U.S. DEPARTMENT OF COMMEHtc
National Technical Information Service
PB-280 959
Source Assessment: Textile
Plant Wastewater Toxics
Study, Phase I
Monsanto Research Corp, Dayton, Ohio
Prepared for
Industrial Environmental Research Lab, Research Triangle Park, N C
Mar 78
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EPA-600/2-78-004h
March 1978 Environmental Protection Technology Series
SOURCE ASSESSMENT:
TEXTILE PLANT WASTEWATER TOXICS STUDY
PHASE I
Industrial Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Research Triangle Park, North Carofina 27711
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development. U.S. Environmental
Protection Agency, have been grouped into five series. These five broad
categories were established to facwitatetfurther development and application of
environmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface irVfelated.fteWft.
The five serf tsar* : •/?•_. •' -' ' "*fl ••••.'•>.
1. ,' Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research "''.;)
4. Environmental Monitoring C
5. Sdctoeconomfc Environmental Studies
This report has been assigned to the ENVIRONMENTAL PROTECTION
TECHNOLOGY series. This series describes research performed to develop ^
This report has been reviewed by 'the U. S. Environmental
Protection Agency, and approved for publication. Approval
does not signify that the contents necessarily reflect the
views and policy of the Agency, nor does mention of trade
names or commercial products constitute,endorsement or
recommendation for use. .-,'
This document is available to the public through the National Technical Informa-
tion Service. Springfield. Virginiap22161.
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
3. REfillE
12.
1. REPORT NO.
EPA- 600/2- 78- 004h
4; TITLE AND SUBTITLE SOURCE ASSESSMENT: Textile Plant
Wastewater Toxics Study--Phase I
5. REPORT DATE
March 1978
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Gary D. Rawlings
8. PERFORMING ORGANIZATION REPORT NO.
MRC-DA-774
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Monsanto Research Corporation
1515 Nicholas Road
Dayton, Ohio 45407
10. PROGRAM ELEMENT NO.
IAB015; ROAP 21AXM-071
11. CONTRACT/GRANT NO.
68-02-1874
12. SPONSORING AGENCY NAME AND ADDRESS
EPA, Office of Research and Development
Industrial Environmental Research Laboratory
Research Triangle Park, NC 27711
13. TYPE OF REPORT AND PERIOD COVERED
Task Final; 1-12/77
14. SPONSORING AGENCY CODE
EPA/600/13
.SUPPLEMENTARY NOTES ffiRL-RTP task officer is Max Samfield. Mail Drop 62. 919/541-
2547. Earlier reports are in the EPA-600/2-76-032 and EPA-600/2-77-107 series.
*s.ABSTRACT, Tne report gives results of the first phase of a study to provide chemical
and toxicological baseline data on wastewater samples collected from textile plants
in the U.S. Raw waste and secondary effluent wastewater samples were analyzed for
129 consent decree priority pollutants, effluent guidelines criteria pollutants, and
nutrients. Level 1 chemical analyses were also performed. Secondary effluent sam-
ples from the 23 plants selected for study in the EPA/ATMI BATEA Study (American
Textile Manufacturers Institute/best available technology economically achievable)
(EPA Grant 804329) were submitted for the following bioassays: mutagenicity, cyto-
toxicity, clonal assay, freshwater ecology series (fathead minnows, Daphnia, and
algae), marine ecology series (sheepshead minnows, grass shrimp, and algae), 14-
day rat acute toxicity, and soil microcosm. The bioassay results indicated that 10 of
the 23 textile plants have secondary effluents sufficiently toxic to proceed to a
second phase of the study. In the second phase, samples will be collected from
these 10 plants to determine the level of toxicity removal attained by selected ter-
tiary treatment technologies.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
Pollution
Textile Industry
Waste Water
Toxicity
Bioassay
Mutagens
Ecology
3 DISTRIBUTION STATEMENT
Unlimited
b.lDENTIFIERS/OPEN ENDED TERMS C. COSATI Field/Group
Pollution Control
Stationary Sources
Toxic Materials
Source Assessment
Baseline Data
Cytotoxicity
Clonal Assay
13B
11E
06T
06A
06E
06F
19 SECURITY CLASS (This Report)
/Unclassified
20 SECURITY CLASS (This page)
Unclassified
- Affil
EPA Form 2220-1 (9-73)
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EPA-600/2-78-004h
March 1978
SOURCE ASSESSMENT:
TEXTILE PLANT WASTEWATER TOXICS STUDY
PHASE I
by
6. D. Rawlings
Monsanto Research Corporation
1515 Nicholas Road
Dayton, Ohio 45407
Contract No. 68-02-1874
ROAP No. 21AXM-071
Program Element No. 1AB015
EPA Task Officer: Max Samfield
Office of Energy, Minerals, and Industry
Industrial Environmental Research Laboratory
Research Triangle Park, North Carolina 27711
Prepared for
U.S. ENVIRONMENTAL PROTECTION AGENCY
Office of Research and Development
Washington, DC 20460
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PREFACE
* v ""•*"
The Industrial Environmental Research Laboratory (IERL) of the
U.S. Environmental Protection Agency" (EPA) has-the responsibility
for insuring that pollution control technology is available for
stationary sources to meet the requirements of the Clean Air Act,
the Federal Water Pollution Control Act, and solid waste legisla-
tion. If control technology is unavailable, inadequate, or
uneconomical, then financial support is provided for the develop-
ment of the needed control techniques for industrial and extrac-
tive process industries. The Chemical Processes Branch of the
Industrial Processes Division of IERL has the responsibility for
investing tax dollars in programs to develop control .technology
for a large number of operations (more than 500) in ilhe chemical
industries. -^^/-
' V^-i'7'"'1"
"*^!t»'>.-
Monsanto Research Corporation (MRC) has contracte.4 'with EPA to
investigate the environmental impact of various industries which
represent sources of pollution in accordance with EPA1s respon-
sibility as outlined above. Dr. Robert C. Binning serves as MRC
Program Manager in this overall program entitled "Source Assess-
ment," which includes the investigation of sources in each of
four categories: combustion, organic materials, inorganic mate-
rials, and open sources. Dr.. Dale A. Denny of the-Industrial
Processes Division at Research Triang'le Park serves- as EPA Pro-
ject Officer. Reports prepared in this program are of three
types: Source Assessment Documents, State-of-the-Art Reports/-
and Special Project Reports.
Source Assessment Documents contain data on emissions from spe-
cific industries. Such data are gathered from the literature,
government agencies, and cooperating companies. Sampling and
analysis are also performed by the contractor when the available
information does not adequately characterize the source emis-
sions. These documents contain all of the information necessary
for IERL to decide whether emissions reduction is required.
State-of-the-Art Reports include data on emissions from specific
industries which are also gathered from the literature, govern-
ment agencies, and cooperating companies. However, no extensive-
sampling is conduc.ted by the contractor for such industries.
Results from such studies are published as State-ofrthe-Art
Reports for potential utility by the government, industry, and
others having specific needs and interests. > h
111
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Special projects provide specific information or services which
are applicable to a number of source types or have special
utility to EPA but are not part of a particular source assess-
ment study. This special project report, "Source Assessment:
Textile Plant Wastewater Toxics, Study, Phase 1,". was prepared
to provide chemical and toxicological data on.wastewater samples
collected from selected textile plants in the United States.
Dr. Max Samfield of the Chemical Processes Branch at IERL-RTP
served as EPA Task Officer.
A second phase of this project is underway to collect samples
of secondary effluents from 10 textile plants.to determine the
level of toxicity removal attained by selected tertiary treat-
ment technologies.
IV
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ABSTRACT
The purpose of this study was to provide chemical and toxicolog-
ical baseline data on wastewater samples collected from textile
plants in the United States. Raw waste and secondary effluent
wastewater samples were analyzed for 129 consent decree priority
pollutants, effluent guidelines criteria pollutants, and nutri-
ents; Level 1 chemical analyses were also performed. Secondary
effluent samples from the 23 plants selected for study in the
EPA/ATMI BATEA Study (American Textile Manufacturers Institute/
best available technology economically achievable) (Grant No.
804329) were submitted for the following bioassays: mutagenicity,
cytotoxicity, clonal assay, freshwater ecology series (fathead
minnows, Daphnia, and algae), marine ecology series '(sheepshead
minnows, grass shrimp, and algae), 14-day rat acute toxicity,
and soil microcosm. Since this was a screening study, samples
of the textile plant intake water were not collected for chemical
analysis.
Based on the bioassay results, 10 of the 23 textile plants were
found to have secondary effluents sufficiently toxic to proceed
to a second phase of the study. In the second phase, samples
will be collected from these 10 plants to determine the level
of toxicity removal attained by selected tertiary treatment
technologies.
This report was submitted in partial fulfillment of Contract
68-02-1874 by Monsanto Research Corporation under the sponsor-
ship of the U.S. Environmental Protection Agency. This report
covers a period from January 1977 to December 1977.
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CONTENTS
Preface
Abstract v
Figures ix
Tables xi
Abbreviations xiv
Acknowledgments - xv
1. Introduction 1
2. Summary 3
3. Results and Recommendations 12
4. Scope of Work I.... 15
Background I. ... 15
Program objective 19
Project organization 20
5. Sampling Procedures 26
Collection technique 26
Sample container preparation 28
Sampling logistics 28
Sample shipping procedures 30
6. Wastewater Chemical Analyses 31
Effluent guidelines criteria pollutants 31
Analysis protocol for the 129 consent decree
priority pollutants 34
Level 1 chemical analysis 48
7. Bioassay of Secondary Effluents 83
Microbiological mutagenicity 84
Cytotoxicity assay 92
Freshwater ecology toxicity 98
Marine ecology toxicity 104
Range-finding acute toxicity 14-day rat test . . Ill
Soil microcosm test 113
8. Discussion of Results 116
Plant ranking by relative wastewater toxicity. . 116
Program outline for Phase II study 118
References 120
Appendices
A. Recommended list of priority pollutants 122
B. Priority pollutant analysis fractions 128
C. Addresses of persons associated with this textile
study i. . . . 131
VII
Preceding page blank
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CONTENTS (continued)
D. Reaction of fathead minnows and daphnia to textile
secondary effluents , 134
E. Reactions of rats to textile plant secondary
effluent 141
F. Protocol to test effects of waste materials on
microbial respiration (carbon dioxide reduction)
in a simple soil microcosm 149
Glossary 150
Conversion Factors and Metric Prefixes . . 152
Vlll
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FIGURES
Number Page
1 Program outline for Phase I: technology assessment
for the ATMI/EPA Grant Study 16
2. Seven tertiary treatment modes for "best available
technology" evaluation 17
3 Overall program approach to determine BATEA 20
4 Program outline for Phase I of the MRC wastewater
I toxicity study 21
5 Program outline for Phase II of the MRC wastewater
toxicity study 21
6 Sampling locations for Phase II of the MRC waste-
water toxicity study 22
7 Interpretation of bioassay test results 22
8 Scope of work for the analysis of textile plant
wastewaters 24
9 EPA-recommended bioassay testing scheme for
toxicity analysis of water samples 24
10 Laboratories and persons involved in sample
analysis of textile plant effluents 25
11 Phase I sampling locations 26
12 MRC sample bottle label 30
13 Analytical scheme for volatile organics analysis. . . 36
14 Sample processing scheme for nonvolatile organics
analysis 36
15 Sample processing scheme for pesticide and PCB
analysis 38
16 EPA-recommended field handling scheme for liquid/
slurry samples 54
17 MRC-modified Level 1 field sampling and analysis
scheme 56
18 Level 1 organic analysis scheme 57
19 EPA, IERL/RTP modification to Level 1 organics
analysis procedure 59
ix
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FIGURES (continued)
Number Page
20 Laboratories and EPA technical advisors involved
in biotesting of effluent samples 86
21 Results of CHO-K1 clonal assay 98
22 Example of graphically interpolated 24-hr, 48-hr,
and 96-hr LC50's for juvenile sheepshead minnows
exposed to textile effluent W 109
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TABLES
Number
1 Summary of Priority Pollutants Found in Raw
Waste Samples, from 129 Total, Showing
Concentration Ranges and Number of Plants
Where the Species were Identified
2 Summary of Priority Pollutants Found in Effluent
Samples, from 129 Total, Showing Concentration
Ranges and Number of Plants Where the Species
were Identified ,
Summary of Metal, Criteria Pollutant, and j
Nutrient Analyses '
4 Purpose of Selected Bioassary Tests in Evaluating
the Potential Toxicity of Secondary Effluents . 8
5 Summary of Biotest Data for Secondary Effluent
Wastewater Samples 10
6 Chemical Compounds as Listed in the Consent
Decree 18
7 Sample Collection and Handling Requirements ... 27
8 Portion of the Field Instructional Form Used by
MRC to Assure Accurate Sample Collection and
Preservation 29
9 Analysis of Wastewater Samples for Effluent
Guidelines Criteria Pollutants 32
10 Summary of Continuous Samples and Volatile
Organic Blank Analyses 39
11 Textile Sample Extraction Dates 40
12 GC/MS Analyses for Base/Neutral Organic Compounds
in Raw Waste and Effluent Samples 41
13 GC/MS Analyses for Volatile, Acid, Pesticide, and
PCB Organic Compounds in Raw Waste and Effluent
Samples 43
14 Number of Priority Organic Pollutants Found in
the Raw Waste and Secondary Effluent Streams. . 47
15 Occurrence of Priority Organic Pollutants t
Combined from Raw Waste and Secondary Effluent
Samples 49
xi
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TABLES (continued) ,
Number : ; Page
~ I
16 Consent Decree Metals, Concentrations in Waste-
water Samples ; 50
i i *
17 Other Metals Given by ICAP Analysis . 52
' i,
18 Lower Detection Limit of Metals Analysis.Systems. 54
19 Selected Parameters of Textile Effluent Samples
from the Inorganic Segment of Level 1 Chemical
Analysis Protocol . ..... 60
20 Field Analysis of Selected Secondary Wastewater
Parameters on Filtered, Unextracted Samples as
per Level 1 Analysis Protocol ......... 61
21 Level 1 Spark Sou'rce Mass Spectrometer Metals
Analysis of the Suspended Solids Collected on
the Filter Paper During Field Filtration. ... 62
' i
22 Level 1 Spark Source Mass Spectrometers Metals
Analysis of Filtered Secondary Effluent .... 67
23 Concentration of Methylene Chloride Extractable
Organics in Filtered Secondary Effluents. ... 72
24 Level 1 Infrared Analysis of the Organic Extracts 73
25 Level 1 Low Resolution Mass Spectrometer Analysis
of Organic Fractions ."...,.... 78
26 Bioassay Tests Used to Evaluate the Toxicity of
Secondary Effluents 85
27 Estimated EC2oiand ECs0 Values for Cytotoxicity
Screening of Filtered Secondary Effluent
Samples 95
28 CHO-K1 Clonal Cytotoxicity Test 97
29 Dilutions Used.for Freshwater Algal Tests .... 99
30 Nutrient Analysis of Secondary Effluent Samples . 100
31 Results of Freshwater Algae Bioassay Tests. . . . 100
32 Relative Ranking of Textile Plants by Tioxic and
Stimulatory Effects of Secondary Wastewater on
S. Capricornutum 102
33 Acute Toxicity Data for Fathead Minnow and
Daphnia Pulex . . . . . . 103
34 Results of Marine Algae Acute Toxicity Tests. . . 105
xia.
