     apparatus in a 80-90°C water bath.

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20.  After cooling, remove the 5 ml graduated tube and transfer the
     hexane extract to an alumina adsorption column washing it in with
     several 5 ml portions of hexane.

21.  Carefully add 100 ml of hexane to the column reservoir and collect
     the total eluent in either a 250 ml volumetric flask or a Kunderna-
     Danish evaporative concentrator.

22.  If the column eluent is collected in a volumetric flask, dilute to
     volume with hexane and proceed with the gas chromatographic
     analysis.

23.  If the column eluent is collected in a Kunderna-Danish evaporative
     concentrator, reduce solvent volume, cool, dilute to volume and.
     proceed with gas chromatographic analysis.
 Electron Capture Gas Chromatographic Procedure

      Instrument;  F&M 402 Biomedical Gas Chromatograph
      Detector:    High Temperature Ni63 Electron Capture Cell
                   6 mm x 61  Glass Column, 4% XE-60 on 80/100 mesh
                   Chromosorb W, HP, AW-DMCS

      Column Temperature:  160°C
      Detector Temperature:  300°C         Flow Rates
      Injection Port Temperature:  195°C   Helium Carrier ^ 60 ml/rain
      Pulse:  150                          Argon-Methane Purge
                                                 ^ 120 ml/min

 Using EC/GC as the determinative step,  inject in duplicate 1-10 yl of
 each solution into the chromatograph.   By comparison with standard
 solutions injected, in duplicate, under the same operating conditions,
 determine the amount and type of Aroclor using the individual or total
 peak height method.

 The electron capture detector should also be used to guide the isolation
 procedures.  Water and sediment extracts can be checked for the presence
 of PCBs and/or interferences by injecting yl portions of the extracts
 at various points in the extraction and concentration schemes.  In this
 manner, it can be determined if the sample needs to be concentrated or
 diluted and if the clean up  procedures  should be employed.
 DISCUSSION

 Extraction

 The extraction of PCB's from water employing hexane as the extractant
 was found to be quantitative and to be sufficiently simple and rapid for
 use as a routine procedure.

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The evaluation of  this method was based on spiking water samples with
standard acetone solutions of PCB's.  The spiking method consisted of
adding  the PCB's in acetone  (25-50 yl) to 500 ml of tap water in a 32
oz. narrow neck screw cap jar.  After thoroughly mixing, duplicate
225-250 ml aliquots were taken and subjected to the proposed sample
preparation and work up as outlined.  The results were quantified by
preparing a calibration curve using standard hexane solutions of the
PCB's used to spike the water samples.  The major isomer peak height
was used to construct the calibration plot.

The average recovery and deviation achieved substantiated the appli-
cability of the method for the quantitative recovery and analysis
of PCB's from water at the ppb-ppm level.

No PCB  recovery experiments  from spiked sediment and soil samples
have been performed.  Instead, several of the residual solids
representative of  some of the types of sediment or soil analyzed were
re-extracted with  hexane/acetone (40/60) in a soxhlet extractor to
test for the efficiency of the acetonitrile extraction step.  The
hexane, after isolation by dilution with distilled water was then
carried through the purification steps.  Recoveries by soxhlet extrac-
tion have indicated that the acetonitrile extraction of PCB's was
essentially quantitative in  the cases checked.
Sample Concentration

Concentration of sample extracts is necessary, prior to clean up by
chromatographic or chemical means, to reduce sample size and increase
sensitivity.  The preferred method of concentrating allows minimum loss
through volatilization or chemical decomposition and requires a minimum
time.  The three methods of solvent volume reduction most commonly used
are evaporation by exposure to a stream of air, evaporation employing
a KunJerna.—Danish evaporative concentrator equipped with a Snyder
column, and evaporation under reduced pressure.  We have used all
three techniques and have not encountered any significant losses from
volatilization or chemical alternation.  However, the Kunderna-Danish
evaporative concentrator and the stream of air were employed because
of the ease of use.
Column Adsorption Chromatography and Chemical Clean Up

Silica gel, Florisil and Alumina deactivated with 0, 1.0, 1.5, 2.0 and
5% water were investigated as adsorbants for the elimination of inter-
ferences.  Alumina (5% water) was found to be more effective and
reproducible than either silica gel or Florisil.  The activity of
alumina varies with age and lot, therefore, 5% water was added to the
alumina, after heating for a minimum of 4 hours at 400°C to insure a
reproducible activity.

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Saponification and subsequent extraction of the sample with sulfuric
acid is an effective way to remove a number of chlorinated hydrocarbon
interferences as well as other matrix interferences.  PCB's are not
affected.
Electron Capture Gas Chromatography

Columns;

Column performance is the key to effective gas chromatographic analysis
and as such the choice of column materials is particularly important.
Ideally, the support employed should be inert, mechanically strong, and
of high surface area.  For these reasons, Chromesorb W, HP, AW-DMCS
was used in all of our work.

