PB87-111738
         Waste/Soil Treatability  Studies for Four Complex
         Industrial Wastes: Mefiodoiog ies and Results
         Volume 1. Literature Assessment, Waste/Soil
         Characterization, Loading  Rate  Selection
         Utah Water Research Lab.,  Logan
         Prepared for

         Robert S. Kerr Environmental  Research  Lab.
         Ada,  OK
        Oct 86
I
    U.S. DEPARTMENT OF COMMERCE
 National Technical Information Service

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                                   TECHNICAL REPORT DATA
                            /FteaterttttInilnicnont on int rt\em bt'.ort compliant/
                                                             RECIPIENT'S ACCESSION NO
1  REPORT NO

  EPA/6Q(l/6-S6/003a
i. TITLE AND SUBTITLE WASTE/SOIL  TREATABILITY. STUDIES FOR
FOUR COMPLEX INDUSTRIAL WASTES:  METHODOLOGIES AND
RESULTS.  Volume 1. Literature Assdssmei.t, Waste/Soil
Characterization, Loading Rare  Selection
              REPORT DATE
                  October 1786
             B PERFORMING ORGANIZATION CODE
 AUTMORISl
   R.'C.  Sins, J. L. Sims,  0.  L.  Sorensen, U. J.
     Doucette. and L.  L. Hastings  	
                                                             PERFORMING ORGANIZATION REPORT NO
I. PERFORMING ORGANIZATION NAMF AND ADDRESS
                                                           10 PROGRAM ELEMENT NO
   Utah State University
   Department of Livil and  Environmental Engineering
   Utah Water Research Laboratory
   Logan, Utah  84322
                      CBUD1A
             II CONTRACT/GRANT NO

                    CR-810979
12. SPONSORING AGENCY NAME AND ADDRESS
  Robert S.  Kerr Environmental  Research Lab. - Ada, OK
  U.S.  Environmental Protection Agency
  Post  Office Box 1198
  Ada.  Oklahoma  74820
             13 TVPE Of REPORT AND PERIOD COVERED
              	Final	
             14 SPONSORING AGENCY CODE
                   EPA/600/15
is. SUPPLEMENTARY NOTES

  Project Officer:  John  E.  Matthews, FTS:  743-2233.
16. ABSTRACT
        This two-volume report presents Information  pertaining to quantitative evalua-
   tion of the soil  treatment potential resulting  from waste-soil interaction studies
   for four specific wastes listed under Section 3001  of the Resource Conservation  and
   Recovery Act  (RCRA).   Volume 1 contains Information from literature assessment,
   waste-soil characterization, and treatablHty screening studies for each selected
   waste.  Volume 2  contains results from bench-scale  waste-soil Interaction studies;
   degradation,  transformation, and Immobilization data are presented for four
   specific wastes:   API separator sludge, slop oil  emulsion solids, pentachlorophenol
   wood preserving waste, and creosote wood preserving waste.  The scope of the  study
   involved assessment of the potential for treatment  of these hazardous wastes  using
   soil as the treatment medium.
                                KEY DWORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                                              b IDENTIFIERS/OPEN ENOEC TERMS  C  CC.ATI FKid.Group
                                                                                   GET
16. DISTRIBUTION STATEMENT


   RELEASE TO PUBLIC.
19 SECURITY CLASS iTtm Rtport)
     UNCLASSIFIED
31 NO Of PA

      17C
20 SECURITY CLASS iThil poftl

     UNCLASSIFIED	
                           22 PRICF
CPA Pm 2210.1 (••.. 4-77)   PUCVIOUS IDITIOM n OBSOLITI

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                                    NOTICE
     The Information in this document has been funded wholly or in part by the
United States Environmental  Protection  Agency under  Cooperative Agreement CR-
810979 to Utah State University.   It  has been subjected to  the Agency's  peer
and administrative review, and  it  has been  approved  for  publication as an EPA
document.  Mention of  trade  names or commercial products does not constitute
endorsement or recommendation for use.

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                                  FOREWORD
     EPA is charged by  Congress  to protect the Nation's  land,  air  and water
systems.  Under a mandate  of  national environmental laws focused on  air and
water quality,  solid  waste management and  the  control of  toxic  substances,
pesticides, noise and radiation,  the Agency strives to  formulate  and imple-
ment actions which lead to a  compatiole  balance between human activities and
the ability of natural systems to support and nurture life.

     The Robert  S.  Kerr  Environmental   Research  Laboratory is  the  Agency's
center of expertise for investigation of the soil  and subsurface environment.
Personnel at the Laboratory  are  responsible for management  of  research pro-
grams to:   (a)  determine  the fate,  transport and  transfer-nation  rates  of
pollutants in  the  soil,  the  unsaturated  and  the  saturated  zones   of  the
subsurface environment; (b)  define  the  processes to  be  used  in  character-
ising the  soil  and subsurface environment  as  a receptor of pollutants;  tc)
develop techniques for  predicting the effect of  pollutants  on  ground water,
soil, and  indigenous  organisms;  and (d) define and  demonstrate the applica-
bility and  limitations  of using  natural processes,  indigenous to the  soil
and subsurface environment, for the protection of this resource.

     'When applicable, enviromentally  acceptable  treatment of hazardous waste
In soil  systems is  a function of  operation  and  management practices  at a
given site.   Successful  operation and management  practices  are dependent on
identifying waste  loading  constraints  for that  particular  site.  There  Is
currently a  lack of  readily  available  Information relative  to  Impact  of
waste loading  rates  and  frequencies  on  transformation and  transport  of
hazardous organic  constituents In  waste-soil  matrices and  to  methodologies
for making  such determinations.   This two-volume  report  1s  Intended  to pro-
pose one set  of methodologies for determining waste loading constraints for
soil systems  and  to  provide an  assessment  of data  collected  using  the pro-
posed set of methodologies for two petroleum refining and two wood preserving
waste streams  applied to two  soil  types.   Volume  1  contains  results  from
literature assessment,  waste/soil characterization  and  treatabillty   screen-
Ing studies;  Volume  2 contains results  from torch-scale degradation, trans-
formation and immobilization studies.
                                       Clinton U. Hall
                                       Director ,
                                       Robert S. Kerr Environmental
                                         Research Laboratory

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                                 ABSTRACT


     This is  Volume  1  of  a  two-volume report  that  presents  information
pertaining  to  quantitative  evaluation  of the  soil  treatment  potential
resulting from waste-soil   interaction  studies for  four  wastes listed under
Section  3001  of the Resource  Conservation  and  Recovery Act  (RCRA).   This
voluise  contains  information  from  literature  assessment,  waste-soil
characterization, and  treatabMity  screening  studies  for  loading rate
selection for  each waste.  The four  wastes  included API separator sludge, slop
oil  emulsion  solids, pentachiorophenol wood  preserving  waste, and creosote
wood preserving  waste.    Chemical  analyses and  bioassays  were used  to
characterize Bastes, soiU.  and w&sie-soil  interactions.

     Objectives of the research reported in this  volume were to:

     (1)  Conduct  a  literature assessment  for each* waste to obtain specific
          information  pertaining  to   so:'   treatment   (degradation,
          transformation,  and  immobilization) of  hazardous constituents
          identified in each waste.

     (2)  Chemically characterize w«.stes for  specific constituents  of concern;
          and  characterize  two  experimental  soils  for  assessment  of specific
          parameters that influence  soil treatabllity.

     (3)  Conduct laboratory  screening  experiments using  a  battery  of
          bioassays to determine waste  loading rates (mg  waste/kg  soil) to be
          used  in  subsequent  longer  term  experiments  designed to  assess
          potential for treatment of each  selected  waste in soil.

     Specific  results and conclusions based on the  objectives include:

     (1)  Literature  assessment of specific  hazardous constituents in each
          waste  indicated a potential for  treatment in  soil systems.

     (2)  Chemical characterization of  the wastes by  GC/MS,  GC, and HPLC
          identified the PAH  class  of semivolatile constituents as  common to
          ?«ch waste.   In  addition,  the PCP  wood  preserving waste contained
          pentachlorophenol   and some dibenzo-p-dioxins  and dibenzofurans;
          however, no tetrachlorcdibenzodioxins were detected at the detection
          limit  of 10 ppb..

     (3)  A comparative  study of the  sensitivity of five micrcbial assays for
          selection of  initial  waste loading rates  indicated  that Microtox,
          soil dehydvogenase,  and  soil nitrification  assays correlated well
          and  were the most cmsitlve to the  presence of hazardous wastes, and
          would  result in  selecting lower   .-.oil  loading  rates.   Soil
          respiration and viable  soil microorganism  plate counts  were  highly

                                       iv

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          variable  and  less  sensitive, ana would  result  in selecting higher
          loading rates.

     {*)   Based on  screening  results  using  the  Microtox  assay,  initial  loading
          rates for petroleum refinery wastes  (6  to 12  cercent)  were  indicated
          to be  an order  to  magnitude higher  than for  wood  preserving wastes
          (0.1 to 1.3 percent).

     (5)   Loading rates selected for the clay  loam soil *ere generally higher
          than rates  selected for  the sandy  loam soil,  thus  indicating  a
          difference with  respect  to  the  effect of soil  type on waste-soil
          interactions.

     Based on  results  obtained  for  the  specific wastes and soils evaluatfid,
the use of chemical  inslyses  alone appears  to  be insufficient  to  characterize
waste-soil  interactions  and  the effects of waste-soil  mixtures on  microbial
activity.  Chemical  and  bioassay characterization of waste, soil, and waste-
soil mixtures provides valuable information concerning  treatment potential  for
industrial wastes.

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                                   CONTENTS
Notice
Foreword
/strart
Figures
Tables
Acknowledgments
     1.  Introduction  .................     1
               Objectives ................     2
               Evaluation Approach  .............     2
               Waste Characterization  ........   ....     4
               Soil Characterization   ............     4
               rfaste Loading Rate Determination    ........     4
               Waste Treatment in Soil    ...........     5
               Mathematical Model for  Soil -Waste Processes  .....     5
     2.  Conclusions   .................     7
     3.  Recommendations  ................     8
     4.  Literature 'Review   ...............     9
               Introduction  ......      . "  .......     9
               Wood Preserving Industry   ...........    10
               Petroleum Refining Industry   ..........    42
     5.  Results and Discussion  ..............    56
               Quality Assurance/Qual it)  Control   .   .......    56
               Waste Characterization     ...........    57
               Soil Characterization      ...........    97
               Waste Loading Rate Evaluation    .........   106
References
                                        vi

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                                   FIGURES

Number                                                                   Page
  1. Rates of degradation of PAH compounds in soil as a function of
     initial soil concentrations (Overcash and Pal 1979)  	    31
  2. Biodegradation of PCP	    38
  3. Scheme for the analysis of waste samples for organic constituents.    62
  4. Schematic of the Ames assay  	    65
  5. Ames assay results for creosote sludge base/neutral fraction  .   .    94
  6. Ames assay resu'ts for pentachlorophenol sludge base/neutral
     fraction	    95
  7. Ames assay results for pentachlorophenol sludge acid fraction .   .    96
  3. Ames assay results for API separator sludge base/neutral fraction.    98
  9. Ames assay results for slop oil emulsion solids base/neutral
     fraction	    99
 10. Soil moisture characteristic curve for Durant clay loam ....   105
 11. Soil moisture characteristic curve for Kidman sand loam ....   105
 12. Toxicity of water soluble fraction measured by the Microtox assay
     with incubation time for creosote waste mixed with Durant clay
     loam soil for loading rate determination, Trial II	119
 13. Toxicity of water soluble fraction measured by the Microtox assay
     with incubation time for creosote waste mixed with Durant cla>
     loam soil for loading rate determination. Trial #2	119
 14. Toxicity of water soluble fraction measured by the Microtox assay
     with incubation time for creosote waste mixed with Durant clay
     loam soil for loading rate determination, Trial 13	120
 15. Soil respiration results for creosote waste mixed with Durant clay
     loam soil	
                                       vii

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                             FIGURES (CONTINUED)
Number
 16. Toxicity of water soluble fraction measured by me Microtox assay
     tor creosote waste mixed with Kidman sand loam soil for 1 Odd ing rat?
     determination 	   122

 17. Toxicity of water soluble fraction measured by the Microtox assay
     with incubation time for PC? wood preserving waste mixed with Ourant
     clay loam soil for loading rate determination	122

 18. Soil respiration results for PCP waste mixed with Ourant clay
     clay loam soil	123

 19. Toxicity of water soluble fraction measured by the Microtox assay
     for PCP wood preserving waste mixed with Kidman sandy loam soil
     for loading rate determination  	   125

 20. Toxicity of water soluble fraction measured by the Microtox assay
     for API separator sludge waste mixed with Durant clay loam soil
     fcr loading rate determination	125

 21. Soil respiration results for API separator sludge mixed with
     Durant clay loam soil	126

 22. Toxicity of waste soluble fraction measured by the Microtox assay
     for API separator sludge waste mixed with Kidma'i sandy loam soil
     for loading rate determination	127

 23. Toxicity of water soluble fraction measured by the Microtox assay
     for slop o5' waste mixed with Durant clay loam soil for loading
     rate determination   .   .     	127

 24. Soil respiration results for slop oil emulsion solids mixed with
     Durant clay loam soil	129

 25. Toxicity of water soluble fraction measured by the Microtox assay
     for slop oil waste mixed with Kidman sandy loam soil for loading
     rate determination	130

 26. Microtox response to PCP waste application to Kidman soil  after
     24+2 !i incubation (LSD=least significant difference) 	   132

 27. Initial ammoniin and nitrite ion oxidatijn  in response to
     treatment of Kidman  soil with PCP waste  after ?4+2 h incubation
     {LSD=',east significant  difference)	"	133

 28. Othydrogenaic response  to PCP apolication  to Kidman soil after
     24+2 h incubation (LSD*°least significant difference) 	   134
                                      viii

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                             FIGURES (CONTINUED)
Number
 29. Respiration response to application of PCP waste to Kidrnar-
     soil :rter '4+p h incubation (LSD=l«»ast significant difference)   .    135

 30. Viable aerobic htterotrophic ba-:i^r»a and  fi-nvjal propagules  in
     Kidman soil treated with PCP waste after 24+2 h  incubation (LSD=
     least significant difference)    ............
 31. Microtox response to slop oil emulsion  solids waste  application
     to Kidman soil  (LSD= least significant Difference)  ......    138

 32. Initial ammonium and nitrite  ion oxidation  in response  to
     treatment application of sloo oil  emulsion  solids  to Kidman  soil
     (LS03least  significant difference)  .   .   .   .  . .......    139

 33. Dehydroqenase  response to slop  oil  emulsion solids waste  applica-
     tion to Kidman soil (LSDHea«t  significant  difference)   ....    140

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                                    TABLES


Number                                                                   Pa9e

  1. Principal constituents of high- temperature creosote (Wii.slow
     1973)   ....................    U

  2. Specific components in creosote oil (Lcrenz and Gjovik 197<>)  .  .    12

  3. Selected physical properties of PCP (Crosby 19811 ......    13

  4. Ring arrangement and relative  stability of PAH compounds (olumer
     1976)   ....................    15

  5. Properties of 16 priority pollutant PAH compounds ......    16

  6. Summary of physical properties for selected phenolic compounds
     {Versar Inc. 1979)  ................    19

  7. Health effects of chemical constituents of creosjte (U.S. EPA
     1984a)   ....................    20

  8. Human health effects of  exposure  to creosote  (U.S. EPA 1974)   .   .    23

  9. Polynuclear  azaarenes  in c^eosote-PCP wood preservation waste-
     water (Adairs and Glam  1984)   .............    24

  10. Kinetic  parameters  Describing  rates of  degradation of PAH and
     phenolic compounds  in  soil systems (Sims  and  Overcash 1983,
     ERT  1985b)  ............   .......    Zb

  11. Summary  of  soil  sorption data  for constituents of creosote  waste
      (ERT,  Inc.  1985b)    ...............    34
  12.  Summary of  bench and  pilot  scale PCP degradation studies  (FRT,
      Inc.  1985a)   .. .................     40

  13.  Refinery wastes known to be land treated and relative percentages
      of  each waste which are l»n.1 treatment (ERT 1<»84> ......     43

  14  Physical  composifion  of refinery wastes (Engineering Science
      1976)   ....................     44

  15.  Composition of 12 API refinory wastes (Overcash and Pal  1979) .   .     44

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                             TABLES  (CONTINUED)
Number
 16. Categories for Appendix VIII constituents in refinery wastes
     which are land treatment (ERT, Inc. 1984)
46
 17. Constituents of petroleum wastes (as approved by U.S. EPA)  ..
 18. Waste characterisation for aggregate of sixteen solid waste
     streams from a category IV petroleum refinery (Pal and Overcash
     1980)    ....................   50
 19. Relative resistance of hydrocarbons to biological oxidation
     (Fredericks 1966)    ................   51
 20. Results of degradation of petroleum wastes at a land-farm after
     25 months (Meyers and Huddleston 1979) ..........   53
 21. Waste characterization parameters   ...........   57
 22. Hazardous wastes selected for evaluation   .........   58
 23. GC/MS analysis conditions   ..............   M
 24. Physical characterization -f wastes ...........   68
 25. Characterization o* residues in hazardous  wastes    ......   69
 26. Characterization of metals  in petroleum refinery  wastes   ....   70
 27. Characterization of metals  in wood  preserving wastes      ....   71
 28. Characterization of metals  in petroleum refinery  wastes:  quality
     control data    ..................   'z
 29. Characterization of metals  in creosote wastes:   quality control
     data for  spiked creosote  waste  samples - high  and low level  ...   73
 30. Characterization of metals  in pentachlorophenol  wastes:   quality
     control data  for spiked  pentachlorophenol  waste  samples - high
     and  low  level      .................   '
 31. Characterization of metals  in creosote and pentachlorophenol
     wastes:   quality control  uata for  EPA quality  control samples   .  .   75
 32. Total  organic carbon  (TOC)  content of hazardous  waste samples   .  .   76
 33. Characterization of oil  and grease in hazardous  waste samples   .  .   76
                                        xi

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                              TABLE (CONTINUED)

Number                                                                   Page
 34. Organic compounds tentatively identified in API  separator sludge
     waste (base/neutral fraction) by GC/MS    .........    77
 35. Organic compounds tentatively identified in slop oil  emulsion
     waste (base/neutral fraction) by GC/MS    .........    80
 36. Organic compounds tentatively identified in creosote waste
     (base/neutral fraction) by GC/MS    ...........    M
 37. Organic compounds tentatively identified in pentachlorophenol  waste
     (base/neutral fraction) by GC/MS    ...........   85
 38. Organic compounds tentatively identified in creosote waste and
     pentachlorophenol waste (acid fraction) by GC/MS   ......   88
 39. Organic compounds tentatively identified in API separator sludge
     and slop oil waste samples (volatile fraction) by GC/MS  ....   90
 40. Organic compounds tentatively identified in PCP and creosote waste
     samples (volatile fraction) by GC/MS   ..........   91
 41. Concentration of individual PAH compounds in wastes determined by
     HPLC  .....................   92
 42. Chlorinated dibenzo-p-dioxins and dibenzofurans in pentachloro-
     phenol wiste by GC/MS   ...............   93
 43. Toxicity of water soluble fraction measured by the Microtox assay
     for hazardous waste samples   .............   93
 44. Soil physical and chemical properties evaluated for soil
     characterization  .................  100
 45. Measurement methods and data quality objectives for soil analyses  .  lul
 46. Characterization of Durant clay loam soil collected from
     hazardous waste land treatment facility, U.S. EPA, Ada, Oklahoma   .  103
 47. Characterization of Kidman sandy loam soil collected from USU
     Agricultural Experiment rarm at Kaysville, Utah .......  104
 48. Soil loading rates for wastes based on Microtox and soil
     respiration results  ................  131

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                              ACKNOWLEDGMENTS
     Technical  contributions  to  this research  report  concerning  chemical
analyses were made by Dr. William  J.  Ooucette  (Organic  Chemst)  and  Ms.  Joan
E. McLean (Inorganic  Soil  Chemist)  of the Toxic and Hazardous Waste Management
Group at the Utah  Water  Research  Laboratory.
                                      xiii

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                                 SECTION 1

                                INTRODUCTION
     Land treatment is the  hazardous waste management technology pertaining  to
application/incorporation of waste Into the upper layers of the  soil for the
purpose of degrading, transforming, and/or immobilizing hazardous constituents
contained in  the  applied waste (40 CFR Part 264).   Land  application systems
have been utilized for a variety of industrial wastes; however,  application  o"
hazardous industrial  waste  utilizing a  controlled  engineering design ana
management approach has not been widely practices.   This is due, in part,  to
the lack of a comprehensive technical information base concerning the behavior
of  hazardous  constituents  as  specifically related to  current  regulatory
requirements  (40 CFR  Part  264) concerning  treatability  in soil,  i.e.,
degradation,  transformation, and  immobilization of  such constituents.   Soil
treatment systems that are designed and managed  based on a knowledge of  soil-
waste  interactions  may represent a  significant  technology for  simultaneous
treatment  and  ultimate   jisposal  of selected   hazardous  wastes  in   an
environmentally acceptable  manner.

     •n  this  research  project,  representative hazardous  wastes  from two
industrial categories, wood preserving  and  petroleum refining,  were evaluated
for potential  for treatment in  soil systems.  A literature assessment for each
waste category was  conducted  as an aid in  the  prediction of  land treatment
potential.  The  literature assessment  also  was  used as a guide  to design  an
experimental  approach to obtain  specific treatability  information pertaining
to  degradation, transformation, and  Immobilization  of  hazardous  constituents
In soil.

     Results  of this  research  project  are contained  in  two volumes.  The two
volumes contain information concerning  an approach  (methodology) and results
for  evaluating  the  potential  for  treatment  of  hazardous  waste  In soil
systems.Volume 1  contains information concerning literature review, results  of
laboratory waste  and  soil  characterization, bioassay results  for soil
microbial  activity  in the  oresence  of hazardous wastes,  and  experimental
approaches and results for  selection of waste loading  rates  using a bioassay
battery.   Volume  2  contains  Information  concerning results  of  treatabllity
studies designed  to  generate  degradation,  transformation,  and  Immobilization
information  for  API   separator  sludge,  slop   oil  emulsion  soHds,
pentachlorophenol  woof preserving  waste, and creosote wood preserving waste.

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OBJECTIVES

     Specific objectives of this research project were to:

     (1)  Conduct  a literature assessment  for  each  candidate  hazardous waste,
          API separator sludge,  slop oil  emulsion  solids,  creosote  wood
          preserving  waste,  and per.tachlorophenol  (PCP) wood  preserving waste
          to obtain specific  land treatability information,  i.e.,  degradation,
          transformation, and immobilization,  for hazardous  constituents
          identified  in each  waste.

     (2)  Characte, ize candidate wastes for  identification  of  specific
          constituents of concern;  and  characterize experimental soils  for
          assessment  of specific parameters that  influence land  treatability
          potential.

     (3)  Conduct  treatability screening experiments  using  a battery of
          microbial assays to determine waste loading rates  (mg waste/kg soil)
          to  be used in subsequent experiments to assess potential  for
          treatment.

     (4)  Develop degradation, transformation,  and  immobilization information
          as a function of loading for each candidate hazardous  waste in the
          soil types.

     (5)  Develop  methodologies for  the  measurement  of  "volatiliza*ion-
          corrected"   degradation  rates  and  for  measurement of n»: tition
          coefficients; use  methodologies  developed  to qe-.criLe  degradation
          kinetics/partition  coefficients  for   a  subset  of  soil/waste
          combinations and for constituents common to all  candidate wastes.

     Information generated relative to the first three objectives is  presented
in this volume (Volume 1)  of  the project report.


EVALUATION APPROACH

     Standards are promulgated  in 40 CFR  Part  264.272  for  demonstrating  land
treatment  of hazardous wastes.   The  standards  require  demonstration of
degradation, transformation,  and/or immobilization of a candidate waste  in the
treatment soil.  Demonstration of degradation  of waste and  waste constituents
is  based  on the  loss  of   parent   compounds  within the  soil/waste matrix.
Complete  degradation  is the  term  used  to  describe the process  whereby waste
constituents  are mir.sralized to inorganic end  products, generally  including
carbon  dioxide,  water, and  inorganic  species of  nitrogen,  phosphorus,  and
sulfur.   The rate  of degradation  may be established  by measuring the loss of
parent  compound from  the  soil/waste  matrix with  time.  Transformation  refers
to  the  partial degradation  in the  soil  converting  a  substance into an
innocuous or  harmless  form,  or problem wastes into environmentally safe forms
(Huddleston  et  al. 1986).  Ward et  al. (1986)  also  discussed the difference
between  rates of  mineralization  (for  complete  degradation)  and  rates of
biotransformation.    Therefore  transformation refers  to the  formation of

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intermediates  during  the  process  of degradation  or the  formation of
intermediates as  refractory compounds  in  the soil matrix.    Immobilization
refers to  the extent of  retardation  of the  downward  transport, or  leaching
potential,   and  upward transport,  or  volatilization potential,  of  waste
constituents.  The mobility potential for waste constituents  to  transport  from
the waste to  later, air,  and soil  phases is affected by the  relative affinity
of the waste  constituents  for  each  phase,  and can  be  characterized  in  column
and batch  reactors.   Therefore,  demonstration of soil treatment requires an
evaluation of degradation,  transformation,  and  immobUizatio".  processes,  and
the quantification of the processes for obtaining an  integrated assessment of
the design  -tnd management requirements for  successful  assimilation of a waste
in a soil system.

     The  requirement  for  demonstrating treatment,  I.e.,  degradation,
transformation,  and/or  immobilization,  can be  addressed  using  several
approaches.   Information  can be obtained  from several sources,  including
literature  data,  field  tests, laboratory  analyses  and  studies,  theoretical
parameter   estimation methods, or, in  the case of  existing   land  treatment
units, operating  data.    Information presented  in  Literature  Review in  this
report addresses  information obtained from  literature  data  and existing  land
treatment  units.   Specific  information obtained  from  literature  sources
included  quantitative  degradation,   transformation,  and immobilization
information for waste-specific hazardous constituents   in soil  systems.    Four
hazardous  wastes   are considered,  Including  API separator  sludge,  slop oil
emulsion solids, creosote sludge and pentachlorophenol sludge.   However, the
U.S. EPA considers the use  of  jnly literature information as insufficient to
support demonstration of  land treatment of  hazardous  wastes  at the p?  ont
time.    A  laboratory experimental  approach used during  this  projec   for
obtaining  additional  information  concerning  treat ability data  for  the  four
hazardous wastes selected for study is presented.  Results using the approach
are also presented.

     The regulations  also  require  that the effect of design   and management
practices  on  soil  treatment be evaluated.   Design and  management  practices
specifically  identified  in the regulations  include  waste  application rate, or
loading rate, and  frequency of  waste application.
     The experimental  approach used  in this study was  to select  waste
rates and  to characterize treatment,  including  degradation,  transformation,
and  immobilization  of four hazardous  wastes in  two  soil  types.   For  each
hazardous waste and each soil  type, treatment was evaluated as  a function of
waste  'oaainq  rate,  soil  moisture,  and  time.   A  combination of chemical
analyses and bioassays  (including  general  toxicity and tnutagenicity  assays)
was  used   to  characterize  treatment  endpoints,   i.e.,  degradation,
transformation,  and immobilization.

     The experimental  approach described above was used to test the  hypothesis
that treatment would  be  achieved  for  each  hazardous  waste In  both selected
soil types.   This  approach was also used to evaluate the  effect of selected
design and management  factors  on treatment.  Therefore, the scope of the  study
was  to  address  the demonstration of  treatment  of hazardous  wast* 'jsi-ij the

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soil  as  the treatment  medium as  expressed  in  the current  federal  HWLT
regulations promulgated July 26,  198?.


WASTE CHARACTERIZATION

     Treatment  of  a  hazardous waste refers  specifically  to tre
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     A  comparative study of  the  sensitivity  of  Microtox,  respiration,
dehydrogenase,  initial  nitrification  activity,  and  soil  plate counts  to
pentachlorophenol (PCP) and slop oil wastes i"  the  Kidman sand.y  loam soil was
performed  to  evaluate the  response of commonly  used  bioasbay:  to  identical
soil/waste mixtures.
WASTE TREATMENT IN SOIL

     The  degradation  potential  of hazardous constituents in waste!s) applied
to  soil  is  critical  since degradation usually represents the primary removal
mechanism for organic constituents in waste(s).  The basis for biodegradation
coefficient  measurements  is the  determination of  specific  constituent soil
concentrations  as  a function of  time.    The experimental  approach to  the
determination of biodegradation was to characterize biodegradation as a  first
order kinetic rate  process.   The first order  reaction rate constant was then
used to  calculate  half-lives for each  parameter.   The half-lives calculated
provided quantitative  information  for  evaluating the  extent and  rate  of
treatment,  and  for  comparing  treatment effectiveness  for  each  waste/soil
combination as a  function  of  design and  management  factors.    Results  and
discussion  concerning degradation of each  hazardous  waste  are  discussed  in
Waste Degradation Evaluation. (Volume 2)

     A  waste cannot  be  applied to  land  unless  it  is  rendered  less  or
nonhazardous as  a  result of treatment.   Therefore, conversion  of hazardous
constituents to less  toxic intermediates within the soil treatment medium was
evaluated.   Information concerning the  toxicity  reduction  in each waste/soil
combination was evaluated using an acute toxicity assay (Microtox  assay),  and
a  mutagenicity assay (Ames  Salmonella  typhimurimn/manmalian'microsome
mutagenicity assay).   Results  and  discussion of the  transformation  of each
hazardous waste are discussed in Waste Transformation Evaluation. (Volume 2)

     Evaluation of treatment also  involved  an investigation  of  the extent  of
migration of each  hazardous waste.   A loading  rate based on biodegradation
potential was  selected  for  each soil/waste  combination.    The  leaching
potential was subsequently characterized for these loading rates  in laboratory
column studies.   Partition coefficients  among waste  (oil), water  and air for a
subset of constituents  were  also determined for evaluation of immobilization
input parameters  required for  the regulatory  investigative  treatment  zone
(RITZ)  model developed by the  U.S.  EPA Robert S. Kerr Environmental Research
Laboratory (RSKERL).  Results obtained for evaluation of the  immobilization of
each hazardous  waste are described in  Waste  Immobilization Evaluation. (Volume
2)


MATHEMATICAL MODEL  FOR S&IL-WAI/.E PROCESSES

     A mathematical description of the  soil/waste system  provides  a unifying
framework for the  evaluation of laboratory  screening  and  field  data that  is
useful  for the determination of  sell  treatment potential  for  a  waste.   While
current  models  cannot be relied  upon for  long-term predictions of absolute
contaminant  concentration:   due to  the  lack of  an  understanding  of  the

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biological, physical, and  chemical  complexity  of the waste/soil  environment,
they represent powerful tools  for  ranking  design,  operation,  and maintenance
alternatives as well  as  for the design of monitoring  programs.

     Short  (1986)  developed a  model  (RITZ) for  evaluating  volatilization-
corrected degradation  and partitioning  of organic  constituents  in  soil
systems.   The  model  is  generally based on  the approach  used by Jury  et  al.
(1983) for modeling pesticide fate in soil.  The RITZ model has been expanded
at Utah State University to incorporate features  that increase its utility for
the planning and evaluation of  treatment for  land/waste systems.

     A  mathematical  description of  soil/waste  systems  provides a  framework
for:

     (1)  Evaluation  of  literature and/or experiment data;

     (2)  Evaluation of  the  effects of  site characteristics  on  treatment
performance (soil type,  soil  horizons, soil  permeability);

     (3)  Determination  of  the effects  of  loading rate,  loading  frequency,
soil moisture,  and  amendments  to increase  degradation  on  soil  treatment
performance;

     (4)  Evaluation of  the  effects of  environmental  parameters  (season,
precipitation) on soil  treatment performance; and

     (5)  Comparison of  the  effectiveness of  treatment  using  different
practices in order to maximize  soil treatment.

     The extended version of the model  is  programmed for the comuuter in  such
a way that  additional enhancements  (such  as unsteady flow and  lime  variable
decay transport/partition coefficients) may be  incorporated into  the  model  in
the *uture.  A summary of  the model is provided  in Appendix B of Volume 2  of
this report.

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                                  SECTION  2

                                 CONCLUSIONS
     Specific conclusions  based  on  project  objectives and  research  results
presented in Volume 1 include:

     (1)  A  literature  assessment  for  each candidate  waste  type  and  for
          specific hazardous constituents  that were experimentally identified
          in each waste indicated a potential for achieving treatment  in soil
          systems.

     (2)  Chemical characterization of  all  four wastes by GC/MS, GC,  and HPLC
          ideitified the  polycyclic aromatic  hydrocarbon  (PAH)  class  of
          semivoiatile constituents as  common to  each waste.  The acid extract*
          fraction of the PCP wood preserving waste contained some dibenzo-p-
          dioxins and  dibenzofurans  in addition  to pentachlorophenol; but no
          tetrachlorodibenzodioxins were detected.

     (3)  A  comparative  study of the  sensitivity  of five  microbial  assays
          including  Microtox,  soil respiration, soil  dehydrogenase,  soil
          nitrification,  and viable   soil  microorganism plate  counts  for
          selection of initial  loading  rates  indicated that the Micro to/., soil
          dehydrogenase, and soil  nitrification  assays were the most sensitive
          to  the  presence  of  hazardous wastes; use  of  these  assays  would
          result  in  selecting  initial   loading  rates  at  lower  levels.   Soil
          respiration (carbon dioxicie evolution)  and viable soil microorganism
          plate counts were less  sensitive  to hazardous waste application; use
          of thesa assays would result  in the selecting higher initial loading
          rates.

     (4)  Soil loading rate studies indicated that the Microtox  assay was more
          sensitive to changes in wa^re loading  rate than  CO2 evolution assay.
          Bu:»ed on  results of nicrobial assays, loading rates  selected  for
          evaluation  in  long-term treatability studies  were  in order of
          magnitude  higher for  petroleum  refinery  wastes than for  wood
          preserving wastes.

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                                 SECTION 3

                              RECOMMENDATIONS


     Based on the results of this research  investigation,  the  following  sets
of recommendatIons are made  pertaining to loading rates and soil treatment for
hazardous wastes:

     (1)  A combination of  data  sources should  be  used  to evaluate  loading
          rates and soil  treatment  potential  for  hazardous  wastes; these  data
          sources  should  include  literature  sources, laboratory analyses of
          the candidate  waste  for  identification and  quantification of
          hazardous constituents, characterization  of  the  proposed  soil  for
          treatment,   and  laboratory studies for  evaluation  of  treatment
          potential.

     (2)  Specific Analyses  for  parameters  that  are  used  in the proposed  U.S.
          EPA treatment  zone  model   for  the  assessment of soil  treatment
          potential  are also recommended  so  that a  common data base can be
          established  for use  in future  assessments of the   potential   for
          treatability of  specific hazardous wastes in soil.

     (3)  A battery of microbial  assays  is recommended for use  in  sel Acting
          initial  waste loading  rates;  the  battery should  include assays  that
          assess specific metabolic  activities  as well  as  gross microorganism
          viability;   a final set of waste  loading  rates   should be  selected
          only  after  identifying  the  detoxification  and  immobilization
          potentials  of soil/waste mixtures in long-term treatability studies.

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                                 SECTION 4

                             LITERATURE REVIEW
INTRODUCTION

     Treatment  in soil  systems  may  represent a  significant engineering method
for control/treatment and ultimate disposal  of  selected hazardous constituents
in  applied waste.   Land  application  for the  assimilation  and  treatment  of
hazardous  constituents  is a  potentially significant cost-effective,
environmentally safe, low energy  technology  that has been  successfully
utilized for  a  multitude of  domestic and  industrial wastes.  Soil systems for
treatment of a a  variety  of industrial  wastes,  including  food processing,
organic chemical  manufacturing, coke industries, textiles, and pulp and paper
have been utilized  for  many years  (Overcash  and Pal 1979).   However, Phung et
al.  (1978) reported that routine  application of  Industrial  hazardous wastes
onto the  soil  surface  and  incorporation into the  soil  for  treatment is not
widely  practiced,  except  for the  oil  refining  industry.   There  are  few
definitive data  in  the literature  quantifying  treatment rates  in full-scale
soil treatment  systems  (Huddleston et al. 1986).

     Land  treatment is  defined  in RCRA  as the  hazardous  waste management
technology pertaining  to application and/or incorporation of waste  into the
upper  layers  of  the  soil  in order  to  degrade,  transform  or immobilize
hazardous constituents  contained  in  the applied  waste  (40 CFR  Part  264,
Subpart Ml.   Land  treatment  also  has been  defined as  the controlled
application of  hazardous wastes onto or into the aerobic  surface  soil  horlzr.i,
accompanied by continued  monitoring  and management,  in order   to  alter the
physical,  chemical,  and  biological  state of  the  waste  via biological
degradation and  chemical  reactions  in  the  soil  so as  to render  such waste
nonhazardous  (Brown et  al. 1983).

     The  current regulatory requirement for  demonstrating  treatment,  i.e.,
degradation,  transformation,  and/or  immobilization  of  hazardous  waste
constituents  in  soil   systems, can  be  addressed  ising several approaches.
Information concerning  each treatment  component can be obtain*! from several
sources including literature data, field tests,  laboratory studies,  laboratory
analyse*  iheo, etical   parameter  estimation methods,  or,  in  the  case  of
existing units, operating  data  (40 CFR  Part  264.272).   It  is suggested that a
combination of data sources should  be  utilized,  e.g., literature  data,
laboratory analyses,   laboratory  studies  and  field  verification  tests,  to
strengthen confirmation of  hazardous constituent treatment demonstration.  The
availability and completeness of existing literature data will   influence the
need for further  collection of  performance  data.   The U.S.  EPA considers the

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use  of  only  literature  data  as  insufficient  to  support a demonstration of
treatment at the present  timt.

     In  this  project,  representative hazardous  wastes  from  two  industrial
categories, wood preserving and petroleum refining,  were  used  to  evaluate  the
impact  of  waste  loading  on  soil  assimilative  capacity  ir,  'and  treatment
systems.   A comprehensive  assessment  of  literature available  for each  waste
type was conducted  as an  aid in making these evaluations.


WOOD PRESERVING INDUSTRY

Introduction

     The  wood  preserving  industry,  as  defined  in  Standard  Industrial
Classification (SIC)  2491,  is comprised of establishments primarily engaged in
treating  wood,  which  are  sawed or  planed  in  other  establishments, with
creosote or other preservatives to prevent decay  and  to  protect  against fire
and  insects.   This industry also  includes  the  cutting,  treating,  and  selling
of crossties, poles,  posts, and piling.   Wood preservation  increases the life
of wood  products  by decades,  which reduces  the demand  for wood  production.
Thus wood preserving  allows time for renewal of timber resources.

Process Description

     Wood  preservation is  a two-stage  process:    1) conditioning the  wood to
reduce  its natural  moisture content  and to  increase permeability  and  2)
treating the wood with the  preservative  (Sikora- 1983).   Several  methods have
been used for conditioning  the  wood, including: seasoning in open  yards;  steam
conditioning; vapor drying; kiln  dry'ng; controlled air  seasoning  and  tunnel
drying.    After the  wood is conditioned,  it is  immersed  in preservative
chemicals,  either  at ambient  or  elevated  temperatures,  and  either  with or
without the use of  pressure.

     The  use  of wood preservatives has  been restricted by the  U.S.  EPA to
certified applicators.

Characteristics of  Wood Preservatives

     Desirable properties of wood  preservatives are:  1) inhibitory effects on
wood-destroying organisms,  2) permanence, i.e., preservation effects should be
sustained  for  lonq  periods  of  time,  and  3)  freedom from objectionable
qualities  (i.e., health  hazards,  fire  hazards,  corrosiveness,  and  reduced
strength of the treated wood).

     Two  major types  of wood preservatives  include  creosote  and
pentachlorophenol  (PCP).     Creosote is  used  primarily  for  railroad ties,
utility poles,  and pilings, and  PCP  for utility  poles,  cross arm  posts,  and
lumber (Sikora 1983).

     Creosote is made  hy high-temperature carbonization of bituminous  coal.
The  high  temperature results  in  a complex  mixture  of organic  compounds
                                        10

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consisting mainly of aromatic hydrocarbons,  tar  acids  (phenolic derivative of
the  aromatic  compounds),  and  tar  bases  (heterocyclic compounds containing
nitrogen  plus  some  neutral  oxygenated compounds).  Principal constituents of
hign  temperature  creosote  wood   preservatives  as reported by  various
investigators  are  shown  in  Tables   1 and 2.   The irajor polynuclear  aromatic
hydrocarbons (PAHs)  present  are two, three,  and  four  ring compounds and their
methyl derivatives.  Creosote may also contain  small   amounts of five and six
ring  PAH  compounds, some of which are suspected or recognized carcinogens as
pure  compounds.  Concentrations of inorganic constituents are  typically low in
creosote.  Creosote jlone  or in combination  with coal   tar or petroleum is the
major preservative  used in  the wood pressure treating industry (Merrill  and
Wade  1985).

            TABLE 1.  PRINCIPAL CONSTITUENTS  OF  HIGH-TEMPERATURE
                          CREOSOTE  (WINSLOW 1973)
     Compound                                    *  by Weight
Naphthalene
Phenanthrene
Acenaphthene
Fluor anthene
Fluorene
Methyl naphthal enes
Pyrene
Carbazole
Anthracene
Oiphenylene oxide
9, 10-Di hydro anthracene
7 - 28
9 - 14
2-5
2-5
2-4
1 - 4
2-3
1.8-2.7
1.2-1.8
0.5-1.0
0.1-0.3
     Commerc -'al  PCP contains  85-90  percent  PCP,  3-8  percent  of
tetrachlorophenols,  2-6 percent other chlorinated phenols,  and  the remainder
consists of  other  chlorinated  compounds  and inert  materials  (Crosby  1981).
Prcperties of PCP  are  shown in Table 3.  When used as a wood preservative,  PCP
is usually mixed  with petroleum  products or added  to  creosote.   PCP  1s of
environmental concern  due  to  its toxicity to  humans  and to aquatic life.  The
level of  impurities  in PCP  may  also oe important, for most  technical  PCP
samples contain  the higher-chlorinated dibenzodioxins and dibenzoflurans.  The
dioxin  usually  present  in the highest  concentration  is the  comparatively
nontoxic  oc tach1orodibenzo-p-dioxin  (OCDD).   The  highly  toxic
tetrachlorodibenzo-p-dioxin  (TCDD)   1s  not  present,  but  the  toxic
hexachlorodibenzo-p-dioxin  (HCOD)  and heptachlorodibenzo-p-dioxin  (HpCDD)
isomers are  usually  present  (Crosby  1981).   Other  impurities may include
predioxins,  isopredioxins,  poly-chlorodiphenyl ethers, cyclohexadienones,  and
chlorinated  hydrocarbons (Crosby 1981).
                                     11

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  TABLE   2.   SPECIFIC COUP'
IN CREOSOTE  OIL  (LORENZ  AND  GJOVIK  1972)
Component
Naphthalene
2-Methylnaphthalene
1 -Methyl naphthalene
Biphenyl
Acenaphthene
Bircethylnapthdlenes
Dibensofuron
Carbazole
Fluorene
Hethylfluorenes
Phenanthrene
Anthracene
9,10-Dihydroanthracene
Methylphenanthrenes
Nethylanthracenes
Fluoranthene
Pyrene
Benzofluorenes
Chrysene
Benz(a)anthracene
Benz(j)fluoranthene
Benz(k)fluoranthene
Benz(a)pyrene
Benzjejpyrene
Perylene
Benzo(b)chrysene
rorr.icla
ClQHs
CllH'.O
CllHlO
C12H10
Cl2H10
Cl2«12
Ci2H80
Ci2H9N
Cl3"lO
Cl4"l2
CUHIO
Cl4H10
Ci4Hi2
C15H12
Cl5«12
C16H10
CI&HIO
C17H12
Cl8"l2
Cl8"l2
ClflHl2
C2QHl2
C20H12
C20ri12
C3QH12
C22H14
Molecular
Weight
128.
142.
142.
154.
154.
156.
168.
167.
166.
180.
170.
178.
180.
192.
192.
202.
202.
216.
228.
228.
252.
252.
252.
252.
252.
278.
Boiling Fraction in
Point, °C Creosote Oil
(wt. pet)
218
241
245
255
279
267-269
287
355
297
318
340
340
312
354-355
360
382
393
413
448
438
480
480
496
493
460
500
3.0
1.2
0.9
0.8
9.0
2.0
5.0
2.0
10.0
3.0
21.0
2.0
-
3.0
4.0
10.0
8.5
2.0
3.0
•
-
-
-
-
-
-
TOTAL
                                                                      W7T
*Values shown are "aoprox. pet. * 0.7»."  Analysis was by gas chromatography
with flame  lonization detection using a reference mixture of compounds  JS a
quantitative and qualitative standard for calibrating the gas chromatograph."
The or  'in of the creosote sample used was not described.
                                       12

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         TABLE  3.  SELECTED  PHYSICAL  PROPERTIES OF PCP  (CRPiBY  1981!
       Property
Melting point, °C
Boiling point, °C
Vapor Pressure, Torr (mm hg)
0°C
20°C
50°C
100°C
200°C
300°C
Solubility in water (g/L)
0°C
20°C
30°C
190.2°
300.6°

1.7 x 10'5
1.7 x 10-*
3.1 x 10-3
0.14
25.6
758.4

0.005
0.014
0.020
     70°C                                         0.085

Solubility in organic solvents (g/L, 25°)
     Methanol                                     180
     Acetone                                      50
     Benzene                                      15

pKA (25°)                                         4-7°

Partition coefficient (Kp), 250
     Octanol-water                                1760
     Hexane-water                                 1.03 x 105
                                          13

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Characteristics of  Mood  Preserving Wastps

     The principal  source of  wastewaters  in the wood preserving  industry  is
from the conditioning process,  while  some wastewaters are produced  when  the
treated wood product  is  removed  and  allowed  to drain.  The steam condensate  is
also a source of wastewater.  The characteristics of the resulting wastewater
are  highly variable  and  depend  on  the  conditioning method,  type  of
preservative^) used, type  of solvent  used with the  preservative  (coal  tar,
oil, etc.), and the extent of  dilution with  nonprocess water (boiler blowdown,
rainfall,   steam  c^ndensate,  etc.).    Wastewaters  trom  creosote  and
pentachlorophenol treatment  often have  high phenolic, chemical  oxygen  demand
(COD),  and  oil  concentrations  and  generally appear  turbid  as a  result  of
emulsified oils.   Their  pH  is in the  acidic range (4.1-6.0).   Compounds  that
are  extracted  from wood  (mainly simple  sugars)  during  wood  conditioning
contribute to the high COD values.  Wastes also result from spills, leaks  and
sludges from  wastewater  treatment processes.   The  amount of  creosote  waste
s'jdge  and  PCP  waste  sludge  produced  annually by the entire  industry is  only
239  to  930  and 600 metric  tons, respectively.  However, the sludge  is  often
allowed to  accumulate for months or  even years before removal  and  disposal
(Sikora 1983).

     Both a  creosote  sludge and a combined  PCP-creosote  sludge were used  in
this experimental investigation.

Treatment of Creosote Wastes in  Soil Systems

     The principal  classes of organic  constituents present in  creosote wastes
are  PAHs and phenolics.

     PAHs are compounds which consist of two or  mor?  fused benzene rings, with
each ring sharing two or more carbon atoms.   The relative stability of PAHs is
related to  the ring  arrangement,  as  described  in  Table  4.   Graphical
representations of  the types of ring arrangements described in  Table 4 may be
seen  in Table  5.   Solubilities of  PAHs  decrease as molecular  weight,  chain
length  and  molecular volume  increase.    Properties   of  the 16  PAH compounds
designated as U.S.  EPA priority pollutants are given  in Table 5.

     Phenol ics  are low-to-moderately  volatile  compounds  which may  have
antiseptic  properties towards environmental  organisms.   PhenolIcs are highly
soluble  in  water but have  low  vapor  pressures and  low sorptive  tendencies.
General physical properties of  several  phenolic compounds are  shown in Table
6.

Toxicological  Significance of Creosote Waste;--

     The  use of creosote  has  been restricted by  the  U.S.  EPA  to  protect
applicators of  the preservative and  users  of the treated wood from  unnecessary
exposure.   Creosote  contains many constituents that  are  reported to  be
mutatjenic,  carcinogenic,  teratogenic,  fetotoxic,  and/or toxic.   Reported
health  effects of  these constituents  are  shown in Table  7.   Descriptions of
documented cases of human health effects of creosote  are shown  in  Table 8.
                                       14

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     TABLE 4.   RING ARRANGEMENT AND RELATIVE STABILITY OF PAH COMPOUNDS
                               (BLUMER 19/6)
Ring Arrangement
Linear
Cluster
Description
all rings in line
at least one ring
surrounded on three
Stability*
least
intermediate
Examples
anthracene
tetracene
pyrene
benzopyrene
                    sides

Angular             rings  in  steps           most                phenanthrene
                                                                chrysene


*Chemica1 stability in  the environment from least to most stable.
     Bos et  al.  (1383,  1J84)  determined that mutagenicity  of creosote was
probably due  to  the  presence  of mutagenic Aromatic  hydrocarbon!,  including
benzo(a)pyrane and benz(a)anthracene.  The authors suggested,  that  since  these
compounds  are  probably not  essential  for  wood-preserving properties of
creosote,  a  more  selective  composition  of the  product   by  control  of
distillation temperature  should be considered.

     Polynuclear  azaarenes,  which  are  polycycllc  aromatic  bases  such as
quinolines, isoquinolines, benzoquinolines,  and alkayl-  and benzo-substituted
azanaphthalenes, have been detected  in creosote-pentachloropheno1  wastewaters
(Table  9).   These compounds have  been reported  to be toxic, teratogenic,
mutagenic,  and/or carcinogenic  (Adams  and Gian.  1984).

     Additional  information concerning health effects of  constituents found in
creosote may be found in  the  U.S. EPA health effects assessment documents  for
PAHs (U.S.  EPA 1984e)  benzo(a)pyrene (U.S. EPA  1984b), and coal tars (U.S.  EPA
1984c).

Degradation and Immobilization of PAH  and Phenolic Compounds—

     Microbial metabolism  of PAHs has  been studied primarily  using  pure
cultures and single-compound, laboratory-scale  systems.  There are  few reports
of PAH  biodegradation  under  field  conditions  and even  fewer  concerning  soil
systems  specifically.

     A wide range of soil organisms, including bacteria, funqi, cyanobacteria
(blue-green algae),  and   eukaryotic  algae,  have  been shown to  nave the
enzymatic  capacity to oxidize  PAHs.    Prokaryotic  organisms, bacteria,  and
cyanobacteria, use  different biodegradation patnways than the eukaryotes,
fungi, and  alqae, but  both involve molecular oxygen.
                                       15

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                              TABLE 5.  PROPERTIES OF 16 PRIORITY POLLUTANT PAH COMPOUNDS
66 U980
JL^J
^-X*%. 178 73
.jjJL .^J
^•N, 178 1.290
Melting
Point
°C
80
92
96
116
216
101
Boll ing
Point*
°C
718
?65
?79
293
340
340
Vapor
pressure Length of
9 20°C , Molecule
torr Log K(,w* A° Kot
4.92x10'? 3.37 8.0 1.300*
2.9>10"z 4.07
Z.OxlO'2 4.33
1.3xlO'z 4.18
1.96»10-* 4.4S 10.5 2,600*
6.80x10'* 4.46 9,5 23.00C*

-------
                                                     TABLE 5.   CONTINUED
Vapor
Aqueous Nell ing Boiling pressu. e Length of
Molecular Solubility* Point Point P 20°C f Molecule
Weight mg/1 °C °C torr Log KOM A° Koc
3. Four Rings
F morantnene
Pyrene
B»nz(a)anthrdcens

Chrysene
4. Five Rings
-^V^S^ 202 260 111 -- 6.0xlO'6 5 33 11.4
111 202 135 149 360 6.85xlO'7 5.32 95 62.700*
f^t^f^ 81.000
1^1 228 14 158 400 5 OulO'' 5.61 11 8
QCu
iXV-X^s 228 2 255 •- 6.3xlO'7 5.61 11 8

Benzo(b)fluoranthene
Benzo(k)fluorantnene
                                           252
0.55    217        480   S.OxlO'7       6 84

-------
                                                            TABLE 5.   CONTINUED
00

8i?n2o(a)pyrene
3 ibenz(j,h)anthracene



5. Sin Rings
Beii20(g,h, i Jpe'ylene

Inaenof 1.2,3-Cd)pyrene


Aqueous
Molecular Solubility
Height mg/1
f^Y^r^Y 25? 3'8
1. II 77ft 7 44
TnT^

WvS^X^s,
f^iT^^lL 0
ulfn ?76 °'26
^\IJ
f^C^ — ^1 276 6Z
kyAy^-Ax^
'«JW
Vapor
Melting Boiling pressure Length of
Point Point* 0 20°C Molecule
OC OC torr Log Kow AO Kot
1/9 496 S.OxlO^7 6.04 4.510.651
262 -- l.OKlO'10 5.97 13 5 2.0?9.000'



222 -- l.OxlO"10 7.23

163 — l.OxlO'10 7.66


         Sims and Overcash  (1983).
        *Karickhoff et al.  (1979).
        'Means et al.  (1980) (mean value  is reported).

-------
                       TABLE   6.    SUMMARY  OF  PHYSICAL  PROPERTIES  FOR  SELECTED  PHENOLIC
                                        COMPOUNDS  (VERSAR  INC.  1979)
Compound
Phenol
2,4-Dimethylphenol
4,6-Dinitro-o-cresol
4-Nitrophenol
2,4-0 Inltrophenol
Melting
Point
(°C)
40.9
£4.5
85.8
114.9
114
Boil ing
Point
(°C)
181.8
210.9
No Data
279
No Data
Aqueous
Solubility
(mg/i)
93,000 (at 25°C)
4,200 (at 20°C)

16,000 (at 25°C)
5,600 (at 18°C)
Log Kow
(octanol/
water parti-
tion coeffi-
cient)
1.46
2.50
2.85
1.91
1.53
Vapor Pressure
(torr at 20°C)
0.53*
0.06*
No Data
2.24*
No Data
*Vapor pressure as a supercooled liquid.
*Vapor pressure at 146°C.

-------
        TABLE 7.  HE-1 'H EFFECTS OF CHCMICAL CONSTITUENTS OF CREOSOTE
                               (U.S.  EPA 1984a)
    Compound
         Effect
1.  Unsubstituted 6-membered aromatic ring systems
    chrysene
    pyrene

    benzo[a]pyrene

    benz'.[e]pyrene
    benzo[a]anthracene
       N
    benzo[a]phenanthrene
    naphthalene
    phenanthrene
    anthracene
    dibenzanthracene
    acenaphthene
    triphenylene
2.   Unsubstituted aromatic ring
         mutagenic initiator, carcinogenic
         co-carcinogen (with fluoranthene
          Denzo[a]pyrene),  mjtagenic
         mutagen-ic carcinogenic, fetotoxic,
          teratogenic
         carcinogenic, mutagenic
         mutagenic, carcinogenic
         initiator, mutagenic
         inhibitor
         initiator, mutagcnlc
         mutagenic
         mutagenic
         mutagenic
         mutagenic
systems containing 5-numbered rings
    fluoranthene
    b"nz[j]f1uoranthepe
    fluorene
         co-carcinogenic, Initiator, mutagenic
         carcinogenic, mutagenic
         mutagenic
                                      20

-------
                              TABLE 7.  CUNTINUEO
    Compound
                                     Effect
 3.  Heterocyclic nitrogen bases
    qu^noline
    indole
    ber.zocarbazoles
    isoquinoline
    1-methyl isoquinoline
             Isoquinoline
             quinoline
             quinoline
             quinoline
             Isoquinoline
             isoquinoline
             isoquinoline
                 isoquinoline
3-methyl
5-methyl
4-methyl
6-methyl
5-methyl
7-methyl
6-methyl
1,3-dimethyl
acridine
carbazole
carcinogenic
mutagenic
carcinogenic
mutagenic
possify carcinogenic
possibly carcinogenic
possibly carcinogenic
possibly carcinogenic,
possibly carcinogenic
possibly- carcinogenic
possibly carcinogenic
possibly carcinogenic
possibly carcinogenic
mutogenic
mutagenic
mutagenic
4.  Heterocyclic oxygen and sulfur compounds
    coumarone
    thionaphthene
5.   Alkyl substituted compounds
    1-methyl  naphthacene
    2-methyl  anthracene
    methyl  fluoranthene
    1-methyl  naphthalene
    2-methyl  naphthalene
    ethyl  naphthalene
    2,6-dimethyl  naphthalene
    1,5-dimethyl  naphthalene
    2,3-dimethyl  naphthalene
    2,3,5-trimethyl  naphthalene
    2,3,6-trimethyl  naphthalene
    methyl  chrysene
    1,4-dimethyl  phenanthrene
    1-methylphenanthrene
                                     No effects  found  in the literature
                                     for this  structural class.
                                     mutagenic
                                     mutagenic
                                     possibly carcinogenic
                                     inhibitor
                                     inhibitor
                                     inhibitor
                                     inhibitor
                                     inhibitor
                                     accelerator
                                     inhibitor
                                     accelerator
                                     initiator
                                     initiator, mutagenic
                                     mutagenic
                                      21

-------
                            TABLE 7.  CONTINUED
    Compound
Effect
6.  Hydroxy compounds
    phenol
    p-cresol
    o-cresol
    m-cresol

7.  Aromatic amines
    2-naphthylamine
    p-toluidine
    o-toluidlne
    2.4-xylidine
    2i5-xylidine

8.  Paraffins and naphthenes
promoter
promoter
promoter
promoter
                                       NH2
carcinogenic
carcinogenic
carcinogenic
carcinogenic
carcinogenic
         L~tH2'J n                       (n Is large, e.g.,  greater than 15)

    No effects found in the literature for this structural  class.
                                     22

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                       TABLE 8.   HUMAN HEALTH EFFECTS  OF  EXPOSURE  TO CREOSOTE  (U.S.  EPA  1974}
       Year
IM
CO
       1896
       1920
       1924
       19'»7
       1956
  Substance Tested
     Occupation of
   Exposed Individual
Handling of Creosote
Handling of Creosote
Handling of Creosote
Handling of Creosote
Painting of Creosote
Worker who dipped railway
ties in creosote
Workers who creoscted
timbers

Creosote factory worker
37 men of various
occupations

Shipyard worker
                                                                                   Type of Tumor Response
Wa>ty elevation on arms;
Papillomatous swellings on
scrotum

Skin cancer
Squamous epitheliomata on
hand; epitheliomatous
deposits in liver, lungs,
kidneys and heart walls

Cutaneous epitheliomata
                                                                                  Malignant cutaneous
                                                                                  tumors of the face

-------
      TABLE 9.   POLYNUCLEAR AZAARENES IN CREOSOTE-PCP WOOD PRESERVATIVE
                      WASTEWATER  (ADAMS AND GLAM 1984)
           Compound*                                             Concentration
                                                                   (mg kg'1)


Quincline                                                            260
Isoquinoline                                                          69
2-methylquincline                                                     55
8-methylquincline                                                     11
Ci-azanaphthalene                                                     95
7-mPthylquinoline                                                     38
Cj-azanaphthalenes                                                    47
2,6-/2,7-dimethylquinoline                                            21
Cz-azanaphthalenes                                                    66
Methylvinylazanaphthalenes                                            14
C3-azanaphthalen2                                                     12
4-azafluorene                                                         16
7,8-benzoquinoline                                                    53
acridine                                                              55
5,6-benzoquinol ine/phene-^thridine                                     71
Cj-benzoazanaphthalenes                                              350
Vinylbenzoazanaphthalene                                               3.0
Azafluoranthenos/dzapyrenes                                           54
Ci-azafluoranthenes/azapyrenes                                         4.4
Dibenzoazanaphthalenes                                                 5.2

Total                                                               1300


*Cj, 03, and £•> - methyl-, dimethyl- or ethyl-, and trimethyl- or propyl
substituents, respectively.

-------
     Two and three-ring P£H  compounds  can  be utilized by soil microorganisms
as  a  sole carbon  source  and  are usually  easily  degraded.    In a  study by
McKenna and  Heath  (1976),  naphthalene  and  phenanthrene were rapidly oxidized
by both Pseudomonas and Flavobacterium, while anthracene was metabolized at a
moderate rate  by Plavobacterium.Ro  appreciable  degradation of  four-  and
five- ring compounds was detected.

     Compounds such  as  naphthalene, phenanthrene,  and  anthracene,  which are
readily metabolized, are relatively water  soluble, while  persistent PAHs, such
as  chrysene  and  benzo(a)pyrene,  have  a lower water  solubility.   Exceptions
exist with pyrcne and  fluoranthene in  that  these compounds  are  more soluble
than anthracene  and  yet have not been  found by some  researchers (Graenewegen
and Stolp 1981)  to  be  appreciably metabolized  by  soil microorganisms.   Other
factors that may result  in  the persistence of PAh  compounds are insufficient
bacterial membrane  permeability  to  the compounds,  lack of enzyme specificity
and lack of aerobic  conditions (Overcash and Pal 1979).

     Two incubation studies  were  performed  by  Bulman  et  al.  (1985)  to  assess
PAH  loss  from soil.   In  the  first,  a mixture of  eight PAH's [naphthalene,
phenanthrene,  anthracene,  fluoranthene,  pyrene,  benzo(a)anthracene, chrysene
and benzo(a)pyrene]  was  added to soil  at  levels of 5 and 50 mg-kg'1 and t£»
concentration  of each  compound was monitored with  time.   In the second, "C
labelled benzo(a)pyrene and  anthracene were  added to  soil in biometer flasks.
The  distribution   of  14C  as volatile,  adsorbed   and degraded  products  was
determined  in  sterilized  and  biologically  active   soil.   These  studies were
performed  using  unacclimated  agricultural  so'l.    Naphthalene, phenanthrene,
anthracene,  pyrene  and  fluoranthene initially disappeared  rapidly  from soil
during  an  initial  period of  200 days  or  less.   A  loss  of  94  to  98 percent
occurred during  this initial  period and approximated  first order kinetics,  in
some  cases  following  a  lag  period.   Within the  initial   period,  with the
exception of anthracene, the  first order kinetic-rate constants were the same
for  5  and 50  mg-kg-l  additions  of PAH.    Following  the initial  period, the
remaining 2-6 percent of the added PAH  was  lost  at a much reduced rate and the
first order  rate constants tended  to  be  higher  with  the 50 tng-kg'* addition
than the  5  mg-kg-*  addition of  PAH.    Losses  of  only 22 to 88  percent were
observed  for benzo(a)anthracene,  chrysene  and  benzp(a)pyene and  only  one
kinetic  period was  identified within  the  400-day  incubation period.   With
chrysene the first order kinetic rate  constants were the  same at the 5  and  50
mg-kg-1 levels of addition, however, for benzo(a)anthracene  and benzo(a)pyrene
the  rate  constants differed.   The   disappearance  of  benzo(a)anthracene
approximated first  order  kinetics; however  a  zero  order model  was generally
appropriate  for  the disappearance of benzo(a)pyrene  and chrysene.

     The mechanisms of disappearance  of  anthracene  and benzo(a)pyrene were
assessed using 14C  labelling.  The  results  indicated  that biological activity
was responsible  for some of the loss of anthracene from soil.   Binding to  soil
solids  and  volatilization  (either   as  anthracene or  as  metabolites) were
identified as  the  major loss, mechanisms.   Identification of loss mechanisms
for benzo(a)pyrene  was   less  successful  due  to the  small  amount  of
benzo(a)pyrene that reacted  within   the   incubation period.   Binding of
                                      ?5

-------
benzo(a)pyrene to  soil  solids  appeared to be  the major mechanism  involved,
while microbial  transformation of the compound was minimal.

     Tursten$son  and  Stenstrom  (1986)  have  cautioned,  however,  that  an
indirect measurement of  disappearance,  such  as  liberated  l^CO?  from a  ^C-
labeled compound  is  not always reliable.   They recommend that  the  rate  of
decomposition of a  substance  should  be defined by direct measurement of  its
disappearance.    Liberation  of C02  may not be  concurrent with  degradation
because of accumulation of  metabolites  in the soil.

     PAHs with  a. greater  number  of  rings are  not  known  to be utilized  as  a
sole carbon  source but  have  been reported  to be cometabolized  with  other
organic compounds.   This process  involves  the  concurrent  metabolism  of  a
compound that a microorganism  is unable  to use as a sole source of energy  with
a carbon source capable of  sustaining growth.  In a study by McKenna  and Heath
(1976), the cometabolism of refractory PAH compounds in the presence of  two-
and  three-ring  PAH compounds  was investigated.   The degradation of pyrene,
3,4-benzpyrene,   1,2-benzanthracene,  and 1,2,5.6-dibenzanthracene  in  the
presence and  in  the absence of phenanthrene  was  measured.   Separate  cultures
of Flavobacterium  and Pseudomonas were maintained  in the presence of each  of
the  PAH compounds.   Both Flavobacterium and  Pseudomonas  exhibited negligible
utilization of  the refractory  PAH  compounds  in the  absence of  phenanthrene.
However,  Flavobacter»um,  in the  presence of  phenanthrene,  was  able  to
significantly degrade  all  four  test  compounds.   Cometabolism by Pseudomonas
was not observed.  In  a similar experiment PAH compound degradation by a mixpd
culture was  measured.    For  each  PAH compound  studied,  one container  of
inoculum received naphthalene as a growth substrate  while  a  second  container
received phenanthrene  as  a growth substrate.   Cometabolism of  pyrene,  1,2-
benzanthracene,  3,4-benzpyrene, and  1,2,5,6-dibenzanthracene  by the  mixed
culture was exhibited  in the presence of either naphthalene or phenanthrene.

     The fate of  PAH  compounds  in  terrestrial  systems have been  reviewed  by
Sins and Overcash (1983), Edwards  (1983), and Cerniglia (1984).  These reviews
present additional information on PAH degradation.

     Phenolics  in  general  are  readily  degraded,  with  most  having
biodegradation  half-lives  of  only days.   The effect  of  phenols  on   soil
microorganisms  is dependent on  the  soil  concentration  or  amount added
(Overcash and Pal 1979).  At  low  doses (0.01-0.1  percent  of soil  weight), the
phenol  serves as an available  substrate, and  there  is an increase in  microbial
numbers.  As the dose level is increased (0.1-1.0 percent  of soil weight),  an
increasingly strong  inhibitory  or  sterilizing  effect  is  noted.   At these
levels,  a  partial  sterilization occurs in   which  there  is  a  depression  in
microbial  numbers,  but not  a  complete die-off.   After  a  period  of time,
microbes  adapt  or  phenol  is  lost  through  sorptive  ^activation  or
volatilization and a regrowth  of population occurs.

     Microbial  degradation of  phenol  has  been observed  in  many laboratory
studies in which phenol represented the primary carbon source for isolated and
adapted  microorganisms.    KappoM and  Key (1932)  were among  the  first  to
demonstrate the bacterial degradation of phpp.ol in  phenolic wastes.  Alexander
and  Lustigman  (1966)  ooserved that  phenol  was degraded  rapidly by a mixed
                                       26

-------
population of  soil  microorganisms.   Their data  suqqested that  the hydroxy
group, compared  to  other  benzene ring  substituents,  facilitated nncrobial
degradation.

     Bayly et al. (1966) reported that Pseudomo«as putida converted phenol to
catechol.   Verschueren (1977) reported complete disappearance of phenol  in a
soil suspension in two days.   The effect  of  temperature variations on the rate
of biodegradation of  phenol  in  the soil  was studied by  Medvedev  arri Davido/
(1972).   At  5°C, phenol  remained in the  soil  after 16 days,  while at  19°C
there  was  complete  loss  after   six  days.   The  ability  to  degrade  phenol
improved with  successive  phenol  doses  (Medvedev et  dl.   1975).   Initial
degradation of  phenols in soils  has been  enhanced  by bacterial  seeding of
Pseudobacterium lacticum and  Pseudomonas  1iquefaliens (Dolgova 1975).

     Other phenolic  compounds such as 2,4-dimethylphenol, 4-nitrophenol, 4,6-
dinitro-o-cresol, and  2,4-dinitrophenol  have also  been  shown  to  be readily
degraded in soil (Medvedev and Oavidov 1972, Verschueren 1977, Overcash  et al.
1982).

     A summary of degradation of PAHs and phenolic compounds is given in Table
10.  The term half-life of the  compounds is  used  to indicate the persistence
of  a  chemical  in the  soil, water, or air  environment.   The  half-life   is the
time required for the concentration  of a compound  to decrease to one-half of
its initial value.  Half-lives may be estimated from first-order kinetics, if
first order rate constants are known  for  waste constituents.   Performance data
indicate that  the degradation of  most  chemicals  in the soil can  be modeled
using a  first-order reaction rate (i.e., dC/dt « -KC,  where  at  any one time,
t,  the  rate  of  degradation  is  proportional  to the  concentration,  C,  of the
chemical  in the soil (ERT 1985b)j. First-order kinetics generally apply where
the  concentration  of  the chemical   being  degraded is  low  relative  to  the
biological  activity in the soil  (Kaufman  et al.  1983).   At very high chemical
concentrations, Michaelis-Henten  kinetics  appear to  apply,  and the  rate of
degradation changes  from being  proportional  to the  concentration to  being
independent  of concentration  (Hamaker  1966;  Hamaker  et  al. 1%7).   For
compounds such as PCP, which  serves both as a growth substrate and, at  higher
concentrations, as a  growth  inhibitor, the  Haldane  modification of the Monod
equation has  been shown to be suitable to describe the  kinetics of degradation
(Klerk* and Maier 1985).

     From information given  in  Table 10, initial  rates of degradation  of PAH
compound- in  soil as a function  of initial  soil concentrations,  assuming first
order  kinetics,  are  presented  ii Figure  1.   These  data are  corrected  Tor
variations in  temperature using  an  Arrhenius  equation  with  coefficients
developed from  PAH  data  to  a temperature of  20°C,  n  = 1.013.   Rates  were
normalized to ug PAH  transformed/g soil-dry wt/hr.   The  general trends shown
in  Figure  1  can be  summarized  as follows:  1)  for a  given  PAH compound the
initial  rate  of  transformation  increases with  increasing  initial  soil
concentration,  2)  within  the  class  of polycyclic aromatic  compounds,  the
initial  >-ate  of transformation  decreases  with  increasing number  of  fused
benzene rings for molecular size).
                                      27

-------
                    TABLE  10.   KINETIC  PARAMETERS DESCRIBING RATES OF DEGRADATION OF PAh  AND PHENOLIC
                              COMPOUNDS IN SOIL SYSTEMS  (SIMS AND OVERCASH 1983. CRT 1985b)
er
Substance
Phenol
Phenol
2, 4-dimethyl phenol
4,6-dinitro-o-cresol
2, 4-dlnitro phenol
2,4-dinitrophenol

4-nltrophenol
Pentachloruphenol
Naphthalene
Naphthalene
Naphthalene
Acenaphthylene
Acenaphthylene
Anthracene
Anthracene
Phenanthrene
Phenanthrene
Benz( a) anthracene
Benz( a) anthracene
Benz( a) anthracene
Benz( a) anthracene
Benz( a) anthracene
Benz( a) anthracene
Ben z ( a ) ant hr acene
Initial
Concentration
(ug/g soil)
500
500
500
-.
5-50
20-25

-.
--
7
7
7
0.57
57
0.041
41
2.1
25,000
0.12
3.5
20.8
25.8
17.2
22.1
42.6
(dayl)
0.693
0.315*
0.35-0.69
0.023
0.025
0.099-0.23

0.043
0.018
5.78
0.005*
0.173
0.039
0.035
0.019
0.017
0.027
0.277
0.046*
0.007
0.003
0.005
0.008
O.r%
O.U03
1/2 Life
(days)
1.0
2.2*
1-2
30
28
3-7

16
28
0.12
125*
4*
18
20
36
42
26
2.5*
15.2*
102
231
133
199
118
252
Reference
Medvedev & Oavldov (1972)
Medvedev & Davldov (1972)
Medvedev & Davidov (1972)
Versa* , Inc. (1977)
Overcash et al. (1982)
Sudharkar-Barik &
Sethunathan (1978)
VerschyoTer (1977)
Murthy et al . (13/9)
Herbes & Schwall (1978)
Herbes & Schwall (1978)
Kerbes & Schwall (1<"8)
Sims (1982)
Sims (1982)
Sims (1982)
Sims (1982)
Groenewegen and Stolp (1976)
Sisler and Zobell (1947)
Herbes & Schwall (1978)
Groenewegen & Stolp (1976)
Gardner et al. (1979)
Gardner et al . (1979)
Gardner et al . (1979)
Gardner et al . (1979)
Gardner et al . (1979)

-------
                                                   TABLE  10.   CONTINUED
vo
Substance
Benz( a) anthracene
Benz( a) anthracene
Benz( a) anthracene
Benz( a) anthracene
Benz( a) anthracene
Fluoranthene
Fluoranthene
Fluoranthene
Fluoranthene
Fluoranthene
Fluoranthene
Fluoranthene
Pyrene
Pyrene
Pyrene
Chrysene
Chrysene
Chrysene
Benz(a)pyrene
Benz'ajpyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Initial
Concentration
(vg/g soil)
72.8
0.07
0.10
0.15
7
3.9
18.8
23.0
16.5
20.9
44.5
72.8
3.1
500
5
4.4
500
5
0.048
0.01
3.4
9.5
12.3
7.6
17.0
32.6
k
(day-M
0.004
0.005
0.005
0.005
0.016
0.016
0.004
0.007
0.005
0.006
0.004
0.005
0.020
0.067
0.231
0
0.067
0.126
0.014
0.001
0.012
0.002
0.005
0.003
0.002
0.004
1/2 Life
(days)
196
134
142
154
43
44
182
105
143
109
175
133
35
10.5
3
_
10.5
5.5
50*
694*
57
294
147
264
420
175
Reference
Gardner et al . (1979)
Sims (1982)
Sims (1982)
Sims (1982)
Sims (1982)

-------
                                            TAtLE 10.  CONTINUED

Substance
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
8enz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Benz(a)pyrene
Oibenz( a ,h) anthracene
Dibenz( a, h) anthracene
Initial
Concentration
(ug/9 soil)
1.0
0.515
0.00135
0.0094
0.545
28.5
29.2
9,100
19.5
19.5
19.5
130.6
130.6
9,700
25.000
k ,
(dayl)
0.347
0.347
0.139
0.002
0.011
0.019
0
0.018
0.099
0.139
0.231
0.173
0.116
0.033
0.039
1/2 Life
(days)
2*
2*
5*
406*
66*
37*
__
39*
7*
5+
3*
4*
6+
21+
18+

Reference
Shabad et al . (1971)
Shabad et al. (1971)
Shabad et al. (1971)
Shabad et al . (1971)
Shabad et al. (1971)
Shabad et al. (1971)
Shabad et al. (1971)
Lijinsky and Quastel (1956)
Poglazova et al . (196/D)
Poglazova et al . (1967b)
Poglazova et al . (1967b)
Poglazova et al . (1968)
Poglazova et al. (1968)
Lijinsky and Quastel (1956)
Sisler and Zobell (1947)
*Low temperature (<15°C)
+High temperature (>25°C)

-------
      102,
   o>
  \
   0>
  a
  u_
  o
      10°:
     ID'1 :
     ID'2 :
UJ
h-
(X

2 10'3
t—
z

   10'4
     10
        -5
          *— ACENAPHTHENE
          •— ACENAPHTHYLENE
          	 ACR1DINE
          <— ANTHRACENE
          •— BENZ (•) ANTHRACENE
          ••— BENZO (b) FLUORANTHENE
          t— BENZO(k)FLUORANTHENE
          *— BENZO («)PYRENE
          Z— CHRYSENE         s'

                                       DIBENZ(..J)ACR1DINE
                                       DlBENZ («.h)ANTHRACENE
                                       DIBENZOFURAN
                                       D1BENZOTM10PHENE
                                       FLUORENE
                                       FLUORANTHENE
                                       NAPHTHALENE
                                       PHENANTHRENE
                                       PYRENE
         10
           -1
                10°
101
102
           INITIAL  CONCENTRATION
103     104     10s
(ug/g-dry  wt .)
Figure 1.  Rates of degradation of PAH compounds in soil  as a function of
         initial soil concentrations (Sims and Overcash 1983).

-------
Immobilization of PAH and Phenolic  Compuuiids--

     Quantitative descriptions of immobilization, or sorption, pnenomena  also
contribute  to  the  assessment  of  the fate  of waste constituents  in  soil
systems.   Equilibrium adsorption may be  des:ribed  quantitatively  using
adsorption isotherms, which represent the  relationship  between  the  amount  of a
solute adsorbed and  the  equilibrium  concentration of  the  solute  in the  soil
solution at a given  temperature.  Specific  adsorption  isotherms commo:ily  used
to describe  i-rnnobilization of organic  constituents  in  soils include:   1) the
Langmuir isotherm, and 2) the Freundlich isotherm.

     The Langrcuir  isotherm adsorption  relationships occur  when there  is no
strong  competition  from the  solvent for  sorption sites on the adsorbent
surface.   The  Langmuir  adsorption  isotherm is expressed mathematically using
the *ollowing relationship:
where S  is the mass of  adsorbed  solute  per  unit  mass  adsorbent,  KI  represents
the maximum mass  of  solute that can be  adsorbed by the  soil  matrix, Ki is  a
measure of the bora strength holding the sorbed  solute on a soil' surface,  and
C  is the  equilibrium  concentration  in  the  soil  solution.   The  Langmuir
isotherm has  been used  extensively for  the description  of  inorganic   and
organic constituent soil adsorption.

     The Frerndlich isotherm is an empirical  formulation  describing  adsorption
phenomenon and can be expressed as:

     S = KG."

where K and N = constants.

     The Freundlich isotherm  provides  flexibility  in  that  the use  of the  two
eouation parameters,  K  and N,  allows  the  fitting of  the equation  to a wide
ranc,j of data.    It also  does not  require  a maximum  limit  for  the  amount of
substance adsorbed.

     The linear form of the Freundlich isotherm may be expressed  as:

     S = kdc

where k
-------
     Koc = K/XOC)*100 (nonlinear Freundlich isotherm)

Tnis  parameter  is  less  variable than  non-normalized coefficients,  and  is
normally independent of soil  type.

     °AHs are noniomc, nonpolar compounds that do not ionize significantly in
aqueous systems.  Adsorption of nonionic  compounds  Is  primarily  a  ^unction of
solubility.   PAHs,  therefore,  participate  in  hydrophobic  sorption in a  soil
system, where  the nonpolar  PAH compounds  partition  out of  the  polar  water
phase onto hydrophobic surfaces in the soil matrix.  Hydrophobic  sites include
waxes,  fats,  and resins  of the  soil organic  matter.   The organic matter
content of  the  soil  thus  is  more  important  in determining  the extent  of
sorption of PAHs in a soil system (Nkedi-Kizza et al.  1983)  than  substrate pH,
soil cation exchange capacity,  soil texture, or  clay  mineralogy  (Means et al.
1980).

     Table  11  summarizes  the  range  of  measured or  estimated  immobilization
constants  for the constituents  known  or  suspected  to be present  in  creosote
wastes.   The estimated  range  of organic  partition coefficients  (Koc)  for  a
given compound  is based  on the octanol/water partition  coefficient  (Kow)  for
the  compound,  according   to  the following relationship:   log  Koc  =  log  Kow-
0.317 (Hassett et al.  1980).   The Koc for certain PAH compounds  was estimated
from reported values  for  PAH compounds  of similar molecular weight  and  ring
structure  if no K0w  data  were  reported.   Constituents with  K0(^ values greater
than 10,000 are ve«-> strongly adsorbed and essentially immobilized  in the soil
environment.  The relative mobility of  PAH compounds  was estimated by 'Jmfleet
(1986)  to  be  as follows:   chrysene < fluoranthene <  pyrene < phenanthrene  =
anthracene < naphtha^ne.

     Phenols and phenolics vary in their ability to be adsorbed by soils.  The
moderate  values  of the  octanol/water partition  coefficients  (Table 11)  for
phenol, 4-nitrophenol, and 2;4-dinitrcphenol  indicate only  a  slight  tendency
for these compounds to be adsorbed to organic matter.   2,4-dimethyl-phenol and
4,6-dinitro-o-cresol have  higher octanol/water partition -"efficients  and
therefore  show a greater potential for adsorption.

Photodecomposition of PAH and Phenolic Compounds--

     Photo-oxidation of PAH compounds has been well documented (Radding et :1.
1976;  Versar  1979).   The PAHs absorb  solar  radiation strongly  and undergo
direct  photolysis.   PAH  compounds can be  transformed  into  reactive cytotoxic
and mutagenic  intermediates  following exposure  to natural  sunlight  and  other
sources of radiation.  Polycyclic aromatic amines (e.g., 2-aminofluorene) have
especially been shown to have photomutagenic properties (Okinaka et al. 1983).

     However, although direct photolysis  occurs  in both the atmosphere and in
aqueous environments,  photo-oxidation of PAHs in the  soil  environment  is not
expected to be significant because of limited exposure to light.

     Y-radiation  has  been shown  to  destroy the phenol structure  in aqueous
solutions  (Overcash  and  Pal  1979).    Solar  radiation  may cause photosensitive
reactions  of  phenolics (e.g.,  photonucleophilic mechanism  (Overcash  and Pal
1979)).   Using  the procedure  given  in  U.S.  EPA  (1994f),  the half-life of
phenol  in air was calculated as 1180 days.


                                        33

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       TABLE  11.   SUMMARY OF  SOIL  SORPTION DATA FOR CONSTITUENTS  OF  CREOSOTE  WASTE (ERT, INC. 1985b)
No. Of
Compound Rings
Molecular
Weight
Organic Carbon Log Octanol /Water
Solubility Partition Coefficient Partition Coefficient
(mg/1) Koc (ml/g) Log Kow
Low Molecular Weight PAH
Naphthalene
Acenaphthylene
Acenaphthene
Fluorene
Phenanthrene
Anthracene
Fluoranthene
Pyrenc
2
2.5
2.5
3
3
3
4
4
128
152
166
178
178
202
202
31.7
3.93
1.29
0.073
0.26
0.135
1.300
1,000-10,000*
1,000-10,000*
1,000-10, GOO*
23.000
26.000
10.000-100,000
63.000-84,000
3.37
4.07
4.33
4.18
4.46
4.45
5.33
5.32
Hiqh Molecular Weight PAH
Benzo(d) anthracene
Chrysene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzol a) pyrene
Dibenzo(a,h)anthracene
Benzo(g,h,i)perylene
Indeno(l,2,3-cd)pyrene
Phenol 1cs
Phenol
4,6,Dinitro-o-creSol
4-Nitro phenol
2,4-Dinitropnenol
2,4-dimethyphenol
4
4
5
5
5
5
6
6



228
228
252
252
252
276
278
276

94
139
184
0.014
0.0018
0.0012
0.0008
0.005
0.0005
0.0002

93,000 (25°C)
16,000 (250C)
5,600 (18°C)
1,871,400
100,000-1,000,000*
>1, 000, 000*
>1, 000, 000*
4,510.650
2.029.000
>1, 000, 000*

10-100*
100-1000*
100-1000*
10-100*
10-1000*
5.61
5.61
6.57
6.84
6.04
5.97
70 -3
. dJ
7.66

1.46
2.85
1.91
1.53
^Estimated from reported  values  for PAH compounds of similar molecular weight and  ring  structure.

-------
Volatilization of PAH and  Phenolic Compounds--

     Volatilization of constituents  in creosote wastes from  a  soil  system ib
dependent upon several  factors  that  include: 1) constituent vapo;  pressure; 2)
concentration of  the constituent in  the  soil  solution;  3)  soil/constituent
sorption reactions;  4)  solubility of  the  constituent  in soil  water;  5)
solubility of the constituent  in  soil  organic matter; 6) soil characteristics,
such  as  temperature,  water  content, organic  content,  clay  mineralogy  and
content,  porosity,  and bulk density;  and 7)  waste application  methon1  and
tilling frequency (Spencer and  Cliath  1977).

     In  general,  since PAH compounds tend to  be  nonvolatile  and  are  also
readily  sorbed  by the  soil,  they do not  represent a significant  source of
emissions from  soil  systems (U.S. EPA 1974).   Acenaphthalene and naphthalene
are the most volatile PAH  compounds.

     Phenolics  are  more  volatile i.han other  constituents  of  creosote,  but
because  of  their low concentrations  in creosote  waste,  phenol  emissions  are
not expected to be significant  from  creosote waste .land treatment facilities.

Bioaccumulation of PAH and Phenolic  Compounds--

     PAHs are  ubiquitous constituents of crops,  plants,  and  algae  in  the1
natural  environment.    In  general,  leaves  exhibit  the  highest  PAH
concentrations  (22  to 88  ppb),  and  underground  vegetables  such as potatoes,
carrots,  onions,  and  radishes, exhibit the lowest PAH concentrations (0.01 lo
6.0 ppb)  (Sims  and Overcash 1983).

     Sources  of PAHs in vegetation  include anthropogenic  activities resulting
in PAH  deposition on plants,  biochemical  synthesis by plants, and plant-uptake
from soil (Sims and Overcash 1983).   Many  higher  plants,  however, may not  take
up PAHs (Blum and Swarbrick 1977).

     PAHs have been  demonstrated   to  act like  plant  hormones, stimulating
growth  and  yield of  higher and  lower plants.    Graf (1965) demonstrateo the
growth-promoting  effects  of  PAHs with  higher  plants.   He  also  demonstrated
that  the growth  promoting  effect  was  proportional  to  the  carcinogenic
potential.

     There  is strong evidence  for plant biosynthesis of  PAHs  (Borneff  et al.
1968).    Biosynthesis  of  PAHs  has  also been  investigated  with respect to
bacterial synthesis  (Bnsou 1969).

     Edwards  (1983),  in  a comprehensive  review  of PAHs  in the  terrestrial
environment,  presented  the following conclusions concerning the uptake  of  PAHs
 in terrestrial  vegetation:

     1)  Some  terrestrial plants can take up  PAHs  through  their roots and/or
          leaves and translocate  them to various plant parts,

     2)  Uptake rates are dependent on  PAH  concentrations, solubility, phase
           (vapor or  particulate), molecular size, support media  anchoring the
          plants, and  plant species,


                                       35

-------
     3)   PAHs  may  conontrate  in  certain plant  pa-'ts  more than  in other
          parts, anJ

     4)   Some PAHs can be catabolized by plants.

     Little  information  is  available  on  the fate of phenolics in terrestirel
systems (Overcash and Pal 1979).

Treatment of Pentachlorophenol Wastes In  Soil  Systems

     Pentachlorophenol (PCP), a versatile biocide,  is primarily used as a wood
preservative  and  may be  added  to creosote to  enhance  the wood  prtservation
potential of creosote.   Pentachlorophenol  may  be  used  in  the  phenol  form
(PCP), as  salts  (e.g., sodium  pentachlorophenate  (Na-PCP)),  or  as  esters
(e.g.,  acetate  or  lauryl).   The  hydroxyl  group of  PCP  forms  esters  with
organic  and  inorganic acids.   Oxidation of PCP results  in  the  formation  of
pentachlorophenoxyl radicals  that combine reversibly to  form dimers.   At low
pH, PCP exists as  a  free  acid and  readily adsorbs to soil  particles.  At high
pH  , PCP  exists  in  the  ionized  form (pKa = 4.7), and is  more mobile.   At  pH
2.7, PCP  is only 1 percent ionized,  while at pH  6.7, it is  99  percent iomzea.
Alkaline  salts  of PCP,  such  as sodium  pentachlorophenate  (Na-PCP)  are  nore
mooile than PCP and less likely to be immobilized  in a soil system.

     The vapor pressure of 760 mm of Pentachlorophenol is  achieved at 300.6CC,
but  even  at  ambient  temperatures,  PCP  is relatively  volatile.   Na-PCP,
however,  is  nonvolatile.   PCP is slightly soluole i,i  water and  is soluble  in
most organic solvents (Table 4), while Na-PCP  is more soluble  'n water.

     Txcept  for hydroxyl reactions, PCP  is  quite stable.   However, it absorbs
ana is  rapidly degraded  by  UV light and would  not be expected to  persist  in
the  open environment  (although  it  remains  unchanged  for  long  periods  in
treated wood).

     The  environmental   chemistry  of  PCP was  reviewed by Crosby  (1981).
Information   concerning  the  uses  of PCP, chemical  and  physical   properties,
biological uptake  and transformation  of PCP  and  'cs  impurities,  analytical
methods for  PCP  and  its  impurities,  and environmental  residues  of PCP  and
associated compounds are summarized  in  this  review.


Toxicological Significance of PCP Wastes--

     The  toxic ity  of  PCP  and  potential  for  uptake  by organisms are  pH-
Jependent,   since PCP  is  a weak  acid  with  a  pKa  of  about  10'^.    Both
bioaccumulation  and  toxicity increase as  pH  decreases,  due to  the  greater
penetratio"  of  cell  membranes  by  un-ionized  PCP  mjlecules  than  by
pentachloropfjnate ions.

     In  general,  PCP  is a  biocide  toxic  to microorganisms  (as  it  is  a
bactericide  and fungicide),  to lower and higher  plants (algicide,  herbicide),
to  invertet-ate  and   vertebrate  animals  (insecticide,  mollusciciHe),  and  is
also toxic     man.   Adverse effects  to  man  include  serum  enzyme  induction
(Klemmer  IS ^.,  low-grade infections  and  inflammation  (Klemmer et  al.  1980),
and depressed kidney function (Begley  et  al. 1977).  Technical grade PCP, with

                                     36

-------
associated  impurities,  dibenzodioxins  and aibenzofurans,  produces chloracne
and  liver  damage  (Crosby  1931).    Additional  information concerning  human
health  effects ire  presented  in  the  U.S.  EPA health  effects  assessment
document for PC (U.S.  EPA 1984d).

     The U.S. EPA (1986a) has  summarised  the effects of PCP on aouatic 1 i i~e in
order to develop ambient water quality criteria for PCP.   The  authors of the
report found that the acute and chronic toxicity of PCP to freshwater animals
increases as  pH and  dissolved  oxygen  concentration  of the  water  de:reases.
Ger.°rally, toxicity also increases with increased temperature.   The estimated
acute sensitivities of  32 species  at  pH =  6.5  ranges f-nm 4.355 ug/'L  for
larval common carp to >43,920  ug/L  for  a cray  fish.   At  pH = 6.5,  the lowest
and highest estimated chronic values of  <1.835  and  79.66  ug/L, respectively,
were obtained with different cladoceran species.   Chronic  toxicity to 'ish is
affected by the presence of impurities, with certain industrial grades «>f PCP
being more  toxic  than a purified (99+  percent.)  form.   Freshwater  algae were
affected by  concentrations as  low as  7.5 ug/L. whereas vascular  plants were
affected at  296  ug/L  and above.  Bioconcentration factors ranged  from 7.3 to
1,066 for three species  of  fish.

     Acute toxicity values  from tests  with 18 species of saltwater  animals,
representing 17 genera,  range  from 22.63 ug/L.for late yelk-sac larvae of the
Pacific  herring,  Clupea  harengus  pall asi, to  18,000 ug/L for  adult  blue
mussels, Mytilus edulis.   Five of these  values are for saltwate^  fish.   The
embryo and larval stages of invertebrates  and the  late larval preinetamorphosis
stage of fish  appear  to be the most sensitive life stages to  PCP.  With few
exceptions,  fish  are  more  sensitive  than  invertebrates  to  PCP.   Salinity,
temperature,  and  pH  have  a  slight effect  on  the  toxicity of  PCP  to  some
saltwater animals.

     The  EC50s  for taltwater  plants  rangp  from  17.40  ug/L for  the diatom,
Skeletonema  costatum,  to  3,600  ug/L  for  the green  algae,   Dunaliella
tertiolecta.

     The chlorinated  dioxin  and dibenzofuran  iir.,iuri*.ies  in  PCP are  also of
concern.  The U.S.  FPA has listed PCP manufacturing wastes as acute hazardous
wastes because of the  presence  of  hexachlorodioxins (U.S.  EPA 1985).


Degradation and Transformation  of  PCP--

     Despite its high  degree of chlorination, PCP  has been shown to be readily
degraded  in  soil.    Microbial  decomposition appears   to  be  the  primary
detoxification  mechanism.   Aerobic microbial  degradation of  PCP  results In
transformation  to the ultimate metabolites, carbon dioxide  and chloride ion,
as  shown  in Figure  2  (Crosby  1981).   Watanabe  (1973)  isolated  a PCP-
decomposing  Pseudomonas from  treated  soil.   Pseudomonas degraded  PCP  and
released carbon dioxide and the intermediate metabolites  (tetrachlorocatechol
and  tetrachlorohydroquinone).   Pentachlorophenol  has been reported  to be
converted into  pentachloroanisole and  tetrachlorohydroquinone  dimethyl  ether
by  a  Bac ill us sp.   (Kirsch  and Etzel  1973).   Several  soecies  of  fungi also
depleted PCP  from  PCP-treated  wood  blocks (Duncan and Deverall  1964).   Slow
chloride release and detoxication  of PCP  occurred  emoloying the fungal enzymes
laccase, tyrosinase,  and peroxidase.   Cserjesi   (1972)  found  that  PCP

                                      37

-------
             OH
             OCH3

             I    _
00
             OH
         1.2.4
             OH

             I
                        ox
                      :OC
                       l.2.*_
OCH5
I   .
                        1.2
                         OCH
             OCH3
             OH
                                     OH
             OH

             I
                       OH
                                           \jf  OH
OX

I
                                                OH
                     I     I

             OH

             I
                                                                        OH
                                                OH
                                                                               coa.<
                                                                  Cl
                                                nrw
                                             i   OCH3
                           ci,
                                                  Cl,
                                                                             'Cl
     Figure  2.   Biodegradation of PCP.  For clarity,  Cl substituents are indicated only by lines.  1 = Micro-
                organisms, 2 = Mammals, 3 = Fish and  aquatic invertebrates,  4 = Green plants  (Crosby 1981).

-------
disappeared during  a 12 day  incubation  with  cultures of fungus  Trichodenna.
The  fungus  was  shown  to methylate  PCP to  pentachloroanisole.    Similarly,
several  funqol  species also caused methylation  of  tetrachlorophenol  to
tetrachloroanisole

     Ability  to degrade  PCP  may  not be  uniform amcng microorganisms.    No
degradation of  PCP  was found in  a  mixed  population  grown  from  a   soil
suspension  (Alexander  and  Aleem 1961); likewise, no degradation was  observed
in acclimated activated sludge  (Ingols et  al.  1966).   However, PCP was  found
to be  readily biodegradable  in water  from an activated sludge plant  (Kirsch
and Etzel 1973).  Adaptation  of microbial  populations to PCP  (along  with  the
control of pH) may play an important role in the  degradation.

     A  summary  of  PCP degradation  studies  is  presented  in  Table   12.
Degradation half-lives  for aerobic  soil  treatment systems ranged from greater
than 30 days for nonacclimated systems to less than  1  day for  fully acclimated
or inoculated systems.   Most  studies jsed  initial PCP concentrations  of  from
10 to 30 mg/kg of soil, dry weight.  However,  one long-term study  by  McGinnis
indicated that  PCP  concentrations of over  2,000 mg/kg soil  could be  rapidly
and completely degraded by  a  well-acclimated soil treatment  system  (McGinnis
1985).

Immobilization of PCP--

     The degree of  adsorption  of PCP  affects  both its  rate of  degradation and
its tendency to disperse by leaching.   PCP is, in general, more mobile in  high
pH soils  than in acidic soils  (Choi  and Aomine  1973. 1974a; Green and  Young
1970, Nose 1966).  At alkaline  pH,  PCP exists  as the  dissociated an ion,  which
is highly water soluble  and   is  not easily  adsorbed to soils  having a  net
negative charge.

     In a study by  Choi and Aomine  (1974a)  using 13  soil  samples with various
clay mineral  species,  organic matter content, and  pH, "apparent  adsorption"
(defined  as  the  amount of PCP  that  disappeared  from  the liquid phase of  the
soil/PCP system) was the greatest  in  the strong  acid soil system  compared  to
the moderate  acid  soil  system, regardless  of the species of clay mineral  or
organic natter  content.   No   adsorption  occurred in  the  slightly  acid  or
neutral soil  system.   Organic  matter was  also  important  in  PCP  adsorption,
since soils higher  in organic  matter  showed a greater adsorption of  PCP  than
soils  lower  in  organic matter.  "Apparent  adsorption"  was  shown to  include
both the mechanisms  of  adsorption or  soil  colloids  and  precipitation in  the
soil  micelle and in  the external  liquid  phase, depending on  the soil  pH  (Choi
and Aomine 137
-------
                   TABLE  12.  SUMMARY  OF BENCH AND  PILOT  SCALE PCP DEGRADATION  STUDIES  (ERT,  INC.  19P5a)
X Ski 1 Average Initial
Ii'ir.Bcratur* boil Moisture Concentration
Seal* (°C) ptt Coilcnt («gAg Oiy toil)
Cilol (4-.41 test ploti) 3- 16 f> 1 li 30
Pi'jt (4'»4- x«jl plots' 8-16 67 IS 30
Sfrvh (10 g toil) 21 7 1 16 10
S«.ith (10 g soil) 2J 71 IE 10
Bcrcn 30 6 7 IS to 20 20
O Bench 30 671!. to 20 20
0
2i-
E«-LI (40 g soil) .. 70 22L3*
Microbial
Conditions
Not acclfmatea.
At; iot) 1C
Inoculated.
Nut acclimated.
Not ace If nated.
Anaerobic
Not acclfitiled.
Aerobic
Inoculated,
AciOUIC
Sjl acclimjted.
Aortbic
Not icclim.tn).
AeictiiC
AcclfnateJ 'or
1 yea-. Aerobic
Degradation Rate
2M, after 1? Ciys
Ha If- life 6 da; j
60X after 160 days
7t after 160 days
Hjlf-life 12 tu 14 days
SOX in 24 to 100 hours'
(HaU-1 ite •>• day)
24X in 30 days
2SX in 30 days
Hjlf-1 ifc 21 hrs
R«fer«iice
£03*11 i 1 anu f if n
Edgeni 1 1 and Firm
baker and MjyMeld
Baker and Hayfield
Edgehi 1 1 and Finn
Edgerii 1 1 and ^ inn

(198
; J9U
(11J
(19£
(1983
(1983
Baker. Hayfietd and
Inniss (I960)

McGinn is (Iiai)


'93 percent degradation achieved in 24. 40.  »•••! 100 liouri tut!1. !"ii.-ilun LunLentralions o( 10r>,  10'. and 104 cells per qran of  soil,  respectively
'tssur.es IS percent let) Moisture content

-------
 Photodecompositinn of PCP--

      Photodecomposi t ion  may be  an  important  route wnereby  a chemical  is
 eliminated  from the environment.   PCP  undergoes  var'ous reactions while  it
 absorbs  light  energy  (the  long-wave absorption maxima  lie near  300 nm  in
 organic  solvents or below  pH 5 (Crosby  1981)).    In  organic  solvents or  in
 water, PCP  is photochemical ly reduced to isometric tri- and tetrachlorophenols
 (Crosby  and Hamadmad 1971).   Nudeophiles such  as  bromide  ion can  displace
 chloride  from  the  excited PCP  ring  and  in  an  aqueous  solution  exposed  to
 sunlight, PCP  undergoes the  replacement  of ring  chlorines by hydroxyl  groups.
 The resulting products  are oxidized by air to quinones, which subsequently are
 dechlorinated (Crosby and Wong 1976).

      Pignatello et  al .  (1983)  showed that  in  an aquatic system,  photolysis
 accounted  for  5 to  28  percent  decline  in  initial  PCP  concentrations.
 Photolysis  was  rapid  at ".he water surface but greatly attenuated with depth.
 Lamparski  et al .  (1980) demonstrated  that  PCP  could  undergo  photolytic
 condensation reactions  to form octachlorodibenzo-p-dioxin on  a  wood  substrate.
 This  effect was greatly reduced by the addition of a hydrocarbon oil.


 Volatilization  of PCP—

      The volatility of  PCP from a soil system is dependent on the soil  pH.   In
 general,  volatilization of PCP  is not  expected to  be significant  from  land
 treatment soil  systems.   PCP  is  relatively volatile  but  Na-PCP  is nonvolatile
 (Crosby  1981).   Therefore, as  soil  pH  is raised  above the PCP pKa of  4.7,
 volatility  decreav»<>  because  the  ionized form  of  PCP  is predominant at  pH
 levels  that ore  optimum for  biological  treatment  of  added organic  wastes,
 i.e., ph of 6 to 7 (Luthy 1984).


 Bioaccumulation of PCP—

      Sioaccumulation of PCP  from water, like  toxlclty,  has  been shown to  be
 inversely related to pH  (U.S. t'PA 1986a).  PCP bioconcentrated  in the  tissues
of fish from 7.3 to 1,066  times  w**.h  test  durations  from 16  to  115  days.  The
gall  bladder concentrated  the  hignest levels  of  PCP, whereas muscle and  skin
contained the lowest concentrations of PCP in rainbow trout exposed  to  0.78  to
1.15 ug/l (U.S. EPA 1986a).

      In  general,  bioaccumul ation >-f  PCP has  been  found  to  be   short-term
because organisms  i<»nd  *p  metaboli.:e and  excrete  these  compounds  (Versar
1979).   Residues of PCP  in  fish have been shown to  drop quite rapidly  upon
termination of  exposure (U.S.  EPA  1986a).   Ninety-six  percent  of  whole  body
14C- label led PCP was eliminated  by   fathead  minnows  within  3.5 days, while
about 85 percent of the  PCP residues  in  blueqill  muscle were eliminated  in 4
days.  A first-order simulation model developed from empirical data  indicated
a half-life of 2.7 days  in rainbow trout, with 95 percent  elimination  in  11.7
     Little  information  exists  on plant  metabolism  of PCP,  although  PCP is
very phytotoxic  (Crosby 1<>81).   Studies  were  performed  on  application of

                                      41

-------
radio-label led PCP to cotton plants (Miller and Aboul-Ela 1969).  The
of  bolls,  which  were  closed  at  spraying   time,  containi-c"  residues  of
radioactivity.   Application of  PCP  to  sugar   cane  leaves  resulted  in almost
complete recovery of the PCP from the  leaves,  while root  application deposited
most of the compound in the roots (Hilton  et dl.  1970).   Studies on the growth
of rice  in  soil  treated with radio-labelled  PCP showed  that  after  one week,
the plants had absorbed about 3 percent  of the applied radioactivity (Hague et
al. 1978).


PETROLEUM REFINING INDUSTRY

Introduction

     There are  approximately  250-300  petroleum  refineries  in  the United
States.   These refineries  vary from complex  plants  producing a  variety of
petroleum products and petrochemical  feedstock to simple  plants  producing only
a small number of products (ERT 1984).  The six major gr'ups of  operations  and
processes  in  a petroleum ref'nery are:  1) storage of crude oil intermediates
and  final  products;  2)  fractionation  such as distillative  separation   and
vacuum  fractionation;  3)  decomposition  such   as thermal  cracking,  catalytic
cracking,  and hydrocracking; 4)  hyd-ocarbon rebuilding and rearrangement suet
as  polymerization,  alkylation. rearming, and  isomeriration;  5) extraction
such  as  solvent  refining and solvenc  dewaxing; and  6)  product  finishing such
as d.-ying  and sweetening, Jube oil  finishing, blending,  and  packing  (Hornick
et al. 1983).

Haste Characteristics

     Crude oil   is  the  raw  feedstock  for   all  of  the  refinery  process
operations.   Portions of the crude oil and the refined products  are eventually
discharged  as  wastes, either  directly  from a refinery process or  to  the
wastewater treatment plant (ERT 1984!

      Those  refinery wastes known to  be  land  treated  are listed in Table  13.
Of the listed wastes, five contrib'ite over 90  percent of  the estimated  oil  and
solids content applied to land treatment facilities.  Thesp wastes include  API
separator  sludge  (K051),  dissolved  air  flotation  float (K048),  slop  oil
emulsion solids  (K049), wastewater treatment  sludge  (nonlisted), and nonleaded
tank bottoms  (nonlisted).

      The two  wiste streams investigated for   land tr<-*tment potential  in  this
study  were API  separator sludge  (K051) and slop oil  emulsion  solids  (K049).
API  spparatur sludge  is the sludge  generated  in  the  oil/waier/solids  (API)
separator.   AF'I  separators, are  usually connected to  the ref nery oily  water
sewer.   Ylop  oil  emulsion solids are  the  residuals  left  in  the  emulsion  laysr
after  treatment   in  the  slop  oil  tank,  i.e., the  emulsion  that cannot be
broken.

      Refinery waf.tes vary considerably in physical  composition,  depending  upon
the pelroleuir product being  produced  and  according to waste type, as  shown In
Table  14.   Overrash and  Pal  (1979)  summarized the chemical composition  of 12
API refinery  wastes  (Table 15).


                                       42

-------
         TABLE  13.  REFINERY WASTES KNOWN TO BE LAND TREATED AND RELATIVE PERCENTAGES OF EACH
                                WASTE  WHICH  ARE LAND TREATED  (ERT  1984)
Listed* Hazardous*
Hazardous Waste
Waste Category Waste Number
Dissolved Air Flotation Float
API Separator Sludge
Slop Oil Emulsion Solids
Heat Exchange Bundle
Cleaning Sludge
Tank Bottoms
(leaded products)
Wastewater Treatment Sludge
Storm Wat?r Runoff Silt
Spent Filter Clays
Tank Bottoms4'
(nonleaded products)
Fluid Catalytic Cracking
Catalyst Fines
Spent Catalysts
Coor.nq Tower Sludge-..*
Chemical Precipitation Sludges
Neutralized MF Alkyletion
Sludge
Yes KQ48
Yes KG51
*es MW9
Yes K050

Yes K052

No
No
No
No

No

No
No
No
No

Estimated X Each
Waste Constitutes
of Totals which
Residue from are Land Treated
Known To Wasteweter
Be Land Treatment
Treated? Process
Yes
Yas
Yes
Yes

Yes

Yes
Yes
Yes
Yes

Yes

Yes
Yes

Yes

Yes
Yes
Yes
Yes

No

Yes
Yes
No
No

No

No
Yes
Yes


Oil
Basis
18.68
40.32
14.57
0.01

0.09

7.18
N.D.
0.36
18.35

0.05

0.01
0.04
N.D.
0.30

Oil and
Solids
Basis
12.66
36.46
9.24
G.08

0.19

17.11
N.D.

17. /O

2.06

0.61
1.22
N.D.
1.81

*40 CFR 261.
^Includes crudes intermediates and oroduct  storage tank*.
'includes once through cooling waters sludge.
N.D. - No Data.

-------
             TABLE 14.   PHYSICAL COMPOSITION OF REFUEHY WASTES
                         (ENGINEERING SCIENCE 1976)
     Waste Type                        Typical  Composition, Percent
                        Oil or Hydrocarbon           HaterSolids
API Separator
Tank Bottoms
Air Flotation Frot'i
Biological Treatment
Sludges
Cooling Tower Sludge
Spent Treatment Clay
Waste Lime Sludge
8
60
7
3

1
17
0
73
37
8P
92

74
9
73
19
3
5
5

25
74
27
              TABLE 15.  COMPOSITION  OF 12 API REFINERY WASTES
                           (OVERCASH AND PAL  1979)
                                  Minimum          Maximum          Average
Sulfides (mg/1)
Phenol (mg/1)
BOD (mg/1)
COD (mg/1)
PH
OH (.ig/1!
1.3
7.6
97
140
7.1
23
38
61
280
640
9.5
130
8.8
27
160
320
8.4
57
     ERT,  Inc.  (1984)  conducted  a  literature  review  of  Appendix  VIII
constituents  that  may  be present  in  petroleum  wastes for  the  American
Petroleum Institute.   They Identified three gtneral  classes of constituents:
1)  Appendix  VIII constituents  known  to be  present;  2}  Appendix  VIII
constituents suspected to  be  present; and  3) Appendix  VIII  constituents
expected not be  present.   The results of  these  investigations  are  shown  in
Table 16.  The  U.S. EPA has defined a  list of Appendix  VIII  compounds expected
to be  present  in petroleum  refinery wastes;  this  list is  presented  in Table
17.

Treatment of Petroleum Refinery Wastes in Soil  Systems—

     The petroleum industry has documented its experience with  land-farming in
the  open literature  more  extensively than most  others  (Corey 1982).   The
technique is preferred by the  industry for the management of waste sludges and
petroleum-containing  solutions because  of  the  minimum energy  required  for
implementation  and operation.   The industry has considered and obtained data
on  decomposition  rate,  vegetative response,  odor,   and  flammability.


                                      44

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           TABLE 16.   CATEGORIES FOR APPENDIX VI11  CONSTITUENTS  IN  REFINERY
                    WASTES WHICH ARE LAND TREATED*  (ERT, INC. 1984)
        Known  to be
          Present
       Suspected  to be
            Present
 Expected not to
   be Present
Arsenic
Benzene
Bis(2-ethylhexyl}phthalate
Butyl  benzyl pht'ialate
Benz(a)anthracen»?
Benzo{a)pyrene
Benzojkjfluoranthrene
Beryllium
Cadmium
Chromium
Chrysene
Copper*
Cyanide
Fluoranthene
Lead
Mercury
Naphthalene
Nickel
Pnenol
Selenium
Toluene
Vanadium
Zinc*
Anthracene
Antimony
Barium
Benz(c)acridine
Benzo{b)fluoranfhrene
Benzoijjfluoranthrene
Cobolt
Di&er»z(a,h)acridine
niberu(a,j)acridine
Dibenz(a,n)antte
Nitrobenzene
4-nitrophe"ol
P-cresol
Phenanthrene
Tetraethyl Lead
All other
EPA Appendix VIII
Constituents
*A list of constituents suspected to be present is currently being developed by
 EPA as Of 5/84.
'Non-Appendix V1I1 constituents.
                                      45

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         TABLE 17.  CONSTITUENTS OF PETROLEUM REFINING WASTES
                      (AS APPROVED BY U.S. EPA)
Metals

Antimony
Arsenic
Barium
Beryl 1i urn
Cadmium
Chromium
Cobalt
Lead
Mercury
Nickel
Selenium
Vanadium

Volatiles

Benzene
Carbon disulfide
Chlorobenzene
Chloroform
1,2-Dichloroethane
1,4-Dioxane
Ethyl benzene
Ethylene dibromide
Methyl ethyl ketone
Styrene
Toluene
Xylene
Semivolatile Base/Neutral
Extractable Compounds
Benzo(b)fluoranthene
Benzo(k )f 1uoranthene
Benzol a)pyrene
Bis{2-ethylhexyl) phthalace
Butyl  benzyl  phthalate
Chrysene
Dibenz(a,h)acridine
Dibenz(a,h)anthracene
Dichlorobenzenes
Diethyl phthalate
7,12-Dimethylbenz(a)anthracene
Dimethyl  phthalate
Di(n)buty]  phthalate
Oi(n)octyl  pi;*halate
Fluoranthene
indene
Methyl  chrysene
1-Methyl  naphthalene
Naphthalene
PhenMtrrene
Pyrene
Pyridine
Quinoline

SemivoTatile Acid-Extract able
Compounds'

Benzenethiol
Cresols
2,4-Dimethylphenol
2,4-Dinitrophenol
4-Hitrcphenol
Phenol
Anthracene
Berzo(a)anthracene
                                  46

-------
 Application  rates generally  range  from  less than 200 barrels/year/acre to more
 than 600  barrels/year/acre.    The  frequency of  application of  oi]y  wastes
 v-iries  widely  from  only  one  application per  year  to  a site  to  multiple
 applications as  frequently as once per week.   The  decomposition  rate is site
 specific  but has been  reported  as high as 50  percent  per  year (Corey 1982).
 Subsurface samples  indicate  that  if  land treatment  units  are  operated
 correctly, neitner heavy metals nor oil will migrate appreciably.  Trace metal
 analysis  of  vegetation growing on  oiled areas  is generally similar to control
 locations.  Odor is  reduced and  minimal  once  the  oily  waste  is  blended with
 the  soil.  After the  wastes are mixed with the soil  they  are generally not
 flammable.  A review of land treatment of refinery wastes, Sludge Farming;   A
 Technique for the  Disposal of Oil  Refinery Hastes (CONCAWE 1980) was prepared
 by the  Oil Companies'  International  Study Group for Conservation  of Clean Air
 and  Water-Europe to  evaluate the  potential  for  land  treatment  of  refinery
 wastes  in Europe.   CONCAWE  concluded that,  provided  simple  safeguards  are
 observed, sludge farming  is  ecologically  the most  suitable and cost effective
 method  for  disposal  of  normal  oil  sludges  and for  soil  that  has  been
 accidentally contaminated with oil.

     PAH  compounds  are  important  constituents  in  petroleum  refinery  wastes
 (Tables  16 and 17}  as  well  as in wood  preserving  wastes, and the reader  is
 referred  to  the  discussion of  the fate  and  significance of PAHs  in  soil
 systems'presented previously for wood preserving wastes.

 Toxicological  Significance of Petroleum Wastes--

     API  separator  sludge/slop oil  emulsion  solids and  oil-containing  storm
 water runoff have  been shown to contain mutagenic  compounds  (Donnelly  et al.
 1985).  Organic  compounds  were extracted  from  each waste with dichloromethane
 and  partitioned by  liquid-liquid  extraction  into acid,  base,   and  neutral
 fractions.   A battery of short term  bioassays were used  to  detect  various
 types of  genotoxic damage.   Each chemical  fraction was  tested  in  four strains
 of Salmonella typhimurium to detect point mutations, six  strains  of  Bacillus
 subtil is  to  detect  ""lethal damage  to ONA, and  haploid  and  diploid forms   of
 Aspergfllus nidulans to detect point mutations  and various types of chromosome
 damage.   Results  of  these  biological  analyses  indicated  the   presence  of
 genotoxic  compounds in all three fractions of each waste.

     Brown and Donnelly (1984) conducted a study of the  mutagenic  potential  of
 runoff  and leachate  water  fror. petroleum API separator  sludge-amended  soils,
 using the Salmonella  microsome  assay  and  the Baci11 us  subtilis DNA  repair
 assay.   Mutagenic  activity  was  detected  in a limited  number  of runoff  and
 leachate  samples, but  greater  amounts  of mutagenic activity were  detected  in
 the  runoff water.    The mutagenic  activity from  leachate and  runoff  water
decreased  with time  following  waste  application  in  two of  the  three  soils
used.   The activity  in the third  soil  did not decrease  over  the 3  years  of
observation.

     The  toxiclty of  petroleum refinery effluents  to environmental organisms
 is highly dependent  upon the waste streams, which may vary  widely in  chemical
composition.    Data  suggest  that  many effluents,  especially those that  have
received  primary treatment only,  are toxic at  their discharge  point  (CONCAWE
1979).  CONCAWE  (1979) summarized  the environmental lexicological effects  of
petroleum  refinery  effluents  and  found that  in general,  oils  increase  in

                                       47

-------
toxicity with  levels  of  low-ooiMng  compounds,  unsaturated compounds,  and
aromatics.  Also  aromatics with  increased  numbers  of  alkyl  substituents  have
higher toxicities,  and  toxicity increases along  the  series alkanes-alkenes-
aromatics.   Cycloalkanes and cycloalkenes  appear to be  more  toxic  than
alkanes.

     Other cmponents of  petroleum refinery effluents, such as phenols, sulfur
compounds, cyanides, and  metals  may also  contribute  to  the toxicity  of  the
effluent.  A review of  the toxicity  of  these compounds as well as the toxicity
of  oils  is presented  in a  report  prepared  for  the Council   of  European
Communities by  CONCAWE's  Water  Pollution  Special  Task  Force No. 8 (CONCAWE
1979).

     Human  health  effects  of  specific compounds  often  found   in  petroleum
refinery  effluents  may be found  in the U.S.  EPA Health  Effects  Assessment
documents for  PAHs  (U.S. EPA 1984e),  benzo(a)pyrene,  (U.S. EPA  1984b),  and
coal tars (U.S. EPA 1984,-).


Degradation, Transformation,  and  Immobilization of
Petroleum Refinery Wastes—

     A summary  of  la.id  treatment  practices in  the  petroleum  industry  was
published by API  (API  1983).   Results  of this study showed  high  oil  removal
efficiencies T"Ar the 14 full  scale and  4  pilot scale facilities reviewed.   Oil
reductions at  the full  scale facilities ranged from  0.09  - 0.86 Ib  of  oil/
ft3/degradation month  and were  directly related  to  the  oil loading  rates,
which  ranged  from  0.16  to  1.12 Ibs  of oil/ft3/degradation month.   Slowly
degradable  fractions  were retained  within the zone  of  incorporation.   The
saturate and  light  aromatic fr art  ions  degraded  at a  faster  rate than  the
heavier  fractions.   Lead  and  chromium  accumulated  above  background  in  the
surface  soils  at  some  of the  land treatment facilities  investigated.   The
metals were  attenuated with  depth  and rarely moved beyond  the zone  of
incorporation, generally reaching background concentrations within 1 to 3 feet
below the zone of incorporation.

     Martin  and  Sims  (1984)  and  Martin et  al. (1986)  investigated  land
treatment  practices  in the  petroleum  refining industry.    Sites  for  land
treatment were  characterized  by a variety  of  climate,  soil,  and physical
characteristics  that were  suitable  for  land  treatment.   Maximum waste
application  rates  ranged  from 0.004  weight  percent  of  oil  in  soil  per
application to 8  percent per application.  Five facilities were  identified as
practicing high intensity land treatment, (defined as a minimum of 4.0 percent
oil/soil   for  climatic  regions  where seasonal fluctuations  cause  the  average
minimum  air  temperature  to  fall  below  9.9°C.  and 8.0 percent  oil/soil  for
climatic regions  where the average minimum air temperature is greater than or
equal  to  9.9°C).   Seventy percent  of  the  facilities  added  amendments
(fertilizer and lime) to  the treatment soil.   Calculations using a predicted
half-life  of  304 days  for  oil   in  soil  showed that  expected maximum weight
percentage of oil  in the  soil after treatment ranged  between 2.9 percent and
19 percent, with an average of  7.9 percent.  Calculations to  predict the total
inorganic constituent  loading  over  a  projected  30 year site  life indicated
that levels would be below suggested limits (U.S.  EPA  1983).
                                       48

-------
     Pal and Overcash (1980), using available data on petroleum refinery  solid
wastes, performed an assessment of land treatment technology for these  wastes.
Using  two  representative soil types  and  the  composite  waste characterization
shown  in Table 18, the land-limiting  waste constituent (LLC) (i.e.,  that  waste
constituent  requiring  the  largest   land  area  for  assimilation  in the  soil
system) was determined to be fluoride.  Elimination of one  waste stream,  the
neutralized  HF  alkylation  sludge,  from  the  land  treatment  unit  eliminated
nearly  all  of the  fluoride, and  selenium,  chromium,  and  spent filter  clay
became the LLCs.

     The addition of oily wastes to a soil may change its chemical,  biological
and  physical  properties.    Initially,  oil  applications  tend  to  produce  a
hydrophobia effect in soil,  resulting in  a decreased infiltration rate.   This
effect  is  due to  the oil   itself  and to  the  accumulation of  hydrophobia
mucilaginous  substances generated  by increased  microbial  growth (Overcash  and
Pal  1979).    Long-term effects  of the  applied  oil may be  beneficial.
Aggregation, soil porosity,  and water holding capacity all increase  while  bulk
density decreases (Hornick et al. 1983).

     As the oil  content  of  the soil  decreases at  a  land  treatment facility,
there  is  an  increase  in heavy aromatics  and  asphaltenes  compared  to the
saturates and  light  aromatic  hydrocarbon fraction of  the  applied  oil
(Huddleston and Creswell  1976).  The  heavy aromatics and asphaltenes do appear
to  degrade  but   at  much slower rates than  the overall  oil   reduction  rate
(Weldon 1982).    Table  19 shows   the  relative  order of  resistance of
hydrocarbons  to  biodegradation,  as reported by Fredericks (1966).   Perry  and
Cerniglia  (1973)  reported  that the  recalcitrance of  various  hydrocarbon
substrated increased in the  following  order:   normal   alkanes CIQ  -  Cig;
straight-chain alkanes C\2  ~ ^18»  gases  C?  -  04;  alkanes €5  - Cg; branched
alkanes to  12 carbons;  alkenes  £3   -  GU;  branched alkenes;  aromatics;  and
cycloalkanes.

     Brown et  al. (1981) conducted   a  study  of degradation of  API  separator
sludge  from  a petroleum refinery in four different soils  at  four moisture
levels.   The  greatest amount  of degradation was seen  in a sandy  clay  soil,
intermediate amounts in a clay and a  sandy  loam soil, and  the  least in a  clay
soil.    In  the  sandy  clay  soil,  the  biodegradation  rate  generally  doubled
between 10°  and   30°C  but  decreased  at 40°C.   At  30°C,  after 180 days, 45
percent of  the  refinery  waste  (measured as  total  carbon or  residual
hydrocarbon)  was  degraded,  but  at 40CC,  after 180 days, only  36 percent  was
degraded.    Addition of  fertilizer   nutrients  (nitrogen,  phosphorus,  and
potassium) did  not  increase biodegradation.   Biodegradation  rate  increased
with  increased  application  rates  of the sludge.   Moisture was a dominant
factor  only  at  excessively wet or  dry conditions.  Moisture content had  a
greater influence on biodegradation at 10°C than at higher temperatures.

     Dibble and  Bartha  (1979)  investigated  the  effects  of  environmental
parameters on  the  biodegradation of  oil sludge (as measured  by CO2 evolution
and  analysis  of  hydrocarbons) in  a  loam  soil.    The  environmental  parameters
investigated  included  incubation  temperature,  pH,  soil moisture,  waste
application rate  and frequency,  and the  addition of  mineral nutrients,
micronutrienti and  organic  supplements (sewage sludge).   They  concluded  that
oil sludge biodegradation was optimal at  a  soil water holding  capacity of  30-
90  percent,  a  pH of  7.5  to  7.8,  C:N  and  C:P ratios of  60:1   and 800:1,

                                      49

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   TABLE   18.   WASTE CHARACTERIZATION rOR AGGREGATE  OF SIXTEEN1 SOLID WASTE
    STREAMS  FROM A CATEGORY IV PETROLEUM REFINERY (PAL  AND OVERCASH  1980)
Parameter
Total Solids (TS)
Oil
Nitrogen (N)
Phosphorus (P)
Potassium (K)
Calcium (Ca)
Sodium (Na)
Magnesium (Mg)
Cyanide (CN)
Pheno1
Selenium (Se)
Arsenic (As)
Mercury (Hg)
Beryllium (Be)
Vanadium (V)
Chromium (Cr)
Concentration*
mg/1
500,000
71,000
200
110
40
300
200
80
0.6'.
3.6
1
1.3
0.26
0.06
15.3
58
Parameter
Manganese (Mn)
Cobalt (Co)
Nickel (Ni)
Zinc (Zn)
Silver (Ag)
Cadmium (Cd)
Lead (Pb)
Molybdenum
Boron
Fluoride
Chloride
8enz[a]pyrene
Spent Filter Clay
PH (S.U.)
Chemical Oxygen
Demand
Volume
Concentration*
mg/1
42
l.B
14
53
0.35
0.17
9.3
1.3
0.015
530
99
0.08
70.000
6.5 - 8.2
130,000
15 x 106 fc
*Uet sludge basis.
                                     50

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        TABLE   19.    RELATIVE  RESISTANCE  OF HYDROCARBONS TO BIOLOGICAL
                         OXIDATION  (FREDERICKS !
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respectively,  and  a  temperature of  20°C  or  above.   The  addition  of
nncronutrienis and organic supplements in the  form  of  sewage sludge did not
enhance  biodegradation.    The  sewaqe  sludge  inhabited hydrocarbon
biodegradation.   Breakdown  of   the  saturated  hydrocarbon  (alkane  and
cyclcalkane)  fraction was  the  highest at low  application  rates,  but higher
application rates  enhanced  biodegradation  of  the  aromatic  and  asphaltic
fractions.   Frequent  small  applications  (four small  loadings)  resulted  in
higher biodegradation rates  and total hydrocarbon biodegradation  tnan  a single
large application.   The authors suggested that a  loading rate of two 100,COO
liters/hectare  or  four  50,000   liters/hectare  oil   sludge  hydrocarbon
applications  per  growing  season  mc.y be  appropriate  for most temperate zone
disposal sites.

     Kincannon (1972) conducted  a  study of degradation of three  types of oily
sludges in a  sandy clay loam soil.   The wastes  were  applied to plots  that had
been previously used  for oily waste disposal.   Residual  oil  levels  before the
beginning of  the study  we-e  about 10 percent, and nitrogen and phosphorus were
added as  amendments.   Degradation  rates  rangpo from 0.167  to 1.79 oounds of
oil per month per cubic  feet of  soil.  For crude oil  tank bottoms (containing
a  variety of hydrocarbon  types) and  a  high molecular  weight  fuel  oil
(containing olefinic  and  aromatic compounds),  both  aromatic and  saturated
hydrocarbons  were  reduced  through time, but  for Lhe  waxy raffinate :ludge
(containing highly paraffinic  components),  only  the  saturate fraction  was
reduced.  The optimum fertilization program  was determined  as  the maintenance
of  10-50  ppm  ammonium  and/or nitrate  and a slight excess  level of  potassium
and phosphorus in the soil.   The major species of microorganisms  degrading the
hydrocarbon substrate were  the genera Pseudomonas,  Flavobacterium.  Nocardij.
Corynebactermm,  and  Arthrobacter.    Neither  thetype  of  oil  sludge,
temperature or addition  of  fertilizer affected the types of  organisms  present,
though both the  type of sludge and fertilizer affected  the  total  number of
aerobic bacteria present in  the soil.

     Meyers and  Huddleston  (1979)  investigated the degradation of  a  combined
oily sludge consisting  of  API separator  sludge,  tank  bottoms,  and slop oil at
a  land-farm.    Three  applications were studied  in plots  with and without
vegetative cover:  a single loading, loading one time each year for  2 years,
and loading one time  each  year for  3 years.   Agricultural ammonium nitrate and
phosphate were  added to all test   plots.   Selected  results of the  study ar>
shown  in  Table 20.  All  three oil fractions  were  shown to degrade.  Also,
tilling was shown to  increase biodegradation, likely due to  increased  aeration
end microbial/oil rontact.

     A 1.2EO-day laboratory simulation of the "landfarm ing"  process  by Bossert
et  al.  (1984; explored  the fate  in  soil   of  PAHs  and  total  extractable
hydrocarbon residues originating  from the  disposal  of an  oily sludge.   In
addition  to the  measurement  of  CO? evolution,  periodic analysis of  ^AHs and
hydrocarbons  monitored  b'odegradafion  activity.   The  estimation  of carbon
balance and of soil organic  matter  assessed the fate of  residual  hydrocarbons.
Seven  sludge  applications  during  a 920-day  active disposal  period  were
followed  by  a  360-day  inactive  "closure"  period with  no further  sludge
applications.    A burst of  CO?  evolution  followed each sludge  addition, but
subscantial amounts  of  undegraded  hydrocarbons remained  at  the end of the
study.   Hydrocarbon  accumu'atiot'  did  not  inhibit  biodegradation performance.
Conversion of hydrocarbons to  CO? predominated  during  active  disposal;


                                     52

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 incorporation into soil organic matter predominated during  the  closure  period.
 In  this sludge,  the predominant  PAHs  were  deqraded  more  completely  (85
 percent) than  total  hydrocarbons.   Both biodegradation and abiotic  losses  of
 three-  and four-ring PAHs contributed to this result.   Some PAHs  with five  and
 six  rings were  more  persistent,  but  these  constituted only a  small  portion  of
 the  PAHs in the sludge.

     TABLE  20.   RESULTS  OF  DEGRADATION OF PETROLEUM WASTES AT A LAND-FARM
                 AFTER 25 MONTHS  (MEYERS AND HIJDDLESTON 1979)
Plot                      	      Degradation (%)  	
                       DTIParaffinsAromaticResins and
                                                                 Asphaltenes
Single loading.
No Vegetative Cover
Two Yearly Loadings,
No Vegetative Cover
Two Yearly Loadings,
58

30

43
71

44

50
47

29

44
37

12

30
 Vegetative Cover of
 Wheat and Bermuda
 Grass
     'jnyder  et  al.  (1976)  studied the  disposal  of  waste  oil  re-refining
residues by land farming.  The residues consisted  of  a sludge  and an oil-water
emulsion  (approximately  60-65 percent  water)  containing  various  metals  at
concentrations of 3 - 400ug/g.  For the plots  treated with oil, the mlcrobial
respiration rates were much higher than '.or  the untreated  plots.

     Snyder et  al.  (1976),  Skujins  and McDonald (1983),  and Skujins  et al.
(1983) reported  on  the degradation of  waste oil  In  a  semi-arid  region  soil
near the  Great  Salt  Lake,  Utah.    An  oil  emulsion  (45.7  percent  oil)  and  a
water phase  (formed  in a waste oil  lagoon  between  the  surface  oil  emulsion
layer and  the  bottom sludge  sediment)  were  treated  by a  land-farming method
employing neutralization of the waste and supplemental fertilization.  By the
end of the  first year following  the application  of  the  oily waste,  the mean
value of oil degradation was 37 percent.  During the  second,  third and fourth
years, 81,  84,  and 91  percent,  respectively,  of the  added  oil  was  degraded
(S'-cujins  and  McDonald  1983).   There  was no  significant  difference  in
degradation rates among  the  various  treatments with  respect to the amount of
oil  and  nitrogen fertilizer  (i.e.,  the C/N  ratio)  applied to  the  soil.
Maximum rates of degradation  occurred  during the  moist,  warm spring  seasons.
The  area  was successfully  revegetated,  but the  plants  contained  elevated
levels of metals  in  comparison to  plants from control  areas (Skujins  et al.
1983).   The  Investigators  suggested that reuse of  the disposal  area  may be
limited by increased  metal  availability to plants.
                                      53

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     A study by Westlake et al. (1977) of the degradation of refined oil in a
fertilized soil  of the  boreal  region of  the  Northwest  Territories,  Canada
showed  that  the  addition  of o-1-utilizing  bacteria to  soil  plots  did not
increase the  number of bacteria  present compared  to plots not seeded with oil-
utilizing bacteria.    All  plots  that  received an  application of  oil  and
fertilizers with or without bacterial  seeding  showed  a two-log  increase  in the
number  of viable bacteria  present  within  22 days  of  the  initiation  of the
experiment.   The high  bacterial  numbers persisted  for almost  three  years
before decreasing to levels present in unfertilized oil-soaked plots.  Little
change was noted in the aromatic contents of both fertilized and unfertilized
plots, but the n-alkar.e components of the saturate hydrocaroon fractions were
shown to be degradable in the fertilized  plots.

     An  industrial  oily waste was applied  to  field   plots  in  New  York  to
determine  the  degradation   and  immobilization of waste  constituents and  to
determine the impact of the wastes on soil biota  (Loehr et  al. 1985).  Wastes
were applied three times *o the test plots at loading rates ranging from 0.17
to 0.5  kg oil/m2  or from 0.09 wt  percent -  5.25 wt  percent oil in soil.  The
waste application increased soil pH and volatile materials.  Half-lives of the
oil  ranged  from 260  to  400 days.   Refractory  fractions of  the applied oil
ranged  from  20  to  50  percent,  but did  not  appear  to  adversely affect soil
biota.    Naphthalenes,  alkanes,  and specific  aromatics were  lost  rapidly,
especially in the warmer months, with half lives generally  less tnan 30 days.
The waste  applications reduced  numbers and  biomass  of  earthworms and numbers
and kinds  of microarthropods, but  both  types of biota  were able to recover.
Earthworms did  not accumulate specific waste  organic compounds.

     Results at  pilot scale  facilities have  shown  that  accumulated  oil
continues to degrade for several years after oil  applications have terminated
and the  land treatment facility is closed,  even with  no  efforts  to enhance
degradation  ;Wei don 1982).   A 50  percent reduction  in soil oil concentration
was observed ever 2-year periods at closed pilot scale facilities that had oil
concentrations  in the  soil  greater  than 3 percent at  the time of closure.

     A  15-month  closure  evaluation  study  was  performed for  three  land
treatment sites that had ceased  application  of refinery  wastes  for 6 months, 9
months,  and  6  years (Streebin et  al.  1984).    Considerable variation  In oil
content  existed  among the  three  sites.   Concentrations of oil  greater than
background levels were found as  deep as  45-50 cm at all three sites.  Average
oil content -emained relatively constant throughout the study, perhaps due to
long periods of wet or dry soil, low soil nitrogen,  and  presence  of persist?nt
hydrocarbons.  Only traces of organic  priority pollutants were  found below the
zone of  incorporation.   Metals  were fixed  and/or sorbed In the  top  25 cm of
soil at all sites.

     A  study of  the distribution  of inorganic constituents in soil following
land treatment  of refinery wastes at five sites was conducted by Brown et al.
(1985).   At  one site, where  soil  pH was less than  6.5, two metals, chromium
and lead,  were  found  below the  zone of   incorporation at concentrations  above
untreated  soil.   Metal levels within the  zone  of Incorporation at all  sites
were at  levels considered common for natural  soils.   A wide variation  In oil
content  In the  zone of  incorporation was noted,  I.e.,  from 3.4 percent to 8
percent  (K.W. Brown and Associates  1981).   The  oil  content below the zone of
incorporation  Decreased  rapidly with depth.   The  maximum extent  of oil

                                      54

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migration for the facilities reviewed  >*as  less  than  1.5 feet below the zone of
incorporation


Volatilization o* Petroleum Refinery Wastes--

     Dupont  (I986b)  evaluated volatile  air  emissions  from  Ian*!  treatment
systems  iiSinq  API  separator  sludie  and  slop oil  emulsion  solids applied to
laboratory  soil  microcosms.   Data  indicate  that  vapor  partitioning  and
retardation  by soil  organic  matter were  of  minor  importance  in  vapor soil
transport processes, and that  volatile  organic  vapor jwi'i diffusion could be
described by t'-» pnysical environment throuqh which the vapor travels.  Using
an air emission predictive model (the Thibodeaux-Hwang AERR model (Thibodeaux
and Hwang  1982)),  subsurface  waste  application  produced a  two-  to ten-fold
decrease in  predicted  emission rates  compared, to  surface application.   This
reduced  emission  rate  persisted  for  the 80  to  100  hours over  which  the
experimental runs were conducted.

     Radian  Corporation  (Wetherold  and  Balfour  1986) has  also  conducted
studies  to  evaluate  air  emissions  from  land treatment of oily  sludges.  The
studies  included   laboratory  land treatment simulation experiments,  field
studies  at  a waste  treatment facility,  and field studies at  a refinery land
treatment site.  Results of these  studies include:   emission rates reach their
maximum  in  a relatively short time  after surface  application  of a volatile
sludge;  the most  significant parameters  affecting the emission  rate from
surface  applications of sludqe are the loading and  the sludge volatility; the
Thibodeaux-Hwang  emission  model  appears »o  agree  reasonably  well  with  the
measured rates for  some  selected compounds;  the   Thibodeaux-Hwang  emission
model  has  not been  generally applicable to multicomponent mixtures, probably
because  of the uncertainty  in  defining accurate multicomponent  parameters for
use in the model; during  land  treatment  a  significant fraction  of the applied
VOC is emitted from  the  surface application  of oily sludge; tilling causes  a
significant, short-term increase in the VOC emission rate from  land treatment
sites.   API  (1983)  has  suggested that  subsurface  injection may be  used to
reduce volatile  air emissions if  tilling does not  occur within  four  to six
days of  application.

     A description of the flux chamber/solid  sorbent monitoring  system used to
evaluate air emissions from land treatment facilities was presented by  Oupont
(1986a).
                                      55

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                                  SECTION 5

                           RESULTS AND DISCUSSION


QUALITY ASSURANCE/QUALITY CONTROL

     A quality  assurance (QA) project  plan  was developed by  the  Utah Water
Research Laboratory (UWRL), approved by the  U.S. EPA,  and implemented by the
UWRL  to  ensure that  data generated  in  this research  investiqation  were of
adequate quality and quantity to support the conclusions being drawn from the
study.  Key elements of this  plan  included the following activities during the
performance of the project:

     (1)  Use  of  U.S.  EPA-approved or  other  standard  methodology  for
analytical measurements and sample preservation  and collection

     (2)  Documentation of modifications of  standard procedures

     (3)  Thorough description of  experimental procedures

     (4)  Use of  replicate analyses  and  positive  and  negative  controls for
          experimental and analytical  procedures

     (5)  Analysis of subsamples of selected samples

     (6)  Use of U.S. EPA quality  assurance  audit samples

     (7)  Calculation of mean values,  standard deviations, and coefficients of
          variation

     (8)  Use of  statistical  procedures for evaluation and interpretation of
          data

     (9)  Use of standardized data collection formats  and  reporting,  including
          the use of bound laboratory notebooks

     (10) Periodic maintenance and calibration of laboratory  instrumentation

     (11) Participation  in U.S. EPA performance  evaluation study

     (12) Systems  audit by  U.S.  EPA RSKERL   QA  project officer  (i.e.,  a
          qualitative, on-«ite review to ensure that data are being  collected
          in accordance with the QA project  plan).
                                       56

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     Descriptions of quality control !(JC) activities for  specific expenir?ntal
and analytical  procedures are included with the description of the procedure
in this report,  or  in  the referenced procedure.   Results of  QC  activities are
included with presentations of results for specific experimental  studies and
analytical  measurements.


WASTE CHARACTERIZATION

Introduction

     Demonstration of  the land treatability of  a waste  beqins  with waste
characterization.    A  waste  characterization  scheme  should  delineate  the
various  waste  components that must  be managed  to  preclude adverse  health.
safety,  or  environmental impact from  land treatment  of  a given  hazardous
waste.    Characterization  provides  the basis both  for  evaluating  thp
feasibility of  using  land  treatment  technology  and assuring that  the
operational system  can  be adequately monitored.

     For  each  hazardous  waste evaluated during this project,  a waste
characterization  program  was  conducted.   Representative composite waste
samples were obtained.   Each waste was characterized for general physical  and
chemical parameters and  for  individual  organic  and  inorganic constituents of
concern.   Table  21 contains  a  1'st  of general waste  characterization
parameters  that   were determined for  each  waste.   Specific  organic  and
inorganic constituents of  concern  for  each waste  were selected as  monitoring
parameters in consultation with  the U.S.  EPA project officer.


                TABLE  21.  WASTE CHARACTERIZATION PARAMETERS
     Density                            Total organic carbon
     Water content                      Volatile organic constituents
     Solids content (Residue)           Extractable organics
     Ash content (Residue)              Metals
     pH                                 OH  and grease
     Waste characterization procedures were based on procedures given in Test
 Methods  for Evaluating  Solid Wastes:   Physical/Chemical  Methods.   Second
 Edition  (U.S.  EPA  1982).   However,  since the  wastes  chosen  were complex
 mixtures  that  exhibited unique  properties,  compositions,  and  problems with
 respect  to  waste characterization, modification of  some  standard procedures
 and  the use  of additional referenced procedures  were  somet-mes necessary.  All
 standard  procedures, modifications of standard procedures,  and additional and
 alternative  procedures used durinq this project  are documented  in this report.
                                     57

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Materials and Methods

Selection of Hastes--

     Four listed hazardous wastes were  selected  for  study  in consultation with
the U.S. EPA project officer.  The wastes  chosen  are produced in high volume
in  the  U.S.,   contain  numerous  organic  and  inorganic  constituents,  and
represent   a  broad  spectrum of  physical,  chemical,  and  lexicological
characteristics.   The  specific wastes  selected  for  study are listed below  in
Table 22.

             TABLE 22.   HAZARDOUS WASTES  SELECTED  (-OR  EVALUATION
     Waste                                        EPA Hazardous Waste No.
Petroleum Refinery Wastes
     API Separator Sludge                                  K051
     Slop Oil Emulsion Solids                              K049
Wood Preserving Wastes
     Creosote                                              K001
     Pentachlorophenol                                     K001
     Petroleum Uastes--

     API  separator  sludge--(KOSl) This  waste  is generated  from  the primary
settling of wastewaters that enter the oily water  sewer.  API separator sludge
typically consists of  approximately 53 percent  water,  23 percent  oil, and 24
percent solids  (Brown, K.  W., and Associates  1980).   The solids are largely
sand and  coarse silt, but  also  may  contain significant  quantities  of heavy
metals  such  as the metals  that  cause this  waste to  be  listed as hazardous
(i.e., chromium and lead).   The heavy oils that  settle  in an  API separator and
become part of the bottom sludge  will  largely be composed of  heavy tars, large
multiple branched aliphatic compounds  (paraffins), polyaromatic hydrocarbons,
and coke fines.  The  proportions of the  oily material  in API separator sludge
which  are  tar-like,  paraffinic or polyaromatic are  largely  dependent on the
source  crude  being  refined.   The  amount of coke fines  in  the sludge should
increase as the amount of thermal cracking used  by the  refinery increases.

     Slop  oil  emulsion solids--(K049) This  waste  is  generated from  skfronno
the API separator.Slop oil  emulsion solids are typically  40  percent water,
43  percent  oil, and  12  percent   solids.   Chromium  and  lead  are  present in
significant concentrations in the solid phase and  are the reason this  waste is
listed as hazardous (Brown, K. W. and Associates 1980).
                                      58

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     Wood Preserving Nastes--

     CreogQte--(KOOl) Creosote  ib  a  distillate  from coal  tar made  by hiqh
temperature  carbonization  of  bituminous coil.   Creosote alone  or  ir.
combinatioi. with coal tar  or  petroleum is a major  preservative used  in wood
treatmen*   (Merrill  and   Wade  1985).    The principal  classes  of  organic
constituents  present  in creosote  wastes  are polyaromatic  hydrocarbons  and
phenolics.

     Pentachlorophenol  (PCP)—(K001)  Pentachlorophenol   Is  widely used  as  a
wood preservative.   PCP has  also  been used for  slime and algae coni.ro!.  The
combined PCP-creosote sludge  used  in  this  experimental investigation contained
polyaromatic hydrocarbons,  phenolics,  and  PCP.


Physical Characterization  of  Wastes--

     Density—

     The  density  of  a  liquid waste  can  be determined by weighing  a known
volume of  the waste in  water or other  liguid.   A water insoluble viscous or
solid  waste is weighed in a calibrated  flask containing  a known volume and
mass  of  watfr.   The water displaced  is  equivalent  to the  volume  of waste
material    added.    A similar technique  is used  for   the  analysis  of water
soluble  wastes by  replacing  water with  a  nonsolubil i zing  liquid  for  the
volumetric  displacement measurement.   In  this case a correction must be made
for the density of the solvent used.

     Water  Content—

     The water  content  of  each  waste was determined using  ASTM Method  D95-70
(Standard  Method  cf  Test  for Water  in  Petroleum  Products  and  Bituminous
Materials by  Distillation).  A summary of  the method is  presented  below.

     The waste  is  heated  under  reflux  with  a  water immiscible solvent which
co-distills  with  the water  in  the sample.  Condensed  solvent and water are
continuously  separated on  a trap.  The  wate»- settles  in the graduated section
of the trap and the solvent is returned to the  still.

     Residue—

     The term "residue" refers to  solid matter that is  suspended  or  dissolved
in water  or waste.   Total residue is  the  term  applied to the material  that
remains  after evaporation  of  a sample and its subsequent, drying  in an oven  at
a  defined  temperature  (103°C).   Total includes  "nonfilterable or suspended"
and the "dissolved  or filterable" residue.

     The  total  volatile suspended residue  is obtained  by-igniting the total
suspended  residue  at SSOoc.   Thp  test is used  to  obtain  an  approximation  of
the  amount  of organic matter present  in the  solid fraction of  the  waste.
                                    59

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Chemica1.  Characterization  of  Wastes--


     Inorganic Const ituents—

     Metals—A digest for  the analysis of the total metal content of slop oil
and API  separator  sludge  samples was  prepared  using  EPA Method 3030 for the
acid digestion  of  oils,  greases,  and waxes  (U.S.  EPA  1982).   Samples were
prepared-in  triplicate.   Standard  quality  control  procedures were followed,
including analysis  of digested EPA reference material  and  reagent blanks.  All
analyses  were  performed  using a  Perkin-Elmer  Model  6000 Inductively Coupled
Plasma (ICP).   Detection  limits for As,  Hq.  Se,  Cd,  Pb,  Ni,  and V  on the ICP
were  not  satisfactory  to define   environmentally  significant  levels.
Therefore, graphite furnace  atomic  absorption spectrophutometry (AA) was used
for the  analysis of Cd,  Pb,  Ni, and  V.   The As, Se, and Hg were analyzed by
atomic adsorption (AA) using  hydride generation.

     The  pentachlorophenol and creosote samples were digested  using  EPA Method
3050  (U.S.  EPA  1982)  for the  acid  digestion  of  sludges.    Quality control
procedures and specific methods for metal analysis  were  performed as described
above.   In addition, triplicate PCP and  creosote samples  were spiked with one
of two EPA reference materials before digestion to  determine  percent recovery.

     Organic Constituents—

     Total organic  carbon—The  total  organic carbon (TOC) conient  ot the API
separator sludge and creosote waste  was determined.   One gram of  waste (dry
weight)  was thoroughly mixed with sand in a  SPEX Ball Mill.   An aliquot  (0.01
to  1 g)  of  th-is waste/sand mixture was accurately  weighed out and placed into
a glass  ampule  along with 1C percent  hydrochloric  acid  (1 ml/gram of mixture)
and 200  mg of combusted copper oxide.   The  ampule was sealed and baked for  5
hrs at  550°C.   After cooling, the  contents  of  the ampule were  analyzed using
an Oceanography International (Model b24B) Carbon Analyzer.

     Oil  and grease—Oil  and grease are defined as  any material  recovered as  a
substance soluble   in  fluorocarbor  113.   The  oil  and grease rontent of each
waste was determined  using Method  413.1  (U.S.  EPA 1979).  In  this  procedure,
the oils and  greases are  extracted by direct contact with an organic  solvent,
fluorocarbon  113.    The  solvent is  separated  from  the  aqueous and/or  solid
phases,  dried  and evaporated  to  determine  the  extractaole  residue by
gravimetric  techniques.

     Volatile  organic constituents—The volatile  fraction  of  each waste was
prepared using  the  purge  and trap (Method 5030, U.S. EfA 1982).  A portion of
solid  or  liquid  waste was  dispersed in methanol  to  d'ssol/e the volatile
organic  constituents.   A portion  of  the methanol  solution  was combined  with
water  in a specially designed purging chamber.  Nitrogen was bubbled  through
the solution  at   ambient   temperature,  and  the  volatile components  were
transferred  from the aqueous phase to the  vapor phase.   The vapor  was swept
through  a TenaxR sorbent column  where  the  volatile  components were  trapped.
After  purging  was  completed, the  sorbent  column  was heated  and  backflushed
                                      60

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with inert gas  to desorb the components onto  a  gas chromatcgraphic  column.
The qas chromatographic  column was heated to elute the volatile constituents,
which  were  detected  by  flame   lonization  detector  (FID)  or  gas
chromatography/mass  spectrometry (GC/MS).

     Extractable  organic constituents—The  sample  preparation  and  analysis
scheme for extractabfe organics  fs shown  in Figure ?.  A  10-g  aliquot of  the
waste  in  a  225-ml  centrifuge  bottle  was  diluted  with  fiO ml  distilled.
deionized  water.    The  sample  was  homogenized  with a Tekmar  TissumizerR
blending  probe for 30 seconds to enhance the  wetting of the sediment.  Each
sample was adjusted  to  pH 11 with  6 N  sodium hydroxide.   Three  sequential
extractions with 80-ml  aliquots  of dicKloromethane were  performed  to  isolate
the base/neutral  compounds and  the  pesticides.   Following  each addition of
dichloromethane,  the sample  was  homogenized  for 30 seconds  with  a  Tekmar
Tissumizer"   blending   probe,  and  centrifuged  for  30 minutes  at  2,500
revolutions per minute to promote phase  separation.    The three  base/neutral
extracts  were  combined  and dried by  passage through a short  column of
anhydrous sodium sulfate prior  to concentration to  5 ml  in a  Kuderna-Oanish
evaporator.   The  waste  samples  were adjusted  to  pH  1 with 6  N  hydrochloric
acid,  and  the extraction, extract  drying,  and  concentration steps  were
repeated  to isolate  the  acidic compounds.

     GC/MS analysis—The waste  extracts were analyzed according to U.S.  EPA
standard  methodoloqy (U.S.  EPA 1982) on  a  Hewlett-Packard  (HP)' 5985B  Gas
Chromatograph/ Mass Spectrometer/Data  System  (GC/MS/DS).    The  mass
spectrometer  was tuned  prior  to the  analyses using  perfluorotributylamine
(PFTBA) and the Hewlett-Packard "Autotune"  program, which optimizes  ion
source,  mass  filter,   and  electron multiplier   parameters  for  optimum
sensitivity,   peak resolution  and  mass  axis  calibration.   An  abundance
normalization  program  .was  also run to meet  U.S. EPA specifications  for
spectral  reproducibility.

     The  dichlorcinethane/waste  sample extracts  were  analyzed  using a 30 m  x
0.32 mm I.D. SPB-5 bonded phase fused silica capillary column.  Helium carrier
gas was set at  a  split  vent flow of  140 mL/min with column flow  set at  1.4
ml/mm (split  ratio  1:100).   A  summary of the GC/MS  analysis  conditions used
is presented  in  Table 23.

     HPLC  u.~. ^lysiS'-Polynuclear  aromatic hydrocarbon compounds (PAHs) were
determined using high  performance  liquid  chromatography  {HPLC)  following
Method 8310,  (U.S.  EPA  1982).   A  Perkin-Elmer  HPLC system equipped with  a
quadruple solvent  delivery system (Series 4),  a UV  detector  (Model  LC-85B),
integrator (Model  LCI-100)  and  reverse phase  column  (HC-ODS SIL-X),  was used
for analysis.

     Chromatography  conditions  were  as follows: isocratic  for 1 minute with
acetonitrile/water  (40/60),  linear  gradient  elution  to 100  percent
acetonitrile  over  7 minutes,   followed  by a  3-minute  hold  at  100  percent
acetonitrile.
                                    61

-------
         Aqueous Phase
         * Solids
Dii:ord
                                Waste
                                tlOg)
                              Add Water
                              (60ml)
                                  —	  Add Spike
                           Adjust topH>ll
                           with 6N NaOH
                        Extract 3X with CH2CI2
                        by Homogenlzation/
                        Centrifugatlon
                       Extract
                                 Dry with Na2S04
                                     Aqueous Phase
                                     + Solidi
                            Adjust to pHS2
                            withGNHCI
Extract 3X  with CH2CI2
by Homogenization/
Centrifuaation
                                Determine Base/
                                Neutrals by GC/MS
                                on SPB-5 Fused
                                Silica Capillary
                                Column
                            Dr> with NazS04
                          Determine Phenols
                          by GC/MS on SPB-5
                          Fused Sllico Column
Figure    3.   Scheme for the analysis  of waste samples for organic constituents.

-------
                     TABLE  23.   GC/MS ANALYSIS CONDITIONS
Instrument:

     Gas chromatograph:
     Mass spectrometer:
     Data system:

Column:




Temperature program:




Injector temperature:

Transfer line temperature:

Carrier gas:

Splitless injection:

Injection volume:

Solvent:
Mass spectrometer operating
  conditions:

     Ion source temperature:
     lo-iization energy:
     Trap current:
     Electron multiplier:
     Scan range:
     Scan speed:
HP 5840
HP 5985B
HP

30 m x 0.32 mm ID SPB-5 bonded phase
  fused silica capillary column
  (Supelco) routed directly into the
  ion source

60°C (2 min) to 300°C at 4°C/min
  (base/neutrals/pesticides)
60°C (2 min) to 300°C at 8°C/min
  (acids)

Z90°C

300°C

Helium at 29 cm/sec

30 sec

4 pi

Oichloromethane (samples)
Methanol/dichloromethane (standards)
280°C
70 eV
200 UA
-1.75 kV
bO-450 amu
1-2 sec/scan
                                      63

-------
Biological Characterization of  Waste--

     A  comprehensive  assessment of  the  hazardous  characteristics  of  a
particular waste involves both chemical and biological analyses.   Biological
anal •'sis provides information on  potential interactions between waste  and  soil
components  that may not  be  shown by  chemical  identification of  waste
components.  Biological  analysis  may also  indicate  the toxic  am1/or mutagenit
potentials of  the  waste  and  waste-soil  mixtures.   In   this  study,  acute
toxicity  of  aqueous  extracts  of  the  waste  samples was determined using  the
Microtox'M  system  (Microbics  Corp.,  Carlsbad,  CA).   The   Ames  Salmonella
typhimurium  mammalian  microsome  mutagenicity  assay  was   used  to assess  the
mutagenic potential of each hazardous waste.

     Microtox Assay--

     This procedure is described  under  "Waste Loading Rate Evaluation."

     Ames Assay—

     The  procedures used  for  the  Ames  Salmonella typhimurium mammalian
microsome mutagenicity assay for  determining mutagenic potential  of"parent  PAH
compounds and soil  metabolites  are based on the methods described by Maron  and
Ames (1983).  A schematic  of the  Ames assay is shown in Figure 4.

     Sample  preparation—The waste  samples  were extracted  according to  the
extractable organics   procedure described  previously.   The  base/neutral
fractions of all samples were  tested  for mutagenicity.  In addition,  the  acid
fraction of the pentachlorophenol  waste was analyzed using the Ames assay.  An
aliquot of the concentrated extract of  each sample was brought to dryness  in  a
preweighed vial using a fine s+ream of purified air.   The residue was brought
up  to  a predetermined volume  in  dimethyl  sulfoxide (OMSO) and  sonicated  for
several minutes to  ensure  thorough mixing.  A set of five  dilutions ranging to
10~?  was  prepared  for  initial mutagenicity  screening.    If necessary,
additional dilutions were  made  for subsequent testing.

     Experimental  apparatus—The  Salmonella typhimurium tester strain  used  for
testing was TA-9&,  which detects  "frameshift  mutagens.   Genetic alterations to
th/-, strain  and others  included   in  the tester  set have  made them unable to
gro* in the absence of histidine.  When the strains are placed on a histidine-
free medium, the only colonies formed  arise from cells that  have reverted to
histidine-independence.   Spontaneous reversion rates  are  relatively  constant
for  each  bacterial  strain,  but addition of  a  chemical   mutagen greatly
increases the mutation value.  Some mutagens require  activation by mammalian
micro somes.   This  activation is provided by addition   of  the  S9  fraction
derived from Aroclor 1254-induced rats.  S9  can be  prepared  in the laboratory
according to  methods  of  Mar^n and  Ames  (1983)  or may be purchased  from
commercial sources.

     Procedures for preparation of  the  S9  microsomal mix and media  are
presented in Maron  and Ames (1983).
                                     64

-------
      INDUCED
           NONINCUCED
              s-9 MIX

              BACTERIAL
              SUSPENSION

SODIUM PHOS-
PHATE BUFFER

BACTERIAL
SUSPENSION
                                    SAMPLE IN'DMSO
                                            SOFT AGAR 45'C
                               WITHOUT H1STIDINE FOR MUTAGENICITY
                               TEST
                     INCUBATE 46HRS AT 37«C
                          AND COUNT
Figure  4.   Schematic of the Ames assay.

                                65

-------
     The  controls  used  in  the Ames  asr.ay included  plates  with no  chemical
addition, solvent controls  with  DMSO added, and positive controls  with  known
mutagens  added.   The known mutagens  used  included dauncmycin-HCI  without  S?
a.id 2-aminofluorene with S9.   Platos  of each control  type were  prepared  with
and without S9.  The bacterial suspension was added in each  case.

     Experimental procedure—For eac'i Ames assay the following parameters  were
evaluated:   1}  mutaqenicTty  of  tt»e  sample, 2)  response of  the  strains  to
positive  controls,  3)  toxicity of  the  sample to  the  tester  strains, and  4)
sterility of samples and reagents.

     Mutagenicity of the sample and of  positive  controls  was  determined  usinq
the plate incorporation method {Ames et  al.  1975).   A preliminary  test  and
confirmatory test were  performed  for each sample  based on  recommendations  of
Williams  and  Preston (1983).    For  each test, five doses of  the sample  were
evaluated both with  and  without S9, one dose  per plate, with  duplicate plates
for each  dose.   For each plate,  a  mixture of 2.0 ml  soft agar  containing a
trace of histidine, 0.1 ml bacterial suspension,  0.1 ml test material, and 0.5
ml of  the S9  mix (if required) was  overlaid  on  minimal media.   The  agar was
allowed to harden and the plates were incubated 48 hours at  37°C.

     Each  confirmatory test  involved  repeating the  assay using  additional
doses for providing the most active mutagenic response.

     Known mutagens were included in each test to ensure that  the strains  were
active  and  that the  S9 preparation was  activating prcmutaoe.is  to proximate
mutagens.

     Toxicity  of the samples  was determined  by checking for  the presence  of
background growth (lawn) on sample  plates.   This  lawn  results  from the  trace
amount of histidine  in  the  overlay  (top)  agar,  and is necessary in  most  cases
for mutagenesic to  occur.    A lawn  which is sparse  or absent  compared  to
control  plates  with  no chemical addition indicates  toxicity  to  the tester
strains,  and visible colonies  that appear are not necessarily revertants.

     The  sterility of all components of the assay—samples,  positive controls,
solvent,  S9 mix  and agar plates--was checked by  plating without addition  of
the bacterial suspension.

     Quality control procedures are "specially important for the Ames assay to
ensure  that  each  component of  the assav,  prepared  at  different  times  and
stored until use,  is functioning  properly at  the tine the assay is  conducted.
Quality  control  tests  are  performed  periodically with resprct  to  Salironelld
strain  validation  and proper  metabolic  activation of  59.   Strain  validation
ensures  that  the strains have retained the mutations  for proper functioning.
Proper metabolic activation ensures that any promutagen  'n tne test material
will be  activated to the proximate mutaqen successfully.

     Data  calculations—Raw data were  obtained  as revertants/plate  and  were
scored  on an  automatic  coiony counter  {New  Brunswick  Scientific Co., Inc.).
Mean counts were calculated for replicate plates  at  each dose  level  and  for
positive  and  negative  controls.   Toxicity of  samples  was  recorded  when
                                       66

-------
presen',  and  results of  sterility  te^ts  were noted.   Mutaqemc  rains
calculated for  ,.11  Sdiiple concentration  results.   Mutcigenic  ratio is defined
as:
     Mutagemc Ratio =  nu"ber °f cQ1oni(?s  w1^  sample
         *             i.umber of colonies without  sa-nple

A  test  compound  or  sample  is considered  negative (nurmiutagernc)  if  the
mutagenit ratio is less than 2.5.

     Recommendations on data production, handling,  and  analysis  by de Serres
and Shelby (1979) were followed in this research project.  The recommendations
concerning data presentation, definition of  positive and negative results, and
comments  on  statistical   analysis  represent a  modification  of  the  original
protocol of Ames et al . (1975).


Results and Discussion

Physical Characterization  of Wastes--

     The water content density, specific gravity,  and flash points of the four
wastes are presented in Table 24.  Results  from  the  residue analysis are shown
in Table 25.

Chemical Characterization  of Wastes--

     Inorganic Constituent--

     Metal s--Tables 26 and  27 present  the  results  of metal  analyses for the
petroleum refinery and wood  preserving  wastes,  respectively.   Quality control
results,  including  spiked samples for  the  metals  Analyses,  are  presented in
Tables ?8 - 11.

     Organic Const ituents--

     Total organic _ carbon and  oil  an*4 greas»--Thp total  ori^nic  carbon (TOC)
and oil and grease content For three  '•"ilicate samples are presented in Tables
32 and 33.

     GC/MS analysis—The  results  of the GC/MS  analyses of the  ijase/ neutral
fractiuns of  waste  extract  are  presented  in  Tables 34 and  35  for petroleum
refinery wastes and in Tables 36 and 37 for wood preserving  wdstes.  Table 38
lists  compounds  identified   in  thf>  acid   fraction of  creosote  and
pcntachlorophenol waste.   No acid compounds were identified  in  thp petroleum
wastes.   The  compounds tentatively  identified  in each  fraction  are presented
in their order of elution  from the SPB-5 fussd silica capillary column.

     Compounds were  identified by  a  comparison of  sample mass  spectra  with
mass  spectra  in  the EPA/NIH  mass   spectral   data  base,  which contains
approximately  25,000 mass spectra  (Heller  and M.lna  1978),  or  by manual
         at ion.   Identifications  wore  considered  tentative because the scope
                                     67

-------
                                  TABLE 24.   PHYSICAL CHARACTERIZATION OF WASTES
Waste
API Separator Sludge
Slop Oil
Creosote
Pentachlorophenol
Water Content*
(I)
47 * 2.8*
O.T* 0
33 + 0~.7
28 ~0.7
Density
(g/*i)
0.986 * 0.006
0.806 + 0.004
1.01 *T).08
0.824"+ 0.096
Specific Gravity
0.990 + 0.008
0.814 + 0.005
1.01 +T).10
0.815"* 0.098
Flashpoint
92 op
<60°F
pH
4.9
3.9
5.5
5.4
      *Method used:  Standard Method of Test for Water in Petroleum Products  and  Bituminous Materials by
      Distillation.  ASTM 095-70.
      ^Average of three replicates *_ standard reviatios.
00

-------
                      TABLE  25.   CHARACTERIZATION OF RESIDUES  IN HAZARDOUS WASTES
                                                             Total  Suspended            Total Volatile
     Waste  Type                Total  Residue  (103°C)           Residue  (103<>C)           Suspended Residue
                                     (mg/g)                       (mq/g»                     (mg/g)
API Separator Sludge
Slop Oil
Creosote
Pentachlorophenol
257 * 32*
227 "27
522 "9
422 +"19
77.0 + 26.6
1.77 "0.19
384 * 47
302 +~14
33.2 + 10.0
1.77 +"0.19
229 + 36
189 * 10
*Aweraqe of three replicates *_ standard deviation.

-------
    TABLE  26.   CHARACTERIZATION OF METALS IN PETROLEUM REFINERY WASTES*4
                      	Concentration1	
Hetal	Separator Sludge                  Slo^ Oil
                            --- iig'Kg (Tor-pete* for blank ---
Chromium
Zinc
Cadmium
Lead
Nickel
Vanadium
Beryllium
Silver
Aluminum
Strontium
Barium
Copper
Arsenic
Selenium
Mercury
Antimony
Thallium

Iron
209
260
0
11
6
1
<0
3
279
IB
^
30
n
<0
1
<4
<5
— y/kg
1


.41


.4
.2



.4

.62
.08
.4


+
+

+
*


+
+
+
*
»
+~

*


3
11

3
2


3
10
1
0.6
2
0.02

0.7


1
5
0
21
2
<0

-------
      TABLE  27.  CHARACTERIZATION OF METALS IN WOOD PRESERVING WASTES**
Metal
            Concfntration*
                         Creosote
                              TCP
Osmium
Thall mm
Arsenic
Mercury
Selenium
Molybdenum
Chromium
Antimony
Zinc
Vanadium
Cadmium
Lead
Nickel
Manganese
Berylliun
Silver
Strontium
Barium
Copper
Iron
Aluminum
                        --- tmj/kg  (correcteofor blank) - —
 <2.5
<12.5
  1.88 i 0.17
<12.5
<12.5
< 1.25 * 0
  4.36
 62.7
  3.26
 <0.5
    40
    70
 57.6
                   46
               1 0.46

               * 5.0
               £0.29

               * 0.89
               ~ 0.6/
               ~ 4.4
          9.92 * 0.79
        252    ~ 27
         15.1  *1.3

— 9/kg (corrected for blank) ---

          2.9? * D.25
          2.62 * 0.17
 <2.5
<12.5
  1.31
<12.5
<12.5
  <1.25
   3.02

  110
   1.72
  <0.5
  13.0
   3.75
 107
  <0.1
  <1.2
    .1
    .5
  0.02
  0.17

  2.12
  0.09

  0.15
  0.57
                             11
                             23
* 7
                              1.93
                              1.24
                              7.89  *  0.34
  0.08
  0.06
 Digestion Procedure:  Method  30bO,  Acid  Digestion of Sludges.   Test Methods
for Evdludting Solid Waste.  Sy-£46, Second Edition  (U.S.  EPA 1982).
*Artdlytical Method:   ICAP  for  
-------
TABLE  28.   CHARACTERIZATION  OF  METALS IN  PETROLEUM  REFINERY WASTES:
                      QUALITY CON1ROL DATA
Metal Digested Quality Control Sample
Measured Value

Chroniuir.
line
Cadmium
Lead
Nickel
Vanadium
Reryll ium
Silver
Aluminum
Strontiu.Ti
Barium
Copper
Arsenic
Selenium
Mercury
Ant imony
Thallium
Iron
(ug/l)
1143
2000
:ao
2230
1391
1090
42<0
100
3730
<5
100
1920
990
120
59
-
-
4710
Actual Value
(ug/i)
1250
2000
350
2000
1500
4250
4500
-
4000
-
-
1750
1500
250
40
-
-
4500
Relative
Error
m
9
0
20
11
7
4
6
-
7
-
-
10
34
52
25
-
-
5
Blanks

(ug/D
<25
o2
0.04
5
20.0
<2.6
<5
<25
<100
<5
7
-
i.fl
9.0
<0.5
-
-
150
Spike Recovery
(Spiked After
Digestion]
(i)
-
-
59
86
102
68
-
-
-
-
-
-
-
-
-
-
-
-

-------
             TABLE  29.   CHARACTERIZATION OF HETALS  IN CREOSOTE WASTES:  QUALITY  CONTROL
                    DATA FOR  SPIKED  CREOSOTE WASTE  SAMPLES - HIGH AND LOW LEVEL
High Level*
Metal
Osmium
Thai 1 lum
Arsenic
Mercury
Selenium
Molybdenum
Chromium
Antimony
Zinc
Vanadium
Cadmium
Lead
Nickel
Manganese
Beryll iui)
Silver
Strontium**
Bar ium**
Copper

Iron
Aluminun
Measured Value
Spiked Waste-
(•nq/kg)'
-..'.5
<12.5
3.53
<12.5
<12.5
<1.25
7.58
<10
61.5
14.?
0.67
11.8
6.74
56.9
10.2
<1.2
9.37
243
16.2
(g/kg)
2.86
2. «9
Theoretical
Value
(mo/kg) *
-
-
4.17
-
-
-
7.97
-
67.5
14.4
0.52
13.8
7.28
62.1
11.0
-
10.1
256.5
17.5
(g^g)
2.98
2.49
Recovery
(*)
-
-
85.8
-
-
-
95. U
-
91.7
97.8
119
86.5
91.4
92.2
92.1
-
93.2
95.5
92.9
(*)
96.8
100
Low Level*
Measured Value
Spiked Uaste-
(mg/kg)«
<2.5
<12.5
2.44
<12.5
<12.5
<1.25
4.51
<10
63.9
5.23
<0.5
<7.5
1.20
58.7
0.74
<1.2
9.74
109.9
15.3
(g/kg)
2.85
2.55
Theoretical
Value^
(mg/kg)
-
-
2.68
-
-
-
4.52
-
60.3
59.3
0.1
8.89
4.71
55.05
0.82
-
9.37
237.5
14.7
(g/kg)
2.76
2.49
Recovery
m
-
-
91.3
-
-
-
99.8
-
100
88.2
-
-
89.15
107
90.25
-
102
46.15
104.0
(*)
103.3
102.5
 Sample No. WP475 *5.   Spiked sample subjected  to digestion  and  analysis  by  ICAP (arsenic by
 AA-grapnite furnace).
*Low level:  Approximately 2 g  creosote waste  spiked  with  1  ml  of EPA  Quality Control Sample No. WP475
 14.  Spiked samplp subjected to digestion and  analysis  by  ICAP  (a.-senic  by  AA-graphite furnace).
'Average of two  replicate  analysis.   All  concentrations are corrected for digested  olank  values.
**Theoretical value calculated as sume of  average measured  concentration  corrected for sample size and
 amount added in QC sample.
"Note included in spike.

-------
         TABLE  30.   CHARACTERIZATION OF METALS IN PENTACHLOROPHENOL WASTES:  QUALITY CONTROL
                 DATA FOR  SPIKED  PENTACHLOROPHENOL  WASTE  SAMPLES  -  HIGH  AND  LOW LEVEL
Metal
Osmium
Thall ium
Arsenic
Mercury
Selenium
Molybdenum
Chromium
Antimony
?inc
Vanadium
Cadmium
Lead
Nickel
Manganese
Beryl 1 ium
Silver
Strontium**
Barium**
Copper

Iron
Aluminum

Measured Value:
Spiked Waste-
(mq/kg)'
<2.5
<12.5
3.59
<12.5
<12.5
<1.25
6.27
<10
141.5
10.95
0.89
16.95
6.57
104
10.45
<1.2
10.55
22.5
9.8
(g/kg)
1.92
1.21
Hiy!' Level*
Theoretical
Valje
(mg/kg)"

-
4.56
-
-
-
6.89
-
125
13.5
0.61
19.25
7.62
114
11.65
-
11.65
24.55
10.6
is/kg)
2.03
1.33
Low Level*
Recovery
(«)

-
78.95
-
-
-
92.05
-
113.2
81.1
147
88.1
84.85
91.25
89.7
-
90.55
91.65
92.3
m
94.6
91.30
Measured Value
Spiked Waste-
(mq/kg)'
<2.5
<12.5
2.17
<12.5
<1?.5
<1.25
3.50
<10
158.5
4.19
<0.5
15.2
3.75
94.7
0.77
<1.2
11.15
37.6
8.03
(g/kg)
1.15
1.15
Theoretical
Value
(mg/kg)**

-
2.21
-
-
-
2.95
-
114.5
3.79
0.1
15.6
4.86
102.3
0.86
-
10.7
22.55
8.07
(g/kg)
1.20
1.20
Recovery
(I)
-
-
98.4
-
-
-
118.5
-
1J7
113.95
-
97.5
77.25
92.8
90.05
-
105.15
171.05
100.0
(*)
96.5
96.5
 "High level:    „          ...
  WP475 15.   Spiked  sample  subjected  to dicostion and analysis  by  ICAP  (arsenic  by  AA-graphite furnace).
 »Low level:   Approximately 2  g  pentachloruphenol waste  spiked  with  1 ml  of  EPA  Control  Sample No.  WP475
  14.  Spiked  sample subjected to digestion and analysis by  ICAP (arsenic  by  AA-graphite furnace).
 IA11 concentrations are  corrected  for digested blank values
"Theoretical  value  calculated as sum of average measured concentration  corrected  for sample
  size and amount added  in  QC  sample.
"Not included in spike.

-------
                   TAPLE    31.   CHARACTERIZATION OF METALS IN CREOSOTE AND PENTACHLOROPHENOL WASTES:
                                 QUALITY CONTROL DATA FOR EPA QUALITY CONTROL  SAMPLES
•vj
Ul
Measured Valjje
QC Sample fl
Metal (ug/1)
Osmium
Thallium
Arsenic
Mercury
Selenium
Nol)bdenum
Chromium
Antimony
Zinc
Vanadium
Cadmium
Lead
Nickel
Manganese
Beryllium
Silver
Strontium
Barium*
Copoer

Iron
Aluminum
<50
<250
23.4
<250
<2*0
«25
<25
<200
12
67
<10
-150
<50
15
20
<25
<2.5
22
10
(mg/1)
<0.02
<0.1
Theoretical
Value
(ug/1)
-
.
22
0.75
6
-
10
-
16
70
2.5
24
30
15
20
-
-
-
11
(mg/1)
0.02
O.C6
Relative
Deviation
m
.
.
6.4
-
-
-
0
-
-25
- 4.3
-
-
-
0.0
0.0
-
-
-
- 9.1
W
-
~
Measured Value
QC Sample »2*
(ng/D
<5u
<250
58.4
<250
<250
<25
83
<200
77
210
11
<150
82
75
245
<25
<2.5
19
<7
(mg/1)
<0.02
0.5
Theoretical Relative
Value Deviation Blank
(US/1) (%} fog/1)
-
-
60
3.5
30
-
80
-
80
250
13
120
80
75
250
-
-
-
50
(mq/1)
0.08
0.45
-
-
-2.7
-
-
-
3.8
-
3.8
" -16
-16
-
2.5
0.0
-2.0
-
-
-
-6.0
(%)
-100
11
<50
<250
<5
<250
<250
<25
<25
<200
12.7 (*4.7)
<25
<10
<1SO

-------
  TABLE 32.  TOTAL ORGANIC CARBON (TOC)  CONTENT OF  HAZARDOUS WASTE SAMPLES
Waste


API Separator Sludge
Creosote Waste
TOC (mq/kg)*


101,000
347,000
Standard
Deviation
(mg/kg)
14,000
48,000
Coefficient
of Variation
(%)
14
14
*Average of three replicates.
         TABLE 33.   CHARACTERIZATION OF OIL AND GREASE* IN HAZARDOUS
                               WASTE  SAMPLES
Waste Type Oil and
Grease
(mg/kg)
API Separator Sludge
Slop Oil
Creosote
Pentathlorophenol
QA/QC Samples:
n Fuel Oil
EPA-API Reference
Oil: Prudhoe
Bay Crude
(WP 681)
3.5x10*
4.6x10*
3.7x10*
5.2x10*

9.4x10*
8.6x10*

Oil

Coefficient
Standard of
Deviation Variation
(mg/kg) (*)
2.5x10^ 7.2
4.9x10* 11
1.2x10* 3.1
1.5x10* 2.9

1.0x10* 1.1
1.0x10* 1.6



Actual
Value*
(mg/kg)
N.A.
N.A.
N.A.
N.A.

>8.8x!05
53.8x10*



'Procedure:   U.S.  EPA  procedure  (Robert  S.  Kerr  Environmental  Research
I iboratory Standard Operating Procedure (SOP)-2i.
+N.A. = Not applicable.
                                       76

-------
TABLE 34.   ORGANIC COMPOUNDS TENTATIVELY  IDENTIFIED  IN API  SEPARATOR
           SLUDGE WASTE (BASF NEUTRAL FRACTION) BY GC/MS
Compound
Heptane
Hexane, 2, 5-Oimethyl,
Heptane, 2-Methyl
Cyclopentane, ethyl -methyl ,
or i1'~on?
Benzene, methyl
Nonane
Benzene, dimethyl
Nonane, 4-methyl,
actane, dimethyl
Benzene, dimethyl
Decane
Decane, 4-raethyl
Benzene, propyl
Benzene, ethyl methyl;
Benzene, trimethyl
Benzene, alkyl substituted
Benzene, trimethyl;
Benzene, ethyl methyl
Benzene, trimethyl ;
Benzene, ethyl methyl
Undecane
Benzene, trimethyl;
Benzene, ethyl methyl
Benzene, diethyl;
Benzene, methyl propyl
Benzene, rtiethyl;
Benzene, methyl propyl
Benzene, diethyl;
Benzene, methyl propyl
Benzene, ethyl dimethyl;
Benzene, tetramethyl; etc.
Benzene, ethyl dimethyl;
Benzfne, tetramethyl; etc.
Codec ane
Benzene, ethyl dimethyl;
Benzene, tetramethyl, etc.
Tridecane
Formula
C&H16
C8»18

CsHlo

C?H8
C9H20
CsHlO
ClOH22

C8H10
ClOH22
CnH24
C9H12
C9H12

C9H!2
C9H12

C9H12

CnH24
C9H12

CIOHH

ClO»14

C1QH14

ClOH14

ClOH14

Cl2"26
CiQHi4

Cl3H9ft
Molecular
Weight
100.
114.

112.

92.
128.
106.
147.

106.
in.
156.
120.
110.

120.
120.

120.

156.
120.

134.

134.

134.

134.

134.

170.
134.

IRA
Retention
Time
(minutes)
0.8
1.0

1.1

2.1
3.C
4.4
4.6

5.4
6.1
6.6
7.2
7.5

7.7
8.1

8.4

9.1
9.4

9.8

10.0

10.2

10.5

10.8

11.4
11.7

11 A
                                   77

-------
TABLE 34.   CONTINUED
Compound
Naphthalene, Azulene
Tet-adecane
Naphthalene, methyl
Naphthalene, methyl
Pentadecane
Tetradecu.^, trimethyl
l,l'-Bipnenyl
Naphthalene, Dimethyl
N&phthalene, Dimethyl
Hexadecane
Naphthalene, Dimethyl
l.l'-Biphenyl. methyl
Heptadecande
Naphthalene, trimethyl
Naphthalene, trimethyl
Octadecane
Nonadecane
Eicosane
Phenanthrene, anthracene
Heneicosane
Oibenzothiophene, methyl;
9H-thioxanthpne
Dibenzothiophene, methyl;
9H-th
-------
                           TABLE   34.    CONTINUED
          Compound            Formula        Molecular      Rpter>Hon
                                               Weight          Time
                                                            (minjtes)
Hexacosane                    ^26^54           366.             30.6
Heptacosane                   t27«56           380.             31.5
Octacosane                    C28Hes           394.             32.5
Nonacosane                    C^gH^           40ti.             33.7
                              C3QH62           422-             35.
                                    79

-------
TABLE  35.    ORGANIC C01POUNDS TENTATIVELY IDENTIFIED IN SLUP OIL
         EMULSION WASTE (BASE/NtUTRAL FRACTION) BY GC/MS
Compound
Dichloromethdne
Hexane, 2,2-dimethyl; or
Butan.', 2,2,3,3 tetra-
methyl
Heptane
Hethyl benzene
Nonane
Benzene, dimethyl
Benzene, dimethyl
Oecane
Benzene, propyl
Benze.ie, ethy1 methyl
substituted
Cyclohexene, butyl, or
thiophtnenc
Benzene, ethyl nethyi; 01
benzene, trimethyl
lenzine, trinethyl; or
bT^ene, ethy1 methyl
Benzene, motnyl propyl,
bcn/ene, ethyl oi-'.'thyl,
or benzene, tetramethyl
Und'?cane
Fenzene, i,2,3-tr imethyl
b^'i^eno, diethyl
Formula
CH2U2
C8H,6
C6H16
C7H8
C9H20
C8"10
C8H10
C1QH22
C9H12
C9H12
c$&
C,»12
CoH * 2
1 0 1 4
CnH24
C9H12
ClOl'l4
Mol'.cular
Weight
85.
114.
100.
92.
123.
106.
106.
142.
120.
120.
140.,
140.
120.
120.
134.
156.
120.
134.
Setent ion
T ime
(minutes)

0.8
1.0
2.3
3.5
5.1
5.9
6.8
7.5
7.9
8.1
8.4
8.8
5.3
9.5
9.7
10.1

-------
                 TABLE   35.   CONTINUED
Compound
Formula
Molecular      Retention
  Weight          Tine
               (minutes)
Benzene, methylpropyl; or
 benzene, tetramethyl; or
 benzene, ethyldimethyl

Benzene, tetramethyl;
 benzene, ethyldimethyl;
 or benzene, m-thylpropyl

Benzene, ethyl-dimethyl
 substituted; benzene, 1-
 methyl-4-(l-methylethylj-;
 or benzene, diethyl;
 acenaphthylene

Allcyl-substituted benzene

Dodecane

Benzene, ethyl dimethyl
 substituted; or benzene,
 methyl-dipropyl

Benzene, diethylmethyl

Benzene, diethylmethyl;
 or benzene, ethyltri-
 methyl

Indane, dimethyl; naphtha-
 lene, or tetrahydromethyl;
 benzene, pentamethyl or
 alkyl substituted benzene

Tridecane

Naphthalene

Tetradecane

Naphthalene, -methyl

Naphthalene, -methyl

Pentadecane
                    ClOH14
                    Cl2"26

                    ClOH14
                    C13H28
                                     134.
                 134.
                                     134. ,152
                 148.

                 170.

                 134.



                 148.

                 148.



                 146.. 148




                 184.

                 128.

                 198.

                 142.

                 142.

                 212.
                                 10.3
                  10.7
                                 10. 0
                  11.1

                  H.7

                  11.8



                  12.1

                  12.5



                  13.3




                  14.2

                  14.4

                  15.4

                  16.2

                  16.6

                  17.1
                            81

-------
TABLE  35.   CONTINUED
Compound
Naphthalene, dimethyl
substituted
Hexadecane
Naphthalene, dimethyl
substituted
Naphthalene, methyl ethyl
Naphthalene, trimethyl, or
naphthalene, methyl ethyl
Naphthalene, alkyl sub-
stituted
Naphthalene, alkyl sub-
stituted
Heptadecane
Naphthalene, trimethyl
substituted
Naphthalene, trimethyl
substituted
Naphthalene, tetramethyl;
or naphthalene, alkyl
substituted
Biphenyl, dimethyl; or
biphenyl ethyl
Octadecane
Naphthalene, methyl,
isopropyl
Naphthalene, dimethyl,
isopropyl; naphthalene,
alkyl substituted
Nonadecane
Formula
Cl2«12
Cl6H34
C12H12
C13H14
Cl3«14
C13H14
C13H14
C17H36
C13H14
C13H14
C14H16
C14H14
Cl8H38
C14H16
Ci4H16'
C19H40
Molecular
Weight
156.
226.
156.
170.
170.
170.
170.
240.
170.
170.
184.
182.
254.
184.
198., 184.
268.
Retention
Time
(minutes)
18.5
18.7
18.8
19.0
19.5

20.1
20.2
20.4
20.7
20.9

21.6
22.2
22.5
23.0
          82

-------
TABLE  35.   CONTINUED
Compound
Eicosane
Phenanthrene/ anthracene
Henelcosane
Anthracene; phenanthrene.
methyl substituted
Anthracene; phenanthrene,
methyl substituted
Docosane
Anthracene; phenanthrene,
methyl substituted
Oibenzothiophene, dimethyl
Dibenzothiophene, dimethyl
Phenanthracene, anthracene,
dimethyl substituted
Penanthrenc. dimethyl
substituted; anthrazene
Benzo[ghi]fluoranthene
Tetracosane
Phenanthrene, trimethyl;
anthrene, trimethyl
Fluoranthene; pyrene
Pentacosane
Hexacosane
Heptacosane
Octacosane
Nonacosane
Formula
C20H42
Cl4H10
C21H44
C15H12
Cl5Hl2
C22H46
C15H12
C14H12S
Ci4Hl2S
Cl6«14
C16H14
Cl8H10
C24H50
CI/HIS
Cl6H10
C25H52
C26H54
C27H56
C28H58
C29H60
Molecular
Weight
282.
178.
296.
192.
192.
310.
192.
212.
212.
206.
206.
226.
338.
220.
202.
352.
366.
380.
394.
408.
Retention
Time
(minutes)
24.2
24.7
25.3
26.1
26.2
26.4
26.6
26.9
27.1
27.4
27.8
28.0
28.4
28.9
29.2
29.5
30.5
31.5
32.5
33.6
          83

-------
TABLE  36.  ORGANIC COMPOUNDS TENTATIVELY IDENTIFIED IN CREOSOTE WASTE
                   (BASE/NEUTRPL FRACTION) BY GO/MS
       Compound
Formula        Molecular      Retention
                 Ueight          Time
                              (minutes)
Cyclohexane
Trichloroethane
Benzene, Ethyl, He
Benzene, Ethynyl. He
Naphthalene. Methyl
l.l'-Biphenyl or
Acenaphthylene-l,2-Dihydro
Naphthalene, Ethyl or
Dimethyl
Naphthalene, Dimethyl
Naphthalene, Dimethyl
Naphthalene, Dimethyl
l.l'-Biphenyl, Methyl or
Benzene, l.l'-Methylenebis
Biphenylene or acenaphthylene
Dibenrofuran
9H-Fluorene
l.l'-Biphenyl, Methyl,
or Naphthalene, l-(2-
propanyl)
Phenanthrene, 9,10-Dihydro
9H-Fluorene, Methyl
Octadecane
Phenanthrene or
Anthracene
Anthracene or
Phenr"»threne
9H-Caroazole
Phenanthrene, Methyl
or Anthracene, Methyl
Phenanthrene, Methyl
or Anthracei.e, Methyl
Naphthalene, Phenyl
Pyrene, or Fluoranthene
Pyrene, or Fluoranthene
HH-Benzo(a)fluorene
or Pyrene, Methyl
HH-Benzo(a)fluorene
or Pyrene, Methyl
HH-Benzo(a)fluorene
or Pyrene, Methyl
Benz{a)anthrancene, or Tri-
phenylene, or Chrysene
Benz(a)anthracene, or Tri-
phenylene, or Chrysene
C6Hi2
C2HC13
C9H10
C9H8
CnH10
Cl2"lO

Cl2"l2

Cl2"l2
Cl2"l2
Cl2"l2
Cl3"l2


C12H80
Cl3H10
Cl3"l2


Ci4Hi2
Cl4"l2
Cl8«38
CuHio

Cl4"lO

Ci2H9N
C15H12

C15H12

C16"12
Ci6«lO
CicHiO
Cl?Hl2

Cl7«12

C17H12

ClpH12

C18H12

84.
130.
118.
116.
142.
154.

156.

156.
156.
156.
168.

153. or 154
168.
166.
168.


180.
180.
Z54.
173.

178.

167.
192.

192.

204.
202.
202.
216.

216.

216.

228.

228.

5.5
6.4
13.3
14.0
18.3
19.5



19.9
20.2
20.4
20.9

21.2
21.5
22.4
22.8


23.6
23.9
24.3
24.9

25.1

25.5
26.2

26.3

26.9
28.0
28.6
29.2

29.5

29.6

32.1

32.2

                                  84

-------
TABLE 37.  ORGANIC COMPOUNDS TENTATIVELY IDENTIFIED IN PENTACHLOROPHENOL
                 WASTE (BASE/NEUTRAl  FRACTION) BY "3C/MS
Compound
Benzene, methyl
Pyridine, methyl
Benzene, ethyl
Benzene, dimethyl
Pyridine, dimethyl
Benzene, ethenyl
Pyridine, dimethyl
Pyridine, dimethyl
Benzene, propyl
Benzene, isopropyl
Benzene, trimethyl
Benzene, ethyl -methyl;
2nd rVfzane, ethenyl -methyl
Benzonitrile;
Pyridine, tr'methyl; and
Benzene, trimethyl
Benzene, trimethyl
Benzene, ethenyl -methyl
Benzene, ethyl-dimethyl
Undecane
Benzofuran, methyl
Benzene, tetramethyl
1 H-Indene, 2,3-dihydro
or Benzene, methyl -
propenyl
1 H-Indene, methyl
Oodecane
Naphthalene (or Azulene)
Quinoline or Isoquinoline
Quinoline or Isoquinoline
Tridecane
Naphthalene, methyl
Naphthalene, methyl
Quinoline, methyl
Tetradecane
Biphenyl
Naphthalene, ethyl
Naphthalene, dimethyl
Naphthalene, dimethyl
Biphenyl , methyl
Acenaphthalene
Dibenzofuran
Formula
C7Hg
C6K7N

Ceriio

C'3H8
C?HgN
f.7HgN
*9.^12
CgH12
C9H12
CgH12
Cghig
C;H5N
C8HUN

CgHj2
CgH8

^•11^24
CgH80
^10H14
ClOH12


CIOHIO

CIOHS
CgH7N
CgH/N
Cl3H28
CnH10
C11H10
CiQHgN
C14H30
Cl2"lO
C12H12
Ci2H12
C12H12
C13H12

Cl2H80
Weiyht
92.
93.
106.
'06.
107.
104.
107.
107.
120.
120.
120.
120.
118.
103.
121.
120.
120.
116.
134.
156.
132.
134.
132.


130.
170.
128.
129.
129.
184.
142.
142.
143.
193.
154.
156.
156.
156.
168.
154.
168.
Retention
Time
(minutes)
8.4
9.5
10.6
10.7
11.0
11.2
11.9
12.3
12.5
12.6
12.8
13.0

13.1


13.8
14.4
14.4
15.0
15.3
15.6
16.0


16.3
16.6
16.9
17.7
18.0
18.2
18.6
18.9
19.5
19.6
19.8
20.0
20.2
20.4
21.2
2i.4
21.8
                                     85

-------
TABLE 37.  CONTINUED
Compound
Naphthalene, trimethyl
Hexadecane
9 H-Fluorene or 1 H-
Phenalene
Fluorene, methyl and
Biphenyl, methyl
Biphenyl, methyl
Xanthene; or Oibenzofuran,
methyl
Phenanthrene, di hydro
9 H - Fluorene, methyl
Phenanthrene
Anthracene
9 H-Carbazole
Dibenzothiophene, methyl
Dibenzothiophene, methyl
Phenanthrene, methyl or
Anthracene, methyl
Phenanthrene, methyl or
Anthracene, methyl
4 H-Cyclopenta[def]phen-
anthrene
Naphthalene, phenyl
Pentadecane; Telrddecane,
methyl; or Tridecane,
dimethyl
Phenanthrene, dimethyl
Pyrene or Fluor anthene
Pyrene or Fluor anthene
9-Anthracene carbonitrile
Pyrene, methyl or benzo-
fluorene
Pyrene, methyl or benzo-
fluorene
Benzothionaphthalene
Tri phenyl ene, Ben z anthra-
cene or chrysene
Benzofluor anthene and
Benzophenanthrenc, Benz-
anthracene, Triphpnylene,
or Chrysene
Formula
C13JJ14

C13H10

Cl4Hj2
C13H12
Cl3«12
C^HjgO

^14^12
C'4^12
f 4^10
v1 ^Hl Q
129
CnKigS
Cl3HloS
C15H12

C15H12

Cl5HiO

Cl6«12



Cl6H14
Cl6H10
Cl6H10
Ci5H9N
^17^12

^17^12

Cis^loS
C^gH\2

Cl8H10
C18H12


Molecular
Height
170.
226.
166.

180.
168.
168.
182.

180.
180.
178.
178.
167.
198.
198.
192.

192.

190

204.
212.


206.
202.
202.
203.
216.

216.

234.
223.

226.
223.


Retention
Time
(minutes)
22.1
22.2
22.7

23.0
23.0
23.1
23.4

23.9
24.1
25.2
25.4
25.8
26.1
26.2
26.6

26.7

26.8

27.2
27.7


28.0
28.3
28.9
29.3
29.8

29.9

31.6
31.7

31.8



           86

-------
TABLE  38.    ORGANIC COMPOUNDS TENTATIVELY IDENTIFIED IN CREOSOTE  WASTE
         AND PENTACHLOROPHENOL WASTE  (ACID FRACTION] BY GC/MS
Compound


THF
Phenol
Phenol, Methyl
Phenol. Methyl
Phenol, Dimethyl
Phenol, Dimethyl
Phenol, Dimethyl
or Ethyl
Phenol, Dimethyl
or Ethyl
Phenol, Ethyl-Methyl
Phenol. Ethyl-Methyl
Phenol, Pentachloro
Formula


C4H80
C6H60
C7H80
C7H80
CjjHigO
C8HjgO
C8H10D

C8H!00

CgHigO
CgHi20
ceHCisO
Molecular
Weignt

72
94
108
108
122
122
1.22

122

136
136
264
Retention
Time
(minutes)
4.8
12.5
13.9
14.3
15.0
15.6
15.9

16.3

17.0
17.3
24.6
                                 88

-------
of this research project  did  not  include confirmatory analysis with authentic
standards.   Many  of the  mass  spectra  were also  manually  interpreted,
especially when  a match via library search  was  unsuccessful.

     Volatile orqan^cs—Tables  39 and 40  present  the GC/MS  analysis  of  the
volatTTe  fraction of  the  separator  sludge,  slop  oil, creosote,  and
pc-ntachlorophenol wastes.   The prominent  peak  in  all   samples  analyzed  was
identified as naphthalene.   Additionally,  various  substituted  naphthalenes,
substituted benzenes, and  hexane were prominent  in  the PCP waste  as were
substituted naphthalenes  in  the creosote waste due to the high oil content of
these samples.   Phenol was also tentatively identified in the creosote waste.

     HPLC analysis— In addition to GC/MS analysis, HPLC analysis was  used  for
identification and  quantification  for  individual  PAH compounds.
Concentrations  of  individual  PAH compounds determined by HPLC  for  three
replicate samples of each waste are presented in Table 41.

     GC/MS Analysis of Polychlorlnated  PI pen 20- p-d1 ox ins   (PCDDs)  ano
DiberizoTurans  (PCDFs)   in  Pentachlorophpnol  Waste—Two   subsamples  of  fluf
pentachlorophenol waste were  analyzed by GC/MS  for PCDDs  and PCDFs by U.S.  EPA
Environmental   Monitoring  Systems  Laboratory  (EMSL), Las Vegas,  Nevada,
following Method 8280 (U.S. EPA 1982).  Results are presented  in Table 42.

BioKxjical Characterization—

     Microtox—

     Each wastd  was characterized for toxicity of the water soluble  fraction
(WSF) using the  Microtox  assay.  Results for each  waste are  presented in Table
43.   Average valjes for  EC50 indicate that wood preserving wastes exhibited
greater WSF twkity than petroleum  wastes.   However  results indicated that
all wastes exhibited a high degree of WSF  toxicity as measured by the Microtox
assay.

     Ames—

     The  mutagenic  potential  of each waste  was  determined using  the Ames
Salmonella test.    Results for  the base/neutral  fraction of  c-eosote  are
presented in  Figure b.   This  fraction  exhibited relatively low level
irutagenicity with S9 activation,  while no  mutagenicity  is  indicated without
addition  of  59.   Toxicity  (as evidenced by  a sparse  background lawn)  was
indicated at the 400  g/plate  dose  with S9  and  at the  240   g/plate dose
without S9.
          sample  extracts  of  the  pentachlorophenol  waste were  tested  for
mutagenicity.   The results  for the base/neutral  fraction  and the acid fraction
of PCP are presented  in  Figures 6 and 7, respectively.

     The base/neutral extract with  added 59 exhibited low level mutagenicity
which decreased at higher doses.  There was an  indication of toxicity to the
Salmonella bacteria at the  641  g/plate dose with definite  toxicity present at
higher doses.   When no 59  was  added  with the  sample,  no  mutagenicity was
                                      89

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TABLE  39.  ORGANIC COMPOUNDS TENTATIVELY IDENTIFIED IN API SEPARATOR SLUDGE
          AND SLOP OIL WASTE SAMPLES {VOLATILE FRACTION) BY GC/MS
Compound
Cyclohexane
2,2,4-trimethylpentane
Hethyl-cyclohexane
Toluene
1,3-dimethyl-trans-cyclohexdne
Octane
Ethyl-cyclchexane
p-xylene
o-xylene
l-ethyl-3-methylbenzene
trlmethyl benzene
l-methyl-4-propyl -benzene
l-methyl-2 or 4(l-methylethyl)benzene
1 -methyl -3 (l-methylethyl)benzene, or
1 -ethyl -2, 4-dimethyl benzene
(l,l-dimethylbuty"l)benzene
Undecane
l-ethyl-3,5- or 2,4- or 1,2-dimethylbenzene
l-ethyl-3,5-dimethyl or l,2,3/4,5-tetramethylben7ene
Octacosane
Naphthalene
1-ethyl-l-methyl-cyclopentane
2,3-dihydro-l,6-dimethyl-lH-indene
Octadecane
Methyl-naphthalene
2-methyl-naphthalene
Pentacosane
l.l'-biohenyl
Ethylnaphthalene
Dimethyl -naphthalene
Ethyl-naphthalenr
?-(l-methylethyl)-naphthalene
Tn'methy ! -naphtha lene
1, 6, 7-trimethyl naphthalene
l-methyl-9HFluorene
Phenanthrene
4-methylphenanthrene
Dimethyl-phenanthrene
Molecular
Weight
84
114
98
92
112
114
112
106
106
120
120
134
134

134
162
156
134
Ii4
394
128
112
146
254
142
142
352
154
156
156
156
170
170
170
180
178
192
206
Retention
Time
(nin)
5.93
6.53
7.45
8.55
8.82
9.23
10.15
10.95
11.5
12.9
13.57
14.6
14.8

15.17
15.3
15.35
15.85
15.93
17.05
17.2
17.83
18.4
18.6
18.98
19.27
20.07
20.2
20.47
20.62
21.4
22.02
22.3
22.83
24.75
25.73
27.02
28.48
                                    90

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      TABLE  40.   ORGANIC COMPOUNDS TENTATIVELY IDENTIFIED IN PCP AND
             CREOSOTE WASTE SAMPLES (VOLATILE FRACTION) BY 3C/MS
           Compound
Molecular
 Weight
Retention
  Time
  (mm)
PCP Waste Data

Hexane
Z-methyl-4,6-octadiyn-3-one
Ethylbenzene
1-propynl-benzene
l-ethynyl-4-methylbenzene
Azulene
Naphthalene
Benzothiazole
2-methylnaphthalene
Dimethyl naphthalene

Creosote Waste Data

2,4,4-trimethylhexane
Phenol
Benzothiazole
Naphthalene
1,2-benzisothiazole
Methylnaphthalene
    86
   134
   106
   118
   116
   128
   128
   135
   142
   156
   128
    94
   135
   128
   135
   142
   1:50
  11:32
  11:32
  18:08
  19:16
  22:56
  22:56
  25:34
  26:05
  29:50
  20:20
  20:29
  21:39
  23:02
  24:24
  25:37
                                      91

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                 TABLE 41.  CONCENTRATION OF INDIVIDUAL PAH COMPOUNDS IN WASTES DETERMINED BY HPLC
Compound
Naphthalene
Acenaphthalene
Acenaphthene
Fluorene
Phenanthrene
Anthracene
Fluor ---it hene
Pyrene
Benzo( a) anthracene
Chrysene
Benzo{ b ) f 1 uor anthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Benzo(ghi)pyrene
Dibenzf a, h) anthracene
Indeno(l,2,3-cd)pyrene

API
Separator Sludge
580 + 87 (15%)
46o ~ 100 (21%)
<12 ~
^ + 33 (114%)
810 +"140 (17*)
110 +"27 (251)
5.500 +"290 (5*)
6,000 +"440 (7%)
1.400 ~58 (4%)
570 "310 (54%)
<3 "
310 + 62 (20X)
170 +"73 (43%)
<10 ~
40 + 11 (28%)
61 +"25 (41%)
Concentration
Slop Oil
2.500 + 700 (28%)
<15 ~
<10
440 + 300 (68%)
3.600 +"2.100 (58%)
480 +"93 (19%)
18.000 +"5.000 (28%)
23.000 "6,700 (29%)
2.000 +"1,100 (55%)
1,100 +"150 (14%)
340 +"140 (41%)
160 +" 42 (26%)
260 +"200 (77%)
59 *• 18 (31%)
15 +"1 (7%)
88 +" 19 (22%)
in Waste (mq/kq)*
Creosote
28.000 + 1,200 (4%)
3.600 +"1.000 (28%)
180.000 +"40.000 (22%)
23.000 +"5,900 (?6%)
76,000 +" 15,000 (20%)
15,000 +"6,800 (45%)
72,000 "17.000 (24%)
64,000 +"12.000 (19%)
7,400 +"1,600 (22%)
8,300 +"2,100 (25%)
3,000 ~ 700 (23%)
2,400 +"460 (19X)
2,700 ~380 (14%)
1,100 +"280 (25%)
<1,200 ~
820 +76 (9%)

Pentachloropheno'
42,000 + 28,000 (67%)
<2,000 ~
<13,000
<22 ,000
52,030 + 6,200 (12%)
11,000 +"6,800 (62%)
46,000 +"6,200 (13%)
56,000 ~ 13,000 (23%)
16,000 +"2.400 (15%)
6,900 +"2,200 (32%)
10,100 +"5,100 (51%)
<300 "
<280
<100

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          TABLE  42.  CHLORINATED  DIBbNZO-P-DIOXINS AND DIBENZOFURANS IN
                        PENTACHIOROPHENOL  WASTE  BY GC/MS*
                                          Concentration (ppb)
                                 Sample P2T-1
                                  Sample P2T-?
 Dibenzo-p-dioxins (DO)

 TetrachloroDD
 Pen*achloro&D
 HexachloroDD
 HeptachloroOD
 OctachluroDD

 Dibensofurans  (DM
none detected (<10 ppb)
none detected (<10 ppb)
         1,714
        25.019
        80,053
none detected (<10 ppb)
none detected (<10 ppn)
        1,532
       27.810
       73,123
TetrachloroDF
PentachloroCF
HexachloroDF
H»ptachloroDF
OctachloroDF
16.3
77.4
1,760
4,418
6.030
8.2
6.1
1.643
4,748
7,074
  Analysis of EHSL Laboratory U.S. EPA, Las  V-:i-,. Nevada,  Method 8280  (SW-846
 EPA).
       TABLE 43.   TOXIC ITY OF  WATER SOLUBLE FRACTION MEASURED I?Y  THE
                MICROTOX  ASSAY TOR HAZARDOUS WASTE  SAMPLES
                   Waste
          Crr>i)',ot«?

          PCP

          API Separator Sludge

          Slop 0)* (vol


                            0.3

                            0.7

                            6.0

                            1.3
*EC'jO(5,15°) denotes the cond   -ons for th
-------
   4 -i
   3-
Z  2-
LU
3
ID
    I-
                              LEGEND
                                 TA-98wi«hS9
                                 TA-98 without S9
                        \
                       100
      200
DOSE (/ig/plate)
     300
       400
                     O.25
    I
   0.5
0.75
0.5
0.25
                                 mg wet wt waste/plate
 ri]ure 5.  Anes assay results Tor creosote sludge base/neutral fraction.

-------
   3-
O
Z
UJ
O
<
=>
    I-
               LEGENO
                  TA-98 withS9
                  TA-98 without S9
                       250
                                   5OO
                                                        750
1000
                                   DOSE
r
0
C.  .Tries
                       P-5                 1.0                 1.5
                                 mg wet wl waste/ plate
                   results for  pentdcnlocoplienol sludje bas«' neutral fraction
                                                                            2.0

-------
            3 -t
         2 2-
                      LEGEND
                        TA-98 with S9
                        TA-98 without S9
          O
          Z
          UJ
          O
vO
en
             H
                       IOOO
          I
       2000
       3000      4000
      DOSE (/tg/plate)
                5OOO
               6000
              7000
               I
               0
          Tijure 7.
I
2
I
4
I
6
8
I
10
I
12
14
               mg wet wt waste/plate
  results for Montachloroil'enol slu.lge nciJ fraction.

-------
exhibited.   The  background  lawn showed  signs of  toxicity  to the bacteria at
the 641ug/plate dose, with increased tomcity evident with larger doses.  No
mutagenirity was detected for  the  acid  fraction of the PCP waste either with
or without S9 activation (Figure 7).  Some indication of toxicity was present
at the highest Jose (6820 ng/plate).

     Results of mutagenicity testing  on  the  base/neutral  fraction of the API
separator sludge are  presented  in Figure  8.   No mutagenicity and no toxicity
were indicated at any of the doses  tested.

     Figure 9 shows Ames assay results  for th* b«e/«ieutral fraction of slop
oil  emulsion  solids.   Generally no  mutagenicity  is evident for the fraction
either with or without  added  S9.    Toxicity to the  bacteria  was definitely
observed  at  1485 ug/.plate dose.  For this reason,  the mutagenic  ratio at this
point,  1.88,  should  not  be  taken as  an   indication  of a  trend  toward
mutagenicity.  Toxicity  was initially indicated at a dose of 297  ug/plate in
waste samples without addition of 59.

     None of the wastes tested  showed  potential  for inutagenicity without S9
activation.   All of vhe wastes, with the exception of  API  separator sludge,
showed low  level mutacjenic  responses with addition of the  S9 microsomal mix.
Toxicity  was generally present to varying degrees in all  of the  samples  except
for  the base/neutral 'raction of the separator sludge.
SOIL CHARACTERIZATION

Introduction

     Critical  to  an  evaluation of the soil treatebillty of  a hazardous  waste
is  an  understanding of the soil that will  act  as  the  treatment medium for the
waste.   Tnerefore,  soil  characterization  tests were  conducted to obtain
specific physical and chemical properties for the two experimental  soils.  The
two soils  included  Ourant  clay  loam  and  Kidman sandy loam.   Criteria  for
selection  of  experimental  soils   included: 1)  general suitability  for   land
treatment  of  waste  (U.S.  EPA 1983, 19S6b), and 2) differences In physical and
chemical properties to allow for evaluation of the potential  influence of soil
type on waste treatment.

     Soil  physical  and chemical parameters measured  were  those identified as
potentially  influencing  degradation,  transformation, or  immobilization  of
hazardous  constituents in  soil  systems (U.S.  EPA  1983,  1984f,  1986b).
Physical  properties  are  those  characteristics,  processes, or reactions  of  a
soil caused by physical  forces.   Physical  properties that were evaluated are
given  in  Table 44.   Chemical reactions  that  occur between waste constituents
and the  soil  must  be  Identified  and  evaluated  with  respect to  treatment
effectiveness.    Chemical  properties  that  were  evaluated  are also  given in
Table  44.
                                       97

-------
            3n
                        LEGEND

                        •  TA-98withS9

                        O  TA-98 without S9
         O

         <


         O

         z
         UJ
         O
SO

00
I-
                                      500
                                                 1000
                                                                      I
                                                                    I5OO
                                            DOSE (/xg/plate)
          Figure  3.
           I               2              3

           1           mgwetwt  waste/plate

A.HCS assay results for API separator sludge base/neutral fraction,
                                                                             I
                                                                            4
                                                                                I
                                                                                5

-------
vO
vC
         O 2H

         H

         K

         U
         Z
         Ul
         (9
                       LEGEND
                         TA-98 with S9
                         TA-98 without S9
                                                   T
                          T
                          500
1000        1500       2000

      DOSE (/ig/plate)
—T	
 2500
	1
 3OOO
                                         I
                               I

                               2
                                         mg wet wt waste/plate

                   .":nes a-jsa> results for  -Jop oii eii-.-lsion sr'. >Js b.isc/ncutral fraction.

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       TABLE 44.  SOIL PHYSIZAL «.ND CHEMICAL PROPERTIES EVALUATED FOR
                            SOIL CHARACTERIZATION
Soil Physical  Properties                          Soil  Chemical Properties


Soil texture                                      Cation  exchange capacity
Bulk density                                      Total organic carbon or
Soil characteristic curve                          organic matter content
Available water capacity                          Nutrients
Porosity (saturated water content)                 Electrical conductivity
Particle density                                  pH
                                                  Buffering capacity
     Specific  soil  parameters measured  and values  obtained  may be  used  in
quantitative  assessment  to  evaluate treatment  and develop management
approaches for a soil/waste mixture.   An in-depth discussion of the  proposed
mathematical  model  to  evaluate the  effect of  site and  soil  properties  on
hazardous waste treatment  in  soil  is presented in t he  Perm it Gu i danc e Han u a1
on Hazardous Waste Land Treatment Demonstrations (U.S.  EPA  I986b).

Materials and Methods

     The experimental soils were chosen  to  represent a  spectrum  of soil types
that  are considered suitable  for land  treatment of  wastes.    The  standard
procedures followed  for  the  determination  of  the parameters  listed   for  each
soil are summarized in Table 45.  Included for each parameter measured are:  1)
standard method reference,  2)  instrumentation, and 3)  precision and  accuracy
objectives using the method.

Results and Discussion

     Soil physical, chemical  and biological  parameters  measured  are presented
given  in  Table 46 for the  Durant  clay  loam  and  in Table 47  for  the Kidman
sandy  loam.  Soil  moisture characteristic curves  for these soils are  given  in
Figures  10  and II for  the  Durant  and  Kidman soils,  respectively.    Soil
characteristics that are used  in the proposed  mathematical model developed  by
U.S. EPA, RSKERL  for evaluating hazardous  waste  treatment potential  in  soil
are noted in Tables 46 and 47.

     An  important soil  characteristic  with respect  to  waste treatabllHy
potential that varied  between  the two  soils  was  the organic carbon  content,
whir*  was spprcxfoat::! y  C'»  »<«••<  higher in the  Durant clay loam than in the
Kidman sandy loam.  Both soils  had active microbial  gopulst'ons, as indicated
by soil plate counts.
                                       100

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             1ABLE 45.  MEASUREMENT METHODS AND DATA QUALITY OBJECTIVES FOR SOIL ANALYSES
     Parameters
     Method
   Measurement Method/
     Instrumentation
Precision
Accuracy
Particle Size             Chapter 43*
  Distribution (Texture)
Water Holding Capacity    Chapter 19*
Bulk Der-;1ty              Chapter 30*
     pH                   Chapter 12+
Chapter 7*
Chapter 21*
Chapter 8*

Chapter 10+
Moisture Content
Total Porosity
Cation Exchange
  Capacity (CEC)
Soluble Salts
Metals
Nitrogen Forms
  NH4-N
Chapters 2, 16-23+
3000 series*
7000 series*
Chapter 33+
Section 350.2
Method 41 7
                                      **
Hydrometer method            +10*

Gravimetric
Core method
Electrometric method;        _
soil suspension/pH           ~~
electrode
Gravimetric                  +20*
Density method               +20*
Displacement method          +15*
Saturation extract;          +_20X
electrical conductivity
with conductivity meter
Extraction; atomic           +102
absorption analysis
Extraction; Nessler-         +10*
ization; titrimetric
                                                   _            Not Applicable

                                                   +20X         Not Applicable
                                                   +20X         Not Applicable
                                                   +0.1 units   +0.? units
             Not Applicable
             Not Applicable
             Not Applicable

             Not Applicable

             +10%

-------
                                         TABLE 45.  CONTINUED
                                                   Measurement Method/
                                                     Instrumentation
Paramet»rs
Method
                                                   Precision
Accuracy
Nitrogen For-ns (Cont.)
  N02-N, NCh-N
  Total Nitrogen

Phosphorus Form.,
  Ortho-phosphate

  Total Phosphorus

Total Organi: Carbon


Oil and Grease

Enumeration of Soil
Microorganisms
  Bacteria, Fungi
Chapter 33*
Section 353.2
Method 418**
Chapter 31*
                     Chapter 24*
                     Method 424**

                     Chapter 24*
                     Method 424**

                     Chapter 29*
                     lethod 505**

                     Section 413.1
                     Method 503
        **
                     Chapter 37*
                                           Extraction; automated        +_ 5%
                                           cadmium reduction            ~~
                 Micro-Dumas method           +_8%
                 (combustion method)

                 Extraction; ascorbic acid    *_6%
                 method
                 Digestion; ascorbic acid     +_9%
                 method
                 Combustion; TOC analyzer     +10*
                 Extraction method for        +15%
                 sludge samples


                 Total plate counts;          +20%
                 spread plate method	
                                                           +11*
                                                                                     +12X
                                                                +12%


                                                                +15X


                                                                +1RX



                                                                Not Applicable
 *Methods of Soil Analysis. Part 1;  Physical and Hireralogical Properties. Including Statistics  of
  Measurement and Sampling.  C. A. Black, Editor.  American Society of Agronomy, Madison, WI  (1965).
 *Hethods of Soil Analysis, Part 2;  Chemical and Microbiological Properties.  Second Edition.  A. L.
  Page (£d.).American Society of Agronomy, Madison, WI (198Z).
 *Test Methods for Evaluating Soliti Waste, Physical/Chemical Methods, SW-846, Second Edition, U.S.
  Environmental Protection Agency, Washington, DC (1982).
**Methods for Chemical Analysis of Water and Waste, EPA-600/4-79-020, U.S. Environmental  Protection
  Agency, Cincinnati, OH (1979).
**Standard Methods for the Examination of Water and Wastewater, Fifteenth Edition, American  Public
  Health Association, Washington, DC (1981).

-------
      TABLE  46.   CHARACTERIZATION OF  DURANT  CLAY  LOAM  SOIL COLLECTED FROM
      PROPOSED U.S. EPA LAND TREATMENT RESEARCH FACILITY, ADA, OKLAHOMA
     Soil Characteristic
    Value
     Physical Properties:
          Bulk density*
          Texture*
          Moisture at
               1/3 atmosphere
               15 atmospheres
               Saturation*

     Chemical Properties:
          pH
          C1C
          Organic carbon*
          Total phosphorus
          Total nitrogen
          Nitrate nitrogen
          Sulfate in saturated extract
          EC of saturated extract
          Iron
          Zinc
          Phosphorus (bicarbonate extractable)
          Potassium
          Ammonium acetate-extractable cations
               Sodium
               Potassium
               Calcium
               Magnesium

          Water soluble cations
               So-1 * urn
               PC  ssium
               Ca.ciuip
               Magnesium

     Biological Properties:
          Soil plate counts
               Bacteria
               Fungi
  1.59 g/cm3
  Clay loam

 41.6X
 12%
 55X
  6.6
 20.5 meq/lOOg
  2.88%
  0.03*
  0.21*
 18 ppm
  0.3 meq/1
  0.5 mmhos/cm
 28 ppm
  3.8 ppm
  3.0 ppm
177 ppm

  O.t meq/1OOg
  0.7 meq/lOOg
 19.4 meq/1OOg
  4.7 meq/lOOg
  0.03 meq/1OOg
  0.01 meq/1OOg
  0.21 meq/1OOg
  0.08 meq/lOOg
  5.1 x 107/g
  2.6 x 105/g
*Soil properties required for use in modeling land treatment of hazardous
waste (U.S. EPA 1986b).
                                       103

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  TABLE   47.   CHARACTERIZATION OF KIDMAN SANDY LOAM SOIL COLLECTED FROM USD
               AGRICULTURAL EXPERIMENT FARM AT KAYSVILLE, UTAH
     Soil Characteristic
    Value
     Physical Properties:
          Bulk density
          Texture
          Moisture at
               1/3 atmosphere
               15 atmospheres
               Saturation

     Soil Classification:

     Chemical Properties:
          PH
          CEC
          Organic carbon
          Total phosphorus
          Total nitrogen
          Nitrate nitrogen
          Sulfate in saturated extract
          EC of saturated extract
          Iron
          Zinc
          Phosphorus (bicarbonate extractable)
          Potassium
          Ammonium acetate-extractable cations
               Sod ium
               Potassium
               Calcium
               Magnesium

          Water soluble cations
               Sod ium
               Potassium
               Calcium
               Magnesium

     Biological Properties:
          Soil plate counts
               Bacteria
               Fungi
  1.49
  Sandy loam

 20%
  7%
 24%

Typic Kaplustoll
  7.
 10,
  0,
  0,
  0,
1 meq/lOOg
5%
06%
07%
  3.7 ppm
  4.8 ppm
  0.2 iiflihos/cm
  9.0
  1.2
  ppm
  ppm
 27 ppm
117 ppm

  0.24 meq/lOOg
  0.42 meq/100;
 13.6 meq/lOOg
  1.7 meq/lOOg
  0.01 meq/IOOg
 <0.01 meq/IOOg
  0.04 meq/IOOg
  O.G1 meq/IOOg
  6.7 x
  1.9 x 104/g
*Soil properties required for use in modeling land treatment of hazardous
waste (U.S. EPA 1986b).
                                      104

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 1
 I
50.
45.
40
35.
30.
25.
20.
15.
10
 5.
 0
                                       8
                                     Bars
                                         10
12
14
16
  Figure 10.  'Soil moisture characteristic curve for Durant clay loam.
    30
     25.
E   20.

1   '
I   1
     5.
                              6       B       10       12      14      16
                                     Bars
  Figure 11.   Soil moisture characteristic curve for Kidman  sandy  loam.
                                       105

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WASTE LOADING RATE EVALUATION


Introduction

     Determination  of  acceptable  wists  application  rates  (mass/area/
application)  is  specifically identified  in federal  HWLT regulations  (40  CFR
Part 264.272) as  a  requirement  for  conducting  a land treatment demonstration
(LTD).  Since the decomposition  of a hazardous waste  and the detoxification of
organic waste constituents in the soil depend to a large extent on biological
activities of soil  microorganisms,  an  important  consideration  in determining
waste  application rites  is  the potential  impact of the waste  on  microbial
activity.  This impact may be measured using a battery of short-term bioassays
that measure acute toxicity.


Possible Assays-

     Appropriate  bioassays should reflect the activity and/or survival of  the
soil microbial population.   This information may  indicate potential effects on
the microbes  responsible for waste degradation..  The tests selected should be
sensitive enough  to indicate potential adverse  impacts of a candidate waste on
the  soil microbial  population,  which is  directly related to the assimilative
capacity of the  soil.   Soil may be  acclimated through  additions  of  low
concentrations  of  some  wastes   (e.g., pentachlorophenol  waste)  so that  the
toxicity of  successive waste applications  to the degradative  populations is
minimized.   Assays  used  to determine  toxicity In acclimated  soils should
reflect  the  response  of  the general  microbial  community (e.g.,  respiration)
and  may  be  designed  to measure  specific  degradative  activity  (e.g.,
dechlorination)  to  waste  addition.   The  objective  is  to  predict  initial
loading rates  that will  allow detoxification  of hazardous  constituents to
occur  within the  defined  waste treatment  soil  as  a  result  of  normal   soil
biotransformation processes.

     The  toxicity  screening tests  to be  used  should  be  easily performed,
rapid,  and  inexpensive.  The tests also should  be validated  for the ability to
demonstrate responses of the soil microbial population to toxic environments.

Materials and Methods

Microtox—

     The Microtos™ system is a simple standardized  toxicity test system which
utilizes  a  suspension  of  marine  luminescent  bacteria (Photobacterium
phosphoreum)  as  bioassay organisms  (Bulich 1979).   The system measures acute
toxicity  in aqueous samples.    An  instrumental approach  is used  in which
bioassay  organisms  are  handled like chemical  reagents.    Suspensions  with
approximately 1,000,000  bioluminescent organisms in each are  "challenged" by
addition of  serial  dilutions of an  aqueous sample.   A  temperature controlled
photometric device  quantitatively measures the  light  output  in  each suspension
before  and after  addition of the sample.   A reduction of light  output  reflects
                                      106

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physiological  inhibition,  thereby  indicating  the  presence  of  tox
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     Experimental  Apparatus--

     A tumbler, wrist-action  or platform shaker  is  used to extract  the  WSF
from  each  sample.   Following  extraction, the  Microtox™  system is  used  to
determine the relative residual  acute toxicity  in each WSF sample.

     Water Soluble Fraction  (WSF) Extraction Procedure—

     A slight modification of the  distilled, deionized  water  (OW)  extraction
procedure as described by Matthews  and  Bulich  (1986)  is used  to generate  WSF
samples.    The following  steps  are followed  to prepare  these  samples  for
toxidty testing:

     a.   Place a 100 g  sample  of each  of  the background soil, waste,  and
selected soil-waste mixtures into i glass  extraction vessel.

     b.   Add  400 ml  of  distilled, deionized  water  (4:1  vol/wt  extraction
ratio) to each vessel  and  seal  tightly.

     c.   The tumbler shaker is the method of choice for mixing.  If a wrist-
action shaker  is  used, place  the vessels  on  the shaker  at  a 180° angle;  if a
platform shaker is used, place the  vessels on their side.   In  all  cases,  the
extraction vessels must be sealed tightly.

     d.   Allow the  extraction  vessels  to shake  for  20 + 4 hrs at approxi-
mately 30 rpm in the tumbler shaker or  60 rpm on the wrisF-action or platform
shaker.

     e.   Following the specified mixing period, remove  flasks fron< the shaker
and allow "  -n to sit for  30 minutes.  Decant the supernatants into high-speed
centnfuqe •  ,es.   Add 0.4 g of  NaCl  for each 20 ml  of sample; shak^,  then
cenr-    -IP a; 2,500 rpm for 10 minutes.

          JVepare a  sample from  each  test unit  for  Microtox™  testing  by
piK       20  ml   of elutriate  from each  centrifuge tube into  a  clean glass
container, sealing and storing at  4°C.   Carp must be  taken to ensure that any
floating material  is  not  transferred.   As soon as  all  samples  are prepared,
oegin Microtox™  testing;  conduct  all  tests ihe  same  day  that  they  are
prpoared.

     g.   Follow  the  test  procedure  outlined  in  the Hicrotox™  System
Operating Manual  (Beckman  Instruments,  Inc.  1982).

     Data Interpretation—

     Relativ? acute toxicity values (EC50 value along with upper and  lower 95
percent confidence limits) are calculated  for each WSF extract.   This  involves
preparing a  log-log  plot  of concentration versus  gamma light  decrease (gamma
is the ratio of light loss to light remaining),  Corrected for effects  of light
drift  based  on a  blank  response.    The ^concentration  of  thp  sample
corresponding to  a gamma light decrease  of 1  is termed the EC50  (t,T), meaning
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it caused a 50 percent decrease  in liqht output at exposure time (t) and test
temperature (T).

Soil Respiration--

     Soil  respiration  is generally  accepted  as  a  measure of  overall  soil
microbial activity (Hersman and Temple 1979)  and has  been used  as ar.  indicator
of the toxicity or  of  the utilization of organic compounds added to the soil
environment  (Pramer  and  Bartha 1972).    Respiration  may  also  act as  an
indicator  for  microbidl  biomass in  soil  because  the transformations  of the
important organic   elements  (C,N,P,  and  S) occur  through  the  biomass
(Frankenberger and Cick 19R3).   Measurement of  (#2  evolution from soil samples
is a commonly  used  indicator of soil respiration,  although measurenent of 02
uptake using  a Uarburg-type respirometer is a viable alternative  for short-
term  respiration.    Evolution of  C02  can be  measured  in  flow-through  or
enclosed  systems.   Flow-through  systems  involve  passing a stream of C02-free
air through  the  incubation chamber  and  then capturing  C02 from the effluent
gas stream in alkali  traps (Atlas  and  Bartha  1972).   The  Biometer  flask
described  by  Bartha and  Pramer  (1965)  is  an  example of the enclosed system.
It consists  of an Erlenmeyer  flask  modified  with a side-arm  iddition  which
serves as an  alkali reservoir for  trapping CO?.    A septum  in  the side-arm
allows for removing samples of the  alkali.   The flask itself is fitted with an
ascarite  trap for  maintenance  of  CO^-free  aerobic  conditions  within the
container.    The  carbon dioxide  produced  by   microbial respiration  is
quanlitated by titration of the alkaline  solution with acid of  known  normality
or by determination of total inorganic carbon in  the  solution through use of  a
carbon analyzer.

     Determination of soil respiration through  C02  evolution is an  inexpensive
and simple method  for  indicating  general  soil microbial  activity  and  acute
effects  of added  substrates  on that  activity.  Description of  the  use of soil
respiration  in the literature  is widespread,  indicating  the general  acceptance
of  respiration as an indicator of soil microbial  activity.  Soil  respiration
is  limited  in  that  results  wil' not necessarily  Deflect changes  in  specific
types  and groups of  microorganisms nor will  it  reflect  the potential for
anaerobic degradation or  degraoation of specific  organic constituents.

     Experimental Apparatus--

     Each  experimental  unit consists  of  a 500 ml Erlenmeyer  flask having  a
one-hole stopper  fitted  with  an ascari.te trap.  A stiff  wire  bent  to an  V
shape  at  the  bottom  is  suspended from the  stopper.   A scintillation vial
attached to  the  wire with a rubber  band contains  0.5 N KOH for  capturing COg
released from  the soil.

     Experimental Procer'ure--

     The method recommended  below  is .nodified  from the  procedure described  by
Bartha and Pramer (1965).
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     a.   Distribute 50  g  of  each of  the  background  soil,  waste,  and
soil '.waste mixtures to 500 ml  flasks, with  triplicates  for each loading.  Also
include three empty flasks as  blanks

     b.   Place a scintillation vial  Billed with 15 ml of a 0.5 N solution of
KOH into each flask and secure the stoppers.

     c.   Incubate the flasks  at  room temperature  (22 +_1°C).

     d.   Monitor  the  evolut'on  of  C02   for a  24-hour  pe.'iod.    For
determination of detoxification potential,  CO? evolution will  be n on Ho red at
specific time intervals.

     e.   The alkali  traps are changed  by  removing th» vial  of KOH from each
flask, capping it, and replacing  the  vial with  one  freshly filled with alkali.

     f .   Determine the amount of CO? in  each trap  using  a carbon analyzer and
testing for total inorganic carbon.  Where  a carbon analyzer is not available.
the amount of CO? evolved  can  be  determined titrimetrically.  Add an excess of
BaClj  to  the  alkaline  solution  to  precipitate  the  carbonate  as  insoluble
BaC03.  with  phenol phthalein  as  an indicator,  titrate  the unreacted KOH with
0.6 N  HC1.   Calcu ate evolved carbon expressed as CO?-C, using the following
fomula (Stotzky 1965):

     mg C02-C  =    (ml  of  HC1  to titrate blanks) -  (m1  of HC1  to  titrate
                    sample) x  normality  of  HC1  x equivalent weight; equivalent
                    weight = 6 if data expressed in terms of carbon.
     q.   Skw'ract the mean amount of C02-C  found  in the blank flasks from the
mean of  the  results from the other  flasks.   This  accounts  for  the C02 which
enters the flasks when samples are taken  and the flasks are  aerated.

     h.   Check  the  moisture  content  of each  unit  once  a week.    The
availability of water may have a large effect on microbial activity.

Dehydrogenase Activity--

     Oehydroqenation is the general  pathway  of biological oxidation of organic
compounds.  Dehydrogenases catalyze the oxidation of substrates which prcduce
electrons able  to  enter  the electron  transport system  of   a cell  (ETS).
Measurement of  dehydrogenase activity  in   soils  h*s  been  recommended  as an
indicator of general metabolic  activity of  soil microorganisms  (Frankenberger
and Dick  1983; Skujins  1973; Casida 1968).    Fret,  dehydrogenases  in  soil  are
not expected  because cofactors  such as NAD and   NAOH are  required, linking
dehydrogenase activity  to  living  organisms  (Skujins  1978).    The  type  and
quantity  of  carbon  substrates, both present  and  introduced,  will   influence
dehydrogenase activity (Ladd 1978; Casida 1977).

     The soil  dehydrogenase  assay involves   the incubation of soil  with 2,3,5-
triphenyltetr.izoUum chloride  (TTC) either  with  or without  added electron-
donating subs' rates.  The water-«oluble, colorless TTC intercepts the flow of
electrons produced by microbial dehydrogenase activity and  is  reduced  to the
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water-insoluble, red 2,3,5-triphenjltetrazol ium formazan (TTC-formazan).  The
TTC-toririazan  is  extracted  from  the  soil  with methanol  and  quantified
colorimetrically.

     The soil  dehydrogenase  activity assay  is  simple and  efficient.   It is
also  a  convenient  test  to run  since the  only major  pieces  of  equipment
required are a spectrophotometer, a centrifuge,  a.id depending on  selected test
conditions, an incubator.   However, since the  assay  indicates general activity
of  the  major  portion  of   the  soil microbial community,  it may not reflect
effects  of  an  added  substrate  or toxicant  on  specific  segments cf  the
community.

     Experimental  Apparatus and Procedure--

     The method for determination of dehydrogenase activity is based on Klein
et al. (19/1).  Activity both with  and without glucose  addition  is  determined.
Sorensen (1982) found that the increase  in  soil dehydrogenase activity due to
glucose  addition  can  be more  sensitive  to stress  than  the activity without
glucose.

     Triplicate  test  units  are  prepared for  each  of the  background  soil,
waste, and soil:waste mixtures.  Color correction  is  accomplished by preparing
one  extra  tube for each   combination  of soil:wa
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Nitrification—

     Cxidation of ammonium nitrogen to nitrite and then to nitrate nitrogen is
called nitrification.   The chemoautotrophic  bacteria  that derive  their energy
for growth  from  the  oxidation  of  ammonium ion (e.g.,  Nitrosomonas)  or nitrite
ion (Nitrobacter) are  sensitive to  environmental  stress  aid  are  not different
from  heterotropMc  bacteria  in  the  soil  in many  of their  requirements  for
metabolic activity and  growth  (Focht  and Verstraete 1977).   Coupled  with  the
fact  the  energy  yielding  substrates and/or  oxidized products of  nitrification
are easily  extracted from the  soil  and measured,  the  process of  nitrification
may be used as a bioassay of microbial toxicHy in the soil.

     A possible disadvantage of using nitrification as a toxicity indicator is
the high  sensitivity of  the bacteria involved.   This is especially  true  of
Nitrobacter (Focht and  Verstraete  1977).  Heterotrophic microbes may  be more
resistant and resilient.

     Experimental Apparatus and Procedure—

     The methods outlined below were  used by Sorensen  (1982) and  adapted from
Belser and Mays (1980).  The intent of the  assays  is  to  measure the potential
activity  of the ammonium  or nitrite oxidizing  bacteria  in  the  soil  over  a
relatively short period of time, and not to measure the ability of the soil  to
support  growth  of  these  organisms over   an  extended  period.   Substrate
concentrations are kept low to avoid toxic effects, and to avoid the necessity
of dilution prior to nitrite analysis.

     Initial potential  NHa* oxidation activity procedure—

     For each sample:

     a.  Weigh 6 g of soil into a 125 ml  Erlenmeyer flask.

     b.   Add  25  ml  of ammonium-phosphate buffer  solution containing 167  mg
K2HPOa/l,  3 mg  lO^PtVl,  and 66  mg (NHa^SOa/l.   The  pH  of this  solution
should  be  8  +  0.2.   Note:   A  bufier close  to the  test soil  pH may  be
desirable.    "~

     c.  Add 0.25 ml  of 1 M NaClOa to each flask to block M>2- oxidation.

     d.  Cover the flask with aluminum foil and shake  on  an  orbital  shaker  at
200 rpm for 22 ^2 h at ?4 +_2°C.

     e.   Clarify the  slurry or portion of  the  slurry  by  centrifugation  or
filtration.

     f.   Analyze the  filtrate or  supernatant  for NO£-N (Kenney  and  Nelson
1982;  APHA  1985).    Each  batch of  ammonium-phosphate buffer  should  also  be
analyzed  for  N02-N  and the  concentration  subtracted  from sample  results.
Chemical  interferences with the Griess-Ilosudy method  for MOj-N are described
by Kenney and  N?lson (1982) and APHA  (1985).  Most  interferences  are  uncommon
but may occur  in some  wastes.   Oil from petroleum wastes may be  present  in
                                      112

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supernatant; and cause difficulties by coating  colorimeter cuvets or tubing  in
automated chemistry  apparatus.   Oil  can usually be  removed sufficiently by
gravity filtration of  the supernatant through medium  speed  filter paper and
removing the filtrate before the last small  portion  (2-4 ml)  of  extract,  which
holds the oil on the surface, passes through the  filter.

     Initial potential  NO?" oxidation activity  procedure--

     a.  Weigh 6 g of soil into a 125 ml  Erlenmeyer  flask.

     b.   Add  25  ml  of  nitrite-phosphate buffer solution containing  167 mg
K2HP04/1, 3  mg  K^PO^/l,  and 4.5 mg NaN02/l.  The pH of this solution should
be 8 ±_0.2.  Note:  A buffer close to the test  soil  pH may be desirable.

     c.   Add  5   1  of   a  20  percent  solution of  nitropyrin (2-chloro-6-
(trichloromethyl) pyridine)  in  dimethyl  sulfoxide  to  each  flask to block the
oxidation of indigenous Nfy* to N02- (Shattuck  and Alexander  1963).
     d.   Process  each  flask and its contents as described for NH^* oxidation
described above in steps d through f.   !n this case  the  NOj-N concentration in
the  nitrite-phosphate  buffer  is  the   initial substrate concentration,  and
substrate usage is monitored.

Soil Plate Counts--

     Total counts of major  microbial  groups in  the soil are intended to show
the  viability  of  the  soil  microbial  community.   Comparison of  counts made
before and  afte'  waste  addition provide  an  indication of  acute microbial
toxicity to the specific microbial  groups and show the effect on the community
as  a whole.   Dominant  species may be  suppressed, allowing for an  increase in
the predominance of less common groups.  The change  in community structure may
be short-lived, but could possibly continue for  a lengthy period of time.

     Ideally, the plate count  procedures should create optimal  conditions for
the microorganisms to  be  enumerated;  therefore  medium composition, incubation
conditions and  Icnqth  of  incubation   are  important  considerations in  plate
count  assays.   It is  improbable  that all  types of microorganisms present in
the  soil  will   be  detected  using  agar  plates,  since   all  media types  arc
selective to a  certain  extent  (Greaves et  al . 1976).  Another disadvantage of
the plate count assay is that comparisons made among enumerations  performed at
different  times will   be  accurate only  if test condi  ions  for  each  set  of
counts sre identical.   In addition, the plate count  method  is not  conducive to
counting numbers of filamentous organisms or  those producing large quantities
of spores.  Also, there is not  necessarily  any  correlation between numbers of
microorganisms  and measured  metabolic  activities (Greaves  et  al .  1976).  The
microbial life  forms suggested  for enumeration, total  bacteria, actmomycetes
and  fungi,  are  the   most   important  soil  organisms effecting biological
degradation and transformation of hazardous waste constituents.
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     Media Preparation--

     The  following  three media are recommended for determining  viable  counts
of the selected microbial types:  tryptic soy aqar for bacteria,  Martin's rose
bengal media  for  fungi,  and  starch-casein  agar  for  actinomycetes.   Details  on
preparation of these media may be found in Moll urn (1982).

     Experimental Procedure--

     a.  Prepare a sufficient quantity of plates of  each media type.

     b.  Prepare dilutions of  the control  soil  and  each soil:waste-mixture  in
triplicate  according  to  section 4.2.2 of  Wollum  (1982).   Three dilutions  of
each  replicate will  be  plated on  each  type  of media.   For bacteria and
actinomycetes  10-6, 10'5, and  10'4  dilutions are  recommended.  For  fungi, the
suggested dilutions are 10-'. 10'4, and 10-3.   The solutions  to be  used  should
encompass the  optimum number of organisms  for counting, i.e.,  30-300  colonies
for  bacterial and  actinomycete  plates  and  10-20  for fungal  plates.   All
dilutions  should  be  prepared  in  the  same manner  since  comparisons across
treatments will be made.

     c.  Prepare spread plates  according  to section  5.2.2  of  Wollum (1982).

     d.  Incubate the plates at a controlled temperature,  generally between  24
and  28°C.    The  period of  incubation depends on  temperature  and  growth
conditions,   for  bacteria and  fungi 4 to  7 days  should be  sufficient,  while
actinomycete plates may have to be incubated 10  to 14  days.

     e.   Average the number  of  colonies  per  plate  for each dilution and
determine  the number of colony-forming  units  per  gram dry weight  of  soil.
Significant differences  in  numbers of colony-form ing  units  from the control
can be  determined using  statistical  tests.  A significant  reduction in the
number  of  colony-forming units  found in  the  soil  treated  with waste  as
compared to control  soil  indicates the degree of acute toxicity.

Preparation of Waste Soil Mixtures fcr Bioassays--

     If air-dried soil  is used,  it should be brought to the  desired  moisture
content  (ninimjm  60  percent of  the  water-holding  capacity of  the  soil,
preferably a moisture content  that  will  prove typical for field conditions).
The soil is acclimated to the increased soil moisture  content for 7  to 10 days
to .*llow  for  growth  of  soil  microorganisms.   After  the acclimation period,
waste which has osen  thoroughly mixed  is  added  to the soil   at the previously
selected loading rates.   W'.ien smell  percent leadings are to be tested, i.e.,  <_
10 percent, it may be difficult to  evenly  disperse  the waste material  in the
soil.   The use of an  organic solvent  as  a  dispersal agent may not be  feasible
in all cases  *ince  some  solvents have toxic  effects  on microbial processes.
The following  method  has  proved  successful  for  providing  a fairly uniform
distribution of small  quantities of waste in soil.  A soil:waste mixture  at  a
concentration nigher than the  upper loading rate is prepared  using air-dried
soil.   The waste  is incorporated  into  the  soil  by mixing  on  a rotary tumbler
for -12  hours  at 30 rpm.   This soil:waste  concentrate  can  be "diluted"  with
                                        114

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 additional  acclimated  soil  so that the final concentration  of  waste is equal
 to  the desired loading rate.   Soil without  wastt  should be added  and mixed
 into the control using  identical procedures as uspJ with soil:waste mixtures.

     Use of air-dried soil encourages sorption cf volatile constituents, since
 the presence  of water  can  displace volatile constituents  from soil.  Although
 some  volatile constituents  may be  lost  from  the  soil   during  the  mixing
 process,  this  preparation process  simulates aoplication of  waste  in  field
 conditions using surface incorporation and mechanical mixing of waste with the
 soil.   An  alternative  method  of waste  application  should  be considered if the
 goal  is to  simulate subsurface  injection  to minimize  loss of volatile
 constituents.

     After the  waste has  been added to the soil  and thoroughly incorporated,
 the soil:waste mixture  and control are allowed to incubate 24+2 hours.  This
 incubation  allows  for  acute  effects of the  waste on  soil  mTcrobiota to  be
 expressed.    After  the incubation  period,  the  selected  toxicity  assays  are
 started.   Except for soil  plate counts,  the assays described  in  this chapter
 require 24 hours for incubation or extraction.

 Preliminary Loading  Rate Investigation--

     In order to use any of  the previously described acute toxicity tests for
 determining an appropriate range of  waste  application rates,  a  set of initial
 rates to test should be chosen.

     Microtox--

     Matthews and Hastings (1985)  described a method  using the  Microtox assay
 to determine  an initial range of waste application rates.   The following steps
 are involved:

     a.   Obtain a 5 kg sample of the site  soil  and a 1  kg sample of the waste
 to be  applied.  Proper  sample collection  procedures should be used  to insure
 that characteristics of soil  and  waste samples  are representative of  those
 anticipated at the site.

     b.   weigh  out  two  100  g  aliquots   of  air-dried  soil  which has  been
crushed and sieved  to  2 mn; weigh out Uo 100  g aliquots of waste  which has
been thoroughly mixed.

     c.   Prepare WSF  samples  for  toxicity testing by  extracting  aliquots  of
the duplicate waste and soil  samp js as  described  in tho Microtox  methods
 section.

     d.   Conduct Microtox™  tests on  each WSF sample  prepared  as previously
described.   Experience suggests that if the EC50 for the  WSF  of  a  given waste
 as defined by the  Microtox™ system exceeds  25 percent,  the  EC50  for the WSF
of any waste-soil  combination will exceed 20  percent  and  toxicity  as measured
by  the MicrotoxTM  system  will  not be  a   significant  factor in  determining
loading rate.  - This  does not  preclude  use  of  the test system to determine  if
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toxicity reduction  of hazardous  organic  constituents  within  the waste-soil
matrix is occurring over time.

     e.   If tha  soil WSF  is  nontoxic,  i.e.,  foe full  strength DH extract
effects <  25  percent decrease in  bacterial  bioluminescence,  the soil has no
apparent "residual  toxicity.    If  soil  residual  toxicity is  indicated  (> 25
percent decrease  in  light  output  in  the full  strength DW  extract),  the
apparent cause should be determined prior  to  further testing.

     f.   Determine  four  loading  rates  to  be  used  in  subsequent   toxicity
screening tests according to the following criteria:

     1)   Calculata  the  EC50  and 95 percent  confidence  limits  for the waste
          WSF.

     2)   Choose the  upper limit of the 95 percent confidence interval as the
          highest loading rate  to be used.   For example, if the WSF of the
          waste  has  an  average  EC50 of  10   percent  and  upper  and  lower 95
          percent confidence limits of  12 percent and 8  percent, the highest
          loading rate would be 12 g of waste per 100  g of soil.

     3)   Use  1/4,   1/2, and  3/4 of the  upper   limit  as  the  remaining three
         • loading rates (in percent wet weight waste per  dry weight  soil) for
          testing.

     g.   Weigh  out  four  300 g samples  of  prepared  soil.   Add prescribed
amount of waste  and mix  thoroughly to  achieve the four loading  rates  (wt/wt)
determined by the criteria described above.

     h.   From  each of the  four samples, remove   three  100 g  (dry wt)
sub samples  and  place  in  a flask  or bottle  for extraction.    Discard  the
remainder of the sample.

     i.   Extract each  of the  12  subsamples with distilled, deionized water
according to the procedure describee previously and conduct Microtox™ test on
the WSF constituents.

     j.   Calculate  the EC50 and 95 percent  confidence limits for each waste-
soil wSF.  Average  triplicate values to obtain EC50 and 95 percent confidence
limits for each loading rate extracted.  Transpose each EC50  value to toxicity
units  (TU) in soil  using the following equation:


     Soil TU.|g,x4


     k.   Prepare  a log-log plot  of toxicity units versus  loading  rates for
use in estimating an  acceptable initial loading rate window.   The Interception
point  for  20 soil  TU 1s the  lower loading  limit  for the  window;  the  upper
limit  is  defined as  twice  the  lower limit.    Experimental  data  generated  to
date suggest that this is a reasonable window for initial  loading.
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     Other Assays--

     When  using  assays  other  than  Microtox™  for  preliminary  initial
application rate estimation, the  following procedure may be useful:

     a.   Choose three or four  loading  rates  that  cover the range from 0  to
the maximum  rate likely to  be  used  based  on  mobility,  soil  hydraulic
conductivity effects,  anticipated  degradation  rates,  or other criteria.
Concentration steps  should increase by approximately a  factor of  10 (e.g.,  0,
0.1, 1, and 10  percent by weight).

     b. •  Perform  the  selec'ed  acute  toxicity  bioassays on  each  of the
soil:waste mixtures.

     c.   Beginning  at the concentration showing little or no toxicity  in  step
b above, prepare a series of loadings  that encompasses the concentration  where
activity  is reduced  approximately 50 percent  relative  to  the untreated
control.   Smaller increments in concentration  should be used than  in  step  a.
above.

     d.   Repeat the  acute toxicity bioassays.   The results of  these  assays
should identify a range of loading rates that  are not highly toxic to the soil
biota.   The potential  for  these  loading  rates  fo allow for  detoxification
should be determined in a longer  term  toxicity reduction study

Selection of Waste Loading Rates-

     Giving greater weight to  *h<  level of toxicity indicated by assays  which
indicate activity among  a broader spectrum of  the microbial  population (e^g..
respiration  and dehydrngenase)  or  indicating general  toxicity (Hicrotoxin),
but considering  all  assay results,  select  a range of  loading  rates that  are
not  likely  to  inhibit  microbial  activity  but  will utilize  the  apparent
assimilative capacity of the soil.

     The  detoxification potential of  soilrwaste  mixtures .loaded at  rates
determined by tesults of  any of the short-term bioassays previously described
can be evaluated  with a six-week toxicity reduction stud-'.   This information
will prove useful in refining  the set of loading  rates  to be used.for  a long-
term land  treatment demonstration.  Matthews  and Bulich (1986)  have described
a  toxicity reduction  experiment  procedure  using  the  Microtox' assay  where
duplicate  samples  of  wasteisoil mixtures  loaded  at  selected rates  are
sacrificed  immediately  following  waste  application  and at two-week intervals
for  a  six-week  period.   At each  sampling time (i.e.,  days  0,  14,  28  and 42)
the soilrwaste mixtures  are extracted with water and analyzed for toxicity on
the Microtox system.  Scil moisture in the  test units is maintained between 40
and 70 percent  of the soil moisture holding capacity for  the duration of the
experiment.

     The detoxification  potential of  a  qiven  loading rate is indicated by the
changes  in acute toxicity of the water extract during the  experimental  period.
A  significant degree  of  detoxification  is shown by a toxicity  reduction trend
with the calculated EC50 for day 42 approaching or  exceeding 100  percent.
                                       117

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Data Interpretation--

     No single  assay  of  soil  micrcbial  activity  is  likely to  indicate  the
activity or viability of  the  broad  spectrum of soil microorganisms or  their
functions.    Measurements  of  respiration nuy represent  the  activity  of  the
broadest community of  microorganisms.   When  information en the toxicily of  a
waste or  its  degradation  or  transformation  product?  is available from more
than one assay, decisions on  acceptable  levels of toxicity for  loading rate
determinations or determination  of detoxification will  be more  reliable.
Broad spectrum  assays (e.g.,  respiration or  dehydrogenase)   and  general
toxicity  (Microtox™) are recommended  for .inclusion  in  a  any battery of
assays, but assays relating to  specific  subgroups  of the micrpbial  community
(e.g., nitrification,  nitrogen fixation,  or cellulose  decomposition) may also
be considered.

     Results  from  assays  measuring  universal  metabolic  activities  (e.g.,
carbon  dioxide  evolution)  or general  toxicity (e.g., Microtox™)  should
normally be give more  weight  in decision making,  but  if other assay  results
indicate severe toxicity,  lower  loading rates should be investigated.

Results and Discussion

     The bioassays used in  this study for  determining  loading  rates  included
the Microtox system and soil respiration.   Using the Microtox system,  a series
of  loading  rates  were  evaluated  for each  soil  and  waste  combination
immediately after  waste  incorporation into  the soil.    For some  soiltwaste
mixtures,  the  process was  extended  to  a  six-week toxicity reduction  study.
Using CO?  respiration,  loading  rates for  the  four wastes mixed  with  Durant
clay loam soil were evaluated  for a  60-day  period.

Selection of Loading Rates for Creosote Waste—

     Toxicity reduction study results  for creosote waste mixed   with  Durant
clay loam soil are found in  Figures  1? through  14.    The tests were performed
in triplicate at three separate times, using the  same  soil  and waste  samples
each time.  Results of the  three tests  are comparable,  showing that  the 0.25
percent loading became essentially nontoxic after 14 days incubation, and with
the exception of  trial  #2,  the  0.5  percent  loading became  nontoxic  after 42
days.   The  highest  loading, 1.0 percent,  showed  a detoxification  trend,  but
the water-soluble fraction of the soilrwaste mixture exhibited  a  'airly toxic
EC50 (average  of  3 reps = 27.3, S.D. = 9.7)  after  42 days incubation.

     Figure 15 illustrates soil  respiration results for creosote waste applied
to Durant clay loam soil.  All waste loadings exhibited greater CO2 production
than the soil control,  with production of  COg increasing with waste loading.
The  highest creosote  loading,  1.0 percent, did  not  appear  to  inhibit
respiration activity at any  time during the 60  days of incubation.

     Based  on  Microtox and  soil respiration  results,  U.e  loading  rates
determined for  creosote waste mixed with  Ourant clay  loam  soil were  0.7
percent, 1.0 percent,  and  1.3  percent waste wet weight/soil dry weight.
                                      118

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      120
      100.


   ~  80.
   I
   "J   40J
       20.


        0
Figure 12.
             •025% Load Rate
                   •05% Load Rale
                   O1% Load Rate
                      10
              15
20     25
Time (Days)
30
35
40
45
Toxicity of  water  soluble  fraction  measured  by  the  Microtox
assay with incubation  time  for creosote waste mixed with Ourant
clay  loam  soil  for  loading  rate  determination, Trial  #1.
EC50(5,15°) denotes the conditions for the test, i.e., reading
light output 5 minutes after sample  addition  at a temperature
of 15°C.
       120
       100
   I   60J
        40.


        20.


         0
Figure 13.
             •025% Load Rate
                   •0.5% Load Rate
                   01% Load Rate
                       10
               15
 20     25
 Time (Days)
 30
 35
 40
 45
Toxicity of  water soluble  fraction  measured by  the Microtox
assay with incubation time for creosote  waste mixed with Ourant
lay  loam  soil  for  loading  rate  determination,  Trial  K.
EC50(5,15°) denotes the conditions for  the test, i.e., reading
light output 5 minutes  after sample  addition  at a temperature
of 15°C.

-------
        120
        100.
     _.   80.
     *   60J
     in
     O
     w   40,
         20.
Figure 14.
              •025% Load Rate
                    •05% Load Rate
                  O1% Load Kate
                        10
                15
20     25
Tune (days)
30
35
40
45
Toxici*.y of  water soluble  fraction  measured by  the Microtox
assay with incubation time  for  creosote waste mixed with Ourant
clay  loam  soil   for loading  rate  determination.  Trial  #3.
EC50(5,15°) denote', the conditions for the test, i.e., reading
light output 5 minutes  after sample  addition  at a temperature
of 15°C.
     Microtox assay results  for  creosote waste mixed with  Kidman  sandy loam
soil are  presented in Figure  16.    Comparison  of these  results with  day  0
results using Durant soil shows that the EC50 values for Kidman soil :creosote
mixtures  were  approximately  one-half of  those  obtained  from  Ourant
soil:creosote mixtures.   For this  reason,  the loading rates  determined  for
creosote waste mixed with Kidman  soil  w»re  less  than  those for Durant:creosote
mixtures.    The  selected loadings  were 0.4 percent,  0.7  percent,  and  1.0
percent waste wet weight/soil dry weight.

Selection of Loading Rates for Pentachlorophenol Waste—

     The results for the 42-day toxicity reduction  study for pentachlorophenol
wood preserving  waste mixed  with Durant  clay  loam soil  (Figure  17)  showed
detoxification of  the waste  at  the 0.2 percent  loading  rate after  14 days
incubation.  A detoxification trend over the 42-day period was present for the
0.4 percent loading, but only slight  changes  in toxicity  were evident at the
highest loading, 0.8 percent.

     Soil  respiration results for  Durant:PCP mixtures are presented in  Figure
18.  Carbon dioxide evolution increases with waste loading, and production of
C02  for  all  waste  loadings was greater than  that  of  the  soil control.
Doubling  the  PCP  loading  from  0.2  percent  to  0.4 percent  increased CO?
production 25  percent by day 55,  while doubling  the loading  again from 0.4
percent to 0.8 percent increased  production by only 16 percent.
                                        120

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o>
CD
E

Ol
e
3
U
    120-
    100-
     80-
60-
     4O-
     20-
                   n

                   6.
                  LEGEND
                 0.25% Creosote
                 0.5%  Creosote
                 1.0% Creosote
                 Soil Control
                10
                                                                             SO
                                   20            30            40
                                        Incubation Time (days)
Figure  15.   Soil  respiration results  for creosote waste mixed with  Durant clay loam soil
—i
 60

-------
        81
        7.

        6.

        5,

        4.

        3

        2

        1

        0
Figure 16.
in
o*
C.

-------
X

E
o
ui
Ol
•n
.TJ
>

+J
10



E
3
O
    90 ^
    80 J
    70-
    ao-
    50-
    30-
    20-
    10-
                     LEGEND
                  o   0.2% PGP

                  o   0.4-%PCP
                  A   Q.B%PCP

                  •   Soil Control
                    10
                                                                              so
                                    20            30             40

                                         Incubation Time (days)

Figure   18.  Soil respiration results for PCP waste mixed with Durant  clay loam  soil
 i
60

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     Therefore, while  increased  loading  rates exerted  a  toxic  effect on  the
Microtox organism,  an obvious  toxic  effect was  not.  observed  with  the  soil
respiration assay.  The soil loading rates selected for  further  study based on
Microtox results  were 0.3  percent,  0.5   percent,  and 0.7  percent  waste  wet
weight/soil dry weight.


     Figure 19 presents  Microtox  results  for PCP wood preserving waste mixed
with Kidman sandy loam.  Aqueous extracts of these soil:waste mixtures at  very
small percentages of PCP proved highly toxic to the Microtox organism.  At the
lowest  loading  tested,  0.05  percent, the  resulting  extract LC50  was   14.2
percent by volume.  A comparable EC50 of  11.6 percent  was  obtained  for the 0.4
percent  loading  of PCP  mixed  with Ourant  clay loam soil,  almost  a  10-fold
increase  in  waste  loading.   The  loadings  chosen for  PCP  mixed with Kidman
sandy  loam were  0.07  percent,  0.15 percent* and  0.3  percent  waste   wet
weight/soil dry weight.

Selection of Loading Rate for API Separator Sludye—

     The toxicity of  aqueous  extracts  of  various  loadings of  API  separator
sludge mixed with Durant clay loam soil   as  determined bv the Microtox system
is shown in Figure 20.  These results show no trend toward 'ncreasing toxicity
with  increased waste  loading.   It appears  that the toxicity  of  this  waste as
indicated by the Microtcx system, was negligible.

     Carbon dioxide evolution results for Durant:separator sludge mixtures are
presented  in  Figure 21.   Although all  loadings  increased  production of  CO?
over  that  of  the soil  control,  there was little  difference between   the
loadings.  Doubling the  loading  from  8 percent separator  sludge to  16 percent
caused  essentially no  change  in  C02 production  again suggesting  negligible
toxicity and that  some resource other than  the decomposable components of the
waste limited respiration activity when more than  8 percent  waste was applied.
Perhaps oxygen moveme:it through the soil  was hampered  as pore space was filled
with oil.

     Loading rates selected based  on the  information obtained  from  Microtox
and  respiration  assays for  API separator  sludge  mixed  with  Durant clay  loam
soil  werp  6 percent, 9 percent  and 12 percent waste  wet weight/soil   dry
wpight.  Twelve percent waste was  selected  as  an  upper  limit  based  on current
industrial oracticr, the  need  for waste retention in the treatment  zone, end
the  lack of respiration stimulation at higher application  rates.

     Microtox results  for API  separator  sludge mixed with Kidrran sandy  loan
soil are presented in  Figure ?2.   It appears that  there may be a trend towards
increasing toxicity with increased loading rate.  Ths  loading rates for Kidman
sandy loam were  identical to those chosen for Durant  clay  leant, 6  percent,  9
percent, and 12 percent waste net weight/soil dry weight.

Selection of Loading Rates for Slop Oil Emulsion Solids—

     Microtox  water  soluble  extract toxicity  results  for slop oil waste
applied to  Durant clay loam soil  ore shown on Figure 23.   The EC50 of  47.8
                                      124

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 n
 iff
 5"
 in
 O
 ui
16.

14.

12.

10

 8.

 6

 4.

 2
              .1
  .4     J5     .6     .7      .8
Loading Rate (%)
                                                                .9
Figure 19.     Toxicity  of  water  soluble fraction  measured  by  the Microtox
               assay  for PC? wood  preserving waste  mixed with  Kidman sandy
               loam soil  for  loading  rate determination.  EC50(5,15°) denotes
               the  conditions for  the test,,  i.e.,  reading  light  output  5
               minutes after sample addition at a temperature of  15°C.
     80.
 o   SO.
 »n
 8
 o
 ui
50.

40

30

20

10
                            6      8     10     12
                                Loading Rate (%)
                                                  14
                              16
18
Figure 20.     Toxicity  of water  soluble fraction  measured by  the Microtox
               assay  for API  separator  sl-.dge waste  mixed with  Durani c'ay
               loan* soil  for  loading rate determination.  EC50(5,15°) denotes
               the  conditions for  the test,  i.e..  leading light output  S
               minutes after  sample  addition at a temoerature of 15°C.
                                       125

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O)
0>
4-1
I/I

E
3
U
     200-1
      175-
     150-
      •03 J
     100-
      75-
        LEGEND
o   4% Separator Sludge
a   8>% Separator Sludge
&  16% Separator Strdge
•  Soli Control
                       10
                 30            30
                     .I ncubcrtion Time (days)
                                                                                so
                                                                           so
Figure  21.   Soi".  respiration results for API  separator s'ludge mixed with Durant  clay  loam  soil.

-------
      100.


   I  *
   «  60
   in
   8  40
   UJ
       20

        0
                            6     8     10    12
                                   Loading Rate (%)
                                               14
16
18
20
Figure 22.     Toxicity  of water  soluble  fraction  measured  by the  Microtox
               assay  for API separator  sludge waste mixed  with Kidman  sandy
               loam soil  for loading rate determination.  EC50(5,15°)  denotes
               the  conditions  for  the  test,  i.e.,  reading  light output  5
               minutes after  sample  addition at  a  temperature  of 15°C.
  in
  i/i
50

45.

40.

35.

30.

25.

20.

15.

10.

 5.

 0
Figure 23.
                 1   1.5   2   2.5   3   35   4   4.5
                                  Loading Rate (%)
                                                  5.5   6   6.5
         Toxicity  of  water  soluble fraction  measured by  the Microtox
         assay for slop oil  waste mixed with  Durant  clay loam soil  for
         loading rate determination.  EC50(5,15°) denotes the conditions
         for the test, i.e., reading light output 5 minutes after sample
         addition at a temperature of 15°C.
                                       127

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percent  for  the  highest  loading, 6  percent, was  less  toxic  th*»!  the  EC50
values for the  lower  loadings.   As observed with the  separator  sludge  waste,
results from the Microtox system may be less helpful  than  wUh wood preservinq
wastes in determining waste  loading  rates for this particular soil  and  waste
combination.

     Figure 24  shows  soil respiration results  for  slop  oil:Durant clay  loam
mixtures.  All loadings tested show an increase in CO2 production over  that  of
the  soil control,  with  the highest  loading,  6 percent,  showing  the greatest
cumulative production of 003.

     The following  waste loading  rates  were selected for  slop oil  emulsion
solids mixed  with Our ant  clay loam  soil  based on  Microtox   and respiration
results:   8  percent,  12 percent, and  14 percent waste  wet   weight/soil dry
weight.

     Figure 25 presents Microtox  results  for  slop oil  mixed with Kidman  sandy
loam soil.   The results are similar to those obtained for  mixtures of slop cil
and  Durant  clay  loam soil.   The  loading  rates selected  for  Oop oi1:Kidman
mixtures are  lower  than those for using  Durant  soil:  .6  percent, 8 percent,
and 12 percent waste wet weight/soil  dry weight.

Summary of Loading Rates—

     A summary of  loading rates for  all   soil and waste types  is presented  in
Table 48.

Acute Toxicity Comparison Study

     The spectrum of microbes  in the  soil   include  organisms that  are  both
procaryotic and eucaryotic and that have autotrophic, neterotrophic,  and  mixed
autotroohic'heterotrophic metabolism.  The autolrophs may  be photosynthetic  or
chemoautotrophic,  and the heterotrophs may be  oligotrophk  or  prefer easily
metabolizable substrates  in  rich abundance.    Other  organisms  are strictly
predatory.   The types of compounds  attacked  and the  rate  of degradation  of
these  compounds  also  varies widely  among the  soil microbes.   The  waste
degradation process  depenas  on the  activities  of  
-------

X
• r—

e

E
e
3
o
     200 n
   LEGEND

a  2% Slop Oil
                                          Incubcrtion Time (Joys)


Figure  24.  Soil respiration results for  slop oil emulsion solids mixed with Durant  clay  loam  soil

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analysis  of  variance  and  the  least significant  difference were  used  to
identify  significant  differences  among bioassay  responses due  to  different
waste loading rates.
   IA
   O
   in
45.

40

35.

30

25.

20

15

10

 5
         01      2345678
                                  Loading Rate {%)

Figure 25.     Toxicity of  water soluble  fraction  measured by  the Microtox
               assay for slop oil waste mixed with Kidman sandy loam soil for
               loading rate determination.   EC50(5,15°) denotes the conditions
               for the test", i.e., reading  light  output 5 minutes after sample
               addition at a temperature of 15°C.
     The responses of the assays  to  the  PCP waste are illustrated in Figures
26 through  30.   The  Micrctox assay was very sensitive to the aqueous extract
of the  soil-PCP  waste mixtures,  with  less  than 13 percent  (vol:vol)  of the
extract of  the  0.05  percent loading rate producing an  ECso  (Figure  ?6).  At
the  0.5 percent  loading rate the ICy) was  about  3 percent  (volrvoi)  of the
extract.  These results indicate  a high toxicity of the soil waste mixture To
bacteria, even at loading rates  of 0.05 percent  or less.

     Severe toxicity  to initial nitrification  activity was  also  observed at
the  0.05  percent loading rate  (Figure  27),  where the average activity  level
was  reduced to nearly 10 percent of the  untreated  control  for both ammonium
and  nitrite ion  oxidation rate.   If  nitrification  activity is to  be  protected
in this soil,  application  rates below 0.05 percent will  be  required.    While
the  treatment  site  is  being used  and  intensively managed,  maintaining the
nitrification process may  not be necessary  since nitrate fertilizers  may be
applied  to  meet  nitrate demands  if  any arise.   When  the site is closed and
returned  to natural   processes,  however, reestabl ishment  of nitrification
processes  will  be  important  to  nitrogen  cycling  processes  in  many
environments.   Perhaps  the greatest  value in the  indication of nitrification
toxicity lies in the implication that other  specific biochemirjl processes may
                                      130

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TABLE  48.   SOIL LOADING RATES FOR UAS1ES BASED ON MICROfOX
                AND SOIL RESPIRATION RESULTS
Waste
Creosote
Pentachloro phenol
API Separator Sludge
Sloo Oil
Loading Rates

Low
0.4
0.075
6
6
Kidman Sandy
Medium
(%
0.7
0.15
9
8
Loam
High
waste wet
1.0
C.J
12
12

Low
weight/soil dry
0.7
0.3
6
8
Durant Clay
Medium
weight)
1.0
0.5
9
12
Loam
High
1.3
0.7
12
14

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    14 -
c

o

o>  tO
QL


-------
    100
                                                              = LSD
                                        0.2             0.3              0.4
                      LOADING RATE (% wet wt. waste/dry wt. soil)
                                                                              0.5
Figure 27.
Initial  ammonium and nitrite ion oxidation in response to treatment of Kidman soil  with PCP waste
after 24*2 h Incubation (LSD= least significant difference).

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also  be  impacted and  that  further  investigation  of  biological  effects  is
warranted.

     Dehydrogenase activity also  showed  a toxic  response  to  PCP waste (Figure
28), with  similar response being  shown  in assays  with  and  without  glucose
addition at all  loading rates  except at the 0.5 percent rate.   The  appar-.Mt
stimulation of activity  at  the 0.5  percent rate  in the  assay, with  glucose
added,  could  reflect  stimulation of  organisms  capable  of degrading  the
hydrocarbon matrix of the PCP waste.

     Respiration results  (Figure  29)  were highly variable,  and  no
statistically significant difference (p <_ 0.05) could  be  found  between  the
loading rates.   However, the mean activity tended to increase with increasing
loading rate, and  then  decreased  slightly  at  the 0.5  percent rate.   The lack
of  precision  in   the  data make  these  results  somewhat ambiguous,  but  no
evidence of severe toxicity was shown with  this assay.

     The stimulation of  activity  in  the  hydrocarbon degrading  portion  of  the
microbial community  that  are  resistant  to PCP and  other toxics  in  the waste
may mask toxicity of  the populations indicated by the Microtox, dehydrogenase,
and  nitrification  assays.   With this  possibility  in  mind,  a  conservative
approach  would  be to  base loading  rate decisions on  the  results  of  the
Microtox,  nitrification, and  dehydrogenase  assays.   In  this case,  loading
rates of 0.05 percent or less  seem justified.

     Viable  counts  of  bacteria and fungi  (Figure 30) do  not  reflect  any
toxicity  of  PCP   waste  at the loading  rates tested.   In fact,  the  fungal
population  increased  significantly  (p  < O.U5J   at  the  0.25  percent  loading
rate.  The slight  increase in  bacterial "counts  with loading  rate  is  probably
not significant.

     Responses  of the same battery of assays were also investigated  with slop
oil mixed with  Kidman sandy loam  soil.   Respiration  rates and counts of viable
aerobic bacteria  and  fungi  in  the Kidman soil  showed  no  significant toxicity
of  slop  oil  fo"   application rates  ranging from 2  to  14 percent.   Microtox
exhibited an EC50 at  less  than   10  percent  of  the  aqueous  extract of  the 2
percent slop oil  in K:dman soil (Figure  31),  indicating considerable toxicity.
Toxic'ty, as indicated by Microtox,  increased significantly (p £0.05) with an
increase in slop  oil  loading  from 2 to 6 percent, but  changes Tn toxicity due
to  increases  in  loading  to  10  and  14  percent  did  not cause  significant
increases in toxicity.  The average Microtox EC50 increased  at the 14 percent
loading rate, and  was  not significantly different  from  the  2 percent loading
rate.

     Nitrification activities  also showed significant toxicity of the slop oil
waste  (Figure  32).   Ammonium  ion  oxidation  was apparently  more  severely
inhibited by the  slop  oil  than nitrite  ion oxidation.   Nitrite ion oxidation
was significantly less  inhibited  at  the  10  and  14  percent  loading rates than
at the 2 and 6  percent loadings.   The mfe  extreme toxicity of slop oil to the
ammonium oxidizing bacteria would inhibit the nitrification process, however.
                                       134

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   100
        0


Figure  28.
           O.I              0.2              0.3             0.4

          LOADING RATE  (% wet wt. waste/dry wt.soil)
                                                                                       0.5
Dehydrogenase response to PCP application to Kidman soil after 24+2 h incubation (LSD=least
significant difference).

-------
   400 -i
    300 -
O
O
LU

jjjj 200

LU
CL
    100 <
                         i
                        O.i
                            I
                           0.2
                                  0.3             0.4

LOADING  RATE  (%  wet  wt.  waste/dry wt. soil)
0.5
 Figure 2S.
Respiration response  to application of PCP waste to Kidman soil  after 24«2 h in.ubation (LSO=
least significant difference).

-------
   500-i
_, 400-
O
or
o
LJ
O
or
LJ
Q_
   300-
    200-
    100
D
	 1 	
O.I
	 1 	
0.2
	 1 	
0.3
	 1 	
0.4
	 1
0.5
 Figure 30.
          LOADING  RATE  (%  wet  wt. waste/dry wt. soil)
Viable aerobic heterotrophic bacteria and  fungal  prooagules in Kidman  soil treated with PCP
waste  after 24+2 t.  incubation  (LSDOeast significant difference).

-------
            10 -i
            8 -
         c
         Q>
         U

         0>
         Q.

         0)

         E
6 -
OJ
oo
          in
           •»
          m

          o"
          in
             2-
T 	 1
0 2
	 1 	
4
	 1 	
6
	 1 	
8
	 1 	
10
	 7—
12
1
14
                        LOADING RATE  (% wet wt. waste/dry wt. soil)

        Figure  31.   Microtox response to slop  oil emulsion solids waste application to Kidman soil
                    (LSD=least significant differenco).

-------
          100
lO
        Figure  32.
     2         4          6          8         10         12         14

     LOADING  RATE  (%  wet wt. waste/dry wt. soil)

Initial ammonium and nitrite ion oxidation in response to treatment  application of slop
oil emulsion solids to Kidman soil  (LSD-least significant difference).

-------
     Dehydrogenase activity,  both  with  and witTiout  glucose  addition,  was
inhibited 30  to 50 percent  at  the  2 percent  loading  rate  (Figure  33).
Dehydrogenase  toxicity  increased  with  increasing  loading and  activity  was
essentially nil  at  the  10 and 14 percent loadings in  the assay without glucose
addition.  The  a
-------
    100
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LJ
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