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TABLES (continued)
Number
35 Physical Description of Effluent Samples as They
Arrived at BMRL for Sheepshead Minnow and Grass
Shrimp Acute Toxicity Analysis 107
36 Acute Toxicity of 14 Textile Effluents to
Juvenile Sheepshead Minnows 109
37 Acute Toxicity of 14 Textile Effluents to
Grass Shrimp 110
38 Physical Examinations in Acute Toxicity in
Rodents 112
39 Results of Soil Microcosm Tests on Secondary
Wastewater Samples • 115
40 Relative Toxicity Ranking by Bioassay Test ..... 116
41 Prioritization of Textile Plants by Toxicity I
of Secondary Effluent 118
42 Sample Schedule at Each of the 10 Textile
Plants 119
Xlll
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ABBREVIATIONS
ATMI
ATP
BATEA
BOD 5
COD
DO
EC
EC50
GC
GCMA
ICAP
IR
LC
LRMS
MS
PCB
ppt
RAM
SSMS
T/C
TDS
TKN
TOC
TSS
V/V
W/V
American Textile Manufacturers Institute
adenosine triphosphate
best available technology economically achievable
5-day biochemical oxygen demand
chemical oxygen demand
dissolved oxygen
electron capture detector on a gas chromatograph
effective concentration at which 50% of the test
species reach the desired effect
gas chromatograph
gas chromatography mass analysis
inductively coupled argon plasma
infrared analyzer
liquid chromatography
lethal concentration which causes 50% mortality
in the test species
lethal dosage which causes 50% mortality in the
test species
low resolution mass spectrometer
mass spectrometer
polychlorinated biphenyls
parts per thousand
rabbit alvelor macraphage
spark source mass spectrometer
ratio of population density in treated samples
to population density in controls
total dissolved solids
total Kjeldahl nitrogen
total organic carbon
total suspended solids
volume-to-volume ratio
weight-to-volume ratio
xiv
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ACKNOWLEDGMENTS
The author would like to express his gratitude to members of the
American Textile Manufacturers Institute (ATMI), Carpet and Rug
Institute (CRI), and Northern Textile Association (NTA) for their
extensive cooperation in this project.
The author would also like to thank Dr. Max Samfield, EPA Project
Leader on this task, for his guidance and project coordination.
xv
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SECTION 1
INTRODUCTION
The Industrial Environmental Research Laboratory - RTF (IERL-RTP)
of the U.S. Environmental Protection Agency (EPA) is currently
engaged in a joint study with the American Textile Manufacturers
Institute (ATMI) (EPA Grant No. 804329) to determine the best
available technology economically achievable (BATEA) for textile
plant wastewaters. A total of 23 textile mills representing
eight textile processing categories and having well-operated
secondary wastewater treatment facilities were selected by EPA
and ATMI for the BATEA study. For that study, two mobile pilot
plants containing four tertiary wastewater treatment*technologies
were constructed to gather technical data to identify the best
available technology applicable to the 23 plants. Two additional
tertiary treatment technologies were tested in the laboratory.
The grant study focused on only a limited number of so-called
criteria pollutants; i.e., 5-day biochemical oxygen demand, chem-
ical oxygen demand, color, sulfides, total suspended solids,
phenol, and pH.
However, on 7 June 1976 the U.S. District Court of Washington,
D.C., issued a consent decree (resulting from Natural Resources
Defense Council et al. v. Train) requiring EPA to enhance devel-
opment of effluent standards. The court mandate focused federal
water pollution control efforts on potentially toxic and hazard-
ous pollutants. In response to the consent decree EPA developed
a list of 129 specific compounds (known as priority pollutants)
that the agency agreed to consider during the standards setting
process. Based on the consent decree, EPA-IERL/RTP decided to
conduct a study parallel to the ATMI/EPA Grant Study of the
textile-industry. The objective of the IERL/RTP study was to
determine both the removal efficiencies for the 129 consent
decree priority pollutants and the reduction in toxicity by the
six tertiary treatment technologies being investigated under
the original grant study.
The overall wastewater toxicity study is divided into two phases.
The first, covered by this report, establishes a baseline data
base concerning toxicity and level of priority pollutants pres-
ent in raw wastewater and secondary effluents at 23 textile
plants. These data are used to screen the 23 plants and to
select those plants with secondary effluents of highest toxicity
for further study. Toxicity tests were designed to evaluate
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only the reduction in wastewater toxicity by control technolo-
gies, not the potential environmental impacts on receiving
waters.
The second phase of the effort, to be covered in a subsequent
report, will determine the reduction in priority pollutant con-
centrations and in toxicity by the mobile pilot plant tertiary
treatment systems. Only those plants selected in the first phase
of the study will be investigated.
Covering the first phase of the toxics study, this report de-
scribes sampling, chemical analysis, and bioassay procedures
used to establish baseline data. Chemical analyses of raw waste
and secondary effluents are presented for the 23 basic plants
and for 9 additional textile plants. Bioassay data arer presented
for secondary effluents from the basic 23 plants.
The plants are ranked according to relative secondary effluent
toxicity, and a number of plants are selected for study in the
second phase of the overall program. Modifications and recom-
mendations for improvements to the sampling, chemical analysis,
and bioassay protocols are also discussed.
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SECTION 2 °
SUMMARY
The purpose of this Phase I study was to provide chemical and
toxicological baseline data on wastewater samples collected
from selected textile plants in the United States. Raw waste
(untreated wastewater) and secondary effluent (wastewater from
secondary wastewater treatment facilities) samples were collected
from the 23 textile plants selected for the ATMI/EPA Grant Study
(Grant No. 804329) and from 9 additional textile plants. Since
this was a screening study, samples of the textile plant intake
water were not collected for chemical analysis.
«'
Samples were analyzed for 129 consent decree priority pollutants
and for effluent guidelines criteria pollutants. The Level 1
chemical analytical scheme developed by EPA-IERI/RTP was employed
to detect other possible pollutants. Nutrient levels were
measured at 23 of'the 32 plants to supplement interpretation of
algal bioassays. The following bioassays were performed on
secondary effluent samples from the 23 plants chosen for the
ATMI/EPA Grant Study: mutagenicity, cytotoxicity, clonal assay,
freshwater ecology series (fathead minnows, Daphnia, and algae),
marine ecology series (sheepshead minnows, grass shrimp, and
algae), 14-day rat acute toxicity, and soil microcosm.
Grab samples and 8-hr continuous samples were collected both
before and after the wastewater treatment system at each of the
32 plants. Samples were stored in ice at 4°C and shipped by air
freight to the laboratories for analysis. Chemical analyses and
bioassays were performed at eight EPA and commercial laboratories
Analysis for the 129 priority pollutants in raw waste and
secondary effluent samples (totaling 64 samples) was performed
by Monsanto Research Corporation (MRC). Analytical procedures
followed those recommended by EPA. However, the recommended
analytical protocol for priority pollutant analysis is still
in the developmental stage and requires further verification
and validation. Consequently, the analytical results of textile
wastewater samples must be looked upon as good estimates of
which priority pollutants are present, with concentrations
accurate to within an order of magnitude. .
The EPA analytical protocol divided the 129 priority pollutants
into 5 fractions for analysis: volatile compounds, base/neutral
compounds, acid compounds, pesticides and polychlorinated
-------
biphenyls (PCB), and metals; EPA recommended that laboratories
not acquire analytical standards for 2,3,7,8-tetrachldrodibenzo-
p-dioxin (TCDD) because of its extreme toxicity. Asbestos was
not analyzed due to the presence of interfering fibrous mate-
rials in textile wastewaters.
t
A summary of the organic compounds found in the 32 raw waste and
32 secondary effluent samples is given in Tables 1 and 2. Of
the 114 organic compounds on the priority pollutant list, a total
of 45 different compounds were found, 39 in raw waste .samples and
34 in secondary effluent samples. On an individual plant basis
the greatest number of organic compounds found in a raw waste
and in a secondary effluent sample were 14 and 8, respectively,
with an average number per plant of 7 in the raw waste and 5 in
the secondary effluent. The predominant compounds were bis(2-
ethylhexyl) phthalate in 54 samples (0.5 mg/m3 to 300 mg/m3),
toluene in 44 samples (0.4 ;mg/m3 to 300 mg/m3), and ethylbenzene
in 30 samples (0.7 mg/m3 to 3,000 mg/m3). '
A summary of the 13 priority pollutant metals and cyanide con-
centrations ^in raw waste and secondary effluent samples is given
in Table 3, 'which also summarizes the criteria pollutant and,
nutrient concentrations for secondary effluent samples. Nutrient
analyses were performed to support freshwater algae bioassays.
On an individual plant basis it was frequently observed,
especially for the metals data, that the concentration of a
specific pollutant was greater in the secondary effluent sample
than in the raw waste sample. This phenomenon is due, in part,
to the hydraulic retention time of the wastewater treatment
facility. Since raw waste and secondary effluent samples were
collected simultaneously, concentrations in the secondary efflu-
ent were due to raw waste loads that entered the treatment sys-
tem 1 day to 30 days prior to sampling. The average'retention
time for the 32 plants was about 5 days.
Level 1 chemical analyses were performed on secondary effluent
samples from 15 of the 23 basic textile plants. Level 1 proto-
col identifies classes of compounds present in environmental
samples and measures the general concentration range. Results
indicate that total concentration of methylene chloride extract-
able organics ranges from 3 g/m3 to 64 g/m3. This value is 5 to
10 times lower than the range for total organic carbon (Table 3).
In the Level 1 procedure each sample was fractionated by a liquid
chromatography column into eight fractions based on polarity.
Infrared analysis of each fraction indicated the presence of
aliphatic hydrocarbons, esters and acids, aromatic compounds,
phthalate esters, and fatty acid groups. Low resolution mass
spectrophotometric analysis of the eight fractions of each sample
detected the following types of compounds: paraffin'ic/olefinic,
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TABLE 1. SUMMARY OF PRIORITY POLLUTANTS FOUND IN RAW WASTE SAMPLES,
FROM 129 TOTAL, SHOWING CONCENTRATION RANGES AND NUMBER OF
PLANTS WHERE THE SPECIES WERE IDENTIFIED
Volatile organic
Compound
Toluene
Benzene
Chloroform
Chlorobenzene
Ethylbenzene
Trichlorofluorome thane
1,1, 1-Tr ichloroe thane
Trichloroethylene
1,1,2, 2-Tetrachloroethylene
Trans- 1 , 2 , -dichloroethylene
1 , 1 -Dichloroethane
1 , 2-Dichloropropane
Cis-1, 3-dichloropropene
Number
of times
found
22
4
12
6
20
2
5
8
8
1
2
1
1
Concentration
range, mg/m3
2 to 300
5 to 200
2 to 500
1 to 300
0.7 to 2,800
30 to 50
2 to 300
2 to 200
15 to 1,100
2
0.6 to 4
2
2
Acid organic
Phenol
Phentachlorophenol
2 -Ni trophenol
p-Chloro-m-cresol
4-Nitrophenol
2,4, 6-Trichlorophenol
2-Chlorophenol
19
8
1
1
1
2
1
0.5 to 100
2 to 70
70
5
70
0.7 to 20
130
Base/neutral
Compound
Naphthalene
Dimethyl phthalate
Diethyl phthalate
Bis (2-ethylhexyl)
phthalate
1 , 4-Dichlorobenzene
1,2, 4-Trichlorobenzene
1 , 2-Dichlorobenzene
Anthracene
Pyrene
Acenaphthene
Di-n-butyl phthalate
Fluorene
Hexachlorobenzene
N-Nitrosodiphenylamine
2 , 6-Dinitrotoluene
Indeno ( 1 , 2 , 3-cd ) pyr ene
organic
Number
of times
found
20
5
12
27
5
8
8
1
1
7
6
2
2
1
1
1
Concentration
range, mg/m3
0.03 to 300
3 to 110
0.2 to 70
0.5 to 300
1 to 210
30 to 440
0.1 to 300
0.1
0.9
9 to 270
2 to 23
5 to 15
0.5 to 2
11
50
2
Pesticide
3-BHC
Heptachlor
„
1
1
0.4
6
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TABLE 2. SUMMARY OF PRIORITY POLLUTANTS FOUND IN EFFLUENT SAMPLES,
FROM 129 TOTAL, SHOWING CONCENTRATION RANGES AND NUMBER
OF PLANTS WHERE THE SPECIES WERE IDENTIFIED
Volatile organic
Compound
Toluene
Benzene
Chloroform
Chlorobenzene
Ethylbenzene
Tr ichlorof luorome thane
Trichloroethylene
1,1,2, 2-Tetrachloroethylene
Cis-1 , 3-dichloropropene
Trans- 1 , 3-dichloropropene
Bromodichlorome thane
Number
of times
found
22
2
5
2
10
6"
2
3
1
2
1
Concentration
range
0.4
0.5
5
4
0.7
2
5
0.4
0.9
, mg/m3
to
to
to
to
to
to
to
to
6
to
2
110
60
60
30 *
3,000
2,100
80
40
4
Acid organic
Phenol
2 , 4-Dimethylphenol
p-chloro-m-cresol
2,4, 6-Tr ichlorophenol
Chloro cresol
2-Chlorophenol
2
2
1
1
1
1
2
8
to
to
2
20
30
10
3
9
Base/neutral
organic
Number
of times
Compound
Naphthalene
Dimethyl phthalate
Diethyl phthalate
Bis (2-ethylhexyl) phthalate
1 , 4-Dichlorobenzene
1,2, 4-Trichlorobenzene
1 , 2-Dichlorobenzene
Anthracene
N-Nitroso-di-n-propylamine
Pyrene
Acenaphthene
Di-n-butyl phthalate
Hexachlorobenzene
Butylbenzyl phthalate
found
5
3
9
27
3
6
5
1
2
4
2
3
3
1
Concentration
range, mg/m3
0.5
0.2
0.5
3-
0.05
2
0.2
2
0.1
0.5
4
0.3
•
to
to
to
to
to
to
to
4
to
to
to
to
to
70
250
1
10
230
2
920
25
20
0.3
2
60
0.8
Pesticides
a-BHC
Heptachlor
1
1
0.3
2
-------
TABLE 3. SUMMARY OF METAL, CRITERIA POLLUTANT, AND NUTRIENT ANALYSES
Element
Antimony
Arsenic
Beryllium
Cadmium
Chromium
Copper
Cyanide
Lead
Mercury
Nickel
Selenium
Silver
Thallium
Zinc
Metal
Concentration
Raw waste
sample
0.0005 to 0.06
0.005 to 0.2
<0.0001
0.0005 to 0.05
0.0002 to 0.9
0.0002 to 2.4
0.004 to 0.2
0.001 to 0.2
0.0005 to 0.004
0.01 to 0.2
<0.005
0.005 to 0.1
<0.005
0.03 to 8.0
range , g/m3
Secondary
effluent sample
0.0005 to 0.07
0.005 to 0.02
<0.0001
0.0005 to 0.01
0.0002 to 2.0
0.0002 to 0.3
0.004 to 0.2
0.001 to 0.2
0.0005 to 0.0009
0.01 to 0.2
<0.005
0.005 to 0.1
<0.005
0.07 to 38
Criteria pollutant3
Concentration
Pollutant range , g/m3
BOD5 <5 to 170
CODC 45 to 1,600
d
Color (APHA) 10 to 2,500
Sulfide 0.01 to 6
Phenol 0.01 to 0.2
TSS 0.02 to 580
pH 5.8 to 10
Nutrient
Concentration
Parameter range, g/m3
Nitrite 0 to 17
Nitrate 0.002 to 40
Ammonia 0.02 to 14
TKN6 2 to 40
o-Phosphate 0.02 to 11
Total phosphorus 0.4 to 15
TOC9 19 to 260
For secondary effluent samples.
5-day biochemical oxygen demand.
"Chemical oxygen demand.