A variety of polar and non-polar liquid phases were investigated.  The
following columns were found to provide adequate separation, etc., for use .
in PCB analysis by electron capture:  4% (w/w) DC-200, SF-96, OV-17, SE-30,
SE-54, XE-60, Apiezon L, and 6% QF-1.  DC-200 and XE-60 or QF-1 have been
found to be the most suitable of these liquid phases.
                                                    i
Another important consideration when working with an extremely sensitive
detector and consequently low levels of materials is column conditioning.
With polar phases such as XE-60 and QF-1, we have found that operating
a new column overnight at a temperature 25-50°C higher than to be used
during analysis results in a more stable column.  A no-flow conditioning
technique is employed to condition non-polar columns.  The column is
purged with carrier gas, heated for 30 minutes at an elevated tempera-
ture without carrier flow and then cooled to room temperature.  At the
end of this cycle the carrier flow is resumed and the conditioning is
completed as in the case of the polar liquid phase.  Two precautions:
during conditioning, the column should not be connected to the detector
and one should not exceed the maximum safe temperature of the liquid
phase.

Since all liquid substrates bleed to one degree or another and columns
eventually degrade, we characterize all new columns with two column
performance indicators - the number of theoretical plates (N) and a tall-
ing factor (T).  p,pf--DDT is employed to check these parameters because
it is known to degrade on "poor" columns.  In this manner, we can deter-
mine if the performance of a new column is satisfactory and when the
column performance begins to fall off.  We consider a column good if the
number of theoretical plates per foot is on the order of 400-500 with
tailing factors of 1.0-1.3.  Calculation of these parameters is shown
in the Appendix.  Additionally, there should be no significant extraneous
peaks upon injection of a pure p,p'-DDT standard.

Other chromatographic conditions that can be adjusted are column
temperature and flow rates.  Although resolution of a mixture increases
with decreasing temperature, a temperature should be chosen that allows

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           the elution of all  components within a convenient time period.  The
           temperatures  given  are optimum for 42% chlorinated biphenyl, tempera-
           tures are  increased when specifically analyzing for the higher chlori-
           nated biphenyls, i.e., 54%, 60%, etc.  The flow rates are optimum for
           our instrument, column and detector system and, of course, should be
           adjusted if better  results can be achieved.

           Two gas chromatographic systems have been used for PCB analysis - F&M
           Model 402  and 5750.  We find that any system of instrument and column
           suitable for  chlorinated pesticides is satisfactory for PCB analysis.
           The bulk of analyses in our laboratories was carried out using the
           system outlined.  The use of the high temperature Ni63 electron capture
           cell is highly recommended.  The ability to operate at higher tempera-
           tures prevents maintenance problems due to contamination from high
           boiling components.  Glass columns should also be employed.


           Detection  and Measurement;

           Quantitative determinations employing the electron capture detector
           are non-stoichiometric measurements made by comparing peak heights
           or areas for known concentrations with those for unknown compositions.
           Except for sharp peaks, peak area measurements are usually more
           reproducible  than peak height measurements but are extremely time
 ,          consuming  unless.a recording integrator is employed.  However, peak
/          height measurements are as accurate as disc integration of triangu-
           lation and if the peak shape represents a gaussian curve, the height
           may be considered independent of the base.  Three variations of the
           peak height quantification procedure were employed.

           Case I    i   EC gas chromatogram of PCB unknown unchanged with respect
                        to standard PCB with no evidence of interferences.

           Case II      EC gas chromatogram of PCB unknown altered with respect
                        to standard PCB with no evidence of interferences.

           Case III     EC gas chromatogram of PCB unknown unchanged with respect
                        to standard PCB with evidence of interference.

           The amount of PCB's in Case I samples were determined by preparing a
           plot of the major peak height vs. concentration; for Case II, a plot
           of the total sum of all major peaks vs. concentration.  With Case III
           samples, a peak free from interference was used.  When dominant inter-
           ferences were present, one or more of the chemical clean up procedures
           was employed.

           In all cases, the response of the electron capture detector must be
           linear for quantitative analysis.  With our instrument any response
           less than  50 at an attenuation of 8 x 10 fell into the linear response
           range at a pluse rate of 150.  This corresponds to approximately 5 x
           10~9g of Aroclor 1242.

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Contamination;

In determining PCB's in water,  soil  and  sediment by electron capture
gas chromatography, laboratory  sources of contamination can be a major
problem.  The samples and extracts should never be allowed to come in
contact with materials other  than glass,  Teflon or metal.   Laboratory
glassware should be thoroughly  washed with hot, soapy water, rinsed with
distilled water, acetone, and then hexane.   All equipment  should also be
rinsed again with hexane just prior  to use and blanks should be frequently
carried through all steps of  the procedures to insure against the possi-
bility of contamination.           -
SENSITIVITY    Two parts per billion
               Absolute sensitivity -  0.5  x 10~9 grams
               Volume injected -  5  yl.
               Final volume of extract - 5 ml.
               Sample size - 250  ml.
                   Monsanto Company
                   R & D Laboratories
                   Applied Sciences Section
                   St. Louis, Missouri
 •U.S. GOVERNMENT PRINTING OFFICE: 1976 - 648-803/10309

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