PB83-163956
Mobility of Organic Compounds from
Hazardous Wastes
(U.S.) Oak Ridge National Lab., TN
Prepared for
Environmental Monitoring Systems Lab,
Las Vegas, NV
Feb 83
U.S. DEPARTMENT OF COMMERCE
National Technical Information Service
urns
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EPA-600A-83-OOI
February 1983
P333-163956
MOBILITY OF ORGANIC COMPOUNDS FROM HAZARDOUS WASTES
by
D. K. Brown, M. P. Maskarinec, F. W. Larimer,
and C. W. Francis
Oak Ridge National Laboratory
Oak Ridge, Tennessee 37830
Interagency Agreement Numbers EPA AD-89-F-1-058
and DOE 40-1087-80
Project Officer
Llewellyn R. Williams
Quality Assurance Division
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89114
ENVIRONMENTAL MONITORING SYSTEMS LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
LAS VEGAS, NEVADA 89114
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1 REPORT NO.
EPA-600A-83-OOI
2.
4. TITLE ANO SUBTITLE
MOBILITY OF ORGANIC COMPOUNDS FROM HAZARDOUS WASTES
7 AUTHORS) D. K. Brown, M. P. Maskarinec, F. W. Larimer,
and C. W. Francis, Oak Ridge National Laboratory,
Oak Ridcre, Tennessee 37830
9. PERFORMING ORGANIZATION NAME ANO ADDRESS
Environmental Monitoring Systems Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Las Vegas, Nevada 89114
12. SPONSORING AGENCY NAME ANO ADO
U.S. Environmental Protect!
Office of Research and Deve
Environmental Monitoring Sy
Las Vegas, Nevada 89114
RESS
on Agency--Las Vegas, NV
lopment
stems Laboratory
3. RECIPIENT'S ACCESSION NO.
p«f 3 ]63956
S REPORT DATE"
February 1983
6. PERFORMING ORGANIZATION CODE
8. PERFORMING ORGANIZATION REPORT NO.
10. PROGRAM ELEMENT NO.
ABSD1A
11. CONTRACT/GRANT NO. Iftg fjQS .
EPA AD-89-F-1-058 and
'DOE 40-1087-80
13. TYPE OF REPORT ANO PERIOD COVERED
14. SPONSORING AGENCY CODE
EPA/600/07
IS. SUPPLEMENTARY NOTES
Project Officer: Llewellyn R. Williams, Quality Assurance Division, Environmental
Monitoring Systems Laboratory, Las Vegas, Nevada 89114
16 ABSTRACT
The objective of this research is to develop a second generation laboratory extraction test to model the mobility of organic and inorganic
constituents from solid wastes co-disposed with municipal waste This test should more accurately and reproduc.bly model 'eachate
production for selected organic as well as inorganic constituents, than the test procedure referred to as EP promulgated by EPA in 1 980
As a first approach the capabilities of five aqueous extraction procedures to remove organ.c compounds from 1 1 solid wastes were
evaluated The extraction procedures investigated were four batch extractions using (1) de.on.zed distilled water adjusted to PH 5 with 0 5 Ji
acetic acid, (2) deiomzed distilled water, (3) detonized distilled water with a sodium cation exchange resin (4) 0 5 fid sodium citrate, and (5)
an upward-flow column extraction using deiomzed distilled water The major conclusions relative to the effectiveness of the extraction
procedures to remove organic compounds were. (1) the column procedure extracted more organic material than any of the batch
procedures, and (2) among the batch extraction procedures, deiomzed distilled water was the most aggressive medium
The most noticeable differences between the column procedure utilized and the batch procedures were the elevated levels of moderately
volatile and the nonpolar organic compounds found in the column extracts Factors contributing to these results are (1 ) the column
procedure is a completely closed extraction permitting direct collection of volatile compounds, and (2) the column extracts were not filtered
through membrane filters which are known to sorb appreciable quantities of nonpolar compounds _._,..._
Two extracting devices (magnetically stirred and rotary extractor* for conducting the EP were also compared Extracts produced by the two
extractors showed significantly different concentrations of As. Cd, Fe. Ni. and Zn. although neither method showed a consistent pattern
The proposed reverse-phase High-Pressure Liquid Chromatography protocol to assess the bioaccu-nulation potential of solid waste
extracts was found to produce only qualitative information because of nonuniformity of detector response. However, the test does provide a
useful screening method for the detection of potentially bioaccumulative organic compounds
In addressing a secondary research objective, comparison of two isolation techniques (i e . resin adsorption technique using Amberlite
XAD-2 resin and a solvent partition technique using methylene chloride) to isolate organ.c mutagens from aqueous solutions for testing in
the Ames Salmonella mutagemcity assay was conducted Although the assay results were not affected by the type of isolation technique
used, the extract.on efficiency of the resin technique was. in general, less dependent on the specific aqueous medium than was the solvent
partition
17.
». DESCRIPTORS
KEY WORDS AND DOCUMENT ANALYSIS
b.lOENTIPIERS/OPEN ENDED TERMS
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (Thu Report!
UNCLASSIFIED
20. SECURITY CLASS (This page)
UNCLASSIFIED
c. COSATI Field/Group
21. NO. Of PAGES
203
22. PRICE
EPA Farm 2220-1 (R«v. 4-77) PMlviou* COITION i» OBIOUBTE
i
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NOTICE
Although the research described In this article has been funded wholly or
in part by the U.S. Environmental Protection Agency through Interagency
Agreement EPA AD-89-F-1-058 to the U.S. Department of Energy, it has not been
subjected to Agency policy review and therefore does not necessarily reflect
the views of the Agency. Mention of trade names or commercial products does
not constitute endorsement or recommendation for use.
ii
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TABLE OF CONTENTS
LIST OF TABLES v
LIST OF FIGURES ix
ABBREVIATIONS AND SYMBOLS xii
ACKNOWLEDGMENTS xiii
1. INTRODUCTION 1
2. OBJECTIVES AND RATIONALE 5
2.1 TASK 1: Evaluation of Aqueous Extraction
Procedures to Remove Nonpolar Organic
Compounds from Solid Wastes 5
2.1.1 Batch 1: Environmental Protection Agency EP . . . 6
2.1.2 Batch 2: Deionized Distilled Water Extractant . . 6
2.1.3 Batch 3: Sodium-Resin Extractant 6
2.1.4 Batch 4: Citrate Buffer Extractant 7
2.1.5 Column 7
2.2 Task 2: A Comparison of Two Sample Preparation Protocols
for Performing the Ames Test on Solid Waste
Extracts and Wastewaters 8
2.3 Task 3: Assessment of a Magnetically Stirred
Extractor Relative to an EPA-Approved
Rotary Extractor for Conducting
the Extraction Procedure (EP) 9
2.4 Task 4: Evaluation of the Proposed Reverse-Phase
High-Pressure Liquid Chromatography (HPLC)
Protocol for Assessing Bioaccumulation
Potential of Solid Waste Extracts 10
3. EXPERIMENTAL 11
3.1 Description of Solid Wastes .' 11
3.2 Methods 14
3.2.1 Isolation of Organic Compounds
from Solid Wastes 14
3.2.2 Solid Waste Extraction Procedures 17
3.2.3 Filtration Study (Supplemental to Task 1) .... 28
3.2.4 Isolation of Organics for Mutagenicity
Testing - Task 2 29
3.2.5 Assessment of Extracting Devices for
Conducting the Extraction Procedure
(EP) - Task 3 32
3.3 Methods of Analysis 33
3.3.1 Inorganic Analyses 33
3.3.2 Organic Analyses 34
3.3.3 Mutagenicity Testing 36
3.3.4 Analysis of Bioaccumulative Materials in
Wastes and Leachates 37
iii
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Page
4. RESULTS 40
4.1 Task 1: An Evaluation of Aqueous Extraction
Procedures to Remove Nonpolar Organic
Compounds from Solid Wastes 40
4.1.1 Inorganic Analyses of Extracts 40
4.1.2 Isolation of Organic Compounds in Solid Wastes . . 46
4.1.3 Organic Analyses of Aqueous Extraction
Procedure Extracts 52
4.1.3.1 Dissolved Organic Carbon Results .... 52
4.1.3.2 Gas Chromatography and Gas
Chromatography/Mass Spectrometry .... 61
4.1.4 Mutagenicity Testing 71
4.1.5 Filtration Study 84
4.2 Task 2: Comparison of Two Sample Preparation Protocols
for Performing the Ames Test on Solid Waste
Extracts and Wastewaters 90
4.3 Task 3: An Evaluation of the Equivalence of a
Magnetically Stirred Extractor Relative to an
EPA-Approved Rotary Extractor for Conducting
the Extraction Procedure (EP) 99
4.4 Task 4: Evaluation of the Reverse-Phase High-Pressure
Liquid Chromatography Protocol for Assessing
the Bioaccumulation Potential of Solid Waste
Extracts 105
5. CONCLUSIONS 108
5.1 Task 1: An Evaluation of Aqueous Extraction
Procedures to Remove Nonpolar Organic
Compounds from Solid Wastes 108
5.2 Task 2: Comparison of Two Sample Preparation
Protocols for Performing the Ames Test on
Solid Waste Extracts and Wastewaters ill
5.3 Task 3: An Evaluation of the Equivalence of a
Magnetically Stirred Extractor Relative
to an EPA-Approved Rotary Extractor for
Conducting the Extraction Procedure (EP) .... 112
5.4 Task 4: Evaluation of the Proposed Reverse-Phase
High-Pressure Liquid Chromatography (HPLC) .
Protocol for Assessing Bioaccumulation
Potential of Solid Wastes 113
6. FUTURE RESEARCH 114
7. REFERENCES 116
APPENDIX A Quality Assurance Project Plan 119
APPENDIX B FY-1982 Workplan 157
iv
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LIST OF TABLES
Table Page
1.1 Hazardous waste definition threshold levels 3
3.1 Description of waste types 12
3.2 Waste characteristics 13
3.3 Extraction procedures: Identification of variable
leaching factors 18
3.4 Equipment and materials used for the extraction
procedures 19
3.5 Batch 1: Extraction Procedure (EP) extraction data ... 23
3.6 Batch 2: Water extraction data 24
3.7 Batch 3: Na-resin extraction data 25
3.8 Batch 4: Citrate buffer extraction data 26
3.9 Measurements of pH and electrical conductivity in
column extracts after passing through XAD-2 resin .... 27
4.1 Selected inorganic analyses of extracts produced
using four extraction procedures for wastes 1-6 41
4.2 Inorganic analyses of water and extraction procedure
(EP) extracts for wastes 7 and 8 '42
4.3 Inorganic analyses of extraction procedure (EP),
water, Na-resin, and citrate buffer extracts
for waste 9 43
4.4 Inorganic analyses of extraction procedure (EP),
water, Na-resin, and citrate buffer extracts for
waste 10 44
4.5 Inorganic analyses of extraction procedure (EP),
water, Na-resin, and citrate buffer extracts for
waste 11 45
4.6 Comparison of Fe concentrations using four
extraction media 47
4.7 Total organics recovered: Comparison of Soxhlet
extraction with the sequential extraction scheme -
gravimetric data on solid waste, organic isolates .... 48
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Table Page
4.8 Organic compounds Identified In solid wastes 51
4.9 Dissolved organic carbon observed In selected
aqueous waste extracts 56
4.10 Comparison of dissolved organic carbon (DOC) In
extracts filtered through glass fiber and
polycarbonate filters 58
4.11 The relative effectiveness of five aqueous
extraction procedures to remove organic
compounds from solid wastes 63
4.12 Relative amounts of individual compounds in solid
waste extracts and relative ranking of the
extraction procedures 66
4.13 Results of mutagenicity testing of waste 1 extracts ... 72
4.14 Results of mutagenicity testing of waste 2 extracts ... 73
4.15 Results of mutagenicity testing of waste 3 extracts ... 74
4.16 Results of mutagenicity testing of waste 4 extracts ... 75
4.17 Results of mutagenicity testing of waste 5 extracts ... 76
4.18 Results of mutagenicity testing of waste 6 extracts ... 77
4.19 Results of mutagenicity testing of waste 9 extracts ... 78
4.20 Results of mutagenicity testing of waste 10 extracts ... 79
4.21 Results of mutagenicity testing of waste 11 extracts ... 80
4.22 Summary of mutagenic activity of solid waste isolates:
Aqueous extract and isolates 81
4.23 Organic compounds found in column extracts, with and
without an in-line filter 86
4.24 Organic compounds remaining in a column extract after
using four solid/liquid separation treatments 87
4.25 Organic compounds remaining in a batch water extract
after using four solid/liquid separation treatments ... 89
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Table Page
4.26 Organic compounds remaining In a standard solution
after using four treatments for separating solid
and liquid phases 91
4.27 Organic compounds remaining in a standard solution
(agitated for 24 hours) after using four solid/liquid
phase separation treatments 92
4.28 Comparison of two recovery techniques for removing
organic mutagens from wastewaters and solid waste
extracts 93
4.29 Mutagenic activity of the positive controls
benzo(a)pyrene and 9-amino acridine 96
4.30 Mutagenic activity in organic isolates: Comparison
of recovery techniques using benzo(a)pyrene 97
4.31 Mutagenic activity in organic isolates: Comparison
of recovery techniques using 9-amino acridine 98
4.32 Distribution of variation associated with the
variables of agitation method, replicate extraction,
and analytical procedure 103
4.33 Mean (x) concentrations and statistical differences
of selected inorganics using a magnetic stirrer
(2 agitation rates) and a rotary extractor 104
4.34 Bioaccumulation data on solid waste isolates: Aqueous
extracts and organic isolates 106
B-l Amount of extract needed for various analyses
and biotests 169
B-2 Major equipment needs and estimated costs for
performing a batch vs column extraction 171
B-3 Description and preliminary information relative to
industrial wastes to be used in the study 173
vn
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LIST OF FIGURES
Figures Page
3.1 Scheme for proximate analysis of solid wastes and
sludges by sequential extraction 15
3.2 Apparatus used for isolation of volatiles 16
3.3 Column extraction apparatus 22
3.4 Extraction procedures for environmental water samples . . 30
3.5 Partition coefficients of known compounds plotted
against log retention times 38
4.1 Chromatograms for waste 2 - Soxhlet extract (A), and
sequential extraction extracts: acids (B), bases (C),
and neutrals (D) 49
4.2 Chromatograms for waste 11 - Soxhlet extract (A) and
sequential extraction extracts: acids (B), bases (C),
and neutrals (D) 50
4.3 Chromatogram of waste 2 (aged four years in a tightly
covered landfill): Volatile organic compounds 53
4.4 Chromatograms of volatile organic compounds for
waste 3 (A) and waste, 9 (B) 54
4.5 Dissolved organic carbon (DOC) found in water
extracts over the 24-h extraction time 59
4.6 Dissolved organic carbon (DOC) found in Na-resin
extracts over the 24-h extraction time 60
4.7 Chromatograms of waste 10 column extracts -
replicate 1 (1) and replicate 3 (2) 65
4.8 Chromatograms of volatile organics from waste 2
extracts: Citrate buffer extract (a) and Na-resin
extract (b) 67
4.9 Chromatograms of volatile organics in water extracts
from waste 2: Open vessel (a) versus closed
vessel (b), and distilled-in-glass water blank (c) . . . . 69
4.10 High and low stirrer speed values for waste 12:
Mean and standard deviation values for four replicate
extractions over the 24-h extraction time 100
Preceding page blank
IX
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Figures Page
4.11 High and low stirrer speed values for waste 13:
Mean and standard deviation values for four replicate
extractions over the 24-h extraction time 101
A-l Project organization and responsibilities 126
A-2 Disciplines and activity areas 137
A-3 The research data management components 140
A-4 Sample and processing analysis 141
A-5 Corrective action plan 149
A-6 Quality assurance program organization chart 151
A-7 Analytical Chemistry Division QA assessment schedule . . 152
B-l Schematic of lysimeter containing residential
and commercial wastes 179
B-2 Flow diagram for lysimeter study 180
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ABBREVIATIONS AND SYMBOLS
AAS Atomic Absorption Spectroscopy
BaP Benzo(a)pyrene
DDE 2,2-bis-(p-chlorophenyl)-l, 1-dichloroethylene
DMSO Dimethyl sulfoxide
DOC Dissolved organic carbon
EP Extraction Procedure (40 CFR 261.24)
EPA U.S. Environmental Protection Agency
GC Gas Chromatography
GC/MS Gas Chromatography/Mass Spectrometry
HPLC High-Pressure Liquid Chromatography
K-D Kuderna-Danish
MeClp Methylene chloride
NEIC National Enforcement Investigations Center
NIPDWS National Interim Primary Drinking Water Standards
ORNL Oak Ridge National Laboratory
PTFE Polytetrafluoroethylene
Pa Pascals
RCRA Resource Conservation and Recovery Act
rpm Revolutions per minute
S Siemens
TCO Total Chromatographable Organic Compounds
xi
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ACKNOWLEDGMENTS
The authors wish to acknowledge the technical assistance of
J. W. Gooch (Environmental Sciences Division), R. W. Harvey (Analytical
Chemistry Division), and S. Severs (SCUU Science Semester
participant). Appreciation is also expressed to N. M. Ferguson
(Analytical Chemistry Division), J. C. Goyert, and R. H. Strand
(Environmental Sciences Division) for their research contributions, as
well as to M. R. Guerin (Analytical Chemistry .Division), S. E. Herbes,
and T. Tamura (Environmental Sciences Division) for their helpful
suggestions for improvement of this report. A special thanks goes to
L. W. Littleton, D. D. Rhew, and others in the Environmental Sciences
Division Word Processing Center for their assistance in preparing this
report. Finally, the helpful suggestions and assistance during the
course of this project from the following individuals are greatly
appreciated: L. R. Williams of EPA, Environmental Monitoring Systems
Laboratory, Las Vegas, Nevada; and D. Friedman and T. A. Kimmell of
EPA, Office of Solid Waste, Washington, D. C.
xii
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SECTION 1: INTRODUCTION
The disposal of municipal and industrial waste in landfills is a
widely used waste management practice in the United States. It has
become evident during the past few years that there has been serious
environmental damage and possible adverse human health effects because
of improper disposal of hazardous waste in landfills. Under the
Resource Conservation and Recovery Act (RCRA) of 1976 (PL94-580),
Congress directed the Environmental Protection Agency (EPA) to
promulgate regulations to protect human health and the environment from
improper management of solid wastes.
Under Section 3001 of RCRA, Identification and Listing of
Hazardous Waste, EPA is charged with identifying which industrial
wastes pose a hazard to human health or the environment if these wastes
are improperly managed. On May 19, 1980 (EPA 1980b), EPA promulgated
regulations that identified a number of properties of hazardous waste,
which, if exhibited by a waste, would indicate that the waste requires
controlled management. Under these regulations, toxicity of wastes is
determined by the Extraction Procedure Toxicity Characteristic
(40 CFR 261.24) which employs an Extraction Procedure (EP) to predict
the degree to which toxic species might leach out of the waste and
contaminate groundwater if the waste was disposed of in a nonsecure
municipal landfill. The EP is a 24-h extraction procedure using 0.5 N^
acetic acid to adjust downward, if possible, the pH of the solid waste
suspension to pH 5.0 (EPA 1980d). Additional information with regard
to the development of the EP can be found in the EP Toxicity
Characteristic Background Document (EPA 1980a).
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The EP results in an extract that is analyzed for the eight metals
(As, Ba, Cd, Cr Pb, Hg, Se, and Ag), four pesticides (Endrin, Lindane,
Methoxychlor, and Toxaphene), and two herbicides [2,4-D and 2,4,5-TP
(Si 1 vex)] for which National Interim Primary Drinking Water Standards
(NIPDWS) (EPA 1979a) have been established. Hazardous waste definition
threshold levels (Table 1.1) have been established for each of the
species taking into account attenuative processes expected to occur
during the movement of leachate through the underlying strata and
groundwater aquifer.
The EP is considered to be a first-order approximation which
primarily models the leaching action of the low molecular weight
carboxylic acids generated in an actively decomposing municipal waste
landfill. Acetic acid is added to distilled water to make up the
extracting medium used in the EP. The acetic acid primarily affects
the leaching of metals from an industrial waste. The higher molecular
weight organics that are expected to be present in municipal landfill
leachates, and thought to affect the Teachability of nonpolar organics,
are not currently modeled by the EP. This perceived limitation is the
impetus behind the current research.
The primary objective of this research is to develop a second
generation test for mobility that will more accurately and reproducibly
model leachate production, for organic as well as inorganic
constituents, in the previously described disposal environment. A
second important objective of this research program is that the mobility
test developed be compatible with subsequent biological testing.
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TABLE 1.1. HAZARDOUS WASTE DEFINITION THRESHOLD LEVELS*
Contaminant
Arsenic
Barium
Cadmium
Chromium (IV)
Lead
Mercury
Selenium
Silver
Maximum
concentration (mg/L)
5.00
100.00
1.00
5.00
5.00
0.20
1.00
5.00
Endrin (1,2,3,4,10,10-hexachloro-l ,
7-epoxy-l ,4,4a,5,6,7,8,
8a-octahydro-l , 4-endo, endo-5,
8-dimethano naphthalene) 0.020
Lindane (l»2,3,4,5,6-hexachlorocyclohexane,
gamma isomer) 0.40
Methoxychlor (l,l,l-trichloro-2,2-bis
[p-methoxyphenyl] ethane) 10.00
Toxaphene (CioHioCls* technical
chlorinated camphene, 67-69% chlorine) 0.50
2,4-0, (2, 4-0 ichlorophenoxy acetic acid) 10.00
2,4,5-TP; Silvex (2,4,5-Trichlorophenoxypropionic
acid) 1.00
Source: 40 CFR 261.24
Concentrations are 100 times the NIPDWS values.
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Work toward these objectives has centered in four separate but
related tasks as follows:
Task 1: An evaluation of aqueous extraction procedures to remove
nonpolar organic compounds from solid wastes.
Task 2: A comparison of two sample preparation protocols for
performing the Ames Test on solid waste extracts and
wastewaters.
Task 3: An evaluation of the equivalence of a magnetically
stirred extractor relative to an EPA-approved rotary
extractor for conducting the EP.
Task 4: Evaluation of the proposed reverse-phase High Pressure
Liquid Chromatography (HPLC) protocol for assessing the
bioaccumulation potential of solid waste extracts.
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SECTION 2: OBJECTIVES AND RATIONALE
2.1 Task 1: An Evaluation of Aqueous Extraction Procedures to Remove
Nonpolar Organic Compounds from Solid Wastes
The primary objective of Task 1 was to assess the capabilities of
five selected aqueous extraction procedures to remove organic compounds
from 11 solid wastes known to contain significant quantities of organic
compounds. Other objectives included (1) analyzing the solid waste
extracts for selected inorganic constituents known to be present in the
samples and (2) mutagenicity-testing of the extracts. Mutagenicity
testing was included to evaluate any differences in aggressiveness
toward organic compounds among extraction procedures that might not be
detected by 6C/MS analyses; for example, the ability of an extraction
procedure to extract high molecular weight mutagens from solid wastes.
A desirable, relevant extraction procedure should accurately and
reliably model leachate production in the landfill situation, be
relatively simple and economical to perform, and yield an extract that
can be applied to biological assays. The five extraction procedures
selected for this study were four batch extractions and an upward flow
column extraction. The following extractants were used:
Batch 1: Deionized distilled water adjusted to pH 5 with
0.5 N acetic acid (EP).
Batch 2: Deionized distilled water.
Batch 3: Deionized distilled water with a sodium cation exchange
resin.
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Batch 4: 0.5 M soc.ium citrate buffer.
Column: Deionized distilled water.
A rationale for each procedure follows.
2.1.1 Batch 1: Environmental Protection Agency EP
The EP (EPA 1980b) is the test procedure promulgated by EPA under
RCRA to determine if an unacceptably high level of groundwater
contamination might result if an industrial waste is disposed of in a
municipal landfill. The high ionic strength of the extracting solution
and/or the method of membrane filtering the aqueous extract limits the
recovery of organic compounds from a solid waste extract. The acetic
acid used in the EP may also confound the interpretation of the
phytotoxicity and aquatic toxicity assays (Epler et al. 1980).
2.1.2 Batch 2: Deionized Distilled Water Extractant
Deionized distilled water, because it introduces no extraneous
interferences, is a reliable extractant to evaluate the toxicity of
solid waste extracts in phytotoxicity and aquatic toxicity assays. In
addition, distilled water at relatively high liquidisolid ratios tends
to solubilize organics from solid wastes more efficiently than other
aqueous extractants (McKown et al. 1980). For example, solutions
containing citrate and acetate salts at high concentrations can enhance
flocculation over dispersion.
2.1.3 Batch 3: Sodium-Resin Extractant
The solubility of organic compounds in groundwater is largely
dependent on the "hardness" or "softness" of the water. Medium- and
long-chain organic acids are solubilized when the "hard" calcium
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(or iron) is replaced by "soft" sodium. This is the basic principle in
the extraction of fulvic and humic acids from soils by soil scientists
(Schnitzer and Kahn 1972) (i.e., create a dispersed condition by
substituting Na+ for the dominant Ca+ and Fe ions). Conventionally,
this is accomplished using 0.1 J4 NaOH. However, the high pH (11-12)
does not represent environmental conditions. A sodium saturated
chelating resin, added to a suspension displaces the dominant cations
with sodium without significantly changing the pH of the suspension.
The organic compounds are dispersed from the solid matrices, in effect
solubilizing them in an aqueous environment.
2.1.4 Batch 4: Citrate Buffer Extractant
For certain solid wastes citrate buffer (0.5 M sodium citrate) was
observed to be more aggressive than acetate or distilled water in
leaching both organic compounds and metals; however, distilled water
was the overall leaching medium of choice (McKown et al. 1980). The
citrate butter was included because of an existing data base with
citrate, the structural similiarity of citrate to other carboxylic acid
esters present in municipal waste leachates, and the known capacity of
citrate for complexing metals.
2.1.5 Column
An upward flow column extraction procedure was developed as a
possible alternative to a batch extraction. Deionized distilled water
is pumped upward through solid wastes packed in a glass column.
Nonpolar organic compounds are collected directly on XAD-2 resin in
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series with the solid waste column; polar organic compounds remain in
the eluent. Advantages of this system include:
(1) no time-consuming filtration step is needed,
(2) the extraction system is completely closed until the organic
compounds are collected, thus loss of volatiles will be
minimized,
(3) concentration of organic compounds is possible,
(4) column extraction of solid wastes more closely simulates the
physical landfill leaching process, and
(5) column give dynamic data.
2.2 Task 2: A Comparison of Two Sample Preparation Protocols for
Performing the Ames Test on Solid Waste Extracts and
Wastewaters
The objective of this work was to compare two techniques for
isolating organic mutagens from solid waste leachates and wastewaters
for testing in the Ames Salmonella mutagenicity assay (Epler et al.
1980). The two recovery techniques compared were a resin adsorption
technique using Amberlite XAD-2 resin and a solvent partition technique
using methylene chloride. The XAD-2 resin technique has been used
extensively at the Oak Ridge National Laboratory (ORNL) (Epler et al.
1980) and at EPA's Health Effects Research Laboratory at Research
Triangle Park, North Carolina, while the solvent partition scheme is
the one recently developed by the National Enforcement Investigations
Center (NEIC), Denver, Colorado (EPA 1980c). The objective of this
task was to determine if significant differences were obtained in the
Ames Salmonella mutagenicity assay when using either of the two
preparation protocols.
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2.3 Task 3: Assessment of a Magnetically Stirred Extractor Relative
to an EPA-Approved Rotary Extractor for Conducting the
Extraction Procedure (EP)
The objective of this task was to compare a magnetically stirred
extractor to an EPA-approved rotary extractor for conducting the EP.
An acceptable extractor to conduct the EP was described in "Test
Methods for Evaluating Solid Hastes - Physical/Chemical Methods"
(EPA 1980) as "...one which will prevent stratification of a waste
sample and extraction fluid and will insure that all sample surfaces
continuously contact well mixed extraction fluid. Among 'the acceptable
extractors are: (1) stirrers and (2) tumblers. Stirrers consist of a
container in which the waste/extraction fluid mixture is agitated by
spinning blades. Rotators agitate by turning a sample container end
over end through a 360° revolution."
The magnetically stirred extractor consists of a glass vessel and
polytetrafluoroethylene (PTFE)-coated stirring bar in concert with a
commercially available magnetic stirrer readily available in most
laboratories. This design appeared to satisfy all of the above
requirements. The magnetically stirred extractor offers certain
advantages in that, in contrast to the rotary extractor design, the pH
of the suspension can be automatically adjusted. In addition, the
glass vessel and PTFE-coated magnetic stir bar eliminates the
possibility of contamination by the spinning stainless steel blades
used in several of the EP extractor designs.