American Public Health Association color standards.
Total Kjehldehl nitrogen.
f • m *~
Total suspended solids.
Total organic carbon.
-------
bis(hydroxy-t-butyl phenol) propane, tri-t-butyl benzene, alkyl
phenols, dichloroaniline, toluene-sulfonyl groups, vinyl
stearate and azo compounds. r
Bioassays used were selected by EPA and include tests for assess-
ment of both health and ecological effects. Health-effects tests
estimate the potential mutagenicity, potential or presumptive
carcinogenicity, and potential toxicity of the samples to mamma-
lian organisms. Ecological effects tests focus on the potential
toxicity of the samples to vertebrates (fish) ,' invertebrates
(daphnids and shrimp), and plants (algae) in freshwater, marine
and terrestrial ecosystems. ;
i
A total of 8 bioassay systems were tested using 21 different
tester organisms to evaluate the toxicity of secondary effluents.
Table 4 lists the bioassays used and the purpose of' each test.
TABLE 4. PURPOSE OF SELECTED BIOASSAY TESTS iIN EVALUATING THE
POTENTIAL TOXICITY OF SECONDARY EFFLUENTS
Bioassay system
Test organism
Purpose of test
Microbial mutagenicity
Cytotoxicity
Freshwater and marine
static bioassay
Freshwater and marine
algal assay
Range finding acute
toxicity
Nine different strains
of bacteria and one
of yeast.
Rabbit alveolar cells
and Chinese hamster
ovary cells.
Fathead minnow, daph-
nids , sheepshead
minnows, and grass
shrimp.
Freshwater and marine
algae.
Young adult rats.
Terrestrial ecology
Soil microorganisms.
To determine if a chemical mutagen
(possibly a carcinogen) is present.
These microbial strains were selected
because of their sensitivity to
various classes of chemical compounds.
To measure metabolic impairment and
death in mammalian cells. These pri-
mary cell cultures have some degree of
metabolic repair capability.
To detect potential toxicity to organisms
present in aquatic environments.
To detect potential growth inhibition
and-stimulation effects on aquatic
plants.
Whole animal test to detect potential
toxic effects to mammals. These live
animals were selected because of the
extensive data base Ion their response
to known chemicals and because they
have several metabolic systems closely
approximating those in humans.
To determine potential inhibition and
stimulation;effects on soil micro-
organisms. These data are useful if
the effluent is used for crop
irrigation.
-------
A summary of the bioassay results is presented in Table 5. Toxi-
city is expressed as the percent of a secondary effluent solution
that will cause the effect specified for each bioassay over the
testing period. For the cytotoxicity, Daphnia and algal bio-
assays an Effective Concentration 20 or 50 (£€20 or ECso) was
calculated. EC2o for the cytotoxicity test means the concentra-
tion of secondary effluent which impairs metabolic processes in
20% of the test cells.
The viability test is a measure of the cells' ability to survive
exposure to the sample, and the adenosine triphosphate (ATP) test
measures the quantity of the coenzyme ATP produced, indirectly
measuring cellular metabolic activity.
ECso for the algal tests means the concentration of secondary
effluent which causes a 50% reduction in algal growth as compared
to a control sample. The freshwater algae test was performed
over a 14-day period and the marine algae test over a 96-hr
period.
For the fathead minnow, sheepshead minnow, and grass shrimp bio-
assays, death was used to measure toxicity, which was expressed
as Lethal Concentration 50 (LCs0). LC50 indicates the calculated
concentration of secondary effluent that is expected to cause the
death of 50% of the test species. Since rats were given a spe-
cific quantity of secondary effluent, toxicity was expressed as
Lethal Dose 50 (LD50). LD50 indicates the quantity of material
fed to the rats that resulted in the death of 50% of the test
animals.
The measure of toxicity to a soil microcosm was the quantity of
carbon dioxide (C02) produced after sample exposure as compared
to a control sample. The quantity of C02 produced over a 3-wk
period after subtracting the quantity produced by the control
was plotted on graph paper. The slope of the curve then repre-
sented the rate of increase or decrease in CC>2 production due to
addition to the sample.
Based on the bioassay results, the EPA Bioassay Subcommittee
ranked the 23 plants according to their overall secondary efflu-
ent toxicity. The following nine textile plants were selected
for further study based on their relatively high ranking: N, A,
L, T, C, P, S', V, and W. Note the low toxicity response for
Plant Y where the effluent samples were collected after the
polishing pond.
During the second phase of this program the secondary effluents
from these plants will be treated using the mobile pilot plants
constructed under the ATMI/EPA Grant Study. Since effluent
samples $ere inadvertently collected between the aeration lagoon
and settling basin at Plant R, it will also be included in the
Phase II program. While each of these 10 plants is being tested
-------
TABLE 5\ SUMMARY OF BIOTEST DATA FOR SECONDARY EFFLUENT WASTEWATER SAMPLES
a ,b
Freshwater ecology series
Cytotoxicity
viability
(24-hr EC20),
% secondary
Plant effluent
A
B
C
D
E
F
G
H
J
K -
L
H
N
p.
R1
S
T
0
V
H
X
Y
Z
NAT
NAT .
16.8
NAT
NAT
NAT •
NAT
NAT
NAT
NAT
NAT
NAT
13.3
NAT
NAT
NAT
NAT
NAT
NAT
NAT
NAT
NAT
Fathead
ATP minnow
(24-hr EC20>, (96-hr LC50)
% secondary % secondary
effluent effluent
NAT
NAT.
6.l"
RAT
NAT
9.4
NAT
NAT
NAT
NAT
4.0
NAT
3.8
NAT
NAT
2.5
NAT
NAT
13.7
4.8
NAT
NAT
19.0
NAT
46.5
NAT
NAT
NAT
"Jc7
NAT
NAT
23.5
NAT
48.8
NAT
16.5
NAT
46.5
NAT
36.0
55.2
NAT
NAT
NAT
Daphnza Algae
(48-hr ECSO), (14-day EC20),
% secondary % secondary
effluent ' effluent
9.0
NAT
41.0
NAT
7.8
81.7
62.4
40% dead at 100%
concentration
NAT
NAT
28.0
60.0
100% dead at all
dilutions
NAT
8.0
NSAm
NAT
12.1
9.4
6.3
NAT
NAT
42.6
76
30
fli
°3
°!
o1
96
o1
42
0'
2
43
°1
°1
°i
94 1
o1
18
Marine ecology series
Sheepshead
minnow
(96-hr KSO) .
% secondary
effluent
62.0
NAT
69.5
_T
NAT
NAT
NAT
NAT
NAT
_f
47.5
™^
_f
NAT
68.0
NAT
37.5
NAT
T
Grass
shrimp
(96-hr LC50).
% secondary
effluent
21.2
NAT
12t8
_f
NAT
- NAT
NAT
NAT
MAT
_T
26.3
™"^
T
NAT
34. S
NAT
19.6
NAT
I
Algae0
(96-hr ECso),
% secondary
effluent
_f
_9
90
Jf
10 to 50
85
!?
77
17
_f
2.3
9*0
_ t
.9
70
-9
94
50
-9
I
Soil microsm,
normalized
relative COs
_rat« change''
-0.032
0.020
-0.005
-0.099
-0.048
-0.039
0.017
-0.083
-0.163
-0.004
-0.020
-0.059
0.059
0.022
-0.062
-0.017
0.020
0.05S
-0.066
0.031
0.047
-0.172
-0.112
No chemical mutagen was detected by the 10 microbial strains.
Ho rat mortality after 14 days due to maximum dosage of 10"5 m3/kg
body weight (LDso). However, six samples (B, F, L, N, and S) showed
potential body weight effects, and sample R resulted in eye irritation.
Effect was algal growth inhibition.
Negative sign indicates inhibition in CO2 generation rate compared to a
control sample; positive number indicates CO2 stimulation.
No acute toxicity.
Analysis not performed on this sample.
'Growth inhibition <50% in 100% solution of secondary effluent.
pH - 9.1 not adjusted before testing.
Sample stimulated algal growth.
•'95% growth inhibition in 2% solution of secondary, effluent.
Diseased batch of fish nullified this analysis.
Sample inadvertently collected prior to settling pond.
NO statistical analysis because heavy solids concentration obscured
the analysis; the sample did not appear to be acutely toxic.
-------
to determine BATEA, samples will also be collected from the
"best" treatment system to evaluate the reduction in acute toxi-
city. Samples will also be collected to measure removal effi-
ciencies for the 129 priority pollutants. Textile plant intake
water samples will also be collected and analyzed for the prior-
ity pollutants.
11
-------
SECTION 3
*
RESULTS AND RECOMMENDATIONS
Several results can be noted from the data presented with respect
to textile plant secondary effluents.
1. None of the secondary effluent samples resulted in
a positive mutagenic response or indicated acute
toxicity to rats.
2. Even though the series of bioassays are unrelated
in terms of toxicity mechanisms, the data did
provide sufficient information to relatively rank
secondary effluents in terms of toxicity and to
select those plants for further study under the
tertiary treatment technology assessment.
3. The effluent sample collected from Plant R was
inadvertently collected between the aeration lagoon
and settling basin. Therefore the sample tested
was that of the aeration lagoon slurry. Note that
for bioassays where'the sample was first filtered
(cytotoxicity and freshwater algae), the toxicity
was slight; for unfiltered samples (fathead minnow,
.daphnia, and soil microcosm) the toxic effects were
significant. This result tends to imply that the
| toxicity for Plant R is associated with the filter-
able solids. Plant R will be resampled in Phase II
of the program to further evaluate the toxicity
response.
4. In terms of priority pollutants, only 56 of the 114
organic species were detected in either raw waste or
secondary effluent sample, with 49 found in the raw
waste samples and 46 in the secondary effluent samples.
The dominant organic species present in secondary
effluent samples include common plasticizer species,
such as phthalates, and common raw materials used
by the textile manufacturing industry; such as
chlorobenzenes.
12
-------
5. Of the 13 priority pollutant metals, beryllium,
selenium, and thallium were below analytical
detection limits in all raw waste and secondary
effluent samples. The dominant metal species
detected were arsenic, chromium, copper, and zinc.
6. The data indicates that secondary treatment by
aeration lagoons and clarifiers produce a signi-
ficant decrease in phenolic compounds and toluene
and a moderate reduction in chlorobenzenes.
7. It is difficult to accurately evaluate the treat-
ment efficiencies because for this program raw
waste and secondary effluent samples were collected
simultaneously. Due to the 1-day to 30-day hydraulic
retention time in the treatment plants, secondary
effluent samples reflect the waste loading 1-day to
30-day in the past. Also, data are not available to
indicate the quantity of organic compounds stripped
from the wastewater due to the action of surface
aerators.
As a result of performing newly developing bioassay tests, prior-
ity pollutant analyses, and Level 1 chemical analyses on environ-
mental samples, several recommendations can be made with respect
to improvements. The major recommendations are discussed below:
1. For bioassay screening purposes, it proved to be
more economical to conduct mutagenicity and rat
acute toxicity tests using the maximum dose. If
no toxicity was indicated, then the test was com-
pleted; when toxicity was detected, then the dose
response testing scheme was used.
2. Toxicity testing should be performed on filtered
and unfiltered samples. Also, due to the detec-
tion limits of the mutagenicity, cytotoxicity, and
rat acute toxicity tests, consideration should be
given to testing concentrated samples.
3. Whenever extractions are performed on environmental
samples, the percent of extractable organics should
be determined. For the Level 1 chemical analysis
scheme, potentially more organics can be extracted
in methylene chloride if the sample pH is adjusted
to pH 10 or 11 as opposed to use of neutral pH.
Since the Level 1 chemical analysis scheme requires
field extractions, a presurvey and preliminary tests
should be performed to determine the extractability
of the sample and to identify potential problems.
13
-------
4. For priority pollutant volatiles analysis, MRC
obtained better results by cryogenic trapping at
-40°C than at room temperature as recommended by
EPA. In addition, to reduce interference effects,
three internal standards were used as opposed to
the one recommended.
5. To identify the source of priority pollutants in
wastewater samples, samples of the intake water
to the industry complex should be analyzed.
14
-------
SECTION 4
SCOPE OF WORK
BACKGROUND
To understand the nature and purpose of the textile wastewater
toxics program it is first necessary to briefly review the
events which formed the study's foundation. The principal event
occurred on 1 October 1974 when the American Textile Manufac-
turers Institute (ATMI) filed a petition with the U.S. Fourth
Circuit Court of Appeals asking for review of the 1983 effluent
guidelines proposed for the textile industry. ATMI's grounds
for the suit were that the best available technology economically
achievable (BATEA) had not been demonstrated for the textile
industry. As a result, ATMI and EPA filed a joint motion for
delay of the petition, stating that additional information would
be developed through a cooperative study by ATMI and EPA (IERL/
RTP) .
The objective of this ATMI/EPA Grant Study was to gather enough
technical and economic data to determine what is the BATEA for
removing criteria pollutants from textile wastewaters. Criteria
pollutants for the textile industry include 5-day biochemical
oxygen demand (BOD5), chemical oxygen demand (COD), color, sul-
fide, pH, chromium, phenol, and total suspended solids (TSS).
On 3 January 1975 the court instructed ATMI and EPA to proceed
as promptly as feasible to a completion and review of the study.
The ATMI/EPA Grant Study was divided into two phases: Phase I,
to determine the best available technology, and Phase II, to
determine which technology(s) was economically achievable. A
generalized program outline of Phase I is shown in Figure 1. To
evaluate the best available technology, two mobile pilot plants
were constructed by ATMI. This strategy allowed for real-time, -
flowthrough treatment studies. Each pilot plant contained four
tertiary wastewater treatment unit operations; one was scheduled
to visit 12 textile plants and the other to visit 11. Two addi-
tional tertiary treatment technologies were laboratory tested.
i
Treatment operations in each mobile unit include a reactor/clari-
fier (using combinations of alum, lime, ferric chloride, and
anionic:and cationic polyelectrolytes), two multimedia filters,
three granular activated carbon columns, ozonation and dissolved
air flotation. Powdered activated carbon treatability tests
15
-------
/"iATMI/EPA GRANT STUDY
I PHASE I
CONSTRUCT TWO PILOT PLANTS
CONTAINING 6 TERTIARY
TREATMENT UNIT OPERATIONS
COLLECT FIELD DATA FOR 4 WEEKS
ON THE POLLUTANT REMOVAL EFFICIENCIES
OF 7 SaECTED TREATMENT SCHEMES
AT EACH OF THE 23 PLANTS
IS THE
DATA SUFFICIENT
TO DETERMINE THE ONE
BEST TECHNOLOGY?
SET UP THE "BEST TECHNOLOGY
' SYSTEM FOR 2 WEEKS
OF OPERATIONAL EVALUATION
IS THE
DATA ADEQUATE FOR
COMPLETE TECHNICAL
EVALUATION ?
IS THIS
THE LAST OF THE
23 PLANTS
SUBMIT DATA
TO PHASE 11 ECONOMIC STUDY
Figure 1. Program outline for Phase I: technology
assessment for the ATMI/EPA Grant Study.
16
-------
were performed in the laboratory instead of -in the field with
the pilot plant. Using these six unit operations ATMI and EPA
selected seven treatment systems for evaluation (Figure 2).
MODE A : REACTOR / CLARIFIER —- MULTIMEDIA FILTER
MODE B : MULTIMEDIA FILTER —*- GRANULAR ACTIVATED CARBON COLUMNS
MODEC: MULTIMEDIA FILTER —- OZONATOR
MODED: OZONATOR
MODE E : REACTOR / CLARIFIER —-MULTIMEDIA FILTER — GRANULAR ACTIVATED
(OPTIONAL) CARBON—-OZONATOR
MODEF: COAGULATION —MULTIMEDIA FILTER
MODEG: DISSOLVED AIR FLOTATION
Figure 2. Seven tertiary treatment modes for "best
available technology" evaluation.
Each of the seven treatability systems was to be set up, and
operational and pollutant data were to be collected over a 2-day
to 3-day period. Based on that data, the "best" system was to
be selected and set up for 2 weeks of operational evaluation.
These data were then to be forwarded to Phase II for economic
evaluation.
The second event that formed the foundation for this project
occurred when a group of environmental action organizations filed
a class action suit against EPA (Natural Resources Defense Coun-
cil et al. v. Train, U.S. District Court of Washington, D.C.) for
not developing and promulgating regulations establishing effluent
limitations and guidelines and new source performance standards
for 21 industrial point sources, including the textile industry.
As a result, on 7 June 1976 the court issued a consent decree
requiring EPA to enhance development of effluent standards.
The most significant result from the court mandate was that it
focused federal water pollution control efforts on potentially
toxic and hazardous pollutants. The original consent decree
required that 38 classes of chemical compounds (Table 6) be
analyzed in wastewater samples. Recognizing the difficulty of
analyzing for all chemical species present in each category Qf
compounds, EPA developed a list of 129 specific compounds (Appen-
dix A) representative of the classes of compounds listed in the
consent decree. These compounds are referred to as the consent
decree priority pollutants, or priority pollutants for short.
17
-------
TABLE 6. CHEMICAL COMPOUNDS AS LISTED IN THE CONSENT DECREE
Acenaphthene
Acrolein
Acrylonitrile
Aldrin/Dieldrin
Antimony and compounds
Arsenic and compounds
Asbestos
Benzene
Benzidine
Beryllium and compounds
Cadmium and compounds
Carbon tetrachloride
Chlordane (technical mixture
and metabolites)
Chlorinated benzenes (other than
dichlorobenzenes)
Chlorinated ethanes (including
1,2-dichloroethane, 1,1,1-tri-
chloroethane, and hexachloro-
ethane)
Chloroalkyl ethers (chloromethyl,
chloroethyl, and mixed ethers)
Chlorinated naphthalene
Chlorinated phenols (other than
those listed elsewhere; in-
cludes trichlorophenols and
chlorinated cresols)
Chloroform
2-Chlorophenol
Chromium and compounds
Copper and compounds
Cyanides
DDT and metabolites
Dichlorobenzenes (1,2-,1,3-,
and 1,4-dichlorobenzenes)
Dichlorobenzidine
Polychlorinated biphenyls (PCB)
Polynuclear aromatic hydrocar-
bons (including benzanthra-
cenes, benzopyrenes, benzo-
fluoranthene, chrysenes,
dibenzanthracenes, and
indenopyrenes)
Selenium and compounds
Silver and compounds
2,3,7,8-Tetrachlorodibenzo-p-
dioxin (TCDD)
Tetrachloroethylene
Thallium and compounds
Toluene
Toxaphene
Trichloroethylene
Vinyl chloride
Zinc and compounds
EPA also developed a sampling and analytical procedures manual to
be used as a laboratory guide for the analysis of priority pol-
lutants (1). The analytical methods recommended by EPA are still
in the developmental phase and require further verification and
validation.
Therefore, in addition to evaluating the removal of criteria pol-
lutants by tertiary treatment technologies, EPA was charged with
the task of evaluating the removals of toxicity and priority
pollutants by the treatment systems.
(1) Draft Final Report: Sampling and Analysis Procedures for
Screening of Industrial Effluents for Priority Pollutants.
U.S. Environmental Protection Agency, Cincinnati, Ohio,
March 1977. 145 pp.
18
-------
The final event which influenced the formation of the present
program was the three-phase .sampling and analytical-strategy for
environmental assessment developed by EPA, Process Measurements
Branch, IERL/RTP. The purpose"of the assessment procedure was
to determine in a stepwise and cost-effective manner all chemical
species being discharged to the environment from a point source.„
Level 1, the first part of-the three-phase approach, is dejgi^f^f.
to focus available resources on emissions that have a;r.highf;p$.t&n^'
tial for causing measurable health or ecological effects.
The second phase, Level 2, has as its goal the identification and
quantification of specific compounds. Level 3 is designed to
continuously monitor indicator compounds as surrogates for a
large number of specific .pollutants.. At the start of this tex-
tile project, only Level 1'analytical and biological procedures
were available (2) . . ' .. ...
In addition to chemical analyses, the Level 1 recommended proto-
col included bioassay testing procedures for evaluating toxicity
removal by control technologies (3). Bioassays are required to
provide direct evidence of complex biological effects suc)i:.aS
synergism, antagonism, and bioayailability. .j;
PROGRAM OBJECTIVE
The fundamental objective of the textile wastewaters program
conducted by MRC in conjunction with the EPA is to determine the
reduction in toxicity and priority pollutant concentrations
achieved by the tertiary treatment technologies under investi-
gation in the ATMI/EPA'Grant Study. The latter study focuses
directly on the treatabiiity of criteria pollutants. Thus, the
overall EPA-IERL/RTP textile program consists of two separate
projects, each with different activities, running parallel in
time, but converging towards the same goal: determination of
the best available technology economically achievable for remov-
ing textile wastewaters (Figure 3).
To evaluate the reduction in toxicity in a cost-effective manner
for the MRC/EPA project, a two-phase approach was developed.
Phase I was designed to collect baseline toxicity data on
(2) Hamersma, J. W., S. L. Reynolds, and R. F. Maddalone. IERL-
RTP Procedures Manual: Level 1 Environmental Assessment.
EPA-600/2-76-160a (PB 257 850'), U.S. Environmental Protection
Agency, Research Triangle Park, North Carolina, June 1976.
147 pp.
(3) Duke, K. M., M. E. Davis, and A. J^ Dennis. IERL-RTP Proce-
dures Manual: Level 1 Environmental Assessment Biological
Tests for Pilot Studies. EPA-600/7-77-043 (PB 268 484),
U.S.-Environmental^Protection Agency, Research Triangle
Park, North Carolina, April 1977. 114 pp.
' 19
-------
DETERMINE
BATEA
FOR CRITERIA,
POLLUTANTS
PHASE I:
TECHNOLOGY
' STUDY
-BAT-
PHASEII:
ECONOMIC
STUDY
-EA-
DETERMINE
REMOVAL
'OFTOXICITY
BY BATEA
secondary effluents from
PHASE I:
PREENGINEERING
SCREENING
STUDY
PHASE 1 1:
TOXICITY
REDUCTION
BY BAT
DETERMINATION
OF BATEA
' FOR TEXTILE
WASTEWATERS
Figure 3. Overall program approach to determine BATEA.
23 selected textile plants and to rank
the plants in descending order of toxicity (Figure 4)• Phase II
was designed to determine the level of toxicity removal attained
by the tertiary treatment systems in the ATMI/EPA Grant Study at
only those plants with relatively high secondary effluent toxi-
city (Figure 5). Sampling locations for Phase II of the study
are shown in Figure 6, and the strategy used for evaluating con-
trol technologies in terms of toxicity removal is illustrated in
Figure 7.
PROJECT ORGANIZATION '
The major effort of the Phase I MRC/EPA screening study was
devoted to the collection, chemical analysis, and biological
toxicity testing of single, 8-hr composited wastewater samples
from the 23 textile plants scheduled for testing in the ATMI/EPA
Grant Study. In addition, samples were collected from nine
other textile plants for 'chemical analyses only. Wastewater
characterization data were therefore assembled for a total of
32 plants. '•
i
The scope of work for Phase I was divided into three separate
task areas, each based on different EPA data requirements, as
shown in Figure 8. CPB (Chemical Processes Branch, IERL/RTP,
Project Officer, M. Samfield) requested chemical and bioassay
data on secondary effluent samples from the 23 textile plants
scheduled to be studied in the ATMI/EPA Grant Study. These data
were used to characterize and compare the relative toxicities of
the plant effluents tested. EGD (Effluent Guidelines Division,
EPA, Washington, J. D. Gallup) requested chemical analyses of the
raw waste streams entering the 23 wastewater treatment plants, as
well as chemical analyses of the raw waste and secondary effluent
streams at 9 additional textile plants. Raw waste and effluent
data were needed to evaluate the pollutant removal efficiencies
20
-------
MROEPA WASTEWATER
raxiciTYsnioYi
PHASE h SCREENING
COUfCT SECONDARY EFRUENT SAMPLES
FROM EACH OF THE Q PLANTS
PERFORM ANALYSES
BIOASSAYS
PRIORITY
POLLUTANTS
LEVEL 1
CHEMICAL
ANALYSES
EVALUATE ANALYTICAL PROCEDURES
AND MAKE RECOMMENDATIONS
FOR IMPROVEMENTS
PRIORITIZE PLANTS
BASED ON BIOASSAY TOXICITY DATA
SELECT THE PLANTS WHICH HAVE
SECONDARY EFFLUENTS SUFFICIENTLY
TOXIC TO EVALUATE THE EFFECT
OF BAT SYSTEMS
Figure 4. Program outline for Phase I of the
MRC wastewater toxicity study.
/MRC/EPA WASTEWATER TOXICITY STUD'
I PHASED:
\JQXICITY REMOVAL BY BAT SYSTEM^
SatCT BIOASSAY TESTS
COLLECT SAMPLE FOR THE PILOT PLANT
DURING THE 2-WEEK SAT EVALUATION PERIOD
REPORT RESULTS FOR BAT EVALUATION
Figure 5. Program outline for Phase II of the
MRC wastewater toxicity study.
21
-------
INTAKE
WATER
Figure 6. Sampling locations for Phase II of
the MRC wastewater toxicity study.
INTERPRETATION OF BIOASSAY TEST RESULTS
Bioassay Results
Inlet Outlet
Toxic Substance Interpretation
Figure 7.
+ Control Technology Is Not Effective
Control Technology Is Effective
+ Control Technology Is Deterimental
Control Technology Is Not Deterimental
Intepretation of bioassay test results,
of current state-of-the-art secondary treatment systems. In
order to detect other possible pollutant species, PMB (Process
Measurements Branch, IERL/RTP, L. D. Johnson) requested that
Level 1 chemical characterization also be performed on the efflu-
ent samples at the basic 23 textile plants (2). Since these data
were requested after the program began, only 15 textile plants
were sampled for Level 1 chemical characterization.
Chemical characterization of wastewater samples involves four
categories of analysis:
129 consent decree priority pollutants analysis (1)
nutrient analysis (4, 5)
effluent guidelines criteria pollutants analysis (4,
Level 1 chemical characterization (2)
5)
(4) Manual of Methods for Chemical Analysis of Water and Wastes,
EPA-625/6-76-003a (PB 259 973), U.S. Environmental Protec-
tion Agency, Cincinnati, Ohio, 1976. 317 pp.
(5) Standard Methods for the Examination of Water and Waste-
water, Fourteenth Edition. American Public Health Associa-
tion, Washington, D.C., 1976. 874 pp.
22
-------
LEVEL OF EFFORT REQUESTED BY:
EPA -EFaUENT GUIDELINES DIVISION (EGO)
-WASHINGTON, D.C.
CHEMICAL PROCESSES BRANCH (CPB)
IERL - INDUSTRIAL PROCESSES
RESEARCH TRIANGLE PARK, NORTH CAROLINA
PROCESS MEASUREMENTS MtANCH(PMB)
INDUSTRIAL PROCESSES DIVISION
RESEARCH TRIANGLE PARK. NORTH CAROLINA
ANALYZE RAW WASTE FOR:
• 129 PRIORITY POLLUTANTS
• CRITERIA POLLUTANTS
ANALYZE EFFLUENT FOR:
• CRITERIA POLLUTANTS
RAW WASTE
23 TEXTILE
PLANTS:
SECONDARY
WASTEWATER
TREATMENT
PLANT
EFFLUENT
ANALYZE EFFLUENT FOR: .
• 129 PRIORITY, POLLUTANTS
• NUTRIENT, SERIES,-,
. BIOASS'AYS~ j r.
• MUTAGENICITY
• CYTOTOXICITY
• FRESHWATER SERIES
• MARINE SERIES
• 14-DAY RAT TEST
• SOIL MICROCOSM
ANALYZE EFFLUENT FOR:
• LEVEL 1 CHEMICAL ANALYSIS
RAW WASTE
ADDITIONAL
TEXTILE
PLANTS
moo
EFFLUENTS
SAMPLE AND ANALYZE RAW WASTE
AND EFFLUENT STREAMS FOR:
• 129 PRIORITY POaUTANTS
• CRITERIA POLLUTANTS
Figure 8. Scope of work for the analysis of textile plant wastewaters.
-------
The 129 consent decree priority pollutants as listed dn Appendix
A are divided into volatile compounds, nonvolatile compounds,
and metals (see Appendix 'B). The nutrient series required to
support algal tests includes analysis of nitrite, nitrate,
ammonia, total Kjeldahl nitrogen (TKN), o-phosphate, phosphorus,
and total organic carbon , (TOC): Effluent guidelines criteria
pollutants include 5-day biochemical oxygen demand (BODs), chem-
ical oxygen demand (COD); sulfide, color, pH, total suspended
solids (TSS), total dissolved solids (TDS), and total phenol.
A detailed description of Level 1 chemical characterization is
given in Section 6.
\
The bioassay scheme established by EPA for evaluating the reduc-
tion in toxicity of water samples by control technologies is
shown in Figure 9. All the tests shown were used in this project.
The marine ecology series and the soil microcosm tests were
requested after the project began, therefore data were obtained
from 15 textile plants as opposed to the basic 23 plants.
MICROBIAL MUTAGENICITY -
SAUMNEUA TYPHIMURIUM
TA 1535. TA 100. TA 1537, TA 98
FRESHWATER.
FATHEAD MINNOW TOX
ALGAE BOTTU
DAPHN1A TOXICITY
MARINE
SHEEPSHEAD MINNOW
ALGAE
GRASS SHRIMP
Figure 9.
EPA-recommended bioassay testing scheme
for toxicity analysis of water samples.
Figure 10 illustrates the distribution of samples among the eight
EPA and private laboratories that performed the chemical analyses
and bioassay tests. Appendix C lists the names and addresses of
all persons involved in ,the textile project.
MRC collected raw wastes and effluent samples at 23 of the ATMI/
EPA-designated plants. Wastewater samples were collected by EPA-
Environmental Research Laboratory (ERL) (Athens, Georgia) at two
of the additional textile plants and sent to MRC for chemical
analysis. Sverdrup and Parcel and Associates, Inc., (St. Louis,
Missouri) collected the remaining samples at the additional seven
plants and sent them to IMRC for chemical analysis.
24
-------
BIOASSAY SCREENING
CHEMICAL SPECIES ANALYSIS
Figure 10.
Laboratories and persons involved in sample
analysis of textile plant effluents.
Sampling, analytical, and bioassay procedures followed those
recommended in EPA reports (1-5). All procedural modifications
instituted to accommodate the textile wastewater samples are
discussed in detail in the remaining sections of this report.
25
-------
SECTION 5
SAMPLING PROCEDURES
COLLECTION TECHNIQUE
Wastewater was collected by composite and grab sampling techni-
ques. Composite samplers (Isco Model 1680) were used to collect
raw waste samples for analysis of nonvolatile organics and metals.
Tygon® sample tubing used was washed with detergent, rinsed
thoroughly, and given a final washing with organic-free water.
A 0.001-m* sample blank was then collected and analyzed for
organic leachates. Organic-free water was prepared by passing
water, distilled in glass, through a 0.6-m-long activated carbon
column. The blank was collected in glass, sealed with a Teflon®-
lined cap, and stored in ice at 4°C until analyzed.
Grab sampling techniques were used to collect raw waste samples
for other analyses, and for all secondary effluent samples,
Figure 11. Eight individual grab samples were collected at
equally spaced time intervals during the normal working day. To
insure that each of the eight laboratories received a sufficient
portion of the same sample, grab samples were collected in a
Teflon-lined, 0.01-m3 stainless steel bucket. A specified
aliquot was transferred to each of the sample bottles from this
container. Care was taken to insure that the sample remained
homogeneous throughout each of the 10-min pouring sessions.
Containers for volatile organics analysis were collected and
sealed first to minimize possible evaporation losses.
TEXTILE PLANT
RAW
WASTEWATER (
SAMPLE
i^—
I
0
o
0
o
o
o
AERATION LAGOON
SECONDARY
EFFLUENT '
SAMPLE
\
/
2) —
CHLORINE
CONTACT
BASIN
1
EFaUENT
Figure 11. Phase I sampling locations.
26
-------
TABLE 7.'
.<10.SAMPLEi COLLECTION: AND HANDLING ^REQUIREMENTS (4)
IrJ j| 1S\ I I ft,,' - .' I >91 O or* o .'". r ?• ,' '
1 I V, -.J 1 _. ' J - ' ------ ' - ' - T- "J - l"^1- —- "" *-- — > - - -' -
j.
•b
••
t V
Analysis'
1 U
a
Total- sample "'
vol required.
i m5' j
t * ;
Consent decree [pollutants
Volatile organics
Nonvolatile
Metals , O
Nutrient* ajhaly
Total brgani
Ammonia K
Total Mcjelda
Nitrate £.
Nitrite ft
Orthophospha
organics
.-i f t) r-
o pi!
i » t
iis!
c carbon
hi nitrogen
i
«l
Total ..phosphorus
. p i a »' *
Criteria' pollutants ' 1
Biochemical 1 5-d'ay
oxyg.en-hdemand
Chemicalvtoxygen
Color '• ' '" •
Sulfid'e -\
Total j-'suspended
Phenol j 7:
r • ,.4
1 c ; 1 !v 3
demand
f
solids
2 '•
.n
Freshwater- -ecology c '
Fathead minnow |
and tDaphnici'' J 'i ^
Algae' ' *-:. ,' J" fj
1
320 x ID"6
'14,000 x 10"»
^
1,000 x 10 &
. '. , ; •"
25 x IO-6
400 x 10"6
500 x 10"6
100 x io-6
50 x 10"6
50 x IO-6
50 x IO-6
-*[; 1 ^:
1
1,000 x IO"6
50 x 10~6
50 x 10"°
500 x l6~6
100 x 10"6
500 x 10"6
t „
^500 x 10"?:
[ f.
'60 , 000~x^lO"6
9,6M.*^<>~*
1 1 Storage
,, -' >J i; r r"; ', . '.', temper-
Container £ & fi S S T. " '! ature,
used' 'j~! Preservation steps C
,-j j i
' Glass*3' -C ' 4d
Glass 4
Plastic, Nitric .acid ,5 x 10~3 m3/m3 4
: n t. ,
a n q c: i-, • 1 '
Glass / ; Sulfuric acid to pH < 2 4
Glas'Sj: n:sulfuric acid to pH <,2 4
Glass " >> Sulfuric acid to pH < 2 4
Glass » Sulfuric acid to pH < 2 4
Glass '° '; •- '
Glass ° ;
Glass |
- \\ C 1 - o j I t
L. 'J ^: •
u Glass". 4.
« Glass -• Sulfuric acid to pH < 2 4
i* Glass ,y ,. 4
S Glass C: 2 x 10~6 m3 zinc acetate 4
'; GlaVsil ,• tj '•
,; Glass Phosphoric acid t'o pH' < 4,
; 10 3 g copper sulfate/m3 4
, Plastic , Sodium hydroxide to pH > 12 4
1
Glass • ' ' 4
Plastic ;...-,,« 4
r "' , ' '
Marine ecology ' ; t | ' ^ -" .. , . ' ' .
Sheepshead minnow
* °~ • : " . KJ n.
and 'grass shrimp
Algae";, f
-^
Soil microcosm ,:'•
Ames test j-i 1 1 jj
,'i
^
Cytotoxi'ci/t'y test's
14-day Rat test
ri M i
3 Eight indavid
b ^5 i/?
ual 140
J Cl
1
6
1 > , *"
, ,.
0,000 x l6"6 ;, Glass „ 4
'2,000 x 10"6
1 .
c
*'
'10'0*5f"UO "?!
V' ^ *"* i _
250 x 10"6
1,000 x 10"6
x IO-6 m3 samples
10" 6 m3. :e'
-------
An additional 0.02 m3 were required to properly apportion samples
into the six 0.02-m3 bottles for the marine and freshwater
ecology tests. Samples were poured into these bottles at the
end of each of the eight grab sampling sessions. There was no
visible change in flow rate during each of the 15-min sample
collection periods. Fluctuations in effluent and raw waste
stream flow rates were usually on a 45-min to 1-hr time schedule.,,
SAMPLE CONTAINER PREPARATION
All glass containers, except the 0.02-m3 bottles, were thoroughly
cleaned with strong acid (50% sulfuric acid +50% nitric acid),
rinsed, and heated in a glass annealing oven at 400°C for at
least 30 min. The 0.02-m3 bottles were detergent washed and
thoroughly rinsed. The glass containers used to store samples ^
for mutagenicity testing'.were rinsed with acetone. 'The rest of ' :"
the glass containers were rinsed with methylene chloride and
dried in the oven at 100°C. All glass bottles had Teflon-lined
caps. :
Plastic sample containers were thoroughly cleaned before use.
Each bottle was washed with detergent and tap water, then rinsed
with 1:1 nitric acid/tapwater, 1:1 hydrochloric acid/tap water,
and, finally, deionized distilled water (1).
SAMPLING LOGISTICS
The type and volume of sample container varied, depending on the
analysis to be made. Some samples required the addition of chem-
ical preservatives in the field to prevent deterioration during
shipment to the laboratory. The volume of sample required, the
container used to hold the sample, and the preservation steps : •'
used in this project are shown'in Table 7 (4).
A field sampling instructional worksheet was designed to facili-
tate the arduous task of filling 37 glass and plastic bottles of
different sizes requiring different sample volumes and preserva-
tives at each plant. Table 8 shows part of this worksheet.
Each sampling day, before sampling, bottle labels were filled out
and affixed to the appropriate sample bottles. Figure 12 shows
the bottle label that MRC has designed for sample identification.
28
-------
TABLE 8. PORTION OF THE FIELD INSTRUCTIONAL FORM USED BY MRC
TO ASSURE ACCURATE SAMPLE COLLECTION AND PRESERVATION
N)
VO
Organics
Type of test
Sample
destination
Bottle
identification
number
Number of
bottles
Type of bottle
Sample size
10-* m3
Sample number
1
2
3
4
5
6
7
8
Volatile Nonvolatile
MRC
1
8
0
40
+3 x ID"6 m3 Sodium
thiosulfate, seal
Same as No. 1
Same as No. 1
Same as No. 1
Same as No. 1
Same as No. 1
Same as No. 1
Same as No. 1
MRC
2
1
i
3,785
None
None
None
None
None
None
None
Seal
Metals
MRC
3
1
Q
240
+5 x ID"6 m3
Nitric acid
None
None
None
None
None
None
Seal
TOC, NH3, TKN
COD, nitrate
MRC
4
1
0
240
Chenicala added
•t-Sulfuric acid.
pH <2
Same as No. 1
Same as No. 1
Sane as No. 1
Same as No. 1
Same as No. 1
Same as No. 1
Sane as No. 1
Marine
algae
Gulf
Breeze
5
1
fl
240
for sample
None
None
None
None
None
None
None
None
Cyanide
MRC
6
1
0-
240
preservation
•f4 x 10~6 m3 Sodium
hydroxide, check
with potassium
iodide paper
None
None
None
None
None
None
PH > 12, check
seal
Phenol
MRC
7
1
P
in
62
pH <4 with
phosphoric acid
0.5 a copper
sulfate
None
Same as No. 1
None
None
None
Same as No. 1
Sulfide
MRC
8
1
P
in
62
+2 x 10~6 m3
Zinc acetate
None
None
None
None
None
None
Seal
Mes
test
SRI
9
1
JR.
-------
Job
Sample or Run No.
Sample Location
Type of Sample
Analyze for
Preservation_
Comments
Log No. Date
Initials
Figure 12. MRC sample bottle label.
SAMPLE SHIPPING PROCEDURES
Each bottle was capped and sealed with tape to prevent leakage.
Glass bottles were individually wrapped to prevent breakage.
Sample bottles were then packed in one-piece, molded, styrene
foam shipping cartons with 3.8-cm walls and fitted tops. Each
such unit was then placed in a corrugated cardboard box.
Each carton was half-filled with sample bottles, filled with ice,
sealed with cellophane tape, and reinforced with 0.05-m duct tape,
Address labels were affixed to box tops. Warning labels—"This
carton contains glass and ice"—"Hold at airport and call "
messages were also put on the box tops.
All samples were shipped by conventional air freight on the day
that they were collected. The airlines selected offered the
most direct route without, carrier changes.
30
-------
SECTION 6
WASTEWATER CHEMICAL ANALYSES
EFFLUENT GUIDELINES CRITERIA POLLUTANTS
Parameters determined under the category of effluent guidelines
criteria pollutants were: 5-day biochemical oxygen demand (BOD5)/
chemical oxygen demand (COD), color, sulfides, total suspended
solids (TSS), pH, and total phenol. As sample shipments arrived
at MRC, they were logged in and distributed to the designated
technicians for analysis.
w
Analytical and supporting procedures followed those described in
References 4 and 5.
Criteria pollutants were determined on the raw waste and second-
ary effluent streams from the basic 23 plants and the additional
9 plants.
Results of the chemical analyses are given in Table 9. The first
row of numbers for each plant represents data obtained on the raw
waste stream, and the second row of numbers corresponds to the
wastewater treatment plant effluent stream. All values except
color and pH are given in g/m3 (ppm). Color was measured using
the American 'Public Health Association (APHA) system.
Effluent values obtained from wastewater treatment facilities in
some,plants were greater than those of the influent raw waste.
This occurred, in part, because the wastewater entered the treat-
ment system 1 day to 5 days prior to leaving the treatment plant.
The hydraulic retention time in textile wastewater treatment
plants ranged from 1 day to 30 days, with an average value of
5 days.
All of the textile plants sampled had a secondary wastewater
treatment facility that included a lagoon with several surface
aerators, followed by a clarifier. Several plants used equaliza-
tion basins prior to the aerated lagoons. Effluent samples were
collected between the clarifier and the finishing pond in plants
that had both. The two exceptions were plant Y, where the
sample was taken after the finishing pond, and plant R, where the
effluent sample was inadvertently collected between the aerated
lagoon and the settling basin. All other plant effluent samples
were collected after the clarifiers.
31
-------
TABLE 9. ANALYSIS OF WASTEWATER SAMPLES FOR
EFFLUENT GUIDELINES CRITERIA POLLUTANTS
Criteria pollutant, g/m*
Plant code
A/raw waste
A/effluent
B/raw waste
B/effluent
C/raw waste
C/effluent
D/raw waste
D/effluent
E/raw waste
E/effluent
F/raw waste
P/effluent
G/raw waste
G/effluent
H/raw waste
H/effluent
J/raw waste
J/ef fluent
K/raw waste
K/effluent
L/raw waste
L/effluent
M/raw waste
M/effluent
N/raw waste
N/effluent
P/raw waste
P/effluent
R/raw waste
R/effluenta
S/raw waste
S/effluent
5-Day
biochemical
oxygen
demand
459
168
1,050
<5
445
25
71
6.6
18
<5
194
69
203
42
288
14
210
25
564
<5
379
13
830
<5
334
36
680
28
450
70
219
59
Chemical
oxygen
demand
1,735
. 1,652
1,264
99
802
396
224
64
2,660
78
583
276
1,340
502
320
300
810
376
'l,725
131
1,117
234
2,265
255
1,140
286
172
45
1,692
830
559
1,035
Color ,
APHA
2,000
2,000
1,400
90
2,600
1,920
1,875
1,625
250
30
150
80
300
300
1,250
500
1,875
1,375
40,000
150
1,300
370
1,000
500
1,050
90
300
250
1,500
2,000
250
75
Sulfide
6.0
4.0
, 1.4 •
0.2
5.2
5.0
<0.02
2.8
<1
<1
2.1
0.1
<1
<1
<0.02
<0.02
1.8
1.8
<1
<1
4.5
3.0
<1
<1
1.1
0.1
6.19
6.09
<1
<1
9.2
<1
t
Phenol
0.092
0.065
0.042
0.015
0.074
0.088
0.024
0.018
0.069
0.014
0.74
0.028
0.028
0.054
0.047
0.019
0.063
0.024
0.067
0.018
0.038
0.026
0.037
0.025
0.156
0.068
0.228
0.032
0.282
0.162
0.107
0.029
, Total
suspended
solids
165 .
228
32
8
49
300
16
154
52
19
23
44 '
37
6
39
43 •
0.01
0.023
69
21
19
78
210
21
68
77
1 6
45
87
225
25
581
PH
10.7
7.3
10.5
7.5
11.2
10
10 .
7.2
10
7.2
9.2
7.4
11
7.5
10
7.6
11
7.8
10
7.2
7.4
5.8
11
7.5
9.2
7.0
10
7.1
10
8.1
10
7.8
Cyanide
<0.004
0.015
0.017
<0.004
0.007
0.013
0.21
0.21
<0.004
<0.004
<~0.004
<0.004
<0.004
0.006
<0.004
<0.004
< 0.004
< 0.004
<0.004
<0.004
<0.004
0.172
<0.004
<0.004
<0.004
< 0.004
0.19
0.14
<0.004
<0.004
0.007
< 0.004
(continued)
32
-------
TABLE 9 (continued)
Criteria pollutant, q/m3
Plant code
T/raw waste
T/effluent
U/raw waste
0/effluent
V/raw waste
V/ef fluent
W/raw waste
W/effluent
X/raw waste
X/effluent
Y/raw waste
Y/effluent
Z/raw waste
Z/effluent
JJ/raw waste
JJ/effluent
KK/raw waste
KK/effluent
LL/raw waste
LL/ef fluent
MM-1
'MM-2
MM- 3
MM- 4
NN/raw waste
NN/effluent
OO/raw waste
OO/effluent
PPC
Y-001/raw waste
Y-001/effluent
C-001C
5 -Day
biochemical
oxygen
demand
501
32
400
24
53
<5
1,920
84
237
15
122
<5
351
<5
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
Chemical
oxygen
demand
500
414
1,464
748
b
128
6,124
837
786
258
457
115
812
105
1,545
510
1,955
447
' 727
155
b
b
b
b
938
236
1,889
635
339
b
b
b
Color,
APHA
1,345
350
3,200
2,480
500
500
2,200
1,900
1,200
10
10,000
250
500
750
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
Sulfide Phenol
7.6 0.073
6.0 0.041
5.6 0.057
3.5 0.007
<1 0.018
<1 0.016
0.5 0.670
0.1 0.232
0.75 0.940
0.01 0.035
<1 0.064
<1 0.022
2.48 0.56
<1 0.023
<1 0.144
<1 0.055
<1 0.150
<1 0.052
<1 0.001
<1 0.094
<1 0.033
<1 0.031
<1 0.036
<1 0.039
<1 0.043
<1 0.014
<1 " 0.082
<1 0.026
<1 0.044
b b
b b
b b
Total
suspended
solids
28 s
35
111
92
54
26
2,300
300
24
18
33
17
20
13 ,
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
PH
9.5
7.4
10
7.3
9.0
7.1
10.4
8.1
10.2
7.2
10.5
8.0
10
8
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
Cyanide
<0.004
<0.004
<0.004
0.212
0.006
0.018
0.015
0.020
<0.004
0.101
<0.004
<0.004
<0.004
<0.004
0.005
0.028
<0.004
<0.004
0.008
0.006
<0.004
<0.004
<0.004
< 0.004
0.04
<0.004
<0.004
<0.004
<0.004
<0.004
0.029
<0.004
'secondary effluent sample was inadvertently collected between the aerated lagoon and
settling ponds.
Analysis not performed on sample.
Secondary effluent sample only.
33
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ANALYSIS PROTOCOL FOR THE 129 CONSENT DECREE PRIORITY POLLUTANTS
Recommended analytical procedures (1) developed by EPA were used
throughout this project. It is important to realize that these
EPA procedures are still under development and require further
verification and validation. Therefore, the data presented in
this section only serve to identify which of the 129 chemical
species are present and to indicate the general concentration
ranges within an order of magnitude.
Adaptations of these procedures to accommodate the special
requirements of textile wastewaters and/or any ambiguities in
analytical techniques are discussed below. Three chemical
species were not determined in this project: endrin aldehyde,
2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and asbestos. EPA-
Environmental Monitoring and Support Laboratory (EMSL) recommended
that TCDD should be omitted because of its extreme toxicity, and
the potential health hazard involved in preparing standard solu-
tions from the pure compound. Pure endrin aldehyde could not be
obtained in time to prepare standard solutions. Asbestos was
eliminated, as recommended by EPA-IERL-RTP and EPA-EGD, due to
the presence of other fibrous materials in textile wastewaters.
The analytical protocol (2) divides the 129 chemical species into
three basic categories: volatile organics, nonvolatile organics,
and metals. Appendix B lists the chemical species which belong
to each category. The following sections outline the analytical
procedures and MRC modifications for each category.
Volatile Organics
i
The recommended analytical method was designed to determine those
chemical species which are amenable to the Bellar purge and trap
method. Eight 40 x 10~6-m3, hermetically sealed glass vials,
stored in ice, were sent to the laboratory from each sampling
site. The vials were composited within 1 day of receipt at the
laboratory. Two vials of composite solution were sealed and
retained at 4°C as reserve samples. Volatiles from 5 x 10~6-m3
samples of composite solution were sparged with helium onto two
Tenax GC-silica-packed sample tubes. (Internal standards were
added to the solutions in the later stages of the program. The
majority of the samples had been sparged and stored before the
protocol (1) was received and appropriate internal standard
could be obtained.) The second Tenax tube was used as a backup
sample. Tenax tubes were sealed under a nitrogen atmosphere in
glass tubes and stored in a freezer at -18°C until analyzed.
Analyses were carried out using a Hewlett Packard 5981 GC-Mass
Spectrometer with 5934 Data System. Sample tubes were heated to
180°C over a 1-min period and held at that temperature for 4 min
to desorb the compounds onto a Carbowax 1500 column held at -40°C.
Cryogenic trapping at -40°C (liquid nitrogen cooling) gave better
34
-------
reproducibility of retention time than using the suggested tem-
perature of 30,°C, for compounds with boiling points below room
temperature. After desorption, the GC column temperature was
raised 8°C/min to 170°C.
The mass spectrometric analysis method involves fragmentation of
molecules using electron bombardment (70 eV). Masses and rela-
tive intensities of the most characteristic molecular fragments
for each compound are listed in the protocol (1). The population
of ion fragments covering the mass range from 35 atomic mass
units to 500 atomic mass units was measured every 6 s, and the
data were stored on magnetic tape.
These data allow the operator to reconstruct chromatograms of
observed intensity for an individual mass during the course of
the scanning. Specific molecules may be detected in the presence
of other compounds by examining the reconstructed intensity time
plots of their characteristic masses.
Qualitative identification of a compound was made using the three
criteria listed in the protocol: 1) retention time must coincide
with known retention times, 2) the three characteristic-masses
must elute simultaneously, and 3) intensities of the character-
istic masses must stand in the known proper proportions.
Quantitation of volatile organics was initially made using peak
area counts and concentration calibration curves. Later in the
program, response ratios using the 1,4-dichlorobutane internal
standard were used in quantifying the concentrations. Base/
neutral and acid organic compounds were quantified using deuter-
ated anthracene and response ratios as prescribed in the
protocol (1) .
Figure 13 is a simplified diagram of the analytical scheme 'for
volatile organic analysis.
Nonvolatile Organics
This method determined the nonvolatile solvent-extractable
organic compounds that could be analyzed by gas chromatographic
methods. The 129 consent decree priority pollutants contain 81
organic compounds classified as nonvolatile organics.
Nonvolatile organics are divided into three groups: base/neutral
fraction, acid fraction (phenols), and pesticides and polychlo-
rinated biphenyls (PCB). A list of compounds that are classified
as nonvolatile organics is given in Appendix B.
The analytical procedure is described in Reference 1. Figure 14
depicts the sample processing scheme for the base/neutral and
acid fractions. The sample solution, 2 x 10~3-m3, was made
alkaline (pH greater than 11) with sodium hydroxide, and then
35
-------
Figure 13. Analytical scheme for volatile organics analysis.
Figure 14.
Sample processing scheme for
nonvolatile organics analysis
36
-------
extracted three times with methylene chloride. Textile raw waste
and effluent samples formed strong emulsions upon extraction with
methylene chloride. The problem was resolved by drawing off
small amounts of separated solvent and pouring the extract
through the sample in the separatory funnel. Separation was
also enhanced by slowly dripping the emulsion onto the wall of
a slightly tilted flask. This approach gives better separation
by providing a greater surface area for the solvent and water
fractions. Some samples required centrifugation at 1,500 rpm
for 1 hr to break the emulsion.
Extracts were dried on a column of anhydrous sodium sulfate, con-
centrated to 10~6 m3 in a Kuderna-Danish (K-D) evaporator with
a Snyder column spiked with deuterated anthracene, sealed in
septum capped vials, and stored at 4°C until analyzed. Analyses
were performed on the GC/MS system using SP 2250 and Tenax GC
columns for base/neutral and acid samples, respectively (1).
A separate 0.001-m3 sample was used for analysis of the pesti-
cides and PCB (Aroclor® fluids). The basic sample processing
scheme is shown in Figure 15. These compounds were extracted
with a 15% methylene chloride and 85% hexane solvent mixture.
The aqueous phase was discarded, and the organic phase was
analyzed by GC with an electron capture detector. Where neces-
sary, acetonitrile partitioning and a Florisil® chromatography
column were used for further celanup of the sample. In 85% of
the samples, additional cleanup was not required.
Confirmation of identify and quantitation were made using two
different GC columns: SP-2550 and Dexil 410. Compound verifi-
cation was made with the MS when the concentration was greater
than 0.01 g/m3. Concentrations of potential pesticides ranged
from 0.0001 g/m3 to 0.01 g/m3; therefore, MS verification was
not possible in this study. Pesticide species identified only
by GC below 0.01 g/m3 were reported only if they met the follow-
ing two criteria: 1) the retention time window between stand-
ards and unknown peaks correlated within ±3 s, and 2} concentra-
tions calculated from both GC columns had to agree within ±20%.
Unknown peaks not meeting these criteria were assumed not to be
the pesticide species.
Metals
In addition to the volatile and nonvolatile organics, the 129
chemical species include 13 metals, asbestos, and cyanide. Each
metal is measured as the total metal. Asbestos was not deter-
mined in this study; cyanide was measured by conventional wet
chemistry techniques (5).
Eight metals were analyzed by the inductively coupled argon
plasma (ICAP) excitation technique: antimony, cadmium, chromium,
copper, lead, nickel, silver, and zinc. Five metals which can
37
-------
Figure 15.
Sample processing scheme for
pesticide and PCB analysis.
not be quantified by ICAP analysis were measured by conventional
atomic absorption techniques: arsenic, beryllium, mercury,
selenium, and thallium (4, 5).
\
ICAP forms an analytical system for simultaneous multielement
determinations of trace metals at the sub-ppm level in solutions,
The basis of this method is atomic emission. Excitation energy
is supplied by coupling a nebulized sample with high temperature
argon gas which has been passed through a powerful radio-
frequency field. Emitted light is simultaneously monitored at
22 wavelengths corresponding to 22 different elements.
All samples for metals analysis were acidified in the field by
adding 5 x
of sample.
10~6
nr
of redistilled nitric acid to each 10~3
nr
Nitric acid blanks were also analyzed for metals.
Analytical Results
Raw waste samples were collected with continuous samplers using
a peristaltic pump which pulled the sample through Tygon tubing
to the sample bottle. Sample blanks were collected by drawing
laboratory-prepared organic-free water through the sampler prior
to sample collection to determine the presence of base/neutral,
acid, and pesticide organic priority pollutants. Results of
these analyses are given in Table 10.
38
-------
TABLE 10. SUMMARY OF CONTINUOUS SAMPLER AND
VOLATILE ORGANIC BLANK ANALYSES
Fraction
Compound found
Concentration
range/ mg/m3a
Base/neutral
Acids
Volatiles
Pesticides and PCB
Naphthalene
Dimethylphthalate
Diethylphthalate
Bis(2-ethyIhexy1)phthalate
Di-n-butylphthalate
Phenol
Toluene
Trans-1,2-dichlproethylene
Trichloroethylene
Ethylbenzene
None
2
16
0.5 to 10.2
1.5 to 46
1.3 to 1.7
0.6 to 1.1
2.6 to 55
3.2
2.4
8.3
1 mg/m3 equals 1 yg/1.
To determine if any volatile organic species were absorbed from
the air to the samples, EPA recommended that laboratory-prepared
organic-free water be carried to the plant site, poured from the
container into a sample vial, sealed, and shipped back to the
laboratory for analysis (1). These results are included in
Table 10.
All secondary effluent samples were collected by grab sampling
techniques. Therefore, these samples were not passed through
Tygon tubing. Samples were shipped and stored in ice at 4°C
until extracted. A special effort was made to initiate methylene
chloride extraction as rapidly as possible. The data on which
each sample arrived at MRC and the date of its solvent extrac-
tion are shown in Table 11. Only 5 out of 64 samples were not
extracted within 24-hr of receipt at MRC.
Results of GC/MS analysis of textile raw waste and secondary
effluent samples for base/neutral organic compounds are pre-
sented in Table 12. GC/MS analyses for the volatile, acid, and
pesticide/PCB organic compounds are presented in Table 13.
Of the 114 organic compounds in the priority pollutant list, a
total of 45 different compounds were identified in textile
wastewaters, 39 in raw waste samples and 34 in secondary efflu-
ent samples. The,number of compounds found at each plant is
summarized in Table 14. On an individual plant basis, the great-
est number of organic compounds detected in a raw waste and
secondary effluent sample were 14 and 8, respectively, with an
average number per plant of 7 in the raw waste and 5 in the
secondary effluent..
39
-------
TABLE 11. TEXTILE SAMPLE EXTRACTION DATES
Plant
A
B
C
D
E
F
G-l-2
G-2-2
H
J
K
L
M
N
P
R
S
T
U
V
W
X
Y
Z
C-001
Y-001
JJ
KK
LL
MM
NN
00
PP
Date received at MRC
5-6-77
4-28-77
4-13-77
3-3-77
3-31-77
4-19-77
4-1-77
4-1-77
3-10-77
3-9-77
4-5-77
4-12-77
3-16-77
4-27-77
3-2-77
3-15-77
4-6-77
4-26-77
5-4-77
3-31-77
4-14-77
4-21-77
3-llT77
3-17-77
5-20-77
5-20-77
6-23-77
6-24-77
6-27-77
7-5-77
6-30-77
7-5-77
7-5-77
Date of initial ' extraction
5-6-77
4-29-77
4-14-77
3-3-77
3-3,1-77
4-20-77
4-1-77
4-4-77
3-10-77
3-9-77
4-6-77
4-12-77
3-17-77
4-28-77
3-2-77
3-16-77
4-7-77
4-27-77
5-4-77
3-31-77
4-14-77
4-21-77
3-11-77
3-18-77
5-20-77
5-20-77
6-23-77
6-27-77
6-29-77
7-8-77
7-1-77
7-6-77
7-8-77
40
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TABLE 12. GC/MS ANALYSES FOR BASE/NEUTRAL ORGANIC COMPOUNDS
IN RAW WASTE AND EFFLUENT SAMPLES
Plant/source
<10 mg/m'
Compounds identified ana concentrations ooserveda
10 to 100
A/raw wajte Naphthalene 0.1
Dimethyl phthalate 3
Olethyl phthalate 1
Bis(2-ethylhexyl) phthalate 0.5
A/effluent 1,2-Dichlorob«nien« 1
1,4-Dichlorobeniene 0.05
Bia(2-ethylhejcyl) phthalate 6
B/raw waste Diethyl phthalate 3.3
Bis(2-ethylhexyl) phthalate 5.7
Anthracene 0.1
B/effluent N-nitroso-di-n-propylamine 2
Bia(2-ethylhexyl) phthalate 3
Pyrene 0.3
C/raw waste 1,2-Dichlorobenzene 1.1
Diethyl phthalate 4.1
C/effluent 1,2-Dichlorobensene 0.3
Acenaphthene 0.5
Bis(2-ethylhexyl) phthalate 3.0
Anthracene 4.4
D/raw waste Naphthalene 0.3
Bla(2-ethylhexyl) phthalate 8.9
D/effluent Diethyl phthalate 1
Bls(2-ethylhexyl) phthalate 5
E/raw waste 1,4-Dichlorobenzene 2
Napthalene 1
Bis(2-ethylhexyl) phthalate 5
E/effluent 1,4-Dichlorobenzene 0.2
1,2-Dichlorobenzene 0.2
Dimethyl phthalate 1
Diethyl phthalate 0.5
Pyrene 0.1
F/raw waste 1,4-Dichlorobenzene 6.5
F/effluent 1,2,4-Trichlorobenzene 6.3
G/raw waste Fluorene 5
G/effluent Acenaphthene 2.0
Hexachlorobenzene 0.8
U/raw waste 1,2-Dichlorobenzene 0.5
Naphthalene 3
Di-n-butyl phthalate 2
H/effluent NPPO
J/raw waste Diethyl phthalate 6.5
J/effluent Di-n-butyl phthalate 3.6
Pyrene 0.1
K/raw waste Naphthalene 0.03
Diethyl phthalate 0.2
K/effluent Naphthalene 0.5
Bis(2-ethylhexyl) phthalate 8
L/raw waste 1,4-Dichlorobenzene 1
Bis(2-ethylhexyl) phthalate 3
L/effluent Bis(2-ethylhexyl) phthalate 2
M/raw waste NPPO
M/ef fluent 1,2 ,4-Tnchlorobenzene 1.8
N/raw waste Diethyl phthalate 5.9
N/effluent 1,4-Dichlorobenzene 1.5
1,2-Dichlorobenzene 6.0
Diethyl phthalate 9.4
P/raw waste Naphthalene 1.9
Diethyl phthalate 1.7
Di-n-butyl phthalate 9.8
P/effluent NPPO
H/raw waste Di-n-butyl phthalate 7.3
R/effluent0 Diethyl phthalate 2
S/raw waste NPPO
S/effluent NPPO
1,4-Dichlorobenxene 11
1,2,4-Trichlorobenzene 90
l,2,4-Trichlorobenr«no 46
Naphthalene 41
HPPO
NPPO
1,2,4-Trichlorobenzene 10.2
Di-n-butyl phthalate 16.2
NPPO
NPPO
Bis(2-ethylhexyl) phthalate IB
1,2-Dichlorobenzene 34.6
Acenaphthene 12.0
Fluorene 14.6
Diethyl phthalate 33.6
Bis(2-ethylhexyl) phthalate 23
Naphthalene 95
Bi>(2-ethylhexyl) phthalate 19
Diethyl phthalate 11.1
Bis(2-ethylhexyl) phthalate 10.3
Acenaphthene 27
Bis(2-ethylhexyl) phthalate 14
NPPO
Naphthalene 79.7
Di-n-butyl phthalate 23.2
Bis(2-ethylhexyl) phthalate 35.2
NPPO
NPPO
Acenaphthene 30
NPPO
Naphthalene 92.9
Di-n-butyl phthalate 58.4
Naphthalene 17
Bis(2-ethylhexyl) phthalate 10.1
Bis(2-ethylhexyl) phthalate 15.7
Dimethyl phthalate 11.6
Bis(2-ethylhexyl) phthalate 30.2
Bis(2-ethylhexyl) phthalate 72
N-nitroso-di-n-propylamine 18.9
NPPO
Bis(2-ethylhexyl) phthalate 12
NPPO
Bis(2-ethylhexyl) phthalate 41
ai rag/m3 equals ug/].
No priority pollutants observed.
csample inadvertently collected prior to the settling pond.
NPPO11
NPPO
NPPO
NPPO
Bis(2-ethylhexyl) phthalate 136
NPPO
NPPO
NPPO
NPPO
NPPO
1,2,4-Trichlorobenzene 120
NPPO
Acenaphthene 273
NPPO '
NPPO
Bisi2-ethylhexyl) phthalate 231
Bis(2-ethylhexyl) phthalate 162
NPPO
NPPO
NPPO
Dimethyl phthalate 111
NPPO
1,2,4-Trichlorobenzene 156
Bis(2-ethylhexyl) phthalate 305
NPPO
1,4-Dichlorobenzene 215
1,2-Dichlorobenzene 287
NPPO
NPPO
Bis(2-etnylhexyl) phthalate 123
NPPO
1,2,4-Trichlorobenzene 190
Naphthalene 143
Bis(2-ethylhexyl) phthalate 135
1,2,4-Trichlorobenzene 916
Naphthalene 255
(continued)
41
-------
TABLE 12 (continued)
Compounds identified and concentrations observed9
Plant/source
T/raw waste
T/ef fluent
U/raw waste
U/ef fluent
V/raw waste
V/effluent
W/raw waste
W/effluent
X/raw waste
X/ef fluent
Y/raw waste
Y/effluent
Z/raw waste
<10 ng/m*
NPPOb
NPPO
1,2-Dichlorobenzene 2.0
Naphthalene 1.5
Die thy 1 phthalate 6.1
NPPO
Acenaphthene 8.7
Bis(2-«thylhexyl) phthalate S.3
Hexachlorobenzene 2.0
Bis(2-ethylhexyl) phthalate 9.5
Hexachlorobenxene 0.5
NPPO
Naphthalene 1
Bi*(2-ethylhexyl) phthalate 1
Diethyl phthalate 3.2
Bi<(2-ethylhexyl) phthalate 2.3
Bexachlorobenzene 0.5
1 , 2-Dichlorobenzene 0.1
Naphthalene 0.4
1,2-Dichlorobenzene 0.6
Naphthalene 0.6
Diethyl phthalate 2.6
Di-n-butyl phthalate 6.7
Hexachlorobenzene 0.3
NPPO
10 to 100 ma/to*
N-nitrosodiphenylanlne 11.3
Bis(2-ethylhexyl) phthalate 23
Bi»(2-«thylhexyl) phthalatt 14.0
Naphthalene 22
1,2,4 -Trlchlorobenzen* 27.9
Dinethyl phthalate 12.9
NPPO
Bis(2-ethylhexyl) phthalate 18.1
Bia(2-ethylhexyl) phthalate 19.0
Acenaphthene S3
NPPO
2,6-Dinitrotoluene 53.5
Bis(2-ethylhexyl) phthalate 87.3
Bi»(2-ethylhexyl) phthalate 25.2
1,2,4-Trichlorobenzene 45
>100 ng/B9
Bis(2-ethylhexyl) phthalate 138
NPPO
NPPO
A
w
Bi»(2-ethylhexyl) phthalate 140
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
Naphthalene 309
Z/effluent
C-001/raw
waste
Y-001/raw
waste
. Y-001/
effluent
JJ/raw waste
JJ/effluent
KK/raw waste
KK/effluent
LL/raw waste
LL/effluent
MM/raw waste
MM/effluent
NN/raw waste
NN/effluent
00/raw waste
00/effluent
PP/raw waste
Bis(2-ethylhexyl) phthalate 2
NPPO
Naphthalene 4
Indeno(l,2,3-cd> pyrene 2
Bis(2-ethylhexyl) phthalate 3
Naphthalene 4.5
NPPO
NPPO
Diethyl phthalate 2.5
Bis(2-ethylhexyl) phthalate 9.3
Pyrene 0.9
Bis(2-ethylhexyl) phthalate 4.1
Pyrene 0.2
1,2-Dichlorobenzene 0.6
Dimethyl phthalate 0.2
Bis(2-ethylhexyl) phthalate 5.2
NPPO
Dimethyl phthalate 0.2
Diethyl phthalate 1.2
Bis(2-ethylhexyl) phthalate 6.9
Bis(2-ethylhexyl) phthalate 2.8
Bis(2-ethylhexyl) phthalate 3.0
NPPO
-NPPO
NPPO
Bis(2-ethylhexyl) phthalate 3.2
NPPO
NPPO
Naphthalene 17.0
Diethyl phthalate 69.0
Bis(2-ethylhexyl) phthalate 23.3
Acenaphthene 13
Diethyl phthalate 15
Diethyl phthalate 11.7
1,2-Dichlorobenzene 11.4
1,2,4-Trichlorobenzene 32.4
Dimethyl phthalate 11.6
NPPO
Naphthalene 51.3
NPPO
Bis(2-ethylhexyl) phthalate 15.4
(HM-2-1) NPPO
(HM-3-1) NPPO
(HM-4-1) NPPO
Bia(2-ethylhexyl) phthalate 23.1
Bia(2-ethylhexyl) phthalate 27.3
Bis(2-ethylhexyl) phthalate 26.0
Di-n-butyl phthalate 61.4
NPPO
Butyl benzyl phthalate 72.8
Naphthalene 44.3
Bis(2-ethylhexyl) phthalate 218
NPPO
NPPO
NPPO
Bis(2-ethylhexyl) phthalate 134
1,2,4-Trichlorobenzene 435
NPPO
NPPO
NPPO
»
1,2,4-Trichlorobenzene 315
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
1 mg/m3 equals 1 vg/1.
No priority pollutant observed.
42
-------
TABLE 13. GC/MS ANALYSES FOR VOLATILE, ACID, PESTICIDE, AND PCB
ORGANIC COMPOUNDS IN RAW WASTE AND EFFLUENT SAMPLES
Compounds identified and concentrations
Plant/source
ft/raw waste
A/effluent
B/raw waste
B/effluent
C/raw waste
C/.effluent
D/raw waste
D/effluent
E/raw waste
Volatiles, mq/ms
NPPOb
Toluene 8.4 -
_ Chloroform 3 . 04
Toluene 3.74
Trichlorofluoromethane 2.60
. Trichloroethylene 17.8
Toluene 236
Ethylbenzene 112
1,1,2, 2-Tetrachloroethylene 26. 4
Toluene 2.6
Ethylbenzene 2.0
Chloroform 3.3
Toluene 2.3
Ethylbenzene 57.3
Toluene 1.67 '
Benzene 5.4
Acids, mg/m*
Phenol 1.2
Pentachlorophenol 71
NPPO
NPPO
NPPO
Phenol 0.5
NPPO
Pentachlorophenol 22
NPPO
Phenol 5.7
observed*
Pesticides and PCB, mq/m3
Heptachlor 6.37
Heptachlor 1.55
NPPO
NPPO
NPPO '
NPPO
NPPO
NPPO
NPPO
Toluene 61.1
Ethylbenzene 20.7
, . . Chloroform 21.5
2>ans-l,2-dichloroethylene 1.8
1,1,1-trichloroethane 16.7
Trichloroethylene 2.0
Chlorobenzene 1.0
E/effluent Toluene 5.5
F/raw waste Trichlorofluoromethane 45
1,1-Dichloroethane 0.59
1,2-Dichloropropane 1.50
1,1,1-Tnchloroethane 11.26
Ct8-l,3-dichloropropene 2.08
Toluene 12.28
F/effluent Trichlorofluororoethane 3.73
3>ar:s-l,3-dichloropropene 3.90
Cfe-l,3-dichloropropene 5.61
Toluene 0.85
Ethylbenzene 2.66
G/raw waste ; Chloroform 5.19
G/effluent Toluene 0.8 .
H/raw waste .Toluene 25.7
Ethylbenzene 5.7
H/effluent Toluene 11.9,
Trichlorofluoromethane 2,138
J/raw waste Toluene 36.1
J/effluent Toluene 8.0
i
J/effluent Toluene 8.0
Ethylbenzene 50.8
Pentachlorophenol 30.1
NPPO
Phenol 8.2
Pentachlorophenol 2.4
2,4-Dimethylphenol 9
Phenol 0.8
Phenol 2
Phenol 63
2-Nitrophenol 60
p-Chloro-w-cresol 4.5
4-Nitrophenol 65
NPPOb
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
al mg/m3 equals 1 ug/1.
No priority pollutant observed.
(continued)
43
-------
TABLE 13 (continued)
Compound« identified and concentrations observeda
Plant/source
volatilea. mq/m3
Acida, mq/m3
Pesticides and PCB, mg7m»
K/raw waste Chloroform 4.8
Toluene 29.3
Ethylbenzene 63.8
K/effluent Chloroform 58.1
Trichloroethylene 4.6
Toluene 24.0
Ethylbenzene 0.7
L/raw waste Chloroform 2.S
Toluene 5.2
Ethylbenzene 2.0
L/effluent Benzene 0.5
M/raw waste NPPOb
M/effluent Toluene 0.4
N/raw waste Trichloroethylene 20.8
Toluene 43.8
Ethylbenzene 1,770
N/effluent Toluene 16.6
Ethylbenzene 75.0
P/raw waste Chloroform 17.3
Toluene 36.1
Ethylbenzene 1,209'
Chlorobenzene 24.8
P/effluent Chloroform 6.9
Toluene 22.4
Ethylbenzene 278
R/raw waste Chloroform 33.2
Benzene 31.0
Toluene 281
Ethylbenzene 2,835
Chlorobenzene 296
1,1,2,2-Tetrachloroethylene 15.1
R/effluent0 Toluene 16.8
Ethylbenzene 28.7
S/raw waste Chloroform 71.1
1,1,2,2-Tetrachloroethylene 38.7
Chlorobenzene 13.6
Toluene 60.7
Ethylbenzene 851.7
S/effluent Toluene 21.4
Ethylbenzene 109
1,1,2,2-Tetrachloroethylene 0.4
T/raw waste Toluene 303
Ethylbenzene 18.4
1,1,2,2-Tetrachloroethylene 6.4
2,4,6-Trichlorophenol 0.7
Pentachlorophenol 3.9
NPPO
NPPO
NPPO
Phenol 12.4
Pentachlorophenol 6.9
NPPO
Phenol 11
2,4-Dimethylphenol 8
Phenol 6.6
NPPO
NPPO
Chloro cresol 32
Pentachlorophenol 56
NPPO
NPPO
NPPO
al rog/m3 equals 1 pg/1.
No priority pollutant observed.
cSample inadvertently collected prior to the settling pond.
NPPO
y-BHC 0.31
NPPO
NPPO
NPPO
NPPO
1 NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
B-BHC 0.35
NPPO
NPPO
(continued)
44
-------
TABLE 13 (continued)
Plant/source
Compounds identified and concentrations observed'
Volattles, mg/m3
Acids, mq/m3
Pesticides and PCB, mq/mj
T/effluent Toluene 33.1
1,1,2,2-Tetrachloroethylene 2.9
U/raw waste 1,1-Dichloroethane 3.67
1,1,1-Trichloroethane 306
U/effluent Chloroform 18.05
Bromodichloromethane 1.54
Trans-1,3,-dichloropropene 0.89
Toluene 13.03
V/raw waste Toluene 8.4
Ethylbenzene 4.9
V/effluent Toluene 1,401
W/raw waste Trichloroethylene 13.1
Benzene 19.4
Toluene 62.2
Ethylbenzene 1.1
W/effluent Toluene 1.7
X/raw waste 1,1,1-Trichloroethane 8.2
Toluene 63.5
Ethylbenzene 369
1,1,2.2-Tetrachlorethylene 414.2
X/effluent Toluene 39.6
Trichlorofluoromethane 35.0
1,1,2,2-Tetrachloroethylene 40.5
Y/raw waste NPPO
Y/effluent Chloroform 4.8
Trichlorofluoromethane 10.1
Z/raw waste Toluene 5.5
Ethylbenzene 0.7
1,1,2,2-Tetrachloroethylene 12.0
Z/effluent Toluene 110.6
Ethylbenzene 3,018
Chlorobenzene 3.5
Trichlorofluoromethane 89.3
C-001/raw waste Toluene 5.7
Trichlorofluororoethane 26.8
Y-001/raw waste
Y-001 effluent
JJ/raw waste
Chloroform 14.3
Chlorobenzene 1.6
Chlorobenzene 1.6
Toluene 11.6
Ethylbenzene 1.9
Toluene 15.1
Trxchloroethylene 187
1,1,2,2-Tetrachloroethylene 1,126
Ethylbenzene 14
1 mg/m3 equals 1 ug/1.
No priority pollutant observed.
NPPO
Phenol 0.7
Pentachlorophenol 1.6
NPPO
NPPO
NPPO
Phenol 100
NPPO
Phenol 3.8
NPPO
Phenol 10.0
NPPO
Phenol 34
NPPO
NPPO
Phenol 19
Phenol 2.9
p-Chloro-m-cresol 1.6
Phenol 41.4
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
NPPO
Not analyzed
(continued)
45
-------
TABLE 13 (continued)
Compounds Identified and concentrations observed*
Plant/source
Volatlles,
Acids, roq/m3
Pesticides and PCS, mq/m3
JJ/effluent Trichloroethylene 84
KK/raw waste Trichloroethylene 52
Toluene 28
Chlorobenzene 42
Ethylbenzene 26
KK/effluent Benzene 64
Chlorobenzene 26
LL/raw waste Chloroform 498
Trichloroethylene 121
1,1,2,2-Tetrachloroethylene 1,108
Ethylbenzene 484
NPPO Not analyzed
2-Chlorophenol 131 Not analyzed
2,4,6-Trichlorophenol 20.2
Pentachlorophenol 20.4
2-Chlorophenol 10 Not analyzed
2,4,6-Trichlorophenol 21.1
Phenol 16.1 Not analyzed
LL/ef fluent
MM-l/raw waste
MM- 2/ef fluent
MM- 3/ef fluent
MM-4/effluent
NN/raw waste
NN/effluent
OO/raw waste
OO/effluent
PP/raw waste
NPPO
NPPO
NPPO
NPPO
Toluene 2
NPPO
NPPO
Chloroform 48
Trichloroethylene 42
Chloroform 10
Toluene 3
Benzene 200
Toluene 83
Ethylbenzene 42
NPPO
NPPO
NPPO
NPPO
NPPO
Phenol 10.1
NPPO
Phenol 22.9
NPPO
NPPO
, Not analyzed
Not analyzed
Not analyzed
Not analyzed
Not analyzed
Not analyzed
Not analyzed
Not analyzed
Not analyzed
Not analyzed
al mg/m3 equals 1 yg/1.
b
No priority pollutant observed.
46
-------
TABLE 14. ;NUMBERTOP PRIORITY ORGANIC POLLUTANTS FOUND IN
THE RAW WASTE AND SECONDARY EFFLUENT STREAMS
Number of! organic compounds detecteda
Plant In raw waste In secondary effluent
A
B
C
b
E
F
G
H
J
K
L
M
N
P
;R
,s
>T
'U
V
-------
The frequency of occurrence of 45 organic species in 64 waste-
water samples is given in Table 15. Dominant compounds were
bis(2-ethylhexyl)phthalate found in 54 samples, toluene found
in 44 samples, and ethylbenzene found in 30 samples.
Results of metal analyses by inductively coupled argon plasma
(ICAP) and atomic absorption (AA) analysis are presented in
Tables 16 and 17.
Table 16 lists concentrations for the metals included in the 129
consent decree priority pollutants list (Appendix A).; Table 17
lists data for the additional elements automatically measured in
ICAP analysis. The upper and lower rows of numbers for each
plant correspond to metal concentrations in the raw wastewater
and secondary effluent, respectively. All metal concentrations
are reported as g/m3 (ppm).
The lower detection limits ;for routine ICAP and AA analyses,
referred to in footnote "a" of Tables 16 and 17; are given in
Table 18.
LEVEL 1 CHEMICAL ANALYSIS
Analysis Procedures
The EPA-Process Measurements Branch, IERL/RTP, has developed a
phased sampling and analytical strategy for environmental assess-
ment programs (2). Level 1 is the first part of a three-phase
approach for performing the assessments. The Level'1 chemical
analysis procedure, including modifications to accommodate the
special requirements of textile wastewaters, are discussed in
this section.
Level 1 chemical analyses were performed on samples from only 15
of the 23 basic textile plants because this task was!implemented
by EPA after the program began. Eight-hour composited grab
samples were collected from the secondary effluent at the 15
plants.
Figure 16 is a schematic diagram for field handling of waste-
water samples as recommended by EPA (2). The procedure specifies
collection of 0.02 m2 of composite sample, which is divided into
two 0.01 m3 portions of organic and inorganic chemical analysis.
Part of the inorganic composite (0.001 m*) is set aside for
determination of BOD5 and COD. The remainder is filtered in the
field with Gelman Spectro-Grade glass fiber filters (or equiva-
lent) . Tared filters are sent to the laboratory for analysis of
inorganic elements, leachable anions, and total suspended solids.
Filtrate is extracted in the field with methylene chloride to
separate organic from inorganic chemical species. The aqueous
portion is divided into three parts for analysis.
48
-------
TABLE 15. OCCURRENCE OF PRIORITY ORGANIC POLLUTANTS COMBINED
FROM RAW WASTE AND SECONDARY EFFLUENT SAMPLES
Number of Samples in which
pollutant was detected9
Priority pollutant
Bis(2-ethylhexyl)phthalate
Toluene
Ethylbenzene
Naphthalene
Diethyl phthalate
Phenol
Chloroform
1,2, 4-Trichlorobenzene
1 , 2-Dichlorobenzene
1,1,2, 2-Tetrachloroethylene
Trichloroethylene
Acenaphthene
Di-n-butyl phthalate
Pentachlorophenol
Dimethyl phthalate
1 , 4-Dichlorobenzene
Chlorobenzene
Tr ichlorof luoromethane
Benzene
1,1, 1-Trichloroethane
Pyrene
Hexachlorobenzene
2,4, 6-Trichlorophenol
N-nitroso-di-n-propylamine
N-nitrosodiphenylamine
Heptachlor
Anthracene
Fluorene
1 , 1-Dichloroethane
Cts-1, 3-dichloropropene
Trans-l, 3-dichloropropene
2 , 4-Dimethylphenol
2-Chlorophenol
a-BHC
B-BHC
2, 6-Dinitrotoluene
Indeno (1,2, 3-cd ) pyrene
Butylbenzyl phthalate
Trans-l , 2-dichloroethylene
1 , 2-Dichloropropane
2-Nitrophenol
4-Nitrophenol
Chloro cresol
Broitiodichloromethane
Total
54
44
30
25
21
21
17
14
13
11
10
9
9
8
8
8
8
8
6
5
5
5
3
3
3
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
Raw waste
samples
27
22
20
20
12
19
12
8
8
8
8
7
6
8
5
5
6
2
4
5
1
2
2
1
1
1
1
2
2
1
0
0
1
0
1
1
1
0
1
1
1
1
0
0
Secondary
effluent samples
27
22
10
5
9
2
5
6
5
3
2
2
3
0
3
3
2
6
2
0
4
3
1
2
2
1
1
0
0
1
2
2
1
1
0
0
0
1
0
0
0
0
1
1
Observed
concentration
range, "*c mg/m3
0.5 to 300
0.4 to 300
0.7 to 3,000
0.03 to 300
0.2 to 70
0.5 to 100
2 to 500
2 to 900
0.1 to 300
0.4 to 2,100
2 to 200
0.5 to 270
2 to 60
2 to 70 .
0.2 to 110
0.05 to 200
1 to 300
2 to 2,100
0.5 to 200
2 to 300
0.1 to 0.9
0.3 to 2
0.7 to 20
2 to 20
2 to 20
2 to 6
0.1 to 4
5 to 15
0.6 to 4
2 to 6
0.9 to 4
8 to 9
10 to 130
0.3
0.4
50
2
70
2
2
70
70
30
2
Out of a total of 64 samples.
Rounded to one significant figure.
cl mg/m3 equals 1 yg/1.
49
-------
TABLE 16. CONSENT DECREE METALS CONCENTRATIONS IN WASTEWATER SAMPLES
en
O
Plant
Code
A
B
C
D
E
F
G
H
J
K
L
M
N
P
RC
s
Metals concentration -
Silver
0
0
0
0
0
0
0
0
0
0
a
a
a
a
a
a
.011
a
.007
a
.10
.08
.0085
a
.041
a
.06
a
.13
a
a
a
a
a
a
a
.03
.008
a
a
a
a
Arsenic Beryllium Cadmium
a
a
a
a
a
a
0.017
0.006
a
a
a
a
a
a
a
a
a
a
0.006
a
a
a
a
a
a
a
< a
a
a
a
0.005
a
b
b
b
b
a
a
a
a
a
a
b
b
a
a
a
a
a
a
a
a
a
a
a
a
b
b
a
a
a
a
a
a
a
a
0.0007
a
0.005
0.006
a
a
0.006
0.001
0.01
0.01
a
a
a
a
a
a
0.004
a
a
a
a
a
0.046
a
a
a
a
a
a
a
Chromium
0.19
0.18
0.012
0.004
0.035
0.031
a
a
0.011
0.004
0.006
0.004
0.004
0.003
0.004
a
0.048
0.025
0.019
0.004
0.003
0.03
a
a
0.88
1.8
0.003
a
0.067
0.14
0.0007
a
Copper
0.021
0.027
0.074
0.03
0.008
0.020
0.031
a
0.84
0.03
0.59
0.13
0.063
0.028
0.022
a
2.4
0.1
0.026
0.015
0.30
0.096
0.009
0.005
0.020
0.008
a
a
0.51
0.29
0.04
0.06
- raw waste, g/ro3
- secondary effluent, a/m'
Mercury
0.004
a
0.0009
0.0006
a
0.0007
a
a
a
a
a
0.0009
a
a
a
a
a
a
a
a
a
a
a
a
0.0004
a
a
a
a
a
a
a
Nickel
0.009
0.14
a
a
0.15
0.14
0.03
a
0.04
0.04
0.10
0.06
0.028
0.013
0.014
a
0.097
0.09
0.1
a
0.054
0.035
a
a
a
0.03
0.10
0.04
a
a
a
Lead
a
a
a
a
0.12
0.12
a
a
0.008
a
0.08
0.0006
0.006
a
a
a
0.029
a
0.03
a
0.036
a
a
a
a
a
0.013
a
a
a
a
a
Antimony Selenium
a
0.03
a
a
0.007
0.004
0.003
0.002
0.008
0.0008
0.001
0.0003
0.052
0.011
0.004
0.006
0.0007
a
0.003
0.0008
0.005
0.003
0.0008
0.004
0.0002
0.002
a
a
a
a
0.057
0.074
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
Thallium Zinc
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
1.3
6.4
0.3
0.17
0.074
0.12
0.21
0.21
7.9
5.1
0.26
0.57
0.45
0.26
3.9
0.96
2.1
0.8
0.15
0.11
1.0
0.72
1.2
0.41
7.5
38.4
0.20
0.14
_ 0.24
0.21
0.12
0.084
Metals concentration below instrument detection limit - see Table 18 for detection limit.
Analysis not performed.
cSecondary effluent sample inadvertently collected prior to the settling pond.
-------
TABLE 16 (continued)
Plant
code
T
U
V
K
X
r
z
y-ooi
C-001
jj
KK
LL
MM-1
-2
-3
-4
UN
OO
PP
Sliver
a
a
a
a
a
a
0.065
0.095
0.017
0.033
a
a
a
a
0.068
0.057
0.033
0.047
0.049
0.022
0.044
0.058
0.056
0.016
0.025
0.028
0.032
0.042
0.033
0.046
0.050
0.048
Arsenic
a
-- a
a
a
a
a
a
0.004
a
a
a
a
a
a
*
a
a
0.20
0.16
0.12
a
0.10
0.07
0.055
0.003
0.007
0.006
a
a
a
a
a
Beryllium
b
.~b
b
b
a
a
a
a
b
b
a
a
a
a
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
Metals
Cadmium
a
a
a
a
o.oos
a
0.009
0.013
0.005
0.007
a
a
a
a
0.006
0.007
0.003
O.OOS
0.005
0.002
0.004
0.004
0.002
0.002
0.002
a
0.002
0.002
0.004
0.004"
0.005
0.005
concentration - raw waste, g/mj
- secondary effluent.
Chromium
a
a
0.027
0.014
0.004
0.003
0.012
0.003
0.024
0.039
0.026
0.001
a
a
0.65
0.29
0.024
0.16
0.08
0.016
0.013
0.011
0.020
0.111
0.058
0.11
0.13
0.023
0.17
0.011
0.012
0.010
Copper
0.12
0 •. 06.
0.040
0.023
0.23
0.17
0.023
0.002
0.084
0.11
0.096
0.11
0.097
0.050
0.041
a
a
0.032
0.031
0.086
0.037
0.038
0.092
0.036
0.059
0.028
0.042
0.047
0.046
0.039
0.037
0.041
Mercury
0.0007
a
0.0004
a
a
a
a
o.ooos
a
0.0009
a
a
a
a
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
Hickel
0.05
0.004
0.008
a
a
0.054
0.060
0.11
0.072
0.012
a
0.011
a
0.20
0.16
0.099
0.10
0.12
0.077
0.11
0.13
0.15
0.044
0.072
0.067
0.081
0.098
0.079
0.11
0.12
0.12
g/m3
Lead
0.025
_a
a
a
a
a
0.018
0.057
0.032
0.026
a
a
a
a
0.16
0.16
0.073
O.OJ4
0.065
0.049
0.044
0.060
0.048
0.011
0.037
0.031
0.035
0.033
0.025
0.043
0.084
0.078
Antimony
0.007
0.001
a
0.004
a
a
0.0003
0.0009
0.016
0.003
0.011
0.012
a
a
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
Selenium
a
_ a
a
a
a
a
a
a
a
a
a
a
a
a
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
Thallium
a
a
a
a
a
a
a
a
a
a
a
a
a
a
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
Zinc
0.29
0.08
0.26
0.19
0.46
0.34
0.19
0.09
0.034
0.078
0.24
0.091
0.11
0.37
0.13
0.10
0.056
0.13
0.32
1.08
0.39
0.067
0.068
0.24
0.19
0.28
0.37
0.084
0.13
0.12
2.3
0.073
Petals concentration below instrument detection limit - see Table 18 for detection limit.
Analysis not performed.
°Secondary effluent sample inadvertently collected prior to the settling pond.
-------
TABLE 17. OTHER METALS GIVEN BY ICAP ANALYSIS
Ul
Plant
code
A
B
C
D
E
F
G
R
J
K
L
M
N
f
RC
S
Metals concentration - raw waste , g/m3
- secondary effluent, q/m3
Aluminum
0.24
0.23
0.16
0.024
0.30
0.19
0.38
0.15
0.028
0.038
0.11
0.06
0.52
0.23
0.093
0.055"
0.95 -
0.01
0.28
a
0.095
0.064
0.33
0.009
0.29
1.2
0.62
0.14
0.28
0.07 -
0.068
0.91
Barium
0.064
0.030
0.037
0.008
0.083
0.073
a
a
0.015
0.012
0.03
0.03
0.008
0.015
a
a
0.12
0.024
0.028
0.018
0.008
0.019
0.013
a
0.018
0.16
a
a
0.003
a -
0.005
0.028
Boron
0.15
0.18
0.07
0.043
0.39
0.15
1.4
0.90
2.36
1.O6
0.84
0.68
O.69
0.22
a
a
a
a
13
1.1
6.1
8.9
0.91
0.53
0.041
0.058
0.20
0.52
0.21
0.21
1.6
1.5
Calcium
18
28
20
12
5.0
4.5
3.6
6.6
5.4
39
4.1
4.2
4.2
3.1
1.3
6.2
4.9
5.2
5.8
3.6
3.4
6.3
9.7
8 8
10
26
1.6
9.5
7.5
. 5.1.
2.8
7.3
Cobalt
0.0042
0 013
0.0056
0.0052
0.0081
0.0056
0.001
0.001
a
a
0.0041
0.012
0.0008
0.011
0.0054
a
0.0084
0.0009
0.009
0.007
0.0012
0.0012
0.0077
0.0045
0.007
0.030
0.007
0.003
0.028
0.026
0.001
0.001
Iron
0.72
2.1
0.32
0.27
1.0
0.22
0.39
0.46
0.12
0.62
0.33
0.28
0.17
0.39
0.16
0.40
0.70
0.52
0.67
0.088
0.18
0.46
0.18
0.06
1.4
4.7
0.72
0.12
0.30
0.13 .
0.12
0.31
Magnesium
3.6
4.0
9.5
4.67
3.9
0.73
3.1
4.6
2.5
3.1
3.1
3.2
0.52
0.49
2.6
3.2
5.8
6.9
4.2
3.7
1.3
2.4
6.7
6.4
1.68
4.2
0.48
1.8
3.6
- 3.0
1.8
2.8
Manganese
0.05
0.07
0.093
0.059
0.029
0.017
0.03
0.07
0.027
0.10
0.007
0.009
0.05
0.038
0.013
O.O09
0.05
0.05
0.038
0.011
0.022
0.021
0.034
0.14
0.46
1.1
0.042
0.02
0.018
0.013
0.021
0.03
Molybdenum
a
a
a
a
0.019
0.024
a
a
a
0.019
0.025
0.023
0.0026
0.011
a
0.0043
a
0.020
0.0006
a
a
a
a
a
0.008
0.002
a
a
a
a
Sodium
MOO
MOO
MOO
MOO
MOO
MOO
MