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.
-------
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.
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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 !
-------
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
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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
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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
-------
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
-------
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
-------
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
-------
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.
-------
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
-------
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
-------
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
-------
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
-------
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
-------
physiological inhibition, thereby indicating the presence of tox
-------
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
108
-------
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).
109
-------
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
110
-------
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
-------
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
-------
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.
113
-------
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
-------
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
115
-------
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.
116
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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
-------
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).
-------
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
-------
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
•»
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o"
in
2-
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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
_J
o
cr
H
Z
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O
LJ
O
-------
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