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2.4 Task 4; Evaluation of the Proposed Reverse-Phase High-Pressure
Liquid Chromatography (HPLC) Protocol for Assessing
Bioaccumulation Potential of Solid Waste Extracts
The objective of this task was to evaluate the analytical
constraints and interpretations associated with using the proposed
reverse-phase High-Pressure Liquid Chromatography (HPLC) protocol
(EPA 1978a and EPA 19785) to assess the bioaccumulation potential of
solid waste extracts. It has been shown that there is a relationship
between octanol/water partition coefficients and bioaccumulation
potential (i.e., compounds with high octanol/water partition
coefficients have the potential to biologically accumulate) and between
octanol/water partition coefficients and the log of reverse-phase HPLC
retention times. Plotting the log of reverse-phase HPLC retention
times of solid waste extracts (from Task 1) against the log of the
octanol/water partition coefficients of known compounds should provide
a method to estimate the potential for the components in the solid
waste extracts to accumulate in biological tissues.
10
-------
SECTION 3: EXPERIMENTAL
3.1 Description of Solid Wastes
Eleven solid wastes from a variety of industries were obtained for
use in Task 1. The particular wastes were selected because they were
thought to contain toxic organic compounds. Table 3.1 contains a short
description of the wastes used for Task 1 and other studies. Wastes 1
through 6 were taken from actual landfills. Corresponding landfill
leachates were also sampled in a related study comparing the solid
waste EP extracts with the landfill leachates (Brown et al. 1981).
Waste samples 7 through 11 are oily in nature and were selected based
on a prediction that this oily component would challenge the extraction
procedures. Wastes 7 and 8 changed physically (i.e., the oily
components separated from the remainder of the sample) during the
course of the study. For this reason, not all extractions were
performed on these wastes.
Two additional waste samples (12 and 13) were selected for use in
Task 3. Both were selected because they contained Teachable inorganic
elements and were expected to pose stirring problems with the magnetic
stirrer. An additional waste (14) was utilized for the supplementary
filtration study in Task 1.
General sample information, such as water content and general
physical description of wastes 1 to 14, is presented in Table 3.2.
Water content was determined by gas chromatography as described by
Shultz and Spears (1966).
11
-------
TABLE 3.1. DESCRIPTION OF WASTES
Waste Waste description
Used for Task 1:
1 Sludge from the production of feed additives and veterinary
Pharmaceuticals
2 Sludge from the treatment of a municipal sewage, a cannery
waste, and tannery wastes
3 Paper mill sludge
4 Municipal refuse and local industrial waste (plastic
laminate and electronics waste)
5 Paper mill, paper trimmings, and related wastes
6 Municipal wastewater treatment sludge mixed with local
industrial waste (plastic and textile mill)
7 Wastewater treatment sludge from a metal processing plant
8 Untreated washing sludge from a metal bearing factory
9 Wastewater treatment sludge from a coal conversion plant
10 Filter cake from a coal conversion biological treatment
plant
11 Centrifuged heavy oil residuals from a coal conversion
wastewater treatment plant
Used for Task 3:
12 Ash from a coal conversion facility
13 Fly ash from a coal-fired power plant
Used for Filtration Study:
14 Vacuum bottoms from a coal conversion facility
12
-------
TABLE 3.2. WASTE CHARACTERISTICS
Waste
Water
content3
(weight %)
Waste characteristics
1
2
33
55
56
Yellowish green, dark; pasty texture
Dark grey sludge containing hair, hide,
etc.; strong odor
Dark brown, nearly opaque, sandy sludge;
noticeable aroma
4
5
6
7
8
9
10
11
12
13
14
77
62
60
ndb
nd
50
41
59
nd
nd
<1.0
Sawdust-like texture; yellow-orange
Like flaked cardboard; dark purple
Very dark gray, clay-like consistency;
strong odor
Watery, dark brown; slightly aromatic
Dark brown, thick, oily liquid; oily odor
Black, oily sludge; rubbery consistency
Charcoal gray, sandy, and granular;
irritating odor
Black, oily sludge; irritating odor
Hard granular solid containing Cd, Fe,
and Ni .
Fine solid, high in magnetite, contains
As, Cd, and Zn
Dark -colored dry powdery solid.
As determined by the method of Shultz and Spears (1966).
3nd = not determined.
13
-------
Many of the wastes, because they cjntained significant quantities
of organic compounds, had the potential to change during the course of
the study due to biological activity. To slow down such changes, each
waste was thoroughly mixed and subsampled into 200- to 500-g
quantities. The subsamples were placed in glass containers and frozen
at -15°C. Samples were thawed in the refrigerator (4°C) the day prior
to extraction.
3.2 Methods
3.2.1 Isolation of Organic Compounds in Solid Wastes
Identification of the major organic constituents from the wastes
was carried out using the following two procedures:
(1) Twenty-four-hour Soxhlet extraction with methylene chloride,
followed by volume reduction in a Kuderna-Danish (K-0)
evaporator (EPA 1980d).
(2) A three-step sequential extraction procedure developed at
ORNL (Fig. 3.1; Maskarinec and Harvey 1982).
The isolates from procedures (1) and (2) were compared by gravimetry as
well as by gas chromatography.
Volatile organic compounds were determined in selected wastes
using the apparatus shown in Fig. 3.2. This apparatus is a simple
modification of the existing purge and trap devices that allows the
purging of solid materials without dissolution in water (Brazell and
Maskarinec 1981).
14
-------
ORNL DWG 81 20042R ESD
VOLUME REDUCED
TO 10 mL IK D)
VOLUME REDUCED TO 10 mL (K D) ACID
REDUCED TO 100 mL (K D)
Fig. 3.1. Scheme for proximate analysis of solid wastes and sludges by
sequential extraction.
15
-------
ORNL-OWG 81-6937
TENAX
HEAT
SHRINKABLE,
TEFLON
HEAT
SHRINKABLE
TEFLON
GLASS WOOL
OR SINTERED FRIT
GLASS WOOL
CONDENSER
Fig. 3.2. Apparatus used for isolation of volatiles.
16
-------
3.2.2 Solid Waste Extraction Procedures
In this study, five aqueous extraction procedures were evaluated
and compared for ability to extract both Inorganic and organic
components. Table 3.3 contains a listing of the five extraction
procedures, with a compilation of variable leaching factors. Table 3.4
contains a listing of equipment and materials used for the five
extraction methods. All extractions were carried out to a liquid to
solid ratio of 20:1.
The EP was performed according to the current regulatory protocol
(EPA 1980d), using 185 g of wastes. Wastes were placed in borosilicate
glass vessels with 2.96 L of deionized distilled water (ASTM, Type I
Reagent Water). The vessel was fabricated at ORNL from 6.35-mm-thick,
152.4-cm-i.d. (inside diameter) flanged glass pipe that could hold up
to 4.0 L of solution. A fitted vessel cover was constructed of
plexiglass, with holes cut in it to admit a pH electrode and tubing
(for adding acid). The solid waste suspensions were agitated for 24 h
using a magnetic stirrer and 76.2-mm PTFE-coated stir bar. During the
24-h extraction, the pH of the solution was automatically adjusted to
pH 5 using 0.5 N acetic acid. A maximum amount of 4 ml of acid per
gram of waste extracted is specified in the protocol. If less than the
maximum amount of acid was used during extraction, the extract was
diluted with deionized distilled water to a final 1:20 solid:liquid
ratio prior to filtering.
The water extractions were performed as in the EP with the
exception that 3.7 L of deionized distilled water (ASTM, Type I Reagent
Water) were initially added to 185 g of waste (1:20 solid:liquid
ratio). There was no pH adjustment during extraction.
17
-------
TABLE 3.3. EXTRACTION PROCEDURES: IDENTIFICATION OF VARIABLE LEACHING FACTORS
oo
Variable factors3
Extraction
1. Batch 1: EP
Initial
leaching medium
Deionized distilled
waterb
Mode of
extraction
Batch:
magnetically
stirred
PH
adjustment
Adjust to pH 5
with 0.5 N
acetic acid -
maximum limit of
2-meq/g sample
Treatment of leachate solution
for extract analysis
Pressure-filtered through
0.4-um Nuclepore filter
2. Batch 2: Water Deionized distilled
waterb
3. Batch 3: Deionized distilled
Na-resln water0 with 1-g
calculated dry wt
Chelex 100/10-g
sample
4. Batch 4: 0.5 M citrate buffer
Citrate buffer
5. Up-flow column Deionized distilled
water0
Batch:
magnetically
stirred
Batch:
magnetically
stirred
Batch:
rotary
extractor
Column:
upward flow
None
Pressure-filtered through
0.4-um Nuclepore filter
Adjust to pH 7 Pressure-filtered through
with 0.1 N HC1 0.4-um Nuclepore filter
None
None
Pressure-filtered through
0.4-um Nuclepore filter
Leachate from column directly
passed through XAO-2 resin
aFactors such as particle size (<9.5 mm), extraction at room temperature, extraction time (24 h for
batch extractions or until an effect 1:20 solid:liquid ratio contact is reached for column extraction),
one extraction on each waste, and effective 1:20 solid:liquid ratio remained constant.
°ASTM, Type I Reagent Water.
-------
TABLE 3.4. EQUIPMENT AND MATERIALS USED FOR THE EXTRACTION PROCEDURES
Item
Manufacture or supplier3
Catalog Number
Magnetic stlrrer
Teflon stir bar
pH controller
pH electrode
Filtration apparatus
Filters 0.40-Qn pore size
0.8-Qn pore size
3.0-Qn pore size
Pre-filters P40
P80
P100
P300
Drain disc
Rotary extractor
Glass column
Pump
Sea sand, sieve distribution:
20 to 35 mesh - 8X;
36 to 50 mesh - SIX;
51 to 60 mesh - 18%;
61 to 70 mesh - 7%;
71 to 100 mesh - 13%;
and 100 mesh - 3X.
Cole-Partner C-4817-00
Cole-Panner C-4612-30
Chemtrix 45AR
Leeds & Northrop 117493
Mil 11pore Corp. YT30-142-HU
Nuclepore Corp. 112207
Nuclepore Corp. 112209
Nuclepore Corp. 112212
Nuclepore Corp. 211703
Nuclepore Corp. 211705
Nuclepore Corp. 211707
Nuclepore Corp. 211708
Nuclepore Corp. 231700
Associated Design & Mfg. Co. 3740-6-BRE
Glenco Scientific 3400-D-25X45
Lab. Data Control, Div. of Milton Roy 2396-57
Fisher Scientific S-25
aUse of a specific manufacturer or supplier does not imply the endorsement by either
Oak Ridge National Laboratory or the U.S. Environmental Protection Agency.
19
-------
The sodium resin displacement extraction was also performed in the
same manner as the EP. Initially, 2.96 L of deionized distilled water
(ASTM, Type I Reagent Water) and 92.5 g of Chelex 100 resin (Bio Rad
Laboratories, 100-200 mesh, sodium form) was added to 185 g of sample.
The suspension was adjusted to pH 7 using 0.1 N. HC1. This pH
adjustment was performed to make the final extract more suitable for
phytotoxicity and aquatic toxicity bioassays. The final extract was
also diluted to a final 1:20 solid:liquid ratio prior to filtration.
The citrate buffer extractions were performed according to the
procedure of McKown et al. (1980). This procedure specified the use of
a rotary extractor which tumbles solid waste suspensions end-over-end
in closed glass containers. A 75-g sample was placed in the glass
container with 1.5 L of 0.5 M citrate buffer. The tumbling rate was
29 rpm.
Following a 24-h extraction using the above-mentioned batch
procedures, the component solid and liquid phases were separated by
pressure filtration at 5.2 X 105 Pa (75 psi), and the final extract
was passed through a 0.4-um polycarbonate filter (Nuclepore). In
most cases, a series of filters having decreasing pore sizes was needed
in the filter stack. Even though the filter stack was changed, as
deemed necessary, some suspensions took considerable time to filter
(as much as 1-2 days).
For column extraction, 100 g of solid waste was thoroughly mixed
with 100 g of acid-washed sea sand to increase the hydraulic
conductivity of the samples. A glass column 2.5 cm i.d. and 45 cm long
(with plungers on each end adjustable to the varying bulk density of
20
-------
the samples) was packed with the solid waste/sand mixture. A 2.5-cm
layer of sand was placed on each end of the column. Figure 3.3
schematically shows the column extraction apparatus. For this study,
deionized distilled water (ASTM, Type I Reagent Water, passed through
XAD-2 resin to remove trace organic material) was pumped upward through
the solid waste/sand mixture until 2 L had passed through the sample.
A relief valve, set at 5.1 x 105 Pa (60 psi) was used to avoid unsafe
stresses on the glass column. The column effluent was passed through a
10-um pore size PTFE cloth (located on the top plunger) and then
directly through an XAD-2 resin cartridge (16.8 ml) for collection of
organic compounds.
The pH, amount of acid added during extraction (for extractions
requiring pH adjustment), and electrical conductivity were recorded on
the 11 samples during each of the five extraction procedures
(Tables 3.5-3.9). Solution pH is an important factor when considering
dissolution/precipitation and sorption reactions that may be occurring
during extraction. These data are provided as accessory information on
the wastes (e.g., the solid waste's buffering capacity).
Following extraction, all samples were stored and preserved
according to the desired analysis as follows:
(1) organic compounds - stored in glass containers and
refrigerated at 4°C,
(2) Hg analysis - placed in glass volumetric flask with
dichromate/nitric acid preservative,
21
-------
ORNL-DWG 81-13761AESO
RELIEF
VALVE
(60 psi)
RESIN FOR
COLLECTION
OF ORGANIC
COMPOUNDS
POSITIVE
DISPLACEMENT
PISTON PUMP
XAD-2 RESIN
CLEANUP)
EXTRACT
H20
Fig. 3.3. Column extraction apparatus.
22
-------
TABLE I.S. BATCH 1: EP EXTRACTION DATA
ro
LO
Initial
solution
pH (1:16
Replication solld:solution
Waste Number ratio)
1 8.
2 1 7.
2 7.
3 7.
4 7.
5 7.
6 S.
7b
8 6.
9 7.
10 1 12.
2 12.
3 11.
7
5
1
1
5
4
3
5
0
0
0
6
11 5.6
Acetic
acid initially
added
(meq/g
sample)
2.
0.
0.
2.
0.
0.
0.
0.
0.
2.
2.
2.
Oa
65
54
Oa
03
04
02
36
23
0"
Oa
Oa
0.02
Total
acetic acid
added
(meq/g
sample)
2.
0.
0.
2.
0.
0.
0.
0.
0.
2.
2.
2.
o«
79
68
Oa
07
11
12
38
38
0"
Oa
0*
0.03
PH
6.
5.
5.
6.
4.
4.
5.
5.
5.
12.
11.
12.
1
0
0
0
8
7
0
1
1
0
9
2
5.2
Electrical
conductivity
(uS/cm)
2750
2440
2150
3300
498
150
100
2750
418
6575
7600
7550
85
Final EP extract
Color
Transparent.
Translucent,
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
dark yellow
colorless
yellow-grey
pale yellow
colorless
yellow
green tint
colorless
pale yellow
pale yellow
pale yellow
pale yellow tint
aMaximuin amount allowed (4 ml acid/g sample).
bNot extracted.
-------
TABLE 3.6. BATCH 2: MATER EXTRACTION DATA
Waste
1
2
3
4
5
6
7
8
9
10
11
Initial
solution
pH
8.
8.
6.
7.
6.
6.
6.
8.
7.
11.
5.
8
0
6
8
8
3
1
1
5
7
9
Final
solution
pH (before
filtering
8
7
7
7
7
6
6
8
6
12
5
.8
.8
.1
.0
.0
.4
.6
.8
.3
.4
.8
1
i
8
8
7
6
7
6
6
8
6
12
6
PH
.8
.0
.0
.8
.4
.7
.0
.2
.5
.0
.0
Final
Electrical
conductivity
(uS/crn)
2000
1620
820
410
252
1500
121
150
307
6620
69
filtered extract
Color
Transparent,
Transparent,
Opaque, tan
Transparent,
Transparent,
yellow-orange
yellow-green
pink
pink
Opaque, yellow-brown
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
colorless
pale-gold
colorless
gold
pale yellow
24
-------
TABLE 3.7. BATCH 3: Na-RESIN EXTRACTION DATA
INJ
in
Waste
1
2
3
4
5
6
7
8
9
10
11
Blank
Initial
solution
PH
8
8
6
7
7
5
6
7
7
11
5
4
.3
.2
.9
.4
.6
.3
.1
.3
.5
.4
.8
.8
Solution pH
after
resin addition
7
7
7
7
8
5
7
8
7
11
6
9
.0
.8
.5
.9
.1
.9
.8
.3
.8
.5
.3
.5
Amount of
HC1 added to
adjust solution
to pH 7
(meq/g sample)
n
0
0
0
.0
.006
.072
.014
0.028
0
.0
0.027
0
0
0
0
0
.032
.009
.4
.028
.022
Final
pH
(before
filterim
6.8
7.1
7.1
7.1
7.2
5.9
7.0
7.1
6.5
12.4
7.0
7.15
Final filtered extract
I) PH
7.0
7.4
7.5
7.2
7.7
6.4
7.2
7.4
7.2
12.1
7.2
7.0
Electrical
conductivity
(uS/cm) Color
2700
2125
1160
428
425
782
460
345
450
7750
450
165
Transparent,
Transparent,
Translucent,
Transparent,
Transparent,
Translucent.
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
Transparent,
yellow-orange
colorless
gold
apple cider colored
pale orange
pale yellow
tan
tan
black tint
pale gold
pale yellow
colorless
-------
TABLE 3.8. BATCH 4: CITRATE BUFFER EXTRACTION DATA
Waste
1
2
3
4
5
6
7a
8
9
10
11
Final pH
filtered
extract
4.5
7.2
8.0
5.1
5.4
5.4
5.0
5.1
12.3
5.3
Electrical
conductivity
(uS/cm)
7555
7600
5300
5200
5800
6100
5800
5850
10800
ndb
Color
Transparent, yellow
Transparent, pale yellow
Opaque, dark brown/black
Transparent, yellow
Transparent, pale orange
Transparent, yellow
Transparent, pale gold
Transparent, pale yellow
Transparent, amber
Transparent, apple cider colored
"Not extracted.
3nd = not determined.
26
-------
TABLE 3.9. MEASUREMENTS OF pH AND
ELECTRICAL CONDUCTIVITY IN COLUMN
EXTRACTS AFTER PASSING
THROUGH XAD-2 RESIN
Electricity
conductivity
Waste Replication pH (uS/cm)
1
2
3
4
5
6
9
10
1
2
1
2
1
2
3
7.9
7.6
7.2
6.9
7.8
6.4
6.8
6.5
7.4
12.1
10.8
10.5
1999
1500
910
1175
741
560
388
1025
550
6150
4900
2460
11 5.2 20
27
-------
(3) all other metals - placed In plastic containers, preserved
with Ultrex nitric acid (solution pH < 2) and refrigerated
at 4°C (EPA 1979b), and
(4) volatile organic compounds - stored in 25-mL glass bottles
with no head space at 4°C.
3.2.3 Filtration Study (Supplemental to Task 1)
A filtration study was conducted to determine if decreased levels
of organic compounds measured in the batch extract resulted from
sorption of nonpolar organic compounds during filtration through a
0.4-urn nominal pore size membrane filters. As described earlier, the
column extracts were passed through a 10-ym pore size PTFE cloth at
the top of the column before the collection of organics on an XAD-2
resin. The two objectives of the filtration study were to determine if
the XAD-2 resin had collected entrained organic particulates (less than
10 urn) from the column that might be filtered out if filtered through
a 0.4-um membrane filter and to examine the recovery of organic
compounds in four solid/liquid phase separation treatments. The four
separation treatments were:
(1) filtration through a precombusted glass fiber filter (Whatman
GF/F, approximately 0.7-um pore size),
(2) filtration through a Nuclepore filter (polycarbonate,
0.4-um pore size),
(3) filtration through a Millipore filter (type HA - mixed esters
of cellulose, 0.4-um pore size), and
(4) centrifugation (in 250-nt glass bottles, 400 times gravity
for 1 h).
28
-------
Using 100-g samples of waste 14, four extractions were conducted
as follows with deionized distilled water as the extractant:
(1) column extraction performed as described in Section 3.2.2
(i.e., extract from the column passed directly through a
16.8-mL XAD-2 resin cartridge),
(2) column extraction performed like (1) above, except a glasc
fiber filter was placed in line prior to the effluent passing
through the XAD-2 resin cartridge,
(3) column extraction performed like (1) above with the effluent
subjected to the above listed four solid/liquid phase
separation treatments, and
(4) batch extraction was performed on the rotary extractor; the
suspension then subjected to the four solid/liquid phase
separation treatments.
In addition, 4 L of a standard solution were prepared with known
organic compounds at concentrations of 10 ppm. Two liters of this
solution were then immediately subjected to the same four separation
treatments. The remaining 2 L of this standard solution from above
were agitated for 24 h with the rotary extractor. The solution was
then subjected to the four solid/liquid phase separation treatments.
3.2.4 Isolation of Organics for Mutagenicity Testing - Task 2
Two recovery techniques (Fig. 3.4), a resin adsorption technique
(Epler et al. 1980) using Amberlite XAD-2 resin (resin cartridges as
used in Task 1) and a solvent partition technique using methylene
chloride (EPA 1980c), were compared for the isolation of organic
mutagens from aqueous solutions. Two known mutagens were used as
29
-------
ORNL-OWG 81 20041 ESO
LITER SAMPLE
DIVIDE INTO 2 EQUAL PORTIONS
BASE NEUTRAL EXTRACTION
ADJUST TO pH 12
EXTRACT EACH ALIQUOT WITH SO. 30 AND 30 mL MeCL,
OR PASS THROUGH XAD 2 COLUMN
1
AQUEOUS PHASE
I
ACID EXTRACTION
ADJUST TO QH 2
EXTRACT EACH ALIQUOT WITH
100. 60. AND 60 mL MeCI, OR
PASS THROUGH XAD 2 RESIN
1
t t
SOLVENT PHASE/RESIN
1
1
T
DRY WITH Na,SO4
1
»
CONC IN K D
1 ,
PHASE
AQUEOUS PHASE SOLVENT PHASE/RESIN PHASE EXCHANGE INTO ACETONE
* 1
DISCARD DRY WITH Na,SO,
1
CONC IN K D
1
T
FINAL VOL • 10 m
f
75 mL
*
L
EXCHANGE ACETONE
FOR 15 mL DMSO
BY
EXCHANGE INTO ACETONE ROTOEVAPORATION
FINAL VOL • 10 mL
I
T
75 mL
*
AMES TEST
EXCHANGE ACETONE FOR
IS mL DMSO BY
ROTOEVAPORATION
1
AMES TEST
*' ACID FRACTION
BASE/NEUTRAL
FRACTION
Fig. 3.4. Extraction procedures for environmental water samples.
30
-------
markers, representing the base (9-amino acridine) and neutral
[benzo(a)pyrene] chemical classes. The mutagens were purified to the
highest possible level and dissolved in dimethysulfoxide (1 mg/mL).
The aqueous media included:
(1) distilled water,
(2) a solid waste EP extract (waste 10),
(3) a "real-world" landfill leachate (from waste 3), and
(4) an industrial wastewater.
All aqueous media and all extraction blanks were tested for indigenous
mutagenic effects. These four aqueous media were chosen to give large
variety, both in terms of physical characteristics and possible
chemical interferences.
Mutagens were added to each aqueous medium in an amount sufficient
to give an unambiguous response in the Ames test (1 mg/L of water). A
sufficient volume (4 L) of each aqueous medium was used for both
recovery techniques (eight 500-ml aliquots: 2 replicates x 2 treatments
x 2 mutagens). Since the required level of BaP was in exess of its
solubility, the mutagen was added directly to the aqueous medium
contained in a separatory funnel. The funnel was shaken once, the pH
was adjusted, and the extraction was begun immediately. The separatory
funnel/resin system was not disconnected until the extraction was
completed. Thus, any components precipitated out by pH adjustment or
sorbed onto the funnel were extracted with the organic solvent. The
same procedure was used for the 9-amino acridine. An analysis of
variance was performed to determine significant differences between the
recovery techniques. The recovery of benzo(a)pyrene (containing tracer
31
-------
levels of C-labeled benzo(a)pyrene below mutagenic ranio-activation
threshold values) was measured by liquid scintillation counting as well
as by reverse-phase HPLC. The 9-amino-acridine recovery was measured
by fluorescence spectrometry at several dilutions using the standard
addition method.
3.2.5 Assessment of Extracting Devices for Conducting the EP - Task 3
Two extracting devices (magnetically stirred and rotary extractor)
were compared for Task 3. The magnetically stirred extractor at both a
high and low mixing rate and the rotary extractor at 29 rpm were
examined for conducting the EP (EPA 1980b and EPA 1980d). The low
mixing rate utilized in the magnetic stirrer was the lowest speed that
could be achieved while still keeping solids suspended, and the high
mixing rate utilized was the highest speed at which the stir bar could
be controlled.
A factorially designed experiment was conducted using the three
mixing rates, two wastes (Nos. 12 and 13), four extraction replicates,
and three analytical determinations of each extract. Two statistical
treatments of the data were performed (SAS Institute, Inc., 1979):
variance component analysis and the Duncan's multiple range test.
Stirrer-speed determinations using the magnetic stirrer were made
using a low-frequency tachometer designed at ORNL. A description of
this device follows. A photon coupled interrupter module was installed
in the magnetic stirrer, beneath the plate. A disc with 64 slits was
attached to the magnet shaft in the stirrer. As the magnet rotates,
the disc over an optical sensor (utilizing infrared) generates
64 pulses for each revolution. The pulses are fed into a three-channel
32
-------
low-frequency tachometer where, using integrated circuitry, each of the
64 pulses is converted to a d.c. voltage (equal to 1 rpm). The output
is then transferred to a strip chart recorder.
3.3 Methods of Analysis
3.3.1 Inorganic Analyses
All extracts analyzed for inorganic constituents were treated
according to standard methods (EPA 1979b) and were directly analyzed
for metals by fTameless graphite furnace atomic absorption spectroscopy
(AAS) with the following exceptions:
(1) extracts for Hg determination were preserved by addition to a
nitric acid/dichromate solution and worked up for cold vapor
flameless AAS (Feldman 1974),
(2) Se was chelated with 5-nitro-o-phenylene diamine and
extracted with toluene before analysis (Talmi and Andren
1974), and
(3) As was determined by an arsine accumulation-helium glow
detector procedure (Feldman 1979).
Samples were analyzed by the method of standard addition (EPA 1979b).
Because the primary focus of this study was organic content of
various solid waste extracts, only selected elements believed to be in
high concentrations were examined in the batch-type extractions. The
elements were generally limited to those found in the NIPDWS (EPA
1979a) because they are used to identify toxic constituents in EP
o
extracts. However, Fe, Ni, SO. , and Zn were measured in selected
extracts as well.
33
-------
3.3.2 Organic Analyses
The organic analysis of the various aqueous batch extracts was
carried out by first adjusting the aqueous extract to pH 6.8 using
phosphate buffer and to a conductivity of 20 mS/cm using sodium
chloride. The adjusted extract (500 ml) was then passed through a
cartridge containing 4.2 ml XAD-2 resin (XAD-2 resin was obtained in
pre-cleaned, pre-filled form from Isolab, Inc., Akron, Ohio). Previous
work (Epler et al. 1980) had shown that these conditions were effective
for the extraction of nonpolar organic compounds, of primary interest
in this work. This extraction technique effectively excludes the
acetic acid used in the EP and the citric acid used as an alternative
test extractant. In addition, the XAD technique avoids the problems
often encountered in the solvent partition techniques where high
concentrations of organic acids (such as acetic and citric acids) tend
to produce stabel emulsions, complicating the separation of the aqueous
and nonaqueous phases.
The column extracts were isolated directly on 16.8-mL XAD-2 resin
(see Fig. 3.3) that had previously been cleaned by successive Soxhlet
extraction with water, methanol, acetone, and water. Each batch of
resin was examined for nonpolar organic contaminants.
Organic compounds were eluted from the XAD-2 resin cartridge (used
for batch extractions) with 15 ml of acetone and 5 ml of methylene
chloride (60 ml acetone and 20 ml methylene chloride was used for the
column cartridge). Solvent containing the eluted material was then
concentrated to 1 mL using a Kuderna-Danish (K-D) evaporator. The
extract containers were rinsed with the eluting solvent prior to
desorption of the resin.
34
-------
An exception to the above procedure was made in the filtration
study (Section 3.2.3). In this case resin from each XAD-2 cartridge
(through which a column extract had passed) was emptied into a 150-ml
beaker. Fifty milliliters of acetone/methylene chloride (50/50
mixture) were added, and the mixture was placed in an ultrasonic bath
for 15 min. A 1-mL aliquot was removed and spiked with azulene as an
internal standard. This aliquot was analyzed by gas chromatography.
Aliquots (500 ml) of all other aqueous extracts were spiked with
azulene, adjusted to pH 12 with NaOH, and extracted three times with
50 mL methylene chloride. The pH was then adjusted to pH 2 with HC1
and extracted three times with 50 ml methylene chloride. The methylene
chloride extracts were combined, dried over anhydrous sodium sulfate,
and reduced in volume using a K-D evaporator. The standard solutions
were reduced to 1 ml and the solid waste extracts to 10 ml, since
higher levels of organic compounds were present in the solid waste
extracts. In all cases, the final concentration of the internal
standard was 10 ppm. The extracts were analyzed by gas chromatography.
One solid waste extract was also analyzed by gas chromatography/mass
spectrometry to confirm the presence of the identified compounds.
The organic concentrates were analyzed by gas chromatography using
fused silica capillary columns (30-m-long x 0.25-mm-i.d. columns coated
with SE-52 stationary phase from J&W Scientific). Gas chromatography
was performed using a Hewlett-Packard Model 5736A gas chromatograph
with a Model 18835B Grob-type split/splitless injection system. The
splitless injection technique was used throughout this work.
35
-------
Quantitation was obtained using a Hewlett-Packard Model 3390A recording
integrator. Gas chromatography/mass spectrometry was performed using a
Hewlett-Packard Model 5985 gas chromatograph/mass spectrometer/data
system in the electron impact mode. Volatile organic compounds in
selected extracts were isolated by purge and trap procedures (Zlatkis
et al. 1981) and analyzed by gas chromatography, as above. The
combustion-infrared method (EPA 1979b) was used to determine dissolved
organic carbon (DOC) concentrations. The ORNL in-house quality
assurance program, which was used throughout this study, indicates a
coefficient of variation of less than 8% by this DOC method.
3.3.3 Mutagenicity testing
Mutagenicity testing was carried out under both Tasks 1 and 2.
Under Task 1, the Salmonella/microsome mutagenicity assay was applied
in an abbreviated screening mode to Soxhlet isolates, acid/base/neutral
fractions obtained through sequential extraction, and the five aqueous
extraction procedure extracts generated from Task 1. The general
methodology for the Salmonel1a/microsome assay has been described
elsewhere (Epler et al. 1980). In a screening mode, this assay is
restricted to two Salmonella typhimurium strains; TA100, with a hisG
base-pair substitution, uvrB and rfa factors, and the pKMlOl plasmid;
and TA98, the hisD frameshift, also carrying the uvrB and rfa factors
and the pKMlOl plasmid. The tested strains were used without metabolic
activation as well as with addition of microsomal preparations from
both phenobarbital and Aroclor-1254-induced rats (henceforth referred
to as phenobarbital activation and Aroclor activation).
36
-------
Organic concentrates were prepared by methods identical to thosa
used for chemical analysis (Section 3.3.2) except that the isolates
were dried under a nitrogen stream. The residue was then dissolved in
2 ml of dimethylsulfoxide (DMSO).
Under Task 2, the two sample preparation protocols (Section 3.2.4,
Section 4.2) were compared using the Ames test (the full five-strain
Ames plate assay procedure, with and without metabolic activation) on
solid waste leachates and wastewaters. The general methodology has
been described elsewhere (Epler et al. 1980). This procedure was
slightly modified to conform to the EPA/NEIC protocol (EPA 1980c) for
conducting the Salmonel1 a/microsome assay. The criterion used of
"positive" test was the modified two-fold rule, i.e. a positive
dose-response with a revertant value—for at least one of the test
material doses—at least twice that of the spontaneous revertant value
(solvent control).
3.3.4 Analysis of Bioaccumulative Materials in Wastes and Leachates
Soxhlet isolates, acid/base/neutral fractions obtained through
sequential extractions of the wastes, as well as organic concentrates
from the five aqueous extraction procedures were tested for
bioaccumulation potential (EPA 1978) in Task 4.
Basically, the method involves chromatography of a series of
compounds of known octanol/water partition coefficients on a
reverse-phase HPLC system, using ultraviolet (UV) detection. When
plotting the partition coefficient of known compounds against the log
retention time, a straight line is obtained (as shown in Fig. 3.5) that
can be used to extrapolate back to the partition coefficients of
37
-------
~ 5
cc
UJ
< 4
0
O
ORNL-DWG 81-9417
I I
I I
PHENANTHRENT
O-DICHLOROBENZENE
TJROMOBENZENE
BENZENE
-^ACETONE
I I I
I
I I I I
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8
LOG RETENTION TIME (min)
0.9 1.0
Fig. 3.5. Partition coefficients of known compounds plotted against log
retention times.
38
-------
unknown compounds. Constituents of an unknown mixture, having partition
coefficients equal to or greater than log P = 3, are defined (EPA 1978a
and 1978b) as being potentially bioaccumulative.
All samples were stored, refrigerated, at 4°C to avoid evaporation
and decomposition. Each sample was injected onto a Zorbax*
octadecylsilane column (4.6 mm x 25 cm) by use of a loop injector with
a 20-uL sample loop. An 85:15 methanol-water mobile phase was degassed
and pumped through the system using a Chromatix constant-flow pump at a
rate of 1 mL/min. A Laboratory Data Control ultraviolet absorbance
monitor at 254 nm was employed.
A standard solution was prepared using acetone, benzene,
bromobenzene, orthodichlorobenzene, phenanthrene, and
2,2-bis-(p-chlorophenyl)-l,l-dichloroethylene (DDE). The quantity of
each compound was varied to produce peaks approximately 25% of the
recorder scale (approximately 100 yg/mL). Preliminary standard runs
were made with each compound in a 3:1 acetone-cyclohexane solution to
determine optimum peak heights. The acetone-cyclohexane solution was
diluted in the final standard with methanol to decrease the size of the
acetone solvent peak while maintaining the solubility of the DDE.
Retention times of standard compounds were recorded each day. The
bromobenzene peak was selected as an external standard for
semi-quantitative comparison with sample peaks to account for day-to-day
variation in instrumental response. Plots were made of log P
(octanol/water partition coefficient) versus log retention time for
extrapolation. The 500 to 1 concentration factor was used for aqueous
extracts, with lower concentration factors applied to the waste isolates.
39
-------
SECTION 4: RESULTS
4.1 Task 1; An Evaluation of Aqueous Extraction Procedures to Remove
Nonpolar Organic Compounds from Solid Wastes
4.1.1 Inorganic Analyses of Extracts
The analytical results for selected inorganic components (NIPDWS
metals) in the four batch extracts (EP, water, Na-resin, and citrate
buffer) for wastes 1 through 6 are presented in Table 4.1. The citrate
buffer extracts contained higher concentrations of all metals except
for Ba in waste 1 and Cr in waste 4. In addition, Ba was present in
the citrate buffer extracts in considerably higher concentrations than
in the other batch extracts for wastes 2 through 6. The EP
consistently produced extracts containing the second highest levels of
metals (except Ba in waste 5). The water and Na-resin extractions
yielded extracts containing comparable levels of metals.
All NIPDWS elements were analyzed in the water extracts of
wastes 7 and 8 and the EP extract from waste 8 (Table 4.2). Neither of
these wastes contained levels of metals that warranted further analyses
of these wastes for inorganic constituents.
Inorganic analyses of the four batch extracts for wastes 9, 10,
and 11 are presented in Tables 4.3, 4.4, and 4.5, respectively. Three
EP extractions were performed on sample 10 for the organic isolation
study (Task 2, see Section 3.2.4) to produce necessary quantities of
extract. The three extracts were analyzed separately for inorganic
elements.
Extracts from wastes 9, 10, and 11 were also examined for Fe, Ni,
and Zn, because such coal-related wastes are expected to contain these
40
-------
TABLE 4.1. SELECTED INORGANIC ANALYSES OF EXTRACTS PRODUCED USING
FOUR EXTRACTION PROCEDURES FOR WASTE SAMPLES 1-6
Extraction procedure3
Waste Element
1 As
Ba
2 Ba
Cr
3 As
Ba
Cr
4 Ba
Cr
5 Ba
6 Ba
EP
65,000
150
144 + 51C
3,940 +_ 1,216C
3.0
660
18
400
180
1,500
650
Water Na-resin
37,000b- 84,000b
79 100
310 210
131 180
0.54 0.8
590 390
8.9 1.3
460 190
163 160
470 190
460 430
Citrate
buffer
156,000b
35
730
17,400
21
990
510
l,100a
34
1,430
3,000
aSingle determination.
Exceeds EP threshold concentrations as in 40 CFR 261.24.
°Mean and standard deviation of replicate extractions (n=2).
41
-------
TABLE 4.2. INORGANIC ANALYSES OF WATER
AND EP EXTRACTS FOR WASTES 7 AND 8
Water extraction3
Element
Ag
As
Ba
Cd
Cr
Hg
Pb
Se
Waste 7
<0.01
4.7
210
0.13
2.6
0.008
4.1
<0.5
Waste 8
<0.01
0.4
520
0.63
2.2
0.004
2.1
<0.5
EPa
Waste 8
<0.01
2.9
370
0.91
13
0.02
11
<1
aSingle determination.
42
-------
TABLE 4.3. INORGANIC ANALYSES OF EP, WATER, NA-RESIN,
AND CITRATE BUFFER EXTRACTS FOR WASTE 9
Extraction procedure3
Element
Ag
As
Ba
Cd
Cr
Hg
Pb
Se
Fe
Ni
Zn
s°;
EP
0.1
1.7
160
0.73
2.6
0.04
2.6
1.8
4.0
5.9
543
69,000
Water
<0.1
0.65
120
<0.02
0.51
nd
0.99
<2
8.1
<1
70
70,000
Na-resin
. 110/1 - - - -
yg/u
<0.1
3.0
130
<0.02
0.56
nd
2.0
2.3
22
<1
15
nd
Citrate
buffer
ndb
13
600
nd
98
nd
nd
5.0
16,300
140
3383
nd
Single determination.
}nd = not determined.
43
-------
TABLE 4.4. INORGANIC ANALYSES OF EP, WATER, NA-RESIN,
AND CITRATE BUFFER EXTRACTS FOR WASTE 10
Element
Ag
As
Ba
Cd
Cr
Hg
Pb
Se
Fe
Ni
Zn
. EPa
(x + S.O.)
38C
8.3 + 1.6
147. + 5.8
<0.1
5.4 + 2.5
0.06b
16.5 + 13.4
4.6 1 0.8
11+7
25 i 4
30 + 27
Extraction
Waterb
. - - iia/l -
<0. 1
0.7
160
<0.1
3.8
ndd
n
4.9
8.8
21
19
procedure
Na-resin
<0.1
0.6
130
0.67
6.7
nd
8.7
4.3
4.4
46
7.5
Citrate.
buffer0
0.28
3.6
170
1.8
42
nd
43
12
16,500
62
63
aThree extractions were performed. Values (means and standard
deviations) reported are based on single determinations on each
of the the 3 extracts.
bSingle determination.
cHighest value reported, all other values less than
detection limit.
dnd = not determined.
44
-------
TABLE 4.5. INORGANIC ANALYSES OF EP, WATER, NA-RESIN,
AND CITRATE BUFFER EXTRACTS FOR WASTE 11
Extraction procedure3
El ement
Ag
As
Ba
Cd
Cr
Hg
Pb
Se
Fe
Ni
Zn
EP
0.03
0.3
335
0.97
<0.7
<0.01
1.4
<0.5
1,830
130
220
Water
- - - - IIO/I
0.05
0.5
306
0.38
<0.7
ndb
1.3
<0.5
320
70
84
Na-resin
0.16
1.2
222
0.24
<0.7
nd
<1
<0.5
45
7
9.6
Citrate
buffer
0.42
225
777
5.2
483
nd
263
1.8
83,000
330
570
aSingle determination.
nd = not determined.
45
-------
metals. Extract data for wastes 9, 10, and 11 showed consistently
higher concentrations of metals in the citrate buffer extracts. In
several cases (Cr in sample 9, Se in sample 10, and As, Cr, and Pb in
sample 11), the citrate buffer extraction produced levels in excess of
the NIPDWS concentrations. In general, higher concentrations of Ba,
Cd, Cr, Pb, and Se were noted in the citrate buffer extracts.
The higher extraction efficiency for metals by the citrate buffer
extraction may be explained in part by the large quantities of Fe
released with this extractant (Table 4.6). The high complexing
capacity of citrate for Fe tends to solubilize Fe from iron-solid phase
matrices, thus releasing associated metals from the solid phase.
4.1.2 Isolation of Organic Compounds from Wastes
The isolates produced by the Soxhlet extraction and the three-step
sequential extraction procedure were compared by gravimetry as well as
by gas chromatography (see Section 3.2.1). The gravimetric data are
shown in Table 4.7. The three-step sequential extraction scheme
consistently extracted more organic material from the wastes than did
the Soxhlet. The major organic components of the wastes were isolated
by both techniques. Figures 4.1 and 4.2 indicate the complexity of the
waste isolates. For identification of the major organic compounds in
the wastes, it was sufficient to analyze only the Soxhlet extracts by
gas chromatography/mass spectrometry. Table 4.8 lists the compounds
which were identified confidently in each of the waste isolates. In
most cases where identifications were made, appropriate standards were
not available, and identification was made by comparison with reference
spectra. The compounds are listed in order of decreasing concentration.
46
-------
TABLE 4.6. COMPARISON OF FE CONCENTRATIONS
USING FOUR EXTRACTION MEDIA3
Waste
1
2
3
4
5
6
9
10
11
EP
0.009
0.12
1.55
0.035
0.77
0.99
0.004
0.01
1.8
Water
0.001
0.059
0.41
0.037
0.036
0.24
0.008
0.008
0.32
Fe
Na-resin
0.001
0.070
0.14
0.013
0.028
0.076
0.022
0.004
0.045
Citrate
buffer
25
10.5
8.7
0.51
3.3
32
16.3
16.5
83
aSingle determination.
47
-------
TABLE 4.7. TOTAL ORGANICS RECOVERED: COMPARISON OF SOXHLET
EXTRACTION WITH THE SEQUENTIAL EXTRACTION SCHEME -
GRAVIMETRIC DATA ON SOLID WASTE ORGANIC ISOLATES
Sequential extraction
Waste
1
2
3
4
5
6
9
10
11
Acids
0.74
2.4
6.5
3.5
1.6
1.0
2.7
0.25
0.48
Bases
4.4
0.15
0.2
0.1
0.2
0.2
0.24
0.4
0.31
Neutral s
C rt 1 1 H lalAC^O
38.4
14.9
24.0
58.2
26.6
60.6
368
16
344
Soxhlet
12.1
3.0
1.3
21.0
14.2
11.2
151
4.1
25.8
48
-------
«*/WB 81-4545
I
JJUL
ulwLjL_A.
78 86 106 126 146 166 186 206 226
1 1 i
I 1 1 .1
Fig. 4.1. Chromatograms for waste 2 - Soxhlet extract (A), ana
sequential extraction extracts: acids (B), bases (C), and
neutrals (D). Injection temperature - 300°C, detector
temperature - 300°C, carrier gas - \\2 at 1.2 mL/min.
49
-------
ORNL-OWG 81-15954
B
C
\iM
TEHP CC)
Fig. 4.2. Chromatograms for waste 11 - Soxhlet extract (A) and
sequential extraction extracts: acids (B), bases (C), and
neutrals (D). Conditions as in Fig. 4.1.
50
-------
TABLE 4.8. ORGANIC COMPOUNDS IDENTIFIED IN SOLID WASTES3
Waste Compounds
1 Nitroanilineb
2 Hydrocarbons (19)c, cresolb, indoleb, methyl indoles (2)c,
phthalates (3)c, nonyl phenols (5)c, decanoic acid,
phenanthreneb
3 Aliphatic hydrocarbons, cresolb, naphthaleneb, hydroxy indole
4 Tributyrin, p_-hydroxy benzaldehyde (2)c, methylene bisphenol
(2)c, hydroxy benzyl alcohol, phenol0, hydroxy benzoic acid,
biphenol (2)c, fluorantheneb, bicresol (Tentative), piperonal
5 Aliphatic hydrocarbons, cresolb, methyl indole, hydroxy indole,
methoxy indole
6 Aliphatic hydrocarbons, siloxanes, dioctylphthalateb, cresolb,
indoleb, dibutylphthalateb, methyl indole, butyric acid,
phenyl acetic acid, octanoic acid, decanoic acid
9 Methyl fluorantheneb, methyl biphenylb, trimethylnaphthalene,b
biphenylb, various aliphatic hydrocarbons
10 Biphenyl^, 05 phenol, C] tetrahydronaphthalene, C] naphthalene
(2)b, tetrahydronaphthalene, minor components, C3 hydroxy
biphenyl ^ hiphenyl (2)c, isopropyl cyclohexane, C3 benzenes
(3)c, indane, naphthalene,b hydroxy anthracene, acenaphtheneb,
CTI biphenyl, dibenzofuran, fluorene, phenanthreneb,
hydroxy fluorene, C] phenanthrene (2)b»c, methoxyphenanthrene
11 Biphenyl ether, biphenyl0, phenanthreneb, tetrahydronaphthalene,
cresol", €3 phenol, xylenols (3)c, phenyl phenol, tplyl phenol
(5)c, Ctj phenol, bipiyridyl, acridine + benzoquinol ines,
dibenzofuran, acenaphtheneb, fluoreneb, dibenzothiopheneb,
phenanthrenec, fluorantheneb, pyreneb
Compounds identified in Soxhlet isolates in order of decreasing
concentration.
Comparison with authentic standards.
cNumber of isomers detected.
51
-------
Comparison of the gravimetric data with the total area of the
chromatogram indicated that 15 to 20% of the mass was eluted under
these chromatographic conditions.
The organic content of the wastes varied considerably, as was
expected, although wastes 2, 3, 5, and 6 contained similar compounds,
predominantly hydrocarbons and carboxylic acids. The acids in these
wastes probably resulted from decomposing municipal refuse (these
wastes were obtained from landfills that also received municipal
waste), while the hydrocarbons may have resulted from oil disposal.
Waste 1 contained nitroaniline, which constituted about 99% of all
organic material present. Waste 4 contained exclusively phenolic
compounds plus tributyrin. These compounds resulted from a phenoxy
resin plastic manufacturing process. Wastes 9, 10, and 11 contained
similar compounds; each waste resulted from a coal conversion process.
Thus, the major constituents of these wastes are phenols, aromatic
heterocycles, and aromatic hydrocarbons.
Volatile organic compounds were determined (see Section 3.2.1) for
wastes 2, 3, and 9 (Figs. 4.3 and 4.4). Volatile compounds included
hydrocarbons, phenols, and sulfur compounds. Figure 4.3 shows that
volatile compounds from waste 2 could be detected after aging 4 years
in a carefully covered landfill.
4.1.3 Organic Analyses of Aqueous Extraction Procedure Extracts
4.1.3.1. Dissolved Organic Carbon Results
Earlier studies of solid waste extracts (from batch extraction
procedures) indicated that a screening procedure for the presence of
water-soluble organic compounds would be desirable (Epler et al. 1980).
52
-------
ORNL-DWG 81-9582
01
40
TIME (min)
Fig. 4.3. Chromatogram of waste 2 (aged four years in a tightly covered landfill) -
volatile organic compounds. Conditions as in Fig. 4.1.
-------
ORNL-OWG 81-9305
B
I
I
I
80 70 60 50 40 30
TIME(min)
20
10
Fig. 4.4. Chromatograms of volatile organic compounds for waste 3 (A)
and waste 9 (B).
54
-------
Such a screening protocol would reduce the laboratory time for
preparing and analyzing extracts for organic constituents. Dissolved
organic carbon (DOC) analysis is an appropriate method for screening
extracts for organic constituents. However, it cannot be used for
extraction procedures that use organic acids (e.g., acetic acid used in
the EP or citric acid) unless very high concentrations of other organic
compounds are present above the acid background levels in the extract.
The water and Na-resin batch extracts were also examined for DOC
concentrations. In addition to the batch procedure extracts, the
column extracts, after passing.through XAD-2 resin, were analyzed for
DOC to screen for organic compounds not retained by the XAD-2 resin
(e.g., very polar organic compounds).
The concentrations of DOC found in extracts of the water,
Na-resin, and column (after passing the XAD-2 resin) procedures for the
11 wastes tested are shown in Table 4.9. All wastes contained
detectable quantities of organic components. There was no significant
statistical difference in quantities of DOC extracted between the water
and Na-resin procedures (SAS Institute, Inc. 1979). Although the
column extracts were not analyzed for the specific organic compound
present, previous studies with XAD-2 resin indicate that polar
compounds, e.g. dicarboxylic acids, nonextractable with organic solvent
accounted for the levels of DOC observed (Epler et al. 1980). Column
extracts from waste 14 did not contain extractable organic compounds
above 10 yg/L (Section 4.1.3.2).
As DOC analysis requires filtration at the submicron level, it was
necessary to evaluate the effect of filter type. Therefore, all water
55
-------
TABLE 4.9. DISSOLVED ORGANIC CARBON (DOC) OBSERVED
IN SELECTED AQUEOUS WASTE EXTRACTS3
Extraction
Waste
1
2
3
4
5
6
7
8
9
10
11
Water
198
457
207
254
92
552
134
42
13
54
46
Na-resin
155
494
178
353
64
406
37
48
13
39
28
Column - after passing
through XAD-2
80
206
246
31 9b
268
423
nec
ne
10
50
ndd
filtered through glass fiber filters (Whatman GF/F,
precombusted 0.7 pm).
Average of two replicates.
cne = not extracted.
nd = not determined.
56
-------
and Na-resin suspensions were filtered using two types of filters: a
polycarbonate membrane filter (Nuclepore, 0.4 ym) and a precombusted
glass-fiber filter (0.7 urn). The glass-fiber-filtered extracts were
expected to be higher in DOC due to sorption of organics on
polycarbonate filters as well as passage of submicron particles
(0.4-0.7 urn) through the glass fiber filter. However, as Table 4.10
shows, this was generally not the case. In cases where large (order of
magnitude) differences were found in DOC values (e.g., the Na-resin
extracts from wastes 3 and 9),'the polycarbonate-filtered extracts
contained the higher level. Statistical analysis (SAS Institute, Inc.,
1979) of data in Table 4.10 showed that there was no difference in
DOC values between glass-fiber- and membrane-filtered extracts.
To determine whether changes fn the DOC values during the 24-h
extractions, warranted consideration of a shorter extraction time,
25-mL aliquots (one aliquot for each sampling time) were drawn from the
extraction vessel immediately after the wastes were thoroughly mixed
and at 1-, 2-, 4-, 8-, and 24-h intervals. Figures 4.5 and 4.6
graphically describe concentrations of DOC found in the water and
Na-resin extracts, respectively. These plots show there was no
consistent pattern of increase or decrease in DOC values with respect
to time. In general, the wastes that contained appreciable levels of
volatile organics (wastes 2 and 3) decreased in concentrations of DOC
over the extraction time. This phenomenon might be expected because
the extraction vessel was not sealed.
57
-------
TABLE 4.10. COMPARISON OF DOC IN EXTRACTS FILTERED
THROUGH GLASS FIBER AND POLYCARBONATE FILTERS
Mater extracts
Waste
1
2
3
4
5
6
7
8
9
10
11
GFb
198
457
207
254
92
552
134
42
13
54
46
NFC
212
505
337
243
72
544
29
45
10
56
49
Na-resin
Displacement extracts
GFb
155
494
178
353
64
406
37
48
13
39
28
NFC
124
437
1740
280
44
480
30
42
no
40
46
aSingle value determinations.
Glass fiber filter (Whatman GF/F, precombusted, «0.7 um).
°Polycarbonate filter (Nuclepore, 0.4 ym).
58
-------
ORNL-OWG 81-13763RESD
500 ^S_2.
O
•6-
•2-
•6
•2
WATER EXTRACTION
I
I
8
12 16
TIME (h)
20
24 26
Fig. 4.5. DOC found in water extracts over the 24-h extraction time.
59
-------
ORNL-DWG 81-13764RESD
1VJUW
500
i
200
|
^
o»
1 100
i «
CD i
% so :
o 1
ORGANIC
r\>
o
DISSOLVED
» ro ui O
1
(
1 1 1 1 1 1
s-s-^rirrrr.'glrrrrrra^.
3^6/"-*-3- ""~~~--6
3 3- — -._, 4
rr*-4~- -^-.-::;.^_^
I^rS-.o..--""6 5
^x« "5"j.v._ — 8 _
o, JO\HO - « ,.1 _ _V.V_Vr _-_ .- — ,10
7X
.9
- 9
9
9-~
/ Na-RESIN
9' EXTRACTION
{-
1 1 1 1 1 1
D 4 8 12 16 20 24 2
TIME (h)
Fig. 4.6. DOC found in Na-resin extracts over the 24-h extraction time,
60
-------
Because the DOC values changed over time for each sample, a linear
regression analysis was performed to determine if these changes were
significant over time (SAS Institute, Inc. 1979). The following waste
extracts showed a statistically significant change (P < 0.05) in DOC
(slope of linear regression different from zero) over the 24-h
extraction time:
(1) the water extracts from wastes 7 and 8 increased in DOC over
time,
(2) the Na-resin extract from waste 4 increased, and
(3) the Na-resin extracts from wastes 2 and 3 decreased in DOC
over time.
Although waste 9 did not show a statistically significant change over
24 h in DOC concentration by water extraction, there was a large drop
in DOC concentrations (from 120 to 20 ug/L) in the first 4 h.
During the first 8-h of extraction, only the Na-resin extract from
waste 9 increased in DOC. Overall, the majority of wastes extracted
did not significantly change in DOC content over the extraction period.
With the exception of the water extraction of waste 7, there were no
significant differences observed between 8- and 24-h extraction times,
suggesting that 8-h extraction may be as effective as the 24-h
extraction, at least for the analysis of DOC.
4.1.3.2 Gas Chromatography and Gas Chromatography/Mass Spectrometry
Using GC and GC/MS analysis, the extracts produced from the five
extraction procedures were compared in two ways. First, the total area
of each of the chromatograms (excluding the solvent peak) was compared,
as a measure of the total mass extracted. Because of the low levels of
61
-------
organic compounds present in most of the extracts, gravimetry was not
useful. Second, the quantities of individual compounds extracted by
the various media were compared. While it was not possible to obtain
absolute quantitative data on the individual compounds, the equivalent
treatment of all extracts ensures a reliable relative comparison.
Blanks were run by carrying the media through all extraction and
analytical operations. Appendix A contains quality control procedures
used at ORNL for organic analyses as well as inorganic analyses.
The data for total chromatographable organic content for each of
the extraction procedure extracts are shown in Table 4.11.
Additionally, an estimate of the total chromatographable organic
concentration is given for each EP extract. This estimate is based on
average integrator response for aliphatic hydrocarbons and should only
be used to make relative comparisons between samples.
To evaluate the relative effectiveness of the extraction procedures
to extract organic compounds, each extraction procedure was ranked from
lowest to highest (1 to 5) for each waste (Table 4.11). This ranking
scheme was used to statistically detect significant differences
(P < 0.05) among extraction procedures across wastes using a
completely randomized design (by extraction procedure). Applying this
ranking scheme, the following conclusions can be made relative to the
effectiveness of the five extraction procedures to remove
chromatographable organic compounds:
(1) The up-flow column was more aggressive than any of the batch
extraction procedures.
62
-------
TABLE 4.11. THE RELATIVE EFFECTIVENESS OF FIVE AQUEOUS EXTRACTION
PROCEDURES TO REMOVE ORGANIC COMPOUNDS FROM SOLID WASTES
Extraction procedure
Waste
1
2
3
4
5
6
9
10
11
Mean
rank
score0
EP
1.0 (1)-
1.0 (3)
1.0 (2)
1.0 (3)
1.0 (3)
1.0 (1)
Ob (1)
1.0 (3)
1.0 (4)
2.33
Water
. . . TTfl i
3.1 (3)
0.79 (2)
0.5 (3)
1.1 (4)
3.6 (4)
37 (4)
1.0 (4)
1.5 (4)
0.49 (2)
3.33+
Na-
resin
/s EP (rank'
8.7 (4)
1.4 (4)
0.32 (1)
0.84 (2)
0.07 (1)
2.6 (2)
0.42 (2)
0.67 (1)
0.40 (1)
2.00
Citrate
buffer
a\
i
2.6 (2.).
0.10 (1)
2.0 (4)
0.45 (1)
0.14 (2)
4.2 (3)
0.44 (3)
0.90 (2)
0.69 (3)
2.33
1
t(
Column
22 (5)
6.7 (5)
7.3 (5)
6.3 (5)
4.1 (5)
49 (5)
4.7 (5)
2.3 (5)
8.9 (5)
5.00*
Estimated
jtal in EP
(yg/L)
328
224
31
340
700
6
37b
754
2740
aRanking of each extraction procedure from lowest to highest (1 to 5)
for each waste sample.
bTotal in distilled water, as no detectable peaks were observed in
the EP extract.
cMean rank scores without the same superscript are significantly
different (P < 0.05) by Duncan's multiple range test.
63
-------
(2) Among batch extraction procedures, deionized distilled water
was the most aggressive extracting medium.
(3) No statistical differences were observed among the EP, citrate
buffer, and Na-resin batch extraction procedures.
The column extraction procedure was replicated three times using
waste 10. The total chromatographable organic compounds (total area of
the chromatographs as measured in integrator counts, a dimensionaless
unit) for the three replicate column extractions were: replicate 1:
2.5 x 107, replicate 2: 2.7 x 107, and replicate 3: 3.2 x 107. The
coefficient of variation associated with these three replicates was 13%.
These data suggest that the extractions were in reasonably close
agreement in terms of organic matter extracted. Chromatograms of the
first and third extractions were quite comparable relative to organic
compounds present (Fig. 4.7). Furthermore, the extract after passing
through the resin cartridge was analyzed by the method described in
Section 3.3.2 (solvent partition at basic and acidic pH). No
extractable organic compounds were detected above 10 ug/L.
For individual organic compounds, data are shown in Table 4.12.
Conclusions 1 and 2 developed above for total chromatographable organic
compounds apply to individual compounds as well. Interestingly, it
seems that the sample matrix is a major factor in individual compound
differences between the four batch extraction procedure extracts.
Cresol, for example, was extracted more effectively by the water
procedure on wastes 2, 3, and 6, but by the EP on wastes 4 and 11.
Volatile organic compounds were also determined on selected
samples. Figure 4.8 shows the results of the analysis of volatiles
64
-------
OR ML DWG 81 21534
1
70
86
106
H—
20
TEMPERATURE (°C)
126 146 166 186
H 1 1 h-
30 40 SO 60
TIME (mm)
206
—I—
226
250
10
70
80
Fig. 4.7. Chromatograms of waste 10 column extracts - replicate 1 (1)
and replicate 3 (2). Conditions as in Fig. 4.1.
65
-------
TABLE 4.12. RELATIVE AMOUNTS OF INDIVIDUAL COMPOUNDS IN SOLID WASTE
EXTRACTS AND RELATIVE RANKING OF THE EXTRACTION PROCEDURES
Extraction procedure3
Na- Citrate
Waste
1
2
3
4
5
W
6
9
10
11
Mean
and =
Compound
Nitroamline
Cresol
Indole
Methyl indole
Decanoic acid
Phenanthrene
Cresol
Naphthalene
Hydroxyindole
Phenol
Cresol
o-hydroxy benz-
aldehyde
Tributyrin
Methyl ene bisphenol
Chrysene
Naphthalene
Methyl indole
Hydroxyindole
Methoxyindole
Phenol
Butyric acid
Cresol
Decanoic acid
Phenyl acetic acid
Indole
Methyl indole
(No peaks identified)
C3 pyridine
Naphthalene
Quinoline
1 -methyl naphthalene
Cresol
Xylenol
Quinoline
£3 phenol
Naphthol
Blphenyl
rank scoreb
not detected.
EP
1.0
1.0
1.0
1.0
1.0
1.0
1.0
nd
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
nd
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0
2.08
Water
2.0
1.6
1.6
0.76
1.1
2.0
1.6
1.0
0.13
2.2
0.72
1.0
1.6
0.87
15
2.0
2.0
0.70
2.3
1
25
12
16
49
16
13
4.7
3.2
5.6
2.0
0.33
0.10
0.52
0.71
0.70
3.5
3.13
resin
5.6
1.0
2.2
1.1
1.0
1.8
3.2
nd
0.07
0.88
0.91
0.20
1.8
0.57
16
nd
nd
0.04
0.11
0.10
6.8
1.3
8.5
1.6
10
1.2
3.1
1.4
1.3
1.2
0.68
0.13
1.1
0.68
0.51
2.8
*T
2.44
buffer
3.6
0.02
1.1
0.98
1.3
0.58
0.28
0.82
0.15
1.6
0.55
0.96
1.6
0.97
7.4
nd
1.4
0.13
0.12
0.63
4.5
6.3
9.7
1.8
7.9
3.0
3.9
1.6
2.3
1.5
1.3
0.51
0.74
0.68
0.58
0.85
2.49
Co 1 umn
13
36
41
14
5.1
21
20
26
1.1
21
12
2.3
3.6
4.6
16
7.5
5.3
0.99
1.9
6.6
nd
1100
22
51
20
15
13
15
6.7
6.7
15
9.6
4.3
8.7
8.8
17
4.87*
bMean rank scores without the same superscript are significantly
different (P < 0.05) by Duncan's multiple range test.
66
-------
(a)
I
ORNL-DWG 81-9581
JuL
II . LUjbJL
60
50
40 30
TIME(min)
20
Fig. 4.8. Chromatograms of volatile organics from waste 2 extracts
citrate buffer extract (a) and Na-resin extract (b).
Conditions as in Fig. 4.1.
67
-------
from the citrate buffer extract and the Na-resin extract from waste 2.
The citrate buffer extract clearly contained higher levels of volatile
organic compounds. Similar results were obtained on wastes 3, 9, 10,
and 11. While the mechanism of extraction of organic compounds using
these two media was expected to be similar, there is a fundamental
difference: the citrate buffer extraction is conducted in a sealed
tumbler while the Na-resin extraction is conducted in a stirred vessel
which is not airtight. To assess if the differences noted were
actually due to the difference between the rotary extractor and the
stirred vessels, or a result of the different extraction media, the
distilled water extract of waste 2 was repeated in a sealed system.
The results are shown in Fig. 4.9. The sealed vessel extract again
contained much higher levels of organic volatiles, although not quite
as high as in the citrate buffer extract, implying that both factors
(vessel type and extracting media) are involved. Further examination
of this phenomenon is recommended.
In summary, the column extraction procedure was found to be the
most effective means of extracting nonpolar organic compounds. The
most noticeable differences are in the levels of moderately volatile
compounds (e.g., phenol and cresol) and in the levels of the nonpolar
compounds (e.g., phenanthrene).
Factors contributing to the observed higher yields of organic
compounds from the column procedure are:
(1) the column procedure is a closed system allowing for sorption
of the moderately volatile compounds directly on the XAO-2
resin, and
68
-------
ORNL-DWG 64-9583
(a)
(c)
ILL
60
50
40 30
TIME(min)
20
<0
Fig. 4.9. Chromatograms of volatile organics in water
extracts from waste 2 - open vessel (a)
versus closed vessel (b), and distilled in
glass water blank (c). Conditions as in
Fig. 4.1.
69
-------
(2) the column extracts were not filtered using membrane filters
which are known to sorb appreciable quantities of nonpolar
organic compounds (evidence relative to this effect is
presented in Section 4.1.5).
It is important to point out that the major objective of this work was
to develop an aqueous extraction procedure that simulated real-world
conditions. The goal was to identify a procedure that was significantly
better, in terms of removal of organic compounds, than the present EP,
and one that could be used for a variety of wastes. The upflow column
procedure appears to be such a procedure. However, the reasons for
removal of greater quantities of organic compounds by this procedure
are yet undetermined. Two of the contributing factors are suggested
above. In a supplemental study, an experiment was conducted, using
waste 14, to evaluate the influence of membrane filtering on column
extracts.
The relative ranking of the extraction procedures to remove
organic compounds from solid wastes (Tables 4.11 and 4.12) indicate
that water is a significantly (P < 0.05) better extracting medium
than the EP (acetic acid), Na-resin (sodium saturated system), or
citrate buffer (0.5 fl sodium citrate). It appears that a medium low in
ionic strength, such as deionized distilled water, enhances
solubilization and mobility of organic compounds from solid wastes.
McKown et al. (1980) observed a similar solvent action with water.
This solvent action in combination with the up-flow column procedure--
which provides a continuous fresh supply of deionized distilled
water—may be the primary mechanism responsible for the greater removal
70
-------
of organic compounds by the column procedure. Work in FY-lfi82 is
intended to evaluate in greater detail the chemical mechanisms and/or
operational processes that are responsible for the observed difference
in quantities of organic compounds extracted by batch and column
procedures.
The majority of the data presented here suggest that the physical
apparatus, i.e. extraction vessel employed, has a greater effect on the
organic content of the resulting extracts than the aqueous medium
tested.
4.1.4 Mutagenicity Testing
For comparative purposes, the Salmonella/microsome assay results
obtained with the various extracts from a given waste are reported
together in Tables 4.13 - 4.21. The results of mutagenicity testing
are summarized in Table 4.22. The modified two-fold increase rule was
used as an indicator of positive mutagenic test response (EPA 1980c).
A sample was considered toxic if in any test the background lawn was
absent at two or more consecutive dose levels. As the presence of
cytotoxicity can alter or mask the apparent mutagenic response, tests
showing toxicity are noted.
Of the nine wastes examined (1-6, 9-11), seven were found to
possess mutagenic activity in at least one extract or isolate. Only
wastes 4 and 6 were without detectable genetic activity.
The results of mutagenic testing of waste 1 isolates and extracts
are shown in Table 4.13. Among the aqueous extracts, a positive
response was elicited, in the TA 98 strain of Salmonella typhimurium
with the water, citrate buffer, and Na-resin extracts. The Soxhlet
71
-------
TABLE 4.13. RESULTS OF NUTAGENICITY TESTING OF HASTE 1 EXTRACTS
-j
ro
Volume 01
DMSO tesi
solution
luL/plale
) EPa
Revertants/plate
TA 98
Uater*
Citrate Na-
buffer* resin"
Column1
Sequential extraction
Soxhlet6 Acid6 8asec Neutral"
EP4
Mater1
Citrate
buffer*
TA 100
Na-
resin Column
So»hletb
Sequential extraction
Acldc Base0 Neutral"
Phenobarbital activation
75
SO
25
10
0
T«
T
52
54
20
T
T
79
111
24
T
59
45
42
20
42
31
38
30
15
S3
54
80
70
44
94
83
63
55
22
T
109
85
50
18
20
27
32
19
26
78
61
35
21
14
T
T
176
191
137
T
377
380
348
282
T
107
109
150
126
114
204
170
192
186
65
79
120
139
162
242
210
201
137
119
106
135
124
102
115
107
121
115
141
115
119
157
145
124
83
Aroclor activation
75
50
25
10
0
75
SO
25
10
0
T
T
T
33
21
T
14
T
T
106
97
14
T
21
30
39
55
27
31
T
20
40
49
44
36
26
28
19
53
54
67
61
53
17
45
70
59
44
41
30
138
62
60
54
20
T
94
72
44
20
150
19
23
29
24
27
24
22
31
60
49
35
26
16
No activation
32
29
T
T
T
167
138
T
163
T
T
326
325
202
T
166
T
T
119
153
78
T
142
T
196
211
196
149
148
177
T
56
104
178
169
T
167
186
173
142
136
108
252
199
167
170
144
102
98
113
108
128
119
107
135
159
142
152
143
106
137
115
141
120
112
91
99
91
a250 |iL of aqueous extract/uL of DMSO test solution.
bl mg solid waste/uL of DMSO test solution.
C3 019 solid waste/uL of DMSO test solution.
d300 ug solid waste/Ml of ONSO test solution.
'T = toxic.
-------
TABLE 4.14. RESULTS OF MUTAGENICITY TESTING OF WASTE 2 EXTRACTS
-•J
C*>
Volume o
OHSO tesi
solution
(uL/plate
F
) EP*
Revertants/plate
TA 98
Water*
Citrate Ka-
buffer* resin"
Sequential
Column*
Soxhlet" Acfdc
extraction
Basec Neutral*1
EPa Water*
Citrate
buffer*
Ha-
res In*
TA 100
Column*
Soxhletb
Sequential extraction
Acid0 Basec Neutral*1
Phenobarbltal activation
75
SO
25
10
0
21
18
15
20
17
168
154
165
137
151
71
71
29
19
21
72
44
42
28
16
IS
16
27
26
16
19
10
20
29
16
T6
16
12
10
22
15
17
17
16
20
32
36
18
18
26
74
81
73
80
94
153
290
308
220
226
107
120
94
83
86
149
169
169
83
112
T
T
148
142
141
59
69
93
103
93
T
T
T
117
150
120
106
94
107
99
113
121
82
106
119
Aroclor activation
75
50
25
10
0
50
0
27
29
23
23
19
19
22
71
99
82
67
74
20
28
52
40
29
32
30
15
8
34
39
32
30
17
18
16
29
27
30
32
18
19
42
20
24
21
18
18
6
14
T
T
13
19
32
T
15
26
IS
14
18
IS
20
16
32
32
26
24
23
No activation
33
14
85
82
94
86
133
93
99
227
193
188
215
178
141
194
T
156
194
125
119
117
104
T
142
92
78
200
94
200
T
T
T
149
125
T
120
T
T
T
T
114
T
119
T
T
T
T
153
T
179
98
107
86
113
122
121
106
120
128
113
124
115
114
141
250 uL of aqueous extract/Ml of DNSO test solution.
bl mg solid waste/wL of DMSO test solution.
3 ng solid waste/uL of DMSO test solution.
d300 ug solid waste/id of DMSO test solution.
er » toxic.
-------
TABLE 4.15. RESULTS OF MUTAGENICITY TESTING OF WASTE 3 EXTRACTS
\lo\ume ol
OMSO tesl
solution
(uL/plate
) EP*
Revertants/plate
TA 98
Water*
curate Na-
buffer* resin*
Column*
Sequential
Soxhletb Acid0 Basec
extraction C i tra te
Neutral*1 EPa Water* buffer
TA 100
Na-
resin* Column* Soxhlet
Sequential extraction
Acidc Base0 Neutral"
Phenobarbltal activation
75
50
25
10
0
T
19
22
31
19
284
284
243
ISO
44
29
27
22
18
20
32
30
31
18
22
29
39
31
33
43
22
23
23
17
24
18
17
24
17
14
11
7
14
18
12
14
21
13
20
16
T
T
130
138
197
488
420
365
350
247
136
142
94
154
137
161
141
180
137
167
106
121
137
120
126
97
98
96
103
83
107
136
109
104
83
85
108
84
98
89
83
133
107
105
112
Aroclor activation
75
50
25
10
0
50
0
T
21
25
20
40
23
32
154
104
89
61
65
122
37
21
26
29
29
21
16
14
52
34
32
29
28
18
58
40
35
29
40
42
14
30
21
18
27
16
22
7
8
10
17
19
15
26
5
12
11
14
16
14
11
13
10
26
20
20
15
23
No activation
16
18
T
42
73
ins
142
8?
171
346
277
221
152
188
247
179
138
161
143
106
138
169
163
174
180
134
91
154
149
110
139
121
119
123
149
108
121
85
93
102
112
89
104
95
ndf
113
113
99
91
91
114
94
77
78
107
81
107
110
102
110
85
106
91
114
134
*2SO pL of aqueous extract/uL of DMSO test solution.
bl mg solid waste/Ml of OMSO test solution.
C3 109 solid xaste/uL of OMSO test solution.
d300 ug solid waste/uL of OMSO test solution.
6T - toxic.
nd • not determined.
-------
TABLE 4.16. RESULTS OF MUTAGEH1CITY TESTING OF WASTE 4 EXTRACTS
Volume oi
DHSO tesi
solution
(uL/plate
f
) EP*
Revertants/plate
TA 98
Water*
Citrate Na-
buffer* resin"
Column"
Sequential extraction
Soxh 1 etb Acidc Base' Neutral"
EP"
Citrate
Water* buffer*
Ha-
res In*
TA 100
Coliwi"
Soxhletb
Sequential extraction
Addc Basec Neutral"
Phenobarbital activation
75
SO
25
10
0
75
50
25
10
0
14
19
14
19
18
20
21
26
31
26
163
175
155
167
112
T
55
57
67
57
38
33
35
57
58
41
36
34
59
43
IB
17
20
21
16
19
19
22
17
21
16
16
17
30
22
19
27
28
16
28
Te
9
25
17
13
T
T
15
8
26
T
17
17
23
26
22
11
31
15
23
12
9
18
12
15
Aroclor
15
20
16
22
12
12
10
14
12
9
activation
17
17
14
20
14
105
91
85
107
85
85
92
113
92
97
T
355
341
363
363
T
260
201
310
204
T
113
160
106
89
T
162
203
179
146
T
162
120
125
82
T
T
135
138
99
T
T
T
165
151
T
T
T
171
149
T
T
85
64
90
T
T
92
72
59
T
T
T
133
119
T
T
111
176
115
111
75
106
112
116
110
116
95
108
108
86
92
97
76
70
109
88
83
89
103
No activation
50
0
13
20
T
24
T
28
15
20
T
29
3
8
11
14
10
13
15
18
93
90
T
106
T
124
T
108
T
122
53
93
T
141
95
105
68
67
*250 uL of aqueous extract/uL of DHSO test solution.
bl ng solid waste/uL of DHSO test solution.
C3 rug solid waste/uL of DNSO test solution.
d300 ug solid waste/uL of DHSO test solution.
8T • toxic.
-------
TABLE 4.17. RESULTS OF NUTAGEN1CITY TESTING OF HASTE 5 EXTRACTS
Volume ol
DMSO tesl
solution
(uL/plate
) EP1
Revertants/plate
TA 98
Citrate Na- Sequential extraction
Water1 buffer1 resin1 Colum1 Soxhlet1 Acid1 Base1 Neutral1
EP1
Water1
TA 100
Citrate Na-
buffer1 resin1 Column1
Sequential extraction
Soxhletb Acidc Base0 Neutrald
Phenobarbltal activation
75
50
25
10
0
15
13
21
12
18
154
165
180
164
85
74
58
42
67
51
50
47
13
30
30
18
22
18
18
32
34
35
35
25
15
22
20
18
16
14
110
63
46
33
22
15
13
11
12
9
75
83
98
74
85
362
335
404
249
322
129
158
188
133
123
185
178
146
163
148
122
160
153
136
141
111
135
113
159
109
94
114
83
115
83
166
178
182
172
150
61
56
68
66
70
Aroclor activation
75
50
25
10
0
50
0
18
22
18
21
26
20
12
85
82
87
91
83
30
29
76
53
44
42
25
24
22
43
21
34
29
37
46
37
34
29
28
30
29
19
29
22
32
22
25
21
17
18
24
24
15
20
26
10
18
24
25
37
32
32
Np_
24
15
15
16
11
10
14
activation
6
IB
99
105
94
107
97
96
90
229
253
215
216
268
180
238
Te
94
71
99
79
119
121
154
135
146
127
113
T
186
162
154
160
148
180
117
165
109
150
95
117
97
110
121
104
99
107
115
91
110
134
150
151
190
137
153
121
179
55
54
72
75
103
62
67
a250 uL of aqueous extract/uL of DMSO test solution.
bl rng solid waste/uL of ONSO test solution.
C3 mg solid waste/Ml of DMSO test solution.
d300 ug solid Haite/uL of DMSO test solution.
eT • toxic.
-------
TABLE 4.18. RESULTS OF HUTAGENICITY TESTING OF WASTE 6 EXTRACTS
Volume c
DNSO tes
solutloi
(uL/plati
,f
s) EP*
Reverlants/plate
TA 93
Water1
Citrate Na-
buffer* resin1 Column8 Soxhletb
Sequential extraction
Acidc Basec Neutral*1 EPa Water*
Citrate
buffer*
Na-
resln*
TA 100
Column'
Soxhlet"
Sequential extraction
Acidc Base0 Neutral*1
Phenobarbltal activation
75
SO
25
10
0
T
T
T
33
19
97
167
151
161
155
T
59
33
41
37
35
33
30
20
22
28
29
31
44
44
18
20
14
20
17
12
16
10
20
18
42
17
22
19
20
no'
nd
nd
nd
nd
T
T
T
138
128
T
T
334
328
352
T
T
T
T
116
169
160
119
191
167
138
137
169
168
220
57
89
86
82
60
63
136
130
86
115
106
126
90
108
118
nd
nd
nd
nd
nd
Aroclor activation
75
50
25
10
0
50
0
T
T
23
22
20
T
28
91
94
106
96
68
T
22
T
40
37
28
37
T
37
T
39
26
14
28
16
58
35
33
35
45
44
30
47
T
35
15
23
9
18
14
22
18
21
24
23
16
15
10
16
12
12
20
11
14
nd
nd
nd
nd
nd
NO activation
nd
nd
T
T
T
135
138
T
116
T
T
219
239
233
T
169
T
T
T
136
113
T
186
T
T
160
180
154
70
110
163
192
240
243
227
118
136
99
82
60
88
61
43
109
T
T
80
103
119
81
123
88
102
103
75
75
81
96
nd
nd
nd
nd
nd
nd
nd
*250 id. of aqueous extrut/uL of DNSO test solution.
bl ag solid waste/uL of DNSO test solution.
C3 mq solid Mtte/uL of DHSO test solution.
d300 ug solid Haste/id, of OHSO test solution.
*T • toxic.
fnd - not deUratned.
-------
TABLE 4.19. RESULTS OF HUTAGENIC TESTING OF WASTE 9 EXTRACTS
oo
Volume
DHSO te
solutic
(uL/plal
it
>n
te) EP*
Revert ants/plate
TA 98
Water*
Citrate
buffer*
Na-
resin*
Column*
Soxhletb
Sequential extraction Citrate
Acid0 Basec Neutral" EP* Water* buffer*
TA 100
Na-
resln* Column* Soxhlet"
Sequential extraction
Acidc Basec Neutral*1
Phenobarbital activation
75
50
25
10
0
75
50
25
10
0
57
40
64
61
48
37
42
39
55
69
TO
41
33
46
40
T
39
44
46
41
43
38
55
44
48
48
39
49
47
42
45
48
39
53
40
45
32
39
42
41
7
13
8
16
17
T
18
15
14
18
41
33
41
47
17
T
T
T
47
27
£5
58
60
52
47
T
T
T
39
28
T
53
41
71
47
Aroclor
46
52
46
59
28
57 206
43 187
35 202
32 191
19 168
activation
54 193
40 177
30 199
32 162
20 188
160
162
181
165
216
187
206
160
165
152
174
215
226
217
201
234
199
174
173
175
161
167
165
175
216
T
167
182
205
152
86
61
70
74
117
T
99
78
91
118
T
T
117
112
104
T
T
T
107
98
T
T
227
220
173
T
T
T
231
182
187
187
150
216
173
207
160
188
20S
182
162
165
163
135
174
173
160
151
163
98
No activation
50
0
35
54
20
34
51
43
30
34
T
6
T
29
T
35
26
35
33 220
28 233
121
191
207
229
163
191
T
82
T
192
204
192
92
103
a250 uL of aqueous extract/uL of OHSO test solution.
bl tug solid Maste/uL of DMSO test solution.
C3 mg solid waste/pL of DHSO test solution.
300 119 solid waste/uL of DHSO test solution.
eT • toxic.
-------
TABLE 4.20. RESULTS Of HUTAGENICITY TESTING OF WASTE 10 EXTRACTS
VO
Volune
OMSO te
solutli
(uL/plat
»n
tej EP*
Revertants/plate
TA 98
Citrate Ma-
Water* buffer* resin*
Colum*
Soxhlet
Sequential extraction Citrate
b Acid0 Basec Neutral" EP* Water* buffer*
Pnenobarbital
75
SO
25
10
0
Te
T
T
44
17
T
T
58
41
52
T
T
T
45
22
T
T
35
33
16
T
T
94
78
42
T
93
94
78
22
T
609
556
97
39
T
T
T
T
22
T
60
61
52
42
activation
T I
T T
T T
156 T
109 1 10
T
T
T
T
154
Na-
resln*
T
T
T
117
111
TA 100
Colum*
T
T
T
149
1)2
Soxhlet"
r
T
T
T
199
Sequential extraction
Acid0 Basec Neutral"
T
670
645
536
167
r
T
T
T
91
T
T
T
T
106
Aroclpr activation
75
50
25
10
0
T
T
T
31
13
T
T
T
42
43
T
T
T
T
28
T
T
119
4B
8
T
T
70
70
31
T
T
156
202
22
T
T
T
T
48
T
T
T
54
42
T
T
T
T
35
T T
T T
T T
T T
96 91
T
T
T
T
167
T
T
T
108
94
T
T
T
117
108
T
T
T
T
192
T
T
T
229
126
T
T
T
I
153
T
T
T
T
123
No activation
50
0
T
7
T
42
T
58
T
17
T
42
T
21
T
40
T
27
T
44
T 109
94 76
T
110
T
71
T
111
ISO
105
T
159
T
153
T
99
*250 uL of aqueous extract/id, of ONSO test solution.
bl Big solid Naste/ut. of ONSO test solution.
C3 ng soltd naste/uL of DMSO test solution.
d300 ug solid waste/uL of DHSO test solution.
eT • toxic.
-------
TABLE 4.21. RESULTS OF HUTAGENICITY TESTING OF WASTE 11 EXTRACTS
00
o
Revertants/Plate
Cone.
(ul/plate) EPa
Water
TA 98
Citrate Na-
a buffer resin" Column2 Soxhlet
Sequential extraction
" Acid0
Basec
Neutrald EPa
Citrate
Water" buffer9
Na-
resina
TA 100
Column9
Soxhletb
Sequential extraction
Acidc Base0 Neutrald
Phenobarbltal activation
7.5
S.O
2.5
1.0
0
110
92
46
30
19
39
52
45
27
16
71
57
54
49
42
Te
45
46
70
22
T
T
17
21
20
T
T
T
13
27
17
17
17
33
26
232
161
57
18
21
T
T
T
28
21
T
T
117
93
85
T
71
136
119
80
163
213
169
152
150
119
109
125
123
106
T
T
T
54
62
T
T
T
T
113
T
120
109
138
107
205
160
176
105
104
T
T
71
Aroclor activation
7.5
5.0
2.5
1.0
0
S.O
0
98
59
53
29
20
7
4
74
52
27
29
18
9
7
80
86
48
39
40
42
30
T
T
110
97
29
27
22
T
T
14
29
26
T
18
T
T
T
14
22
T
3
26
46
20
34
20
T
15
125
68
35
19
41
21
15
T
T
T
T
15
No
T
8
T
T
T
78
65
activation
T
40
T
T
78
79
74
95
30
156
165
214
176
135
120
95
151
109
182
143
110
T
85
T
T
58
48
73
T
46
T
T
T
T
45
T
122
T
T
119
140
94
T
65
T
T
T
T
202
T
90
T
T
T
T
47
T
116
a2SO pL of aqueous extract/luL of DHSO test solution.
bl mg solid waste/uL of OMSO test solution.
C3 mg solid vaste/uL of DNSO test solution.
d300 ug solid waste/uL of OMSO test solution.
eT • toxic.
-------
TABLE 4.22. SUMMARY OF MUTAGENIC ACTIVITY OF SOLID UASTE
ISOLATES: AQUEOUS EXTRACTS AND ORGANIC ISOLATES*
Aqueous extracts
Organic isolates
Sequential
extraction
Waste
1
2
3
4
5
6
9
10
11
Q.
LU
T
NE
T
NE
NE
T
NE
T
M
s_
V
•»->
£
M
NE
M
NE
NE
T
NE
T
M
-------
isolate, as well as the acid and neutral fractions from the sequential
extraction, also gave a mutagenic response in TA 98. The Soxhlet
isolate and the acid fraction were direct mutagens, whereas the
remainder of the mutagenic extracts of this waste required metabolic
activation.
Waste 2 displayed mutagenic activity in the citrate buffer and
Na-resin extracts (Table 4.14). The positive response was confined to
TA 98 and required metabolic activation. The column extract and
Soxhlet isolate and the acid fraction from sequential extraction were
toxic, particularly toward TA 100.
Mutagenic activity detected in waste 3 was restricted to the water
extract (Table 4.15). Significant activity in TA 98 was noted with the
phenobarbital-induced activation system, as well as in the absence of
metabolic activation. Results obtained with TA 98 (Aroclor activation)
and with TA 100 (with and without activation) treated with water extract
support the positive observation noted with TA98/phenobarbital
activated. No other extract or isolate elicited a mutagenic response.
As noted above, no mutagenic activity was observed with any of the
isolate or extracts from waste 4 (Table 4.16). Toxicity was seen,
however, with the Na-resin and column extracts, Soxhlet isolate, and
the acid fraction from sequential extraction, which made mutagenicity
impossible to detect.
Two samples from waste 5 gave a positive response with TA 98: the
citrate buffer extract (with Aroclor activation) and the base fraction
from sequential extraction (with phenobarbital activation) (Table 4.17).
No other activities were noted; waste 5 extracts and isolates were
essentially nontoxic in these assays.
82
-------
No mutagenic response was observed with extracts and Isolates of
waste 6 (Table 4.18). Significant cytotoxicity was noted with the EP,
water, citrate buffer extracts, and the acid fraction. The neutral
fraction from sequential extraction was not available for testing.
Only one extract or isolate of waste 9 (the neutral fraction from
sequential extraction) gave a positive mutagenic response
(Table 4.19). This isolate was mutagenic in the presence of metabolic
activation (either microsome preparation) in strain TA 98. The Soxhlet
isolate and acid fraction were toxic.
Several samples from waste 10 were genetically active
(Table 4.20). The Na-resin extract was mutagenic in TA 98 with Aroclor
activation, while the Soxhlet isolate and column extract were active in
TA 98 with phenobarbital activation. The data suggest that either
activation system will accommodate the mutagenic agent(s) in these
samples. A striking response was obtained with the acid fraction from
sequential extraction. This fraction was mutagenic (with phenobarbital
activation) in TA 98 and TA 100. All of the extracts and isolates from
this waste were toxic at high dose levels.
The samples of the final waste assayed, waste 11, were
sufficiently toxic that the dose range was lowered tenfold to bring
survival up to a reasonable level for a meaningful assay (Table 4.21).
The EP, water, Na-resin extracts, and the base fraction gave positive
mutagenic responses in TA 98 with metabolic activation. The column
extract, Soxhlet isolate, acid fraction, and neutral fraction were
toxic. Even at an additional 10-fold reductions, the column extract
was still toxic at the higher dose levels.
83
-------
In summary, seven of the nine wastes possessed detectable
mutagenic activity in one or more extracts or isolates (Table 4.22).
It is interesting to note that the organic isolates produced by
methylene chloride Soxhlet extraction and the three-step sequential
extraction procedure showed mutagenic activity in two and five of the
wastes tested, respectively. On the other hand, six of the nine wastes
were found to possess mutagenic activity by using the less aggressive
aqueous extracting procedures. The Na-resin extracts accounted for
four of the seven positives. Water and citrate buffer revealed three
of the wastes (but not the same three) to contain mutagenic activity
while the EP and the column (which extracted the greatest quantities of
chromatographable organic compounds) procedures revealed only one of
the eleven wastes to contain mutagenic activity (again not the same
waste). These data demonstrate the difficulties in establishing a
single procedure or media that will be universally successful (with a
broad variety of wastes) in providing an extract that allows
unambiguous testing of mutagenicity. The sequential extraction
procedures has the advantage of separating active components into
separate fractions. In this way the confounding effects of
cytotoxicity and mutagenicity are frequently resolved and information
about the nature of the mutagen is obtained.
4.1.5 Filtration Study
One of the major objectives of the filtration study was to
determine if the filtration of batch extracts (through 0.4-pm pore
size membrane filters) was responsible for the large differences in
concentration of organic compounds noted between the batch and column
84
-------
extraction procedures (the column extracts were no* filtered using the
0.4-um pore size membrane filters). To do this, a series of
experiments were conducted as outlined In Section 3.2.3.
Small differences In amounts of organic compounds were found In
the column extracts produced from experiments 1 and 2 (Table 4.23). In
this experiment, the extracts were passed directly through XAD-2 resin,
with and without an in-line glass fiber filter. These data suggest
that entrained organic particles that can pass through the PTFE cloth
i.e., < 10 urn, but would be retained by a glass-fiber filter i.e.,
> 0.7 ym, either do not represent a significant input to the XAD-2
resin or pass through it without being adsorbed with the waste tested
(waste 14).
A column extract was also subjected to four separation treatments
(Table 4.24). In this case, the glass fiber filter retained less
organic material, judged by analyzing GC-elutable compounds in the
filtrate, than did any of the other treatments. Compounds such as
naphthol, phenanthrene, and benzo(a)anthracene were not detected in
the extract filtered through the Millipore filter, nor was
benzo(a)anthracene detected in the extract filtered through the
Nuclepore filter. Thus, Millipore filters appear to be more sorptive
for some classes of nonpolar organic compounds than do the Nuclepore
filters. Sorption in this case appears to be related to filter type
(material) rather than nominal pore size.
A comparison of the quantities of organic compounds found in the
glass-fiber-filtered extracts collected directly from the column on
XAD-2 resin (Table 4.23, approximately 25,500 ug/100 g) versus those
85
-------
TABLE 4.23. ORGANIC COMPOUNDS FOUND IN
COLUMN EXTRACTS, WITH AND WITHOUT
AN IN-LINE FILTER3
Compound
Without
filter
With
filterb
Naphthalene
Quinoline
Naphthol
Dibenzothiophene
Phenanthrene
Benzo(a)anthracene
Totaic
- ug/100 g waste
945 1275
50 305
70 80
30 50
40 65
300 155
21,900 25,500
aUsing waste sample 14, extracts collected directly
from column on XAD-2 resin.
bGlass fiber filter (precombusted, Whatman GF/F,
0.7 urn) placed on top of column.
cTotal = all elutable compounds using GC.
86
-------
TABLE 4.24. ORGANIC COMPOUNDS REMAINING IN A COLUMN EXTRACT
AFTER USING FOUR SOLID/LIQUID SEPARATION TREATMENTS3
Compound
£-cresol
Naphthalene
Qu incline
Naphthol
Dibenzothiophene
Phenanthrene
Carbazole
Benzo( a) anthracene
Totald
Glass fiber
(0.7-um)
9648
5052
1376
392
336
400
104
208
91,200
Filtered
Nuclepore
(0.4-um)
- - - no/inn
1948
2756
412
80
64
76
76
nd
66,320
Mi Hi pore
(0.45-ym)
1632
2228
428
ndc
68
nd
168
nd
68,200
Centrifugedb
1496
2108
400
72
28
44
40
nd
60,640
Using waste 14.
3Centrifuged in open 250-mL glass bottles, 400 times gravity for 1 h.
"nd - not detected.
Total = all elutable compounds using GC.
87
-------
collected from the column and Isolated using a solvent partition
technique (Table 4.24, ca. 60,000 to 90,000 ug/100 g), suggests that
the in-line XAD-2 resin cartridge (16.8 ml) may not be the most
effective means of collecting organic compounds. For example, for the
waste tested (No. 14) approximately three to four times more elutable
compounds were detected using the solvent partition method to isolate
and concentrate organic compounds from the aqueous effluent. The
treatment of the internal standard (azulene) was different for these
two extracts (in the case of the column/XAD-2 system, the internal
standard was added after resin extraction; with the solvent extraction
system the azulene was added to the aqueous phase prior to
extraction). However, in view of the high recoveries (> 75%)
normally obtained for azulene, it is unlikely that the large
differences in organic chemical levels noted between the two isolation
techniques can be attributed to differences in the handling of the
internal standard.
Amounts of organic compounds observed among the batch extracts
after the four separation treatments can be found in Table 4.25.
Phenanthrene was again not detected in any of the extracts filtered
through the Millipore filters. It is interesting to note that
benzo(a)anthracene was not found in any of the batch extracts,
regardless of the filter type used, while it was observed in the
glass-fiber-filtered column extract (see Table 4.24). In all cases,
where extracts were filtered through glass fiber filters, the column
extract contained greater quantities of organic compounds than did the
extracts from the batch procedures.
88
-------
TABLE 4.25. ORGANIC COMPOUNDS REMAINING IN A BATCH WATER EXTRACT
AFTER USING FOUR SOLID/LIQUID SEPARATION TREATMENTS3
Compound
£-cresol
Naphthalene
Qu incline
Naphthol
Dibenzothiophene
Phenanthrene
Carbazole
Benzo(a) anthracene
Total d
F
Glass fiber
(0.7-um)
3056
4564
100
96
60
168
64
nd
43,240
Mltered
Nuclepore
(0.4-um)
_ iin/lflf
4380
3032
100
80
48
60
56
nd
45,280
Millipore
(0.45-un)
3376
4784
108
60
36
nd
48
nd
46,000
Centrifugedb
3856
4520
104
132
64
168
64
nd
44,400
aUsing waste 14.
bCentrifuged in open 250-mL glass bottles, 400 times gravity for 1 h.
cnd = not detected.
Total = all elutable compounds using GC.
89
-------
When £ standard solution containing known concentrations of
selected organic compounds was subjected to the different solid/liquid
separation treatments, differing amounts of organic compounds were also
found (Table 4.26). In general, the glass fiber filter exhibited lower
sorption properties. Again, certain organic compounds
[dibenzothiophene, phenanthrene, and benzo(a)pyrene] were not found in
the Millipore-filtered sample from any extract (<1 yg/L). Additional
losses of organic compounds were seen when the standard solution was
agitated for 24 h (Table 4.27). This loss is probably due to sorption
of the compounds on the glass container walls. This may also explain
the loss of organics in the centrifuged samples (i.e., samples were
centrifuged in glass bottles). Volatilization may also have occurred
in the centrifuged samples.
4.2 Task 2: Comparison of Two Sample Preparation Protocols for
Performing the Ames Test on Solid Waste Extracts and
Wastewaters
The results of the recovery study comparing isolation of two
organic compounds [benzo(a)pyrene (BaP) and 9-amino-acridine] from
extracts and wastewaters using the XAD resin adsorption and solvent
partition techniques are shown in Table 4.28. The BaP recovery values
are somewhat confounded by the fact that the added BaP was well in
excess of its water solubility (1 mg/L added relative to a water
solubility of below 10 yg/L). Thus, the recovery data for solvent
partition showed an unexpected distribution of BAP into the acid
fraction, implying that three extractions with methylene chloride were
not sufficient to remove all of the BAP into the base/neutral
90
-------
TABLE 4.26. ORGANIC COMPOUNDS REMAINING IN A STANDARD SOLUTION
AFTER USING FOUR TREATMENTS FOR SEPARATING SOLID
AND LIQUID PHASES3
Compound
Phenol
p_-cresol
Naphthalene
Qu incline
Naphthol
Dibenzothiophene
Phenanthrene
Carbazole
Fluoranthene
Benzo(a)anthracene
Glass fiber
(0.7-um)
7.8
8.6
5.8
6.3
6.6
9.2
9.7
11.6
7.4
4.4
Filtered
Nuclepore
(0.4-um)
7.8
8.8
1.3
6.7
7.7
9.7
11.5
2.0
8.8
3.8
Millipore
(0.45-um)
7.7
8.1
1.0
5.2
7.0
ndc
nd
1.3
nd
nd
Centrifugedb
• -
6.0
9.2
6.1
7.0
7.9
6.7
5.9
6.6
7.8
5.7
Approximately 10 ppm of each compound added to prepare standard
solution.
Centrifuged in open 250-mL glass bottles, 400 times gravity for 1 h.
cnd = not detected at 1 mg/L.
91
-------
TABLE 4.27. ORGANIC COMPOUNDS REMAINING IN A STANDARD SOLUTION
(AGITATED FOR 24 HOURS) AFTER USING FOUR SOLID/LIQUID PHASE
SEPARATION TREATMENTS*
Compound
Phenol
£-cresol
Naphthalene
Qu incline
Naphthol
Dibenzothiophene
Phenanthrene
Carbazole
Fluoranthene
Benzo(a)anthracene
Glass fiber
(0.7-um)
7.2
8.1
5.2
5.3
6.3
4.6
4.7
4.8
3.4
2.1
Filtered
Nuclepore
(0.4-um)
5.9
8.3
2.9
4.9
7.4
4.3
4.3
4.5
2.6
1.8
Millipore
(0.45-um)
4.2
6.7
3.2
5.4
5.3
ndc
nd
1.8
nd
nd
Centrifugedb
4.3
8.2
3.4
7.0.
8.2
4.8
5.1
5.5
4.6
3.2
Approximately 10 ppm of each compound added to prepared standard
solution; standard solution was agitated for 24 h.
Centrifuged in 250-mL glass bottles, 400 times gravity for 1 h.
cnd - not detected at 1 mg/L.
92
-------
TABLE 4.28. COMPARISON OF TWO ISOLATION TECHNIQUES FOR REMOVING
ORGANIC MUTAGENS FROM WASTEWATERS AND SOLID WASTE EXTRACTS
XAD-2 resin
Aqueous medium
Water
EPC
Leachate
Wastewater6
Water
EPC
Leachate
Wastewater6
Base/
neutral
77.0
87.5
85.0
83.0
78.0
83.5
78.5
85.5
Acid
1.5
3.0
1.5
4.5
0.0
2.0
1.0
0.0
% recovery3
Solvent partition
Base/
Total neutral
78.5 50.0
90.5 55.5
86.5 27.5
87.5 83.0
9 -ami no acridine^ -
78.0 93.5
85.5 72.5
79.5 74.0
85.5 96.5
Acid
22.0
24.5
7.5
7.5
0.0
0.0
0.0
0.0
Total
72.0
80.0
35.0
90.5
93.5
72.5
74.0
96.5
al mg/L of mutagen added, average of two replicates.
bRecovery measured using both HPLC and scintillation counting
(results agreed +_2%).
cSolid waste extract using the EPA-EP with waste 10.
^A waste leachate collected at a landfill containing waste 3.
eCoal conversion wastewater.
^Recovery measured using fluorescence spectrophotometry.
93
-------
fraction. However, for the soluble 9-amino-acridine, the recovery
distribution was excellent.
Using Duncan's multiple range test, the only significant
difference (P < 0.05) in recovery of the BaP or 9-amino-acridine
between the two isolation techniques was noted with BaP in landfill
leachate. Significant precipitation and subsequent emulsion formation
took place using the solvent partition technique, which could account
for the difference noted. There were also some significant differences
(P < 0.05) in the recovery of BaP and 9-amino-acridine by the solvent
partition technique, depending on the type of aqueous medium spiked
with these chemicals. For example, in addition to the very low
recovery of BaP in the landfill leachate (35%), the recovery of BaP
from the water and EP extract (72 and 80% recovery, respectively) was
significantly lower than its recovery by solvent partition from coal
conversion wastewater (90.5% recovery). The nature of the aqueous
medium also affected the recovery of 9-amino-acridine. In this case,
the recovery from the EP extract and landfill leachate (72.5 and 74.0%,
respectively) was significantly (P < 0.05) lower than that recovered
from the water or coal conversion wastewater (93.5 and 96.5%,
respectively). In contrast, there were no observed differences in the
recovery of either BaP (range 77.0% to 90.5%) or 9-amino-acridine
(78.0% to 85.5%) among the aqueous media using the XAD-2 resin
technique. These data suggest that the XAD-2 resin technique is
preferable to solvent partition if a wide range of aqueous media are to
be investigated.
94
-------
Mutagenicity testing results of the positive controls and the
organic isolates from the recovery study are shown in Tables 4.29
to 4.31. The mutagenic activity was directly related to the chemical
distribution of benzo(a)pyrene and 9-amino-acridine. There was no
indication of interference from other organic species present in the
extracts.
Aqueous media spiked with 9-amino-acridine were directly mutagenic
when tested with TA1537. When assayed with metabolic activation
applied, activity was also seen with TA1538, TA98, and TA100.
The benzo(a)pyrene-spiked extracts showed no mutagenic activity
with any strain in the absence of metabolic activation. Strong
positive responses were observed in TA1537, TA1538, TA98, and TA100
when metabolic activation was applied. Qualitatively, the mutagenic
activity of the spiked extracts was identical to the results obtained
with the positive control samples of 9-amino acridine and
benzo(a)pyrene.
In summary, there was no major difference in the ability of the
two techniques to recover BaP or 9-amino-acridine from wastewaters.
The single observed difference was the very poor recovery of BaP in the
landfill leachate by the solvent partition technique (35% recovery
compared to 86.5% recovery by XAD-2). However, in the solvent
partition technique, recovery appeared to be dependent on the aqueous
medium. Recovery by the XAD-2 technique, on the other hand, was
observed to be independent of the aqueous matrix. Furthermore, the
resin isolation procedure (XAD-2 resin) is superior, from a practical
standpoint, for the following reasons:
95
-------
TABLE 4.29. MUTAGENIC ACTIVITY OF THE POSITIVE CONTROLS
BENZO(A)PYRENE AND 9-AMINO ACRIDINE
Compound
Benzo(a)pyrene (25 ng/uL)
With activation
Without activation
9-amino acridine (25 ng/yL)
With activation
Without activation
Strain
TAT 535 TA1537 TAT 538 TA98
0 9.5 13.1 7.4
0000
0 1.4 2.0 1.7
0 3.1 0 0
TA100
4.2
0
0.9
0
96
-------
TABLE 4.30. MUTAGENIC ACTIVITY IN ORGANIC ISOLATES: COMPARISON
OF RECOVERY TECHNIQUES USING BENZO(A)PYRENE
Aqueous medium
Water
EP
Leachate
Wastewater
Resin
Base/neutral
TAT 537
7.0
8.1
6.9
7.4
Revertants/uLa
Solvent partition
Acid Base/neutral
, Aroclor activation
0 4.0
0 5.6
0 4.7
0 4.4
Acid
0.3b
1.2
1.4
1.3
TA1538, Aroclor activation
Water
EP
Leachate
Wastewater
11.5
11.5
10.2
11.2
0 4.5
0 5.4
0 4.6
0 4.7
3.8b
3.0
2.6
2.2
TA98, Aroclor activation
Water
EP
Leachate
Wastewater
Water
EP
Leachate
Wastewater
8.0
8.7
8.1
8.9
TA100.
5.7
4.6
8.2
3.8
0 5.4
0 6.4
0 6.9
0 7.3
Aroclor activation
0 3.8
0 2.9
0 5.0
0 3.5
1.5"
1.9
2.6
2.4
0.8b
1.8
2.1
1.6
aAverage of two replicates.
bSingle determination.
97
-------
TABLE 4.31. MUTAGENIC ACTIVITY IN ORGANIC ISOLATES: COMPARISON
OF RECOVERY TECHNIQUES USING 9-AMINO ACRIDINE
Revertants/uL3
Aqueous medium
Resin
Base/neutral
Solvent partition
Acid Base/neutral
Acid
TA1537, No activation
Water
EP
Leachate
Wastewater
Water
EP
Leachate
Wastewater
Water
EP
Leachate
Wastewater
Water
EP
Leachate
Wastewater
1.7
2.2
2.7
1.6
TA1538,
0.7
0.8
1.3
1.1
TA98,
0.8
0.9
1.4
1.2
TA100,
1.0
1.3
1.2
0.9
0
0
0
0
Aroclor
0
0
0
0
Aroclor
0
0
0
0
Aroclor
0
0
0
0
2.4
2.4
2.1
2.2
activation
0.9
1.2
1.1
0.8
activation
1.7
0.5
1.0
0.9
activation
1.4
0.6
1.7
0.9
0
0
0
0
0
0
Ob
0
0
0
Ob
0
0
0
0
0
aAverage of two replicates.
bSingle determination.
98
-------
(1) It is technically simpler, and a series of XAD columns can be
used to facilitate handling large numbers of samples.
(2) There are no problems with emulsion formation. Emulsions may
pose severe problems in solvent partition techniques with
"dirty" water samples, e.g. the real-world landfill leachate.
(3) It is more economical; much less solvent is consumed.
However, the resin procedure has limitations. If significant
precipitation occurs during pH adjustment, flow of water through the
column can be impeded, increasing extraction time. Also batch-to-batch
variability in the resin, as well as contamination, requires that
extensive clean-up of the resin is necessary to ensure reproducibility.
4.3 Task 3; An Evaluation of the Equivalence of Magnetically Stirred
Extractor Relative to an EPA-Approved Rotary Extractor
for Conducting the EP
A magnetically stirred extractor and a rotary extractor were
compared for conducting the EP. Using the low frequency tachometer
(see Section 3.2.5), the stirring rates for the magnetically stirred
extractor were recorded every hour during the 24-h extraction for
wastes 12 and 13. Figures 4.10 and 4.11 present the mean speed and the
standard deviation found for the four replicate EP extractions during
the extraction period. Waste 12 was a very hard granular solid, and
427 rpm represented the lowest mean rate (over the 24 h) required to
keep the solids suspended. Waste 13 was a powdery solid, high in
magnetite, which was expected to load the magnetic stirrer. Even so, a
lower mean 24-h stirring rate of 342 rpm was obtained. In both cases,
the stirring speed gradually increased for 1 to 2 h after adding the
solids to the extractor. The stirring rate was reproducible between
99
-------
ORNL-DWG 81-21532 ESD
700
600
500
,>••••{
-«~l f
200
100
-------
ORNL-DWG 81-21533 ESD
Q.
Q
ill
LU
cc
tt
CC
700
600
500
400
300
200
100
12 16
TIME (h)
20
24
Fig. 4.11. High and low stirrer speed values for waste 13 - mean and
standard deviation values for four replicate extractions
over the 24-h extraction time.
101
-------
the four replicate runs of each 24-h extraction; for example, the
coefficients of variation (calculated each hour for the four
extractions) for wastes 12 and 13 were 6 and 3% for the highest speed
and 12 and 5% for the lowest speed, respectively.
The percent variation associated with the variables of agitation
method, replicate extractions, and analytical procedure was determined
using variance component analysis (Table 4.32). The major source of
variation was associated with the agitation method (39-95%), although
the amount differed depending on the waste and element. The amount of
variation associated with extraction replicates and analytical
procedure also differed according to waste and element, ranging from
5.0 to 39.1% for extraction replicates and 0.3 to 22.0% for analytical
procedure.
Mean inorganic concentrations for wastes 12 and 13 and statistical
differences found between agitation methods are shown in Table 4.33.
In all cases, the magnetic stirrer agitation method produced
statistically (P < 0.05) different inorganic concentrations than the
rotary extractor. Neither method showed consistently higher
concentrations. For waste 12, the rotary extraction method extracted
greater amounts of Cd, while Fe and Ni concentrations were higher using
the magnetic stirrer at either stirring rate. For waste 13,
concentrations of As were highest using the rotary extractor; Cd
concentrations were highest using the magnetic stirrer, and Zn values
were lowest using the high-speed magnetic stirrer. It is interesting
to note that concentrations of a more volatile element such as As are
considerably reduced when using the open magnetic stirrer system versus
102
-------
TABLE 4.32. DISTRIBUTION OF VARIATION ASSOCIATED WITH
THE VARIABLES OF AGITATION METHOD, REPLICATE EXTRACTION,
AND ANALYTICAL PROCEDURE3
Sample
12
13
Variable
Agitation method
Agitation replicates
Analytical procedure
Agitation method
Agitation replicates
Analytical procedure
_Cd_
38.9
39.1
22.0
As_
94.7
5.0
0.3
Element
%_ _
_F_e__
78.7
17.1
4.2
_Cd_
59.4
26.7
13.9
Ni
71.9
10.9
17.2
In
71.9
6.0
22.1
aDetermined using variance component analysis (SAS Institute, Inc.,
1979).
103
-------
TABLE 4.33. MEAN (x) CONCENTRATIONS AND STATISTICAL DIFFERENCES
OF SELECTED INORGANIC COMPOUNDS USING A MAGNETIC STIRRER
(2 AGITATION RATES) AND A ROTARY EXTRACTOR3
Waste
12
13
Agitation
Magnetic stirrer - high speed
Magnetic stirrer - low speed
Rotary extractor
Magnetic stirrer - high speed
Magnetic stirrer - low speed
Rotary extractor
Cd
66*
70+
76
As
99*
343+
548
Element
iin/l
fe_
36+
47
22*
Ci
91 +
91 +
78*
Ni
207 7+
2042+
1777*
_Zn
564*
1084
880"""
aValues within a sample for a particular element that have different
superscripts are significantly different (P < 0.05), as determined
using Duncan's multiple range test (SAS Institute, Inc., 1979).
104
-------
the closed rotary system. This is also supported by the fact that
high-speed stirring produced lower As concentrations than did the
slower stirring speed. Concentrations of the other elements
investigated also differed between the high- and low-speed stirring
rates; where statistical differences were found (for Cd, Fe, Zn), the
high-speed magnetic stirrer yielded lower amounts.
In summary, differences can be expected in inorganic contaminant
concentrations when using a magnetic stirrer extractor versus rotary
extractor. Neither method will yield consistently higher
concentrations than the other. However, because the concentrations of
elements in these samples were very low (ppb-range), it is recommended
that additional samples containing higher levels of extractable
constituents be studied. If volatile constituents are of interest, the
closed rotary extractor may be the preferred method, although this
system does not allow for automatic pH adjustment.
4.4 Task 4: An Evaluation of the Reverse-Phase High-Pressure Liquid
Chromatography (HPLC) Protocol for Assessing the
Bioaccumulation Potential of Solid Waste Extracts
Nine wastes were tested for bioaccumulative materials using the
bioaccumulation potential test as described in Section 3.3.4. The
materials tested were the organic isolates from the Soxhlet and
sequential extraction fractions, and the organic concentrates from the
five aqueous extraction procedures on the nine wastes. Table 4.34
illustrates data collected in terms of the number of chromatographic
peaks found with retention times equal to or greater than log P = 3.0
as well as the total peak area with respect to bromobenzene. All
wastes, with the exception of wastes 1 and 3 were found to contain
105
-------
TABLE 4.34. BIOACCUMULATION DATA ON SOLID WASTE ISOLATES:
AQUEOUS EXTRACTS AND ORGANIC ISOLATES*
Aqueous extracts
Waste
1
2
3
4
5
6
7
8
9
Q.
0
0
0
0
0
0
ned
0
0
i.
Ol
-u
ID
0
0
0
0
0
0
0
0
0
c
in
ia
_ _ _ M,,n
0
0
0
0
0
0
0
3/12.3
0
t-
-------
bioaccumulative materials, as Identified in the Soxhlet isolates and
sequential extraction fractions, but none of these materials were
observed in the organic concentrates from the aqueous batch extraction
tests. An exception was waste 8 which contained by far the highest
levels of these components. The low quantities of bioaccumulative
compounds observed in the batch extracts are probably due in large
measure to the low water solubility of the bioaccumulative compounds
which promotes their loss by adsorption on the membrane filters
utilized in the batch extractions.
Considering the highly lipiphilic nature of bioaccumulative
materials, it is interesting to note that significant levels of these
materials were isolated in the column extracts. In almost all cases,
the materials found to be bioaccumulative in the solid wastes (by
Soxhlet and sequential extraction) were also found in the column
extracts. Among the aqueous extraction procedures, only the column
procedure is effective for the detection of bioaccumulative materials,
using the current methodology.
The bioaccumulation potential test itself has several drawbacks.
No UV detector is not universal (therefore, nonabsorbing materials such
as chlorinated aliphatic compounds may be missed) nor is the response
of the UV detector uniform (highly absorbing materials may be given too
high a ranking). Therefore, information obtained is essentially
qualitative. However, the test does provide a useful screening method
for the detection of potential bioaccumulative organic compounds in the
wastes as well as in the extracts.
107
-------
SECTION 5. CONCLUSIONS
5.1 Task 1; An Evaluation of Extraction Procedures to Remove
Nonpolar Organic Compounds from Solid Wastes
The primary objective of Task 1 was to assess the capabilities of
five selected extraction procedures to remove organic compounds from
11 solid wastes. Other objectives included mutagenicity testing of the
extracts and examining the solid wastes for organic compounds (Soxhlet
and sequential extraction) and the solid waste extracts for selected
inorganic constituents. The five extraction procedures used were four
batch extractions and an upward flow column extraction. The following
extracting media were used:
(1) Batch 1: EP, acetic acid, pH 5.0,
(2) Batch 2: deionized distilled water,
(3) Batch 3: deionized distilled water with a sodium
displacement resin, and
(4) Batch 4: 0.5 M citrate buffer,
(5) Column: deionized distilled water.
The following conclusions are based on data presented for Task 1:
• For the four batch extractions, the citrate buffer extracts
consistently showed highest concentrations of inorganic
compounds.
• Of the two methods compared for identification of organics in
the waste samples (Soxhlet extraction with methylene chloride
and a three-step sequential extraction procedure developed at
ORNL), the three-step sequential extraction scheme consistently
extracted more organic material than did the Soxhlet extraction.
108
-------
No significant differences in organic concentrations (based on
DOC levels) were found between water and Na-resin extracts.
No significant differences were found in DOC values between
glass fiber and Nuclepore-filtered extracts from the water and
Na-resin extracts.
Total chromatographable organic (TCO) content data for the
five extraction procedures revealed that the column procedure
extracted more organic material than did any of the batch
extraction procedures.
When the five extraction procedures were ranked from lowest to
highest (1 to 5) for the ability to extract total
chromatographable organics (TCO), the column procedure
produced extracts higher in TCO and selected organic compounds
than batch extracts (P < 0.05).
Of the four batch extraction procedures, the water procedure
extracted the highest levels of TCO and selected organic
compounds (P < 0.05).
No significant differences in the ability to extract organic
compounds were observed among the EP, Na-resin, citrate buffer
extractions.
In general, the column leaching procedure was the most
aggressive means of extracting organic compounds from solid
wastes. The most noticeable differences between the column
procedure and the batch procedures were the levels of
moderately volatile compounds and the nonpolar compounds found
in the extracts.
109
-------
The five extraction procedure extracts, as well as the Soxhlet
solid waste Isolate and the solid waste Isolates produced from
the sequential procedure, were compatible with the
Salmonella/microsome assay.
Of the nine wastes examined, seven yielded organic Isolates
that possessed detectable mutagenic activity. The
Salmonel1 a/microsome mutagenicity assay performance comparison
with the nine wastes Indicated that, of the five aqueous
extraction procedures, the Na-resin extraction was the most
effective extraction, revealing four of the mutagenic
activities detected.
Six mutagenic waste components were identified (from five
wastes) by the sequential extraction method. This method has
the great advantage of (1) indicating the class fractions
harboring the active components and (2) revealing multiple
mutagenic components.
Only one of the nine EP extracts examined exhibited mutagenic
activity.
The coefficient of variation associated with replicating the
column extraction procedure was 13%.
The use of glass fiber filters for the separation of solid and
aqueous phases allows higher recovery of organic compounds
than filtering with Nuclepore and Millipore filters.
For a single waste, the column extraction procedure extracted
greater quantities of total chromatographable organic
compounds than a closed batch extraction, even when the
extracts were filtered through the same filter type.
110
-------
5.?. Task 2: Comparison of Two Sample Preparation Protocols for
Performing the Ames Test on Solid Waste Extracts and
Wastewaters
The objective of Task 2 was to compare two techniques for the
isolation of organic mutagens for performing the Ames test on solid
waste extracts and wastewaters. The two recovery techniques were a
resin absorption technique using Amberlite XAD-2 resin and a solvent
partition technique using methylene chloride. Two mutagens were
examined, benzo(a)pyrene and 9-amino acridine. The following was
concluded:
There were no major differences.in the ability of the two
techniques to recover BaP or 9-amino acridine from wastewater.
One exception was the very poor recovery of BaP in the
landfill leachate by the solvent partition technique.
The recovery of BaP and 9-amino acridine was dependent on the
aqueous matrix using the solvent partition technique.
Recovery of the mutagens by the XAD-2 technique was
independent of the aqueous matrix.
The various aqueous matrices employed did not appear to modify
the amount of mutagenic activity attributable to
benzo(a)pyrene and 9-amino acridine.
From a laboratory performance standpoint, the resin isolation
procedure was considered superior.
The resin procedure, has its limitations. If significant
precipitation occurs during pH adjustment, flow of water
through the column XAD-2 cartridge can be impeded.
Ill
-------
5.3 Task 3; An Evaluation of the Equivalence of a Magnetically
Stirred Extractor Relative to an EPA-Approved Rotary
Extractor for Conducting the EP
The objective of Task 3 was to compare a magnetically stirred
extractor with an EPA-approved rotary extractor for conducting the EP.
The conclusions were as follows:
• The high and low stirring rates obtained using the
magnetically stirred extractor between replicate extractions
of two wastes showed low variability. Coefficient of
variation values for the two wastes were 6 and 3% for the high
speed and 12 and 5% for the low speed.
• The percent variation (determined using variance component
analysis) associated with the variables of agitation method,
replicate extractions, and analytical analysis differed
depending on the waste and metal. The major source of
variation, however, was associated with the agitation method,
ranging from 39 to 95%.
The magnetic stirrer agitation method produced statistically
different inorganic concentrations than the rotary extractor.
Neither method showed consistently higher concentrations.
If volatile constituents are of interest, the sealed rotary
extractor is the preferred method, although this system does
not allow for automatic pH adjustment.
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5.4 Task 4; Evaluation of the Proposed Reverse-Phase High-Pressure
Liquid Chromatography (HPi.C) Protocol for Assessing
Bioaccumulation Potential of Solid Waste Extracts
The objective of Task 4 was to evaluate the proposed reverse-phase
High-Pressure Liquid Chromatography (HPLC) protocol (EPA 1978) to
assess the bioaccumulation potential of solid waste extracts. Soxhlet
extracts, acid/base/neutral fractions from the sequential extraction
procedure, and the organic isolates from the aqueous extracts of the
five extraction procedures produced in Task 1 were tested for
bioaccumulative material. The following was concluded:
Of the five aqueous extraction procedures, only the column
extraction produced extracts that contained bioaccumulative
material.
A majority of the Soxhlet isolates and sequential extraction
procedure fractions of the solid wastes contained
bioaccumulative organics.
The bioaccumulation test itself has several drawbacks; the
UV detector is not universal (therefore, nonabsorbing
materials may be missed) and the response of the UV detector
is not uniform.
The present bioaccumulation protocol can provide a screening
method for the detection of potentially bioaccumulative
organic compounds in the wastes as well as in their leachates.
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SECTION 6. FUTURE RESEARCH
Further research should be conducted to determine if the up-flow
column extraction procedure Is superior to batch-mode extractions for a
variety of wastes and extracting media. It appears that one of the
major faults in the batch-mode extractions is the filtration step:
(1) it is very time consuming for some samples, and (2) the filtration
using 0.40- to 0.45-ym membrane filters appears to sorb significant
quantities of certain organic compounds. It was demonstrated that
up-flow column extracts not filtered through membrane filters contained
significantly greater quantities of total chromatographable organic
compounds than batch extractions which were filtered through 0.4-ym
pore size filters. When water was used as the common extracting medium
for a single waste in both the up-flow column and a batch-mode
extraction, the total quantity of elutable organic compounds by gas
chromatography was 1.4 to 2.1 times higher using the up-flow column.
The quantity of organic compounds varied according to the method of
liquid/solid phase separation and type of filter. Further experiments
should be conducted to confirm if this is typical for a variety of
wastes and possibly for different classes of extracting media.
Regardless of which type of extraction procedure is found to be
superior for extracting organic compounds (either up-flow column or
batch extraction), its relevance to simulating the leaching of organic
and inorganic constituents from wastes co-disposed in a municipal
landfill environment needs to be established. The present EP was
conceived as a first-order approximation to simulate leaching action of
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the low molecular weight carboxylic acids generated In an actively
decomposing municipal landfill. It uses acetic acid, the carboxylic
acid most prevalent In municipal waste landfill leachate. The acetic
acid, because of Its weak acidic character, plays a dominant role in
the leaching of metals; however, its role relative to the removal of
polar and nonpolar organic compounds from wastes is unclear. Acetic
acid also was not compatible with a variety of biotesting procedures
(Epler et al. 1980). Thus, the limitations of acetic acid as an
extracting medium were the impetus behind this past year's research.
Future research needs to (1) confirm the aggressiveness of the
up-flow column to remove organic and inorganic constituents from a
variety of wastes using selected leaching media, and (2) test the
effectiveness of the up-flow and batch-mode extractions to simulate
leaching by municipal waste leachate. The objective of this research
is to develop a second generation test for mobility, henceforth known
as EP-III, that will more accurately and reproducibly model leachate
production, for inorganic as well as organic constituents, in a
co-disposal municipal landfill. A workplan with these objectives is
appended (Appendix B).
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7. REFERENCES
Brazell, R. S., and M. P. Maskarinec. 1981. Dynamic Headspace
Analysis of Solid Waste Materials. J. High Resolut. Chromatogr.
Chromatogr. Comm. 4:404-405.
Brown, D. K., C. W. Francis, M. P. Maskarinec, and F. W. Larimer.
1981. Toxicity of Leachates, Comparison of Extraction Procedure
Extracts and Landfill Leachates. ORNL/TM-7563. Oak Ridge
National Laboratory, Oak Ridge, Tennessee.
Environmental Protection Agency (EPA). 1980a. Background Document.
Section 261.24 - EP Toxicity Characteristic. Office of Solid
Waste, Environmental Protection Agency, Washington, D.C.
Environmental Protection Agency (EPA). 1980b. Identification and
Listing of Hazardous Waste. _IN Environmental Protection Agency
Hazardous Waste Management System. 40 CFR 261.24.
Environmental Protection Agency (EPA). 1980c. Microbial Bioassay for
Toxic and Hazardous Material. EPA-330/9-80-002, Office of
Enforcement, EPA, Denver, Colorado.
Environmental Protection Agency (EPA). 1980d. Test Methods for
Evaluating Solid Waste, Physical/Chemical Methods. SW-846,
U.S. Environmental Protection Agency, Washington, D.C.
Environmental Protection Agency (EPA). 1979a. National Interim
Primary Drinking Water Regulations. 40 CFR 141.
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Environmental Protection Agency (EPA). 1979b. Methods for Chemical
Analysis of Water and Wastes. EPA-600/4-79-010, Environmental
Monitoring and Support Laboratory, Office of Research and
Development, U.S. Environmental Protection Agency, Cincinnati,
Ohio. 460 pp.
Environmental Protection Agency (EPA). 1978a. Hazardous Waste:
Proposed Guidelines and Regulations and Proposal on Identification
and Listing. Fed. Regist. 43:58966.
Environmental Protection Agency (EPA). 1978b. Identification and
Listing of Hazardous Waste. Advanced Notice of Proposed
Rulemaking. Fed. Regist. 43:59025-59026.
Epler, J. L., F. W. Larimer, T. K. Rao, E. M. Burnett, W. H. Griest,
M. R. Guerin, M. P. Maskarinec, 0. A. Brown, N. T. Edwards,
C. W. Gehrs, R. E. Millemann, B. R. Parkhurst, B. M. Ross-Todd,
D. S. Shriner, and H. W. Wilson, Jr. 1980. Toxicity of
Leachates. EPA-600/2-80-057, Office of Research and Development,
U.S. Environmental Protection Agency, Washington, D.C. 134 pp.
Feldman, C. 1974. Perchloric Acid Procedure for Wet-Ashing Organics
for the Determination of Mercury (and Other Metals). Anal. Chem.
46:1606-1609.
Feldman, C. 1979. Improvements in the Arsine Accumulation - Helium
Glow Detector Procedures for Determining Traces of Arsenic. Anal.
Chem. 51:664-669.
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Kam, R., M. A. Anderson, R. Stegman, and R. Stanforth. 1979.
Background Study on the Development of a Standard Leaching Test.
EPA-600/2-79-109, Office of Research and Development.
U.S. Environmental Protection Agency, Cincinnati, Ohio.
Maskarinec, M. P., and R. W. Harvey. 1982. Screening of Sludges and
Solid Wastes for Organic Compounds. Int. J. Environ. Anal. Chem.
11:53-60
McKown, M. M., J. S. Warner, R. M. Riggin, M. P. Miller,
R. E. Heffelfinger, B. C. Garrett, and G. A. Jungclaus. 1980.
Development of Methodology for the Analysis of Solid Wastes.
Draft Final Report to U.S. EPA, Washington, D.C.
SAS Institute, Inc. 1979. SAS User's Guide. Cary, North Carolina.
Schnitzer, M., and S. U. Kahn. 1972. Humic Substances in the
Environment. Chapter 2, pp. 9-27. IN Extraction, Fractionation,
and Publication of Humic Acids. Marcel Dekker, Inc., New York.
Shultz, F. J., and A. W. Spears. 1966. Determination of Moisture in
Total Particulate Matter. Tob. Sci. 10:75-76.
Talmi, Y., and A. W. Andren. 1974. Determination of Selenium in
Environmental Samples Using Gas Chromatography with a Microwave
Emission Spectrometric Detection System. Anal. Chem. 46:2122-2126.
Zlatkis, A., R. S. Braze!1, and C. Poole. 1981. The Role of Organic
Volatile Profiles in Clinical Diagnosis. Clin. Chem.
27(6):789-797.
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APPENDIX A
QUALITY ASSURANCE PROJECT PLAN
For
"Toxicity of Leachates Project"
Oak Ridge National Laboratory
Oak Ridge, Tennessee 37830
Revision No. 1
April 15, 1982
Interagency Agreements
DOE 40-1087-80
USEPA AD-89-F-1-058
APPROVAL:
ORNL Project Manager: Date:
C. W. Francis
ORNL QA Coordinator (ESD): Date:
M. H. Shanks
ORNL QA Coordinator (ACD): Date:
L. T. Corbin
EPA Project Officer: Date:
Llewellyn R. Williams
EPA QA Officer: Date:_
John Santolucito
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Table of Contents
Section Description
1.0 Project Description
2.0 Project Organization and
Pages Revisions Date
3 1 4-15-82
2 1 4-15-82
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0
11.0
12.0
13.0
Responsibility
QA Objectives in Terms of
Precision, Accuracy, Complete-
ness, Representativeness, and
Comparability
3.1 Organic Analyses
3.2 Inorganic Analyses
Sampling Procedures
Sample Custody
Calibration Procedures and
Frequency
Analytical Procedures 3
7.1 Extraction and Concentration
7.2 Analytical Chromatography
7.3 Gas Chromatography/Mass Spectrometry
7.4 Atomic Absorption (AA) Spectrometry
7.5 Inductively Coupled Plasma Atomic
Emission Spectrometry (ICP)
Data Analysis, Validation, and 6
Reporting
Internal Quality Control Checks 1
Performance and Systems Audits 1
Preventative Maintenance 1
Specific Procedures to be 2
Used to Routinely Assess
Data Precision, Accuracy, and
Completeness
Corrective Action 3
4-15-82
4-15-82
4-15-82
4-15-82
4-15-82
4-15-82
4-15-82
4-15-82
4-15-82
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Section Description Pages Revisions Date
14.0 Quality Assurance Reports to 5 1 4-15-82
Management
15.0 References 1 1 4-15-82
16.0 Distribution List 1
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1. Project Description
The objective of the research is to develop a laboratory extraction
test for solid wastes that: (1) models the leaching action a waste
would undergo when disposed of, along with a municipal waste, in a
municipal waste landfill following a 95/5 (weight fraction of municipal
and industrial waste) co-disposal scenario, (2) is compatible in aquatic
toxicity and phytotoxicity testing protocol, and (3) is relatively
inexpensive to conduct in terms of time, equipment, and personnel. The
extraction test will be henceforth referred to as EP-III.
The project involves research in both the laboratory and field.
The laboratory work is centered on evaluating the extraction conditions
that are most aggressive in removing inorganic and organic constituents
from four selected industrial wastes by a particular municipal waste
leachate (MWL). Municipal waste leachate has been obtained from a
lysimeter located at the U.S. Army Corps of Engineers Waterways
Experiment Station (WES), Vicksburg, Mississippi, for this work. The
extraction procedure (column aerobic, column anaerobic, or rotary
batch) found to be most aggressive will serve as baseline data to be
simulated by the laboratory method developed for EP-III. The
laboratory variables to be investigated in this phase of the work will
consist of the type of extraction procedure (up-flow column or batch
rotary) and type of extraction media (acetate buffer, CCL-saturated
water, distilled water, or synthetic leachate). The experiment will
consist of a factorial arrangement of 11 treatments and four industrial
wastes in a randomized block design with two blocks (time) per
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treatment-waste combination. The 11 treatments will consist of the
eight (procedure x media) laboratory methods plus the three WES/MWL
extractions.
The field work will test the ability of various laboratory
extraction methods to simulate the leaching characteristics of
industrial wastes in large-scale field lysimeter experiments containing
wastes in a 95% municipal to 5% industrial scenario and will evaluate
the aggressiveness relative to age of a municipal waste leachate to
extract contaminants from industrial wastes.
Municipal waste leachate from two large-scale lysimeters (1.8 m in
diameter and 3.6 m in height) will be diverted to 16 columns per
lysimeter containing four industrial wastes (the same wastes used in
the laboratory studies). For each lysimeter, one-half of the columns
(4 wastes x 2 replicates) will be used to compare the 95% municipal and
5% industrial waste scenario to the laboratory extraction studies. The
remaining columns will be used to evaluate the relative aggressiveness
of the municipal waste leachate as a function of age of the leachate.
The projected work schedule is presented in Table 1.
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TABLE 1. PROJECTED WORK SCHEDULE
Milestone Projected Date
1. Laboratory study
WES municipal leachate procured Jan. 27, 1982
Industrial wastes (8) procured May 15, 1982
Proximate analysis of wastes initiated May 15, 1982
Select sastes (4) for laboratory and
field studies May 31, 1982
Laboratory extractions started
(88 extractions) June 1, 1982
Laboratory extractions completed Sept. 1, 1982
Inorganic and organic analysis of
extracts completed Sept. 15, 1982
Statistical analyses completed Oct. 15, 1982
2. Field Study
Municipal refuse obtained and lysimeters packed Feb. 23, 1982
Industrial waste columns packed for laboratory
Comparison and aggressiveness of MWL, begin
leaching June 15, 1982
Laboratory comparison, leaching completed Aug. 10, 1982
Inorganic and organic analyses of leachates
from laboratory comparison completed Sept. 1, 1982
Aggressiveness of MWL, leaching completed Sept. 15, 1982
Inorganic and organic analyses of leachates
from aggressiveness of MWL completed Oct. 1, 1982
Statisical analyses completed, aggressiveness
of MWL Oct. 15, 1982
3. Draft report on laboratory and field studies Jan. 1, 1983
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2. Project Organization and Responsibility
The project directly involves personnel from two Divisions at
Oak Ridge National Laboratory (ORNL): (1) the Environmental Sciences
Division (ESD) and (2) the Analytical Chemistry Division (ACD). The
Environmental Sciences Division is responsible for conducting the solid
waste extractions, developing experimental design, performing
statistical analysis, and interpretating environmentally related data.
The Analytical Chemistry Division is responsible for inorganic and
organic analyses of the leachates and extracts. The project is managed
from the Environmental Sciences Division (Fig. A-l).
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ORNL-OWG 82-11556
LABORATORY SPECIALIST
J W GOOCH, JR
ORNL
PROJECT MANAGER
C W FRANCIS
CHEMICAL ANAYSIS
R W HARVEY'
N M FERGUSON*
0 R HEINE*
ENVIRONMENTAL SCIENCES
EARTH SCIENCES SECTION HEAD
T TAMURA
SOLID WASTE
EXTRACTIONS
CHEMICAL
ANALYSIS
QA OFFICER
ESD
M H SHANKS
QA OFFICER
ACO
L T COR8IN*
PRINCIPAL INVESTIGATOR
D K BROWN
PRINCIPAL INVESTIGATOR
M P MASKARINEC*
DATA
MANAGEMENT
PRINCIPAL INVESTIGATOR
JC GOYERT
•ANALYTICAL CHEMISTRY DIVISION
Fig. A-l. Project organization and responsibilities.
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3. QA Objectives in Terms of Precision. Accuracy, Completeness,
Representativeness, and Comparability
3.1 Organic Analyses
Standard methods are in place for the classes of organic compounds
expected to occur in the solid waste extracts: PAH's, phenols,
purgeable halocarbons, and aliphatic hydrocarbons. The isolation of
the organic compounds is performed according to EPA methods 624
and 625. By using check standards (externally prepared) and periodic
repetitive analyses of selected extracts as well as National Bureau of
Standards (NBS) prepared samples, QA objectives for these parameters
have been established as follows:
A. Precision - The relative standard deviation of repetitive
measurements on a sample should not exceed that achieved in
methods development/evaluation studies. This value has been
determined to be ±10% for organic analyses.
B. Accuracy - The analytical data obtained from the application
of each analytical inethod to a check standard (of different
concentration than the calibration standards) should agree
within ±15% of the true value defined by its gravimetric
preparation. Analytical results for samples also examined by
other EPA contractors should agree statistically at the 90%
confidence limits.
C. Completness - At least 80% of all possible analytical
measurements should meet QA objectives.
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3.2 Inorganic Analyses
Elemental inorganic analyses will be conducted by atomic
absorption (AA) spectroscopy (equipped with fTameless graphite furnace)
and Inductively Coupled Plasma (ICP) methodologies. All samples by AA
will be conducted according to standard methods (USEPA 1979) except for
Hg, Se, and As analyses which will be conducted as described in Section
7.4. For AA and ICP analyses the QA objectives are as follows:
A. Precision - The relative standard deviation of repetitive
measurements will vary from element to element and as a
function of detection (the closer to the detection limit the
higher the deviation). For samples containing elements ten
times the detection limit the objective is < ±15%, while for
samples near the detection limit the objective is ±100%.
B. Accuracy - Using known standards furnished by EPA, the
accuracy for inorganic analyses is expected to be ±15% at
levels ten times the detection limit. Near the detection
limit the accuracy may decrease to 100 to 200%.
C. Completeness - At least 90% of all possible analytical
measurements should meet QA objectives.
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4. Sampling Procedures
Municipal refuse leachate has been obtained from the Corps of
Engineers Waterways Experimental Station (WES) in Vicksburg,
Mississippi. The municipal refuse leachate was collected in 3.4-L
Teflon*-lined cans under pressure (using helium) in a sealed glovebox
with a helium atmosphere. This leachate (approximately 125 L) will be
kept anaerobic and will be refrigerated at 4°C until use. Aliquots of
the municipal refuse leachate will be analyzed throughout the
extraction experiments to assess any changes in the chemical nature of
the samples.
The industrial wastes are being obtained through EPA's Washington
Headquarters, Office of Solid Waste, Todd Kimrnell (coordinator). The
wastes are being shipped to ORNL in 208-L (55-gal) drums. Wastes in
the drums will be thoroughly mixed before subsampling for individual
extractions. A universal sampling procedure will not be applicable in
obtaining representative subsamples from the drums. The types of
wastes range from a total liquid to slurry and dry solid. Therefore,
each waste will be examined, and a specific sampling procedure will be
documented. When applicable, the drum will be placed on a drum roller
to obtain a thorough mixing of the waste sample. Subsample from the
drums will be analyzed for physicochemical properties to ensure
adequate mixing and sampling procedures.
Municipal wastes will be collected at local municipalities and
sorted to desired composition for the large-scale lysimeter studies.
Sampling history, conditions, and procedures will be recorded in
ORNL-registered laboratory notebooks.
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5. Samples Custody
The bulk industrial wastes [208-L (55-gal) drums] and municipal
refuse leachate will be stored in a locked, refrigerated cargo trailer
(ORNL Bldg. 7038). The trailer is located in a locked fenced area;
only authorized personnel are allowed access. The trailer was
purchased and is operated for the EPA Synfuels Repository Project
(DOE Activity No. 40-740-78). It is maintained at a constant 4°C
temperature. The air temperature is continually recorded in the
trailer with the recorder paper being changed monthly; the chart papers
are kept in a permanent file by W. Griest (ACD).
All wastes, leachates, laboratory-derived extracts, extraction
procedures used to obtain extracts, and important laboratory variables
will be identified by a unique six-digit sample identifier code. For
example, sample code 1CIN21 may represent: reconnaissance study using
oil-reclaiming clay extracted aerobically in a column extraction -
extraction replicate 2 and aliquot 1 analytical analysis. All persons
involved with the experiments will use the same identifier code for a
particular sample. The sample identifier code will be established by
D. K. Brown who is in charge of the repository of waste samples.
D. K. Brown also initiates sample extractions. All sample information
will be logged into the data management system using the Project
Results Tracking System (PRTS) (Strand et al. 1981; and Strand et al.,
in review). Information will be kept concerning sample delivery, dates
of extraction, dates for submission for analytical analyses, completion
dates of the analyses, and results of analyses. The PRTS is a complete
record-keeping system that allows information to be obtained concerning
130
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the status of the project; the system also allows statistical analyses
of the data without any data transferring (see Section 8).
In addition, a registered ORNL technical notebook will be kept for
the solid wastes and leachate samples. A record will be kept as
subsamples are taken from the storage area in terms of what samples
were taken and by whom, quantities removed, and for what purpose.
In terms of waste disposal, separate labeled containers will be
kept for liquid (extracts, washwater contacting solids, etc.) and solid
(spent wastes, filters, etc.) wastes. Wastes will be handled and
disposed of according to ORNL's Environmental Protection Manual and
Hazardous Materials: Management and Control. These manuals are
in-house reference manuals that are updated as needed by ORNL's
Department of Environmental Management (T. W. Oakes, Department Head).
Waste disposal will be handled upon advice by each Division's
Environmental Protection Officer.
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6. Calibration Procedures and Frequency
Each analytical Instrument will be recalibrated by the operating
Analytical Chemistry personnel before samples are analyzed. Flame
atomic absorption will be checked after analysis of six samples.
Standard addition techniques will be used for graphite furnace analysis.
Commercial standards prepared for atomic absorption analysis by Fisher
Scientific Co. will be used for preparation of calibration solutions.
For inductively coupled plasma-atomic emission spectroscopy the
calibration and analysis of the samples will be performed according to
EPA method 200.7 [Martin and Kopp (ed.)» Inductively Coupled
Plasma-Atomic Emission Spectrometric Method for Trace Element Analysis
of Water and Wastes, Environmental Monitoring and Support Laboratory,
Cincinnati, Ohio]. Single and multielement standards obtained from
SPEX Industries (Metuchen, New Jersey) as well as certified reference
samples (e.g., EPA or NBS) will be used to check the instrument
calibration.
Instrument standardization will be conducted by subjecting an
aliquot of the standard solution to the analytical procedures in the
same manner as a sample aliquot and by calibrating the observed
instrument response for each standard constituent to the known
concentration of that constituent. Matrix effects will be checked by
spiking samples and alternate method checks. Any observed matrix
effects will be corrected.
Standardization of organic analysis will be conducted using
appropriate NBS materials (PAH's, phenols) on a biweekly basis.
Periodic quality control samples will be used for a wet chemical
operations check.
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7. Analytical Procedures
7.1 Extraction and Concentration
All solvent partition/concentration steps are carried out using
internal standards. The internal standards (azulene, fluorophenol, and
C-cresol) are added prior to extraction and are used as a measure
of recovery as well as for instrumental calibration. A sample blank
(distilled water + internal standards) is run with each set of sample
analyses (up to a maximum of eight).
7.2 Analytical Chromatography
A standard mixture (containing the above-mentioned internal
standards) is analyzed with each sample set (all samples received on a
given date, maximum of eight samples) to verify the response of the
instrument, the retention time of the internal standard, and the
integration parameters. If any parameter varies more than 10% from the
previous standard analysis, corrective action is taken. Such action
might include recalibration, detector cleaning, column replacement, etc.
7.3 Gas Chromatography/Mass Spectrometry
Instrumental variables (tuning parameters, etc.) are checked and
recorded daily. Calibration curves are checked with each sample set
(as above).
7.4 Atomic Absorption (AA) Spectroscopy
All aqueous samples to be analyzed by atomic absorption (AA)
spectroscopy will be analyzed by a Perkin-Elmer Model 403 or 603
AA instrument. Flame atomic absorption will be used if the sample
contains high enough concentrations of the elements to be determined
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accurately by this method. If concentrations are lower than this,
graphite furnace AA will be used. All samples will be preserved and
treated according to standard methods (USEPA 1979) with the following
exceptions:
(1) extracts for Hg determination will be preserved by addition
to a nitric acid/dichromate solution and worked up for cold
vapor fTameless AAS (Feldman 1974),
(2) Se will be chelated with 5-nitro-o-phenylene diamine and
extracted into toluene before analysis (Talmi and Andren
1974), and
(3) As will be determined by an arsine accumulation-helium glow
detector procedure (Feldman 1979).
External reference solutions and EPA multielement standards will be
used for instrument calibration. All samples will be analyzed by the
standard addition method (USEPA 1979).
7.5 Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP)
Multielement analysis will be performed with an Instruments SA,
Inc. model JY85 inductively coupled plasma-atomic emission spectrometer
(ICP-AES), which contains a 1-m polychromator equipped with exit slits
centered at the most sensitive emission line (except for Na, Li, and K)
of 35 different elements for simultaneous determinations. A scanning
"n + 1" channel is also used either for the determination of an
additional element or as an alternate emission line of one of the
existing 35 elements. This instrument is controlled by a PDP-11/23
minicomputer, which also handles all data manipulations.
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The computer software provided by the manufacturer includes
background correction, interelement correction, and blank subtraction
algorithms, which are included in the program that automatically
changes samples, takes data, etc.
Dilute aqueous solutions containing the analyte are pumped into a
cross flow nebulizer, which produces a fine aerosol. The aerosol is
carried into the ICP, an extremely hot, flame-like electrical discharge
in argon gas, where the analyte species are thermally excited and
spontaneously emit radiation characteristic of the elements present.
The radiation is measured with the polychromator and monochromator to
determine the concentrations of elements in the solution.
Multielement standards from SPEX Industries (Metuchen, New Jersey)
are used to calibrate this instrument. At the present time,
calibration is performed once each day and periodically checked with an
EPA or an NBS standard.
The simultaneous multielement capability of this instrument
permits rapid, cost-effective analyses to be performed. Accuracy is
generally better than 10% for elements whose concentrations are more
than ten times their detection limit, while precision is usually better
than 5% relative standard deviation (RSD).
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8. Data Analysis. Validation, and Reporting
All data analysis, validation, and reporting conducted for the
"Toxicity of Leachates" project will be accomplished through the
establishment of a Research Data Management System which incorporates a
number of computerized systems (Fig. A-2). Data (inorganic and organic
analyses) will be entered into this data management system in
accordance with the sample identifier code as described under
Section 5. The ICP data will be programmed directly by the analyst;
other data will require formatting and entry by the data management
group.
A software package entitled Statistical Analysis System (SAS) is
leased by ORNL to conduct statistical analyses' of the data obtained in
the project. This is a well-documented heavily used package produced
by SAS Institute, Box 8000, Gary, North Carolina 27511.
The Decision Support System (OSS) is one component of this system
and is designed for management-level integration of environmental
research data. The DSS includes a plan of study documentation
component for determining ongoing research among different sites, a
status component for determining the current and proposed research
within research sites, and a criteria component for documenting the
numeric standards or limits (e.g., EPA/RCRA guidelines) within a
research project.
To cope with the problem of accounting for and summarizing the
status and results of all samples taken during the project, a Project
Results and Tracking System (PRTS) will be included in the validation
procedures. The PRTS is designed to ensure that each sample, matched
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ORNL-DWG 82-7384 ESO
ENVIRONMETRICS
ENVIRONMENTAL SCIENCES
I
RESEARCH DATA MANAGEMENT
STATISTICS
EXPERIMENTAL DESIGN
SAMPLING DESIGN
ANALYSIS
INTERPRETATION
M
COMPUTERIZED SYSTEMS
DATA BASE MANAGEMENT
MANAGEMENT SUPPORT
SAMPLE TRACKING
QUALITY ASSURANCE
GRAPHICS
STATISTICAL COMPUTATION
Fig. A-2. Disciplines and activity areas.
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with its corresponding characterization and effects data, represents an
appropriate estimate of a component of the research design. A PRTS, by
automating the bookkeeping procedures, reduces the time and effort
often needed to track large numbers of samples across research groups,
and it can provide an initial check on validity, completeness, and/or
accuracy of the results from each sample. Formal and detailed quality
assurance controls are also established in the Quality Assurance
System. The Quality Assurance System is a computerized program that
checks for the proper syntax, coding, and variable values using
criteria developed specifically for the type of data collected by the
investigators.
Implementing quality assurance controls on developing and/or
extant data bases is necessary to ensure the validity of the results of
the research program. The Quality Assurance System for the Toxicity of
Leachates project has been developed to assist scientists in detecting
potential errors in both qualitative and quantitative data. All major
data sheets will have a program written to ensure that the proper
syntax and variable values have been recorded and keypunched. The
program will document all outlier and syntax errors in the raw data
files and will provide an annotated output for resolving other
potential errors, such as values outside a variable's existing range,
improper lab identification, improper dates, etc. All error
definition, recognition, and correction will be accomplished by staff
within the program.
138
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A generalized figure representing the data flow for the Toxicity
of Leachates project through the Research Data Management System is
illustrated in Fig. A-3. As stated earlier, a number of generic
computerized systems have been developed to support management
inquiries* determine the status of sample analyses and results, detect
possible errors in the data, establish computer file structure and
manipulation techniques, support graphical displays, and provide
statistical computation procedures. A flow diagram showing sample
processing and analyses is presented in Fig. A-4. All files are
automatically backed up weekly by staff in Computer Sciences, UCCND.
The criterion for flagging possible erroneous data is developed by
the investigator and is based on known or possible ranges of values and
techniques and methodologies employed. Outliers or extreme data points
are identified using an empirical approach rather than any formal
statistical decision rules. The investigator first examines outliers
for possible measurement or transcription errors. If these are not
present, the analyses are conducted with and without the outliers.
Data are graphed to investigate residual patterns and are only deleted
at the request of the investigator.
139
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ORNL-DWG 82-7382
MANAGEMENT
SUMMARIES
RESEARCH PROGRAM
EXPERIMENTAL
SAMPLE DESIGN
DECISION
SUPPORT
SYSTEM
QUALITY
ASSURANCE
SYSTEM
MANAGEMENT
DOCUMENTATION
TRACKING
SYSTEM
UPDATED. ERROR
FREE FILES
DATA BASE \
MGMT SYSTEM I
I ORGANIZED DATA
V SETS
(STATISTICAL
IMPUTATION
SYSTEM
GRAPHICS
SYSTEM
ENVIRONMENTAL
SCIENCES
INTERPRETATION
PROGRAM SYNTHESIS
Fig. A-3. The research data management components.
140
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MIKE * MASKARINEC
JONATHANC COVERT
R WALLACE NAAVEV
JOE w GOOCH
D R HEINE
N MARION FERGUSON
L R WILLIAMS
Fig. A-4. Sample and processing analysis.
141
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9. Internal Quality Control Checks
Internal quality control checks will be conducted In AA and ICP
analyses of each set of solid waste extracts to assess precision and
accuracy of instrument calibration. The checks consist of:
(1) re-analysis of calibration standards to confirm calibration, and
(2) analysis of check standards (these are standards of different
concentrations from the standards used to calibrate the instrument)
with each sample set. Flame AA results should agree within ±5% of
the true value. For samples analyzed by graphite furnace containing
elements ten times the detection limit, the objective is ±10%, while
for samples near the detection limit the objective is 100 to 200%. If
the results do not agree within these limits, matrix effects will be
checked by spiking samples.
Internal quality control checks for organic analyses will be
conducted by inspection of the chromatographic profiles of each
standard, solid waste extract, and sample chemical fraction. Any
unexpected/unusual chromatographic peaks or features will be noted and
interpreted with consideration of instrument malfunction, sample
contamination, or true (non-artifactual) occurrence. The use of NBS
standard materials will be used to verify instrument performance.
Solid waste extracts and municipal waste leachate analyzed by ICP
will be conducted by analyzing two replicate extracts (two aliquots
from the same solid waste extraction or leachate samples). Periodic
blanks of the relevant extraction medium will be subjected to the same
procedures to determine if the samples are receiving external
contamination. Additional quality control methods used during
analytical analyses are reported in Sections 3, 6, and 7.
142
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10. Performance and System Audits
The system audit will be conducted as a part of the final methods
development and validation work, before the routine sample analyses are
initiated. The audit will ensure that sample handling, solid waste
extraction, fractionation, and analysis and quality control measures
are conducted properly and that data calculation, interpretation,
storage, and reporting flow correctly.
The performance audit will include the analysis of test materials
of concentrations unknown to the chemists performing the analysis.
This audit will be performed at least quarterly. To implement this
audit, ORNL has requested the EPA project officer to provide a
reference solid waste extract containing known concentrations of
organics and inorganics for analyses quarterly. Additionally, the ORNL
QA office has scheduled a QA audit of this project for May of 1982 in
accordance with ORNL Procedure QA-L-8-100.
143
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11. Preventative Maintenance
The chemists assigned to the project are responsible for dally
maintenance and upkeep of the analytical instrumentation. Preventive
maintenance measures include periodic cleaning of instrument housing
ventilation filters, replacement of support gas filters/impurity traps,
lubrication of moving parts, etc., as specified by the instrument
manufacturers. Instrument repairs are conducted by the manufacturer
for those instruments still under warranty, or by the ORNL
Instrumentation and Controls Division (I&C) for those no longer under
warranty.
Spare parts for many of the instruments are kept in supply by the
I&C Division. The I&C Division also provides routine preventative
maintenance on electronic equipment in accordance with IPD2,
Maintenance Information System Instrumentation Manual. The Plant and
Equipment (P&E) Division provides routine preventative maintenance on
mechanical equipment in accordance with QA-PE-18/D.1.14.
144
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12. Specific Procedures to be Used Routinely to Assess
Data Precision. Accuracy, and Completeness
A. Precision - Precision will be assessed by calculation of the
relative standard deviation of repetitive measurements carried
out on selected samples, as shown below:
RSD.
where RSD. = relative standard deviation for measurements of
component i,
X. = individual measurements of component i,
X. = the average of the measurements of component i, and
n. = the number of measurements.
The relative standard deviations should not exceed those
established in the course of analytical methods of
development/validation studies.
B. Accuracy - Analytical measurement accuracy will be assessed in
the Performance Audits, the Internal QC Checks, and in the
comparison of data on samples shared with other laboratories.
In the first two of these measures, the analytical result will
be compared with the true result, and the accuracy of
measurement of each constituent (A.) will be calculated as
the percentage of difference between the measured value (X.)
and the true value (T.):
A. = 100 (XrT.)/T
i i i i. ^
145
-------
Re-analysis of calibration standards and analyses of check
standards should agree to within ±15% of the true value.
C. Completeness - Completeness of data will be evaluated by
multiplying the fraction of samples analyzed (number of
samples actually analyzed/number of samples planned in
expected design which could have been analyzed) by the
fraction data generated (number of actual data points
generated per sample) and expressing the result as a
percentage. A result such as "ND" (component not detected) is
considered a data point, while lack of an analysis due to
instrument malfunction or sample loss would be classified as a
lack of data points. At least 90% of the expected data should
be generated. Where possible, reruns of sample material or
reanalysis of extracts will be performed to minimize the
occurrence of missing data.
146
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13. Corrective Action
The corrective action to be taken depends on the type of
performance deficiency. For deficiencies uncovered in routine internal
quality control checks, the actions outlined in Section 9 will be
taken. For deficiencies discovered in data evaluation, outlined in
Section 8, the following correction actions will be taken:
A. Accuracy - Insufficient accuracy may result from factors such
as instrument malfunction, improper instrument calibration,
degradation or other changes in the calibrating standards, or
improper sample aliquotting. The first three factors may be
identified by analysis of a freshly prepared standard. The
last cause may be checked only be re-aliquotting the sample.
(Aliquots will be held specifically for this purpose.)
Improper measurement of sample recovery also can lead to
inaccuracy and can be determined only by remeasurement.
Samples should be re-analyzed where loss of accuracy is
uncovered.
B. Precision - Loss of acceptable precision may be attributed to
factors such as instrument malfunction or irreproducible
aliquotting. These causes could be checked by repetitive
analysis of a standard and distinguished by comparing results
obtained in two instruments. Re-analysis of at least selected
samples would be required to remedy this deficiency.
C. Completeness. - Insufficient data may be caused by sample loss
(either sample mishandling or isolation procedure failure),
instrument breakdown, or failure to perform the analysis. The
147
-------
potential for sample loss is very low. In the case of
instrument breakdown or failure to perform the analysis, the
sample analysis should be rescheduled. Corrective action
relative to sample loss will be dealt with by making an
extraction on original waste.
Significant quality deficiencies are reported to upper ORNL management
throughout the ORNL QA program in accordance with procedure QA-L-6-103
(Fig. A-5).
148
-------
OAK RIDGE NATIONAL LABORATORY
CORRECTIVE ACTION PLAN
lam MUMIKM
rOLLOW U» A.MIGNID TO
CORRECTIVE ACTIOMISI
RECOMMENDED
CORRECTIVE ACTIOW1SI
PLANNED
ACTION
ASSIGNED TO
CQUfLCTlOH OAfE
SCHEDULED
ACTUAL
11 All
I
tn
-t
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-------
14. Quality Assurance Reports to Management
QA information will be summarized to the EPA Project Officer in
each bimonthly data report. Internally in ORNL each division has a
quality assurance program (see Fig. A-6). Procedures relative to
projects and programs in the respective divisions are outlined in the
Divisions' Quality Assurance Program: Environmental Sciences
Division's Quality Assurance Program (QA-ES-1-100) and Analytical
Chemistry Division's Quality Assurance Program. Each project will be
assessed and reviewed periodically. The QA assessment review date
schedule is included for the Analytical Chemistry Division (see
Fig. A-7).
150
-------
:
o..t
1
1
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1
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|
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Fig. A-6. Quality assurance program organization chart.
151
-------
ua
in
ro
DIVISION DOCUMENT NUMBER
AC AC-A1-1
AC AC-BO-1
»<
ft
n
-•
0
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U.
i/i
rh
-J
2
i/i
o
3
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01
I/I
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O>
ui
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n>
3
ft-
I/I
o
3"
n>
Q.
n>
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC-BO-2
AC-BO-3
AC-BO-4
AC-BO-5
AC-GAL-1
AC-GAL-12
AC-GAL-1 3
AC-GAL-14
AC-GAL- 15
AC-GAL-16
AC-GAL- 17
AC-GAL-2
AC-GAL-3
AC-GAL-4
AC-GAL-5
AC-MES-1
AC-MES-10
QA ASSESSMENT REVIEW DATE SCHEDULE
FOR PERIOD ENDING 12/31/1981
QA ASSESSMENT TITLE
IN-LINE ANALYTICAL SENSORS
ANALYSIS OF OCCUPATIONAL/ENVIRONMENTAL
SAMPLES
GASIFIERS FOR INDUSTRY
GENERATION AND CHEMISTRY OF AEROSOLS
SMOKING AND HEALTH
SPECIALIZED ORGANIC ANALYTICAL CHEMISTRY
BAUSCH AND LOMB SPECTROPHOTOMETER X 11783
ATOMIC ABSORPTION SPECTROPHOTOMETER(S)
P-E 403, P-E 460
TRACOR MODEL 222 GAS CHROMATOGRAPH
ASH FUSION FURNACE
INDUCTIVELY COUPLED PLASMA SPECTROMETER
LECO OXYGEN DETERMINATOR, R018
ION CHROMATOGRAPH, DIDNEX MODEL 16
FISHER TITATOR
TECHNICON AUTOANALYZERS
FLUOROPHOTOMETER - SINTERING FURNACE
(FLUOROMETRIC URANIUM EQUIP)
TEKTRONICS 4025 COMPUTER TERMINAL
OPTICAL EMISSION SPECTROMETER I - PASCHEN
DUPONT 21-490B/21-094B GAS CHROMATOGRAPH-
MASS SPECTROMETER
SCHEDULED
ORIGINATOR
STRAIN
CATON
CATON
CATON
CATON
CATON
RICKARD
RICKARD
RICKARD
RICKARD
STEWART
LAYTON
KELLER
RICKARD
RICHARD
RICKARD
RICHARD
CHRISTIE
RAINEY
ISSUE DATE
8/28/1 981
6/03-1981
6/03/1981
6/03/1981
6/03/1981
6/03/1981
9/07/1979
10/23/1979
11/14/1979
11/12/1979
2/02/1981
2/05/1981
2/18/1981
9/07/1979
10/22/1979
10/22/1979
10/23/1979
11/08/1979
REVIEW DATE
9/01/1982
6/03/1982
6/03/1982
6/03/1982
6/03/1982
6/03/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
2/25/1982
2/05/1982
2/20/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
STATUS
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
-------
DIVISION DOCUMENT NUMBER
AC AC-MES-11
-n
\o
>
•vl
a
3'
CD
•
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC-MES-12
AC-HES-13
AC-MES-14
AC-MES-15
AC-HES-2
AC-MES-3
AC-MES-4
AC-MES-5
AC-MES-6
AC-MES-7
AC-MES-8
AC-MES-9
AC-NRAS-1
AC-NRAS-2
AC-NRAS-3
AC
AC-NRAS-4
QA ASSESSMENT REVIEW DATE SCHEDULE
FOR PERIOD ENDING 12/31/1981
QA ASSESSMENT TITLE
ORNL SINGLE-STATE GAS CHROMATOGRAPH-MASS
SPECTROMETER
SPARK SOURCE MASS SPECTROMETER - 2026
AEI MS-SO/DS-50 MASS SPECTROMETER-DATA
SYSTEM
ORNL THREE STAGE MASS SPECTROMETER -
ORG.
MICROMASS 1201 GAS MASS SPECTROMETER
OPTICAL EMISSION SPECTROMETER II -
WADSWORTH
ION MICROPROBE MASS SPECTROMETER
SPARK SOURCE MASS SPECTROMETER - MS-7
SPARK SOURCE MASS SPECTROMETER - MS-702R
2-STAGE MASS SPECTROMETER - RAL
SINGLE-STAGE MASS SPECTROMETER - RAL
2-STAGE MASS SPECTROMETER - TRU
3-STAGE MASS SPECTROMETER - RAL
COMPUTER BASED PULSE HEIGHT ANALYSIS
SYSTEM ND6620
OAK RIDGE RESEARCH REACTOR PNEUMATIC TUBE
COMPUTER BASED PULSE HEIGHT ANALYSIS
SYSTEM. BLDG 3042
DATA ACQUISITION SYSTEM, 110 6603, G-49.
4500S
SCHEDULED
ORIGINATOR
RAINEY
CHRISTIE
RAINEY
RAINEY
SITES
CHRISTIE
CHRISTIE
CHRISTIE
CHRISTIE
SMITH
SMITH
SMITH
SMITH
EMERY
EMERY
EMERY
ISSUE DATE
11/08/1979
11/08/1979
11/08/1979
11/08/1979
11/08/1979
11/08/1979
11/08/1979
11/08/1979
11/08/1979
11/09/1979
11/09/1979
11/09/1979
11/09/1979
2/20/1980
5/01/1980
5/01/1980
REVIEW DATE
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
STATUS
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
SCOTT
5/01/1980 1/01/1982
OPEN
-------
QA ASSESSMENT REVIEW DATE SCHEDULE
FOR PERIOD ENDING 12/31/1981
DIVISION DOCUMENT NUMBER
ta
i
o
el-
s'
n>
CL
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC
AC-NRAS-S
AC-NRAS-6
AC-PM-1
AC-RMAL-1
AC-RMAL-2
AC-RHAL-3
AC-RHAL-4
AC-RMAL-5
AC-RMAL-6
AC-TRU-1
AC-TRU-2
AC-TRU-3
AC-TRU-4
AC-TRU-5
AC-TRU-6
AC-TRU-7
AC-TRU-8
AI-2
Al-3
QA ASSESSMENT TITLE
HIGH FLUX ISOTOPE REACTOR PNEUMATIC TUBE
COMPUTER BASED PULSE HEIGHT ANALYSIS
SYSTEM AT BLDG 7900
PHYSICO-CHEMICAL ANALYSES GROUP
IN-CELL PIPET
IN-CELL ANALYTICAL BALANCE
IN-CELL FURNACE
IN-CELL SPECTROPHOTOMETER
HOT-CELL MANIPULATORS
X-RAY FLUORESCENCE ANALYZER
PULSE-HEIGHT ANALYZER
PROPORTIONAL COUNTERS
TITRATOR
BAUSCH AND LOMB SPECTROPHOTOMETER
REMOTE PIPETTOR
GEIGER-MUELLER COUNTER
GAMMA SCINTILLATION COUNTER
NEUTRON COUNTER
LASER APPLICATIONS
REMOTE PIPETTOR AND TITRATOR
ORIGINATOR
SCHEDULED
ISSUE DATE REVIEW DATE
STATUS
BATE
BATE
RICCI
LAING
LAING
LAING
.LAING
LAING
STEWART
COOPER
COOPER
COOPER
COOPER
COOPER
COOPER
COOPER
COOPER
WHITTEN
KLATT
7/02/1980
7/02/1980
11/05/1979
11/08/1979
11/08/1979
11/08/1979
11/08/1979
11/08/1979
2/02/1981
11/13/1979
11/13/1979
11/12/1979
11/12/1979
11/12/1979
11/13/1979
11/13/1979
11/13/1979
9/02/1981
9/10/1981
7/31/1982
7/31/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
2/02/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
1/01/1982
9/01/1983
10/01/1982
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
OPEN
-------
15. References
Feldman, C. 1974. Perchloric acid procedure for wet-ashing organics
for the determination of mercury (and other metals). Anal. Chem.
46:1606-1609.
Feldman, C. 1979. Improvements in the arsine accumulation - helium
glow detector procedures for determining traces of arsenic. Anal.
Chem. 51:664-669.
Strand, R. H., M. P. Parrel!, T. K. Birchfield, C. W. Gudmundson,
M. E. Vansuch, and H. N. Polovino. 1981. Environmetrics for
synfuels: II. A Computer-based coding scheme for coal conversion
research data. ORNL/TM-7525. 25 pp.
Strand, R. H., M. P. Parrel!, K. L. Daniels, and J. C. Goyert.
Environmetrics of Synfuels: IV. Project Results Tracking System
(PRTS). Oak Ridge National Laboratory, Oak Ridge, Tennessee (in
review).
Talmi, Y., and A. W. Andren. 1974. Determination of selenium in
environmental samples using gas chromatography with a microwave
emission spectrometric detection system. Anal. Chem. 46:2122-2126.
U.S. Environmental Protection Agency (USEPA). 1979. Methods for
chemical analysis of water and wastes. EPA-600/4-79-010.
Environmental Monitoring and Support Laboratory, Office of
Research and Development, U.S. Environmental Protection Agency,
Cincinnati, Ohio. 460 pp.
155
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16. Distribution List
S. I. Auerbach, ORNL-ESD, Division Director
D. K. Brown, ORNL-ESD, Principal Investigator
L. T. Corbin, ORNL-ACD, QA Coordinator
C. W. Francis, ORNL Project Manager
J. C. Goyert, ORNL-ACD, Principal Investigator
M. R. Guerin, ORNL-ACD (Bio/Organic Section), Section Head
M. P. Maskarinec, ORNL-ACD, Principal Investigator
F. H. Neill, ORNL, Director of ORNL Qualtiy Assurance Program
0. E. Reichle, ORNL-ESD, Associate Division Director
John Santolucito, EPA QA Officer, EMSL-Las Vegas
M. H. Shanks, ORNL-ESD, QA Coordinator
W. D. Shults, ORNL-ACD, Division Director
T. Tamura, ORNL-ESD (Earth Sciences), Section Head
L. R. Williams, EPA Project Officer, EMSL-Las Vegas
156
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APPENDIX B
FY-1982 Work Plan
Toxicity of Leachates
Revised 3/25/82
Oak Ridge National Laboratory
Oak Ridge, Tennessee
Project Manager
C. W. Francis, Environmental Sciences Division
Principal Investigators
D. K. Brown, Environmental Sciences Division
J. C. Goyert, Environmental Sciences Division
M. P. Maskarinec, Analytical Chemistry Division
Project Officer
Llewellyn R. Williams
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada
Interagency Agreements
Between
U.S. Department of Energy
DOE No. 40-1087-80
and
U.S. Environmental Protection Agency
AD-89-F-1-058
157
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Table of Contents
Summary 159
1. Introduction 165
2. Development of EP-III 166
2.1 Leaching Media Requirements 166
2.2 Extraction Procedures 168
3. Performance Criteria for EP-III 170
3.1 Laboratory Studies 170
3.1.1 Waste Selection 172
3.1.2 Approach 172
3.1.3 Quality Control 176
3.1.4 Supplementary Study: Compatibility of
Extraction Media to Toxicity Tests 177
3.2 Field Studies - (95/5) Scenario 177
3.2.1 Objectives and Approach 177
3.2.2 Laboratory Comparisons 178
3.2.3 Aggressiveness of Municipal Waste
Leach ate over Time 181
3.2.4 Experimental Design and Analysis 181
3.3 Basis of Calculation for Large-scale
Lysimeter Research 182
4. Work Schedule 185
5. References Cited 186
Attachment A - Protocol for Evaluating the Aggressiveness
of Three Treatment Procedures 187
Attachment B - Description of Lysimeter Study 189
158
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OAK RIDGE NATIONAL LABORATORY
"TOXICITY OF LEACHATES"
Summary of FY-1982 Workplan
(Revised 3/25/82)
INTRODUCTION
The objective of this research is to develop a second generation
laboratory extraction procedure, henceforth known as EP-III, to test
mobility of inorganic as well as organic constituents of industrial
wastes co-disposed with municipal wastes in a landfill environment.
The desirable characteristics of EP-III include:
1. Ability to simulate leaching in a landfill containing
municipal (95%) and industrial (5%) wastes.
2. Compatibility with biological toxicity tests (mutagenic,
aquatic, and terrestrial).
3. Relatively inexpensive to conduct in terms of time, equipment,
and personnel.
STRATEGY
The strategy proposed by Oak Ridge National Laboratory (ORNL) for
the development of EP-III will focus on:
1. Determining in the laboratory the quantities of inorganic and
organic constituents leached from a variety of industrial
wastes by a particular municipal waste leachate.
2. Comparing the extraction characteristics of the above municipal
waste leachate to those of laboratory methods (combinations of
extraction procedures with artificial extraction media), and
159
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r.. Testing the ability of the laboratory extraction methods to
simulate the leaching of industrial wastes under field
conditions using large-scale lysimeters containing municipal
(95%) and industrial (5%) wastes.
The proposed research will involve both laboratory and field
work. The initial laboratory effort will be directed toward evaluating
the extraction conditions that are most aggressive in removing
inorganic and organic constituents from selected industrial wastes by a
particular municipal waste leachate. The municipal waste leachate
selected for this study will be obtained from a lysimeter located at
the U.S. Army Corps of Engineers Waterways Experiment Station (WES),
Vicksburg, Mississippi. The extraction procedure (column aerobic,
column anaerobic, or rotary batch) found to be most aggressive will
serve as baseline data to be simulated by the laboratory procedure
developed for EP-III. The laboratory variables to be investigated in
this phase of the work will consist of the type of extraction procedure
(uo-flow column or batch rotary) and type of artificial extraction
media (acetate buffer, C02-saturated water, distilled water, or
synthetic leachate).
Concentrations of inorganic and organic constituents found in
waste leachate from field demonstration large-scale lysimeter
experiments containing waste in a 95% municipal to 5% industrial
scenario will be compared to those observed in the laboratory
extractions. Results from both the laboratory and field studies will
be combined and compared so that the relationship between laboratory
extraction methods and actual field validations can be determined. By
160
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comparing laboratory extraction tests with field validations, the
potential toxicity of industrial wastes co-disposed with municipal
wastes can be examined and evaluated.
LABORATORY STUDIES
A. Objective:
Establish the laboratory extraction method (procedure and medium)
that most closely simulates the leaching characteristics of a municipal
waste leachate (MWL) in terms of organics and inorganics extracted.
B. Approach:
1. Determine the amount of organic and inorganic chemical
_ constituents removed from four different industrial wastes
using three extraction procedures. MWL from WES will be used
as the extraction media and will serve as a baseline control
to compare against all artificial extraction media in part 2.
The fou»- industrial wastes w'ill consist of two organic
wastes, a waste containing both organic and inorganic
contaminants, and a waste predominantly inorganic in character.
The three extraction procedures will consist of:
(a) up-flow column - aerobic
(b) up-flow column - anaerobic
(c) rotary extractor - aerobic
2. Compare the results from part 1 with the amount of organic and
inorganic chemical constituents removed from the same four
industrial wastes using two extraction procedures with each of
four artificial extraction media.
161
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The two extraction procedures will consist of:
(a) an up-flow column and
(b) a batch rotary extractor.
The four extraction media will consist of:
(a) 0.1 J^ sodium acetate buffer pH 5,
(b) C02-saturated deionized distilled water,
(c) deionized distilled water, and
(d) synthetic MWL that simulates WES/MWL.
C. Experimental Design:
The experiment will consist of a factorial arrangement of
11 treatments and four industrial wastes in a randomized block design
with two blocks (time) per treatment-waste combination. The
11 treatments will consist of the eight (procedure x media) laboratory
methods plus the three WES/MWL extractions.
D. Analysis:
An analysis of variance will be conducted on the ranked sums
(Attachment A) to determine if there are any differences in the amount
of organic and inorganic chemical constituents removed by the various
extraction procedures and media. Additional analyses will be conducted
to determine if there are any overall differences in: (1) the batch
versus column extraction procedure, (2) the distilled water, synthetic
leachate, and WES/MWL media, and (3) the interaction among the two
procedures and the three extraction media. The eight laboratory
methods (2 procedures x 4 media) will be compared to the three WES/MWL
extractions using a multivariate pattern analysis to determine which of
162
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the eight laboratory extraction methods is most similar in chemical
composition to the three WES/MWL extractions. An analysis will also be
conducted to determine if there are any differences in the amount of
organics and inorganics removed by the WES/MWL using the three
extraction procedures.
FIELD STUDIES
A. Objectives:
1. Test the ability of various laboratory extraction methods to
simulate the leaching characteristics of industrial wastes in
large-scale field lysimeter experiments containing wastes in a
95% municipal to 556 industrial scenario.
2. Evaluate the aggressiveness relative to age of a municipal
waste leachate to extract contaminants from industrial wastes.
B. Approach:
Municipal waste leachate from two large-scale lysimeters (1.8 m in
diameter and 3.6 m in height) will be diverted to 16 columns/lysimeter
containing four industrial wastes (the same wastes used in the
laboratory studies). For each lysimeter, one-half of the columns
(4 wastes x 2 replicates) will be used to compare the 95% municipal and
5% industrial waste scenario to the laboratory extraction studies. The
remaining columns will be used to evaluate the relative aggressiveness
of the municipal waste leachate as a function of age of the leachate.
163
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C. Experimental Design:
The experiment will consist of a randomized block design with four
treatments and two blocks (lysimeters). The experiment will be
replicated twice to test the block by treatment Interaction.
D. Analysis:
An analysis of variance will be conducted to determine if there
are any differences in the amount of organic and inorganic chemical
constituents removed from the four industrial wastes by the municipal
waste leachate. An analysis will also be conducted to see if there are
any differences in leachate quality between the two lysimeters. A
multivariate pattern analysis will be used to determine which of the
laboratory extraction methods most closely simulates the chemical
constituents derived from the field lysimeters. The results from the
laboratory studies and the field studies will be combined and
statistically analyzed to determine the laboratory method (EP-III) that
most closely simulates the mobility of inorganic as well as organic
constituents of industrial wastes co-disposed with municipal wastes in
a landfill environment.
164
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1. INTRODUCTION
To evaluate the potential toxicity of a leachate from an
Industrial waste that has been co-disposed with a municipal waste, a
laboratory extraction procedure is needed that models the leaching
action the waste would undergo when disposed of with a municipal
waste. Currently, under RCRA, toxicity is determined by the EP
toxicity test procedure (40 CFR 261.24). Toxicity criteria are based
on the concentrations of eight elements in the Primary Drinking Water
Regulations (As, Ba, Cd, Cr, Pb, Hg, Se, and Ag), four pesticides
(Endrin, Lindane, Methoxychlor, Toxaphene), and two herbicides (2,4-D,
and 2,4,5-TP Si 1 vex) observed in the extract. The EP is used as a
regulatory test to classify a waste relative to a landfill management
scenario. The EP has a number of limitations, the most important being
that a waste containing other toxicant materials is not included in the
present criteria. For example, the EP does not model the effects that
the higher molecular weight organics present in a municipal leachate
can have on the leaching of a solid waste. Other experimental factors
include the effectiveness of extraction, the deficiency of expressing
kinetic relationships of components extracted, relevance to real-world
leachates, etc. In terms of applying biological testing to EP
extracts, the EP is severely limited in that the acetic acid used in
the procedure interferes with aquatic toxicity and phytotoxicity
testing protocols (Epler et al. 1980).
The objective of the FY-1982 research at ORNL is to develop an
extraction test for solid wastes that: (1) models the leaching action
a waste would undergo when disposed of, along with a municipal waste,
165
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in a municipal waste landfill following a 95/5 (weight fraction of
municipal and industrial waste) co-disposal scenario; and (2) is
compatible in aquatic toxicity and phytotoxicity testing protocol.
This proposed extraction procedure will be henceforth referred to as
EP-III.
2. DEVELOPMENT OF EP-III
The major characteristics of EP-III are:
1. It should simulate leaching in a landfill containing municipal
(95%) and industrial (5%) wastes (by weight).
2. It has to be compatible with biological testing protocol
(mutagenic, aquatic, and phytotoxic).
3. It should be relatively inexpensive to conduct in terms of
time, equipment, and personnel.
2.1 Leaching Media Requirements
A synthetic municipal landfill leaching medium is needed to model
leaching that occurs in a landfill where an industrial waste is
co-disposed with municipal refuse. The characteristics of a municipal
landfill leachate will vary widely depending on the municipal refuse
composition and state of decomposition. Even for individual samples of
municipal refuse it would be impossible to duplicate the leachate
composition over time because of its complexity and varying stability
to biological degradation. Laboratory leachates using municipal refuse
and water are possible to produce; however, reproducing such leachates
and maintaining stability are impractical for a test protocol. Rather
166
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than attempting to define the leachate characteristics emanating from a
defined standard landfill, it is more realistic to develop a synthetic
leachate that models leachate of maximum aggressiveness. Such a model
leachate will also represent likely mismanagement scenarios, thus
adding a degree of conservatism in screening wastes for toxicity.
No one synthetic leachate will model all municipal solid waste
leachates; however, some of the characteristics of the more aggressive
leachates can be used as a model leachate. Typical characteristics
simulating that of a municipal landfill are: (1) low pH (4-6), (2) low
redox potential (anoxic environment), (3) mild to high ionic strength
(0.05-0.1 Mj, and (4) high complexation ability.
A low molecular weight carboxylic acid, acetic acid, was used in
EP-I to acidify the solid waste suspension to pH 5. The EP-I satisfied
items 1 and 3 of the above listed requirements, and for most wastes it
has been shown to be more effective than water for removing toxic
metals from waste. However, for most wastes, acetic acid is not as
effective as water in removing nonpolar organics, and the acetic acid
interferes in the phytotoxicity and aquatic toxicity tests. Thus,
there is a need to develop a model synthetic extractant that
aggressively removes inorganic and organic contaminants from wastes and
one that can also be used in biotesting of the resulting leachate. Use
of a biodegradable organic acid in the EP-III has to be eliminated from
consideration in that such an acid will inherently interfere in the
biotesting protocol. Use of inorganic acids such as ^SO^ and HC1
to acidify solid waste suspensions to pH 5 have also interfered in the
phytotoxicity and aquatic toxicity tests. One extracting medium that
167
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appears to be a viable alternative is carbonic acid. It is an
aggressive leachate from the standpoint of acidity, it is more
representative of the acidity source in landfills than mineral acids
such as HC1, HNO-, or f^SO., and under standard conditions of
temperature and pressure it should not be inhibitory in any of the
biological testing protocols. Bause and McGregor (1980) proposed and
evaluated this extractant using the same liquid-to-sol id ratio in a
batch-type extraction as the EP (20:1) and found that the carbonic acid
medium and EP extracted similar quantities of toxic trace metals from
four fossil energy solid wastes. Its effectiveness in removing organic
compounds from solid wastes has been untested.
2.2 Extraction Procedures
The agitation procedures for conducting the EP have varied from
using stirring paddles in stainless steel vessels (glass vessels on
rotary agitators) to stirring in glass vessels using commercially
available magnetic stirring devices. However, the basic premise has
remained the same; that is, the procedure is a batch-type extraction
using a final liquid-to-sol id ratio of 20:1 during the agitation period
of 24 h. The basic principle is to ensure adequate mixing between the
solid and aqueous phase. The high liquid-to-sol id ratio (20:1)
promotes dispersion of many finely textured wastes (organic-laden
sludges, etc.) which make filtering a slow and time-consuming effort.
To conduct a variety of toxicity tests and the required inorganic and
organic analysis, it is necessary to collect as much as 6 L of effluent
(Table B-l). This may take as long as 1 to 2 d to filter some of the
168
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TABLE B-l. AMOUNT OF EXTRACT NEEDED FOR
VARIOUS ANALYSES3 AND BIOTESTS
Analyses: Extract needed (ml)
Inorganics (NIPDWS) 750
Organics 1,000
Bioaccumulation 500
Biotestsb
Mutagenicity 1,000
Aquatic (D_. magna acute) 1,000
Phytotoxicity (root elongation, 2 species) 1,000
Total 5,250
Assumes single determination.
Assumes no dilution.
169
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samples through the 0.45-um membrane filter (even using a pressure
filter apparatus at 75 psi). Recent work at ORNL has shown that the
up-flow column extraction procedure removed more nonpolar organic
compounds (extracts were collected directly on XAD-2 resins) than the
20:1 batch-type extraction (the common extraction medium being double
distilled water). Other work has indicated that volatile constituents
are not as easily lost in a closed extraction system (such as the
rotary extractor) versus an open system. The column extraction
procedure merits more research, and, contrary to previo is beliefs, it
is not as time-consuming nor is it as nonreproducible relative to the
batch extractions as implied in previous investigations. Major
equipment costs for column extractions appear to be comparable to batch
extractions with a rotary extractor (Table B-2). The important fact is
that not many investigators have experimented with an up-flow column as
an alternative.
3. PERFORMANCE CRITERIA FOR EP-III
The principal performance criteria for EP-III will be (1) how well
it simulates the leaching characteristics of a municipal waste leachate,
and (2) its compatibility with biological toxicity testing protocols
(mutagenic, aquatic, and phytotoxic).
3.1 Laboratory Studies
The objectives of the laboratory studies are twofold:
1. Establish a data base that represents the leaching
characteristics of a municipal waste leachate (MWL) on
industrial wastes, and
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TABLE B-2. MAJOR EQUIPMENT NEEDS AND ESTIMATED COSTS
FOR PERFORMING A BATCH VS COLUMN EXTRACTION*
Batch extraction
Item
Estimated
cost
Column extraction
Item
Estimated
cost
Rotary extractor $2,000
Glass vessels (32-L
capacity) 45
Filtration apparatus
(pressure) 1,400
Filters (per sample)0 15-150
Pressure pumpb $ 500
Glass column with
adjustable plungers 130
aAssume both are closed extractions, with no pH adjustments, using
organic-laden wastes, to produce 6 L of extract.
Average cost for pump, prices vary from $200-2,000.
°Disposal item, number needed will vary with sample.
171
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2. Develop a laboratory extraction method using a selected
procedure and a synthetic medium that best simulates the
leaching of industrial wastes by MWL's.
3.1.1 Waste Selection; Four industrial wastes will be utilized for
all research. The four wastes will be selected from those listed in
Table B-3 and will consist of two organic wastes (tentatively Nos. 1
and 5): a waste containing both organic and inorganic contaminants
(either No. 3, 6, or 7 depending on bulk analyses, physical properties,
and availability), and a waste predominantly inorganic in character
(either No. 2, 4, or 8). Initially major organic and inorganic
constituents will be identified in each waste to target the important
potential contaminants. This information will be used to select the
four most appropriate wastes based on (1) classes of hazardous organic
compounds, (2) concentrations of toxic inorganic constituents
(predominantly those elements listed in the interim primary drinking
water standards), (3) combinations of toxic inorganic and organic
compounds, and (4) potential contaminants amenable to target analyses
(i.e., those contaminants found in the industrial waste that are
different or contained at concentrations sufficiently higher than that
observed in the municipal waste leachate). Wastes will not be selected
relative to their compatibility with any type of extractor.
3.1.2 Approach; Baseline experiments relative to the extraction
capacity of municipal waste leachate (MWL) will be conducted using the
leachate obtained January 27, 1982, from the Waterways Experimental
Station (WES), Corps of Engineers, Vicksburg, Mississippi.
172
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-.4
CO
TABLE B-3 DESCRIPTION AND PRELIMINARY INFORMATION RELATIVE TO THE INDUSTRIAL WASTES TO BE USED IN THE STUDY*
ORNL DWG 8311534
MUTE
•0
,
2
3
4
S
6
I
8
MUTE DESCRIPTION
WASTE OIL
RECLAIMING CLAY
PETROLEUM REFINING
INCINERATOR ASH ID007]
PAINT PRODUCTION
SLUDGE (D001. F003 F005I
ELECTROPLATING
WASTEWATEH SLUDGE
IF006I
HEAVY ENDS AND COLUMN
BOTTOMS FROM PRODUCTION OF
TRI AND PERCHLOROETHYLENE IKO30I
APISEPARATOR SLUDGE
AND OAF FLOAT IKO4B AND KO5II
AMMONIA STILL LIME SLUDGE
FROM COKING OPERATIONS (KO60I
CAUSTIC FILTER SLUDGE IO009)
RCflA CLASSI
FIMTION"
NH
H
H
H
H
H
II
H
|i « 1 a
§ « ii il il |l 11
Al B> U Ci Ft H| Si A| III b > S iJ z go So x8 38
XX XX
xxx x
(MAY ALSO CONTAIN HIGH Pb AND Cr) X X XX
XX XX
XXX X X
XX X X X X
X X X X X
H
Si
H!
X X
X
X
X
X
X
xxx
X
REMARKS
10-5 0*
HgBY
WEIGHT
•COLLATED FROM INFORMATION PROVIDED BY TOOD KIMMELL. OSW/EPA HOT
"NH - NONHAZARDOUS UNDER FCRA. H - HAZARDOUS
-------
Approximately 150 L of MWL in 37 3.8-L (1 gal) containers were
collected under anaerobic conditions. This leachate is currently being
stored under refrigerated conditions. It will be used to extract the
four industrial wastes. Extractions will be conducted under aerobic
conditions in a rotary extractor at a liquid to solid ratio of 20:1 and
under both anaerobic and aerobic conditions in an up-flow column using
the same quantity of waste and the effluent collected when the liquid
to solid ratio of 20:1 is reached. The extracts will be filtered
through glass fiber filters for organic analyses (effective pore
diameter approximately 0.7 urn) and 0.45-um membrane filters for
inorganic analyses.
The quantities of inorganic and organic constituents removed from
the four industrial wastes with the WES/MWL by the most aggressive of
the three extraction procedures will be considered most representative
of leaching in a co-disposal environment. Ue have established, a
priori, a four-step protocol for choosing the extraction procedure that
is most aggressive across all constituents tested. The first step of
the protocol ranks the concentrations of each constituent from least
aggressive to most aggressive. The second step groups all the
constituents into target categories; NIPDWS inorganics and/or toxic
organics (hazardous constituents listed in Appendix VIII, 40 CFR
261.24) and then sums the ranks within a category. The third step
ranks the sums within each category. The fourth step sums the ranks
across categories, resulting in an overall rank sum that is then
statistically analyzed. A short example of such ranking is provided in
Attachment A. These data will provide the following information:
174
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1. Establish if significant occur between aerobic and anaerobic
column extractions with municipal waste leachate.
2. Determine if differences occur between batch and column
extractions with a number of artificial extraction media.
3. Establish baseline data relative to the extraction
capabilities of a municipal waste leachate.
The pattern (relative to concentration and distribution) of
inorganic and organic constituents extracted by the most aggressive
extraction procedure (either anaerobic or aerobid'up-flow column or
aerobic rotary) using the WES/MWL will be compared to those patterns
obtained from the eight laboratory extraction methods (two extraction
procedures and four artificial extraction media). The same four
industrial wastes will be used.
The two extraction procedures will consist of:
(a) an up-flow column and
(b) a batch rotary extractor.
The four extraction media will consist of:
(a) 0.1 ^ sodium acetate buffer pH 5,
(b) C0?-saturated deionized distilled water,
(c) deionized distilled water, and
(d) synthetic MWL that simulates WES/MWL.
The rationale for selecting the extracting media is as follows:
EP-I: The acetic acid pH 5 extraction is a regulatory protocol
that has never been compared to the leaching capabilities of a
municipal waste leachate. It is possible that the EP-I does simulate
the extraction of municipal waste leachates and thus will be included
for comparative purposes in the experimental design.
175
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C0?-Saturated Deiom'zed Distilled Water; This medium best
simulates the pH and anoxic conditions of landfill leachates without
incorporating biodegradable organics into the medium. It should also
be compatible with biotesting protocols.
Oeionized Distilled Water; This medium is included for
comparative purposes.
Synthetic Municipal Waste Leachate; This medium which will
contain the major organic and inorganic components of the WES municipal
waste leachate is intended to simulate the extracting capabilities of
the collected leachate. Incorporation of this medium will provide
information relative to the importance of organic and inorganic
components of the WES leachate to remove various contaminants from the
five industrial wastes. Tentatively, the organic components suggested
are 500 ppm each of C2> C4, and Cg carboxylic acids.
3.1.3 Quality Control; A description of the quality control for the
project is available in the April 15, 1982, "Quality Assurance Project
Plan - Toxicity of Leachates Project" (Appendix A of this report).
Specifically, each 3.8-L (1 gal) aliquot MWL collected at WES will be
analyzed for organic (principally carboxylic acids by high pressure
liquid chromatography) and inorganic constituents (inductive coupled
plasma-atomic emission spectrometry, atomic absorption spectroscopy,
and anionic chromatography). Preliminary analysis of the WES/MWL
indicates that target organic compounds can be used to assess
Teachability. In cases where concentrations of the target constituents
are observed in the MWL (both organic and inorganic), these
concentrations will be subtracted from the concentrations found in the
176
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leachate produced on interacting the MWL and industrial waste.
Duplicate analyses for organic and inorganic constituents will be
conducted on the leachates (such analysis may not be conducted if
analytical accuracy and precision for that analysis is considered to be
less than 10%).
3.1.4 Supplementary Study; Compatibility of Extraction Media to
Toxicity Tests: A supplementary study will be carried out to determine
which of the extraction media can be used with toxicity tests. The
toxicity tests to be considered include: (1) an alga growth inhibition
test, (2) Daphnia magna acute immobilization test and reproduction
test, and (3) a fish acute toxicity test; these tests are found in the
document "OECD Guidelines for Testing of Chemicals." Extraction media
posing potential inherent toxicity characteristics will be tested in a
series of dilutions (including 1:100 and 1:000), with at least one of
the toxicity tests mentioned above. This information will aid in
determining which of the proposed extractants satisfy the
characteristic of being compatible with bioassays.
3.2 Field Studies - (95/5) Scenario
3.2.1 Objectives and Approach; The major objectives are to
(1) determine at what point in time and at what volume of leachate the
concentration of the inorganic and organic contaminants the highest in
the leachate from the industrial wastes, (2) compare these
concentrations and the total extracted to that extracted under
laboratory conditions, and (3) evaluate in what stage (early, mid, or
late) the municipal leachate is most aggressive in removing inorganic
177
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and organic contaminants from the Industrial wastes. The work Involves
the use of two large-scale lysimeters (Fig. B-l, 1.8 m in diameter and
3.6 m in height) containing municipal waste. Volume of municipal waste
is approximately 7.5 m , weighing approximately 1.5 Mg. The
municipal waste was obtained from the City of Oak Ridge, Tennessee, on
February 23, 1982, and consisted of a residential waste (collected from
households) and a commercial waste (collected from fast-order
restaurants, etc.). These wastes are layered in the lysimeter in
layers approximately 71 cm thick (Fig. B-l). The bottom of the
lysimeter contains pea gravel and silica sand to prevent clogging the
outlet with waste. A detailed description of the design is presented
in Attachment B. Leachate from each of these lysimeters will be used
as influent to individual columns containing each of the four previously
studied wastes. There will be four columns for each industrial waste,
making a total of 16 columns for each lysimeter (2 experiments x
industrial wastes x 2 replications) or a total of 32 industrial waste
columns for the total experiment. Blank sand columns will also be used
for quality control.
For one-half of the columns, the quantity of waste in each of the
columns will correspond to the 95/5 scenario. Leachate quality from
these columns, in terms of inorganics and organics, will be compared to
laboratory-derived extractions (Section 3.1). The remaining columns
will be used to evaluate the stage of aggressiveness of the MWL as a
function of the age of the leachate.
3.2.2 Laboratory Comparisons: For each lysimeter, one-half of the
columns (4 wastes x 2 replicates) will be used to compare the 95%
178
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ORNL-OWG 82.11532
(T) THERMOCOUPLE
W LOCATED AS
SHOWN
38 cm LAYER SOIL
COMPACTED
RESIDENTIAL
COMMERCIAL
WASTE (LAYERED)
EACH LAYER
ABOUT 71 cm THICK
^ffi^^_T-S0«
SILICA SAND - 5 cm
0.13 m3 (277 kgl
PEA GRAVEL
- 38 L
TO COVER DRAIN
PIPE
GLASS WOOL
PYREX - 0.11 kj
Fig. B-l. Schematic of lysimeter containing residential and
commerical wastes.
179
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OFWL-OWG 82-11533
PUMP-COLE PARMER
10-CHANNEL MASTER-
FLEX
TANK-U.S. PLASTICS CORP.
POLYETHYLENE NO. 10120
2082 L
LYSIMETER
GLASS MANIFOLD
f
r—° — '
"~?~
lu"
LS-
-g-
Q
_
--
.--
--
I — O—
— 0—
— 0—
-o-
I
I
] I
1
-raJ
fl
PUMP-COLE PARMER
1-CHANNEL MASTERFLEX
PUMPS-TECHNICON
COLUMNS FOR
INDUSTRIAL
WASTE
OVERFLOW
TO DITCH
-OVERFLOW
COLLECTOR
Fig. B-2. Flow diagram for lysimeter study.
180
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municipal and 5% industrial waste scenario to the laboratory extraction
studies. These columns will contain 3.6 kg of waste and will be leached
with the MWL until a 20:1 liquid to solid ratio is achieved (see
Section 3.3). At the selected flow rate of 0.8 mL/min (equivalent to a
p I
flux of 7 mL m min ) this will require eight weeks (59 d).
Samples of leachate will be collected twice weekly the first four weeks
and once weekly the remaining four weeks. Aliquots will also be
composited to ascertain a concentration of inorganics and organics in
the final 20:1 volume.
3.2.3 Aggressiveness of Municipal Waste Leachate over Time; The
remaining columns (4 wastes x 2 replications) in the lysimeter design
will contain 0.86 kg of waste. Leachate will be collected after 15 d
and fresh industrial waste will be substituted in the column. Again
the final quantity of leachate collected will correspond to the 20:1
liquid to solid ratio. This portion of the experiment is planned to
continue for 90 d (three to four changes of waste). The quantity of
inorganic and organic constituents leached from each column of fresh
waste should provide an evaluation relative to the aggressiveness of
the municipal waste leachate with age of leachate.
3.2.4 Experimental Design and Analysis: The experiment will consist
of a randomized block design with four treatments and two blocks
(lysimeters). The experiment will be replicated twice to test the
block by treatment interaction. An analysis of variance will be
conducted to see if there are any differences in leachate aggressiveness
due to age of leachate. A multivariate pattern analysis will be used
181
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to determine which of the laboratory extraction methods most closely
simulates the chemical constituents derived from the field lysimeters.
The results from the laboratory and field studies will be combined and
statistically analyzed to determine the laboratory method (EP-III) that
most closely simulates the mobility of inorganic as well as organic
constituents of industrial wastes co-disposed with municipal wastes in
a landfill environment.
3.3 Basis of Calculation for Large-scale lysimeter Research
The large-scale lysimeters (1.8 m in diameter and 3.6 m in height,
Fig. B-l) contain approximately 7 m of waste weighing on the order
of 1.5 Mg. The wastes were compacted and covered with 0.4 m of soil to
simulate a landfill. Distilled water will be continuously added to
-21 1 1
each lysimeter at a flux of 7 mL m min (ca. 26.5 L d lysimeter ).
Leachate from each lysimeter will be collected in a 27-L sump from
which leachate will be pumped to 16 individual industrial waste columns
containing the eight industrial wastes replicated two times. Leaching
by the municipal waste leachate will be conducted in a downward
trickling fashion. The flow rate will be the same as that applied to
-2 1
the surface of the large-scale lysimeters (7 mL m min ).
Acid-washed sand will be mixed with the industrial waste (1:1) to
increase the hydrologic conductivity. Leaching will continue until a
20:1 liquid: solid ratio is achieved from these columns. Leachate
will also be pumped from the sump for special laboratory extraction
studies. Any excess leachate will be diverted to ORNL waste treatment
facilities. The basis of calculation is as follows:
182
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2
Area of large-scale lysimeter = 2.54 m
-1 -2
Flux = 7 ml min m
Assume 1.5 Mg of municipal waste/lysimeter, for a 95/5 scenario
(78.9 kg of waste is required).
A borosilicate glass column will be used for the industrial wastes
[15.2-in. (38.7-cm) i.d., 12 in. (30.5 cm) high, cylindrical
jar with a glass outlet being placed on the bottom] will be
2
used. Then the area of each column (0.118 m ) times 16
o
(1.88 m ) represents 74.1% of the area of the lysimeter
(1.88/2.54 x 100). Therefore, 74.1% of the leachate from the
lysimeter will be required to "feed" the 16 columns containing
the industrial wastes. The weight of the industrial wastes in
each column will be 3.61 kg (78.9 kg x 0.741 * 16) for the
laboratory comparison study and 0.8 kg for the columns used to
evaluate the aggressiveness of the MWL with age.
The flow rate to each industrial waste column will be 0.8 ml/min
12 2
(7 ml min m x 0.118 m). Thus, it will require at
this flow rate 59 d to reach a 20:1 solution to solid ratio
(72 L) of column leachate for the comparative study.
Total quantity of each industrial waste required will be as
follows:
Laboratory studies - 11 x 0.1 kg = ~23 kg
Field studies - 3.3(4) + 0.8(16) = ~37 kg
Reserve = "40 kg
Total = ~82 kg
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The above calculations and procedures are basetJ on best available
Information relative to the physical and chemical characteristics of
the wastes and anticipated characteristics of the municipal waste
leachate. Alterations to these procedures may be necessary to obtain
the desired results. For example, mixing the industrial wastes with
acid wash sand is Intended to (1) increase porosity of the waste/sand
mixture so that downflow of the municipal waste leachate can be
conducted and (2) decrease the possibility of "hydraulic short
circuiting" of the leachate through the relatively thin layer
(approximately 2 to 4 cm) of industrial waste. The plan is to mix the
wastes with sand on a 1:1 weight basis. Laboratory tests will be
conducted prior to installation of the field columns relative to the
applicability of the 1:1 mixture to facilitate downward flow of
municipal waste leachate. If 1:1 mixture is not a sufficient quantity
of sand to sustain flow, a 2:1 sand mixture will be used for that
waste. Experiments will also be conducted to evaluate the capacity of
the sand to attenuate the mobility of inorganic and organic toxic
materials in municipal waste leachate.
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4. WORK SCHEDULE
Milestone Projected Date
1. Laboratory study
WES municipal leachate procured Jan. 27, 1982
Industrial wastes (8) procured May 15, 1982
Proximate analysis of wastes initiated May 15, 1982
Select wastes (4) for laboratory and
field studies May 31, 1982
Laboratory extractions started
(88 extractions) June 1, 1982
Laboratory extractions completed Sept. 1, 1982
Inorganic and organic analysis of
extracts completed Sept. 15, 1982
Statistical analyses completed Oct. 15, 1982
2. Field study
Municipal refuse obtained and lysimeters packed Feb. 23, 1982
Industrial waste columns packed for laboratory
comparison and aggressiveness of MWL,
begin leaching June 15, 1982
Laboratory comparison, leaching completed Aug. 10, 1982
Inorganic and organic analyses of leachates
from laboratory comparison completed Sept. 1, 1982
Aggressiveness of MWL, leaching completed Sept. 15, 1982
Inorganic and organic analyses of leachates
from aggressiveness of MWL completed Oct. 1, 1982
Statistical analyses completed, aggressiveness
of MWL Oct. 15, 1982
3. Draft report on laboratory and field studies Jan. 1, 1983
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5. REFERENCES CITED
Bause, D. E., and K. T. McGregor. 1980. Comparison of four leachate-
generation procedures for solid waste characterization in
environmental assessment programs. EPA-600/7-80-118. U.S.EPA,
Washington, D.C.
Epler, J. L. 1980. Toxicity of Leachates. EPA-600/2-80-057.
U.S.EPA, Washington, D.C.
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ATTACHMENT A TO APPENDIX B
PROTOCOL FOR EVALUATING THE AGGRESSIVENESS OF
THREE TREATMENT PROCEDURES
1. Rank the concentrations of each constituent extracted from least to
most aggressive within each waste.
2. Group the constituents into target analysis categories and then sum
the ranks within a category:
NIPDWR inorganics
Toxic organics.
3. Rank the sums within each category.
4. Sum the ranks across categories to obtain an overall rank sum.
5. Repeat 1-4 for all wastes.
6. Statistically analyze the rank sum.
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EXAMPLE OF PROTOCOL FOR EVALUATING THE AGGRESSIVENESS OF THREE
TREATMENT PROCEDURES FOR A PARTICULAR INDUSTRIAL WASTE
TREATMENTS TOXIC ORGANICS NIPPUR INORGANICS
Phenol BAP Hg Cd Pb
1 0.1 0.04 0.5 10 43
2 8.0 0.09 1.6 20 25
3 6.0 0.01 2.0 12 68
STEP 1 - RANK THE CONCENTRATIONS OF EACH CONSTITUENT EXTRACTED
1 12 112
2 33 231
3 21 323
STEP 2 - GROUP THE CONSTITUENTS INTO CATEGORIES AND SUM THE RANKS
TREATMENTS TOXIC ORGANICS NIPPUR INORGANICS
1 3 4
26 6
33 8
STEP 3 - RANK THE SUMS WITHIN EACH CATEGORY
1 1.5 1
23 2
3 1.5 3
STEP 4 - SUM THE RANKS ACROSS CATEGORIES TO OBTAIN AN OVERALL RANK SUM
TREATMENT RANK SUM
1
2
3
2.5
5
4.5
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ATTACHMENT B TO APPENDIX B
DESCRIPTION OF LYSIMETER STUDY
A one-line diagram of the lysimeter study setup is depicted in
Fig. B-2. A detailed explanation follows. The two large-scale
municipal waste lysimeters will be continuously watered using distilled
water contained in a 2080-L polyethylene storage tank (U.S. Plastic
Corp. Cat. No. 10120). Water out of the storage tank will flow from
0.5-in. (1.3-cm) PVC pipe into a glass manifold that will direct the
water into 10 separate master-flex pump channels (Cole Parmer Cat.
No. C-7568-00); the pump will be equipped with pump heads and silicone
tubing (Cole Parmer head #C-7013-20 and tubing #C-6411-41) to
accommodate a flow rate of 3.68 ml min channel . Five channels
will continuously provide water to each lysimeter at 3.68 mL/min
(a total of 18.4 mL/min or 26.5 L/d). Each lysimeter will be covered
with a 0.015-cm thick polyethylene tarp.
Municipal waste leachate (MWL) out of each lysimeter will be
delivered to separate 27-1 glass sumps (30.5 cm o.d. x 45.7 cm high,
Corning No. 6942) through 2.54-cm PVC pipe. A 2.0-cm-thick plexiglass
cover with the underside covered with Teflon* overlay (with holes cut
for inlets and outlets) will be fabricated to fit each sump jar. An
overflow pipe will be connected to the sumps to divert any excess MWL
to ORNL waste treatment facilities; a sampling port will be connected
to the overflow pipe to collect MWL for characterization and/or
laboratory studies. MWL for industrial waste leaching will be pumped
from the sump (30 mL/min) to a glass manifold using a masterflex pump
(Cole Parmer pump no. 7545-10, pumphead no. C-7016-00); excess MWL not
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used for industrial waste leaching will return to the sump. The MWL
will contact Teflon* tubing except through the pump which utilizes
silicone tubing [0.125-in. (0.32-cm) i.d., 0.251-in. (0.64-cm) o.d.,
Cole Farmer Cat. No. C-641102]. The glass manifold will contain inputs
to monitor the pH and Eh of the MWL and will also be fitted with
22 ports (Note: All may not be used) to aliquot the MWL to the
industrial waste columns. Each industrial waste column (16 per
lysimeter plus 2 sand controls) will receive MWL at a flow rate of
0.8 mL/min. The MWL will be pumped, using Technicon pumps (2-speed
proportioning pump) from each manifold port (each channel in the pump
will connect to a port in the manifold) through 0.125-in. (0.32-cm)
o.d. Teflon* tubing connected to Technicon solvent flexible manifold
tubing [0.045-in. (0.11-cm) i.d. Technicon No. 1160533]; 0.125-in.
(0.32-cm) o.d. Teflon* tubing will be connected out of the pump and
will introduce the MWL to the industrial waste columns. The industrial
waste columns are purchased borosilocate glass jars (32-L capacity,
38.7-cm i.d. and 30.5 cm high, Corning No. 6942) with an outlet placed
on the jar bottom. A 2.0-cm-thick plexiglass cover with the underside
covered with Teflon* overlay will be fabricated to seal the jar top.
Leachate out of the industrial columns will flow through Teflon*
tubing into Tedlar sampling bags.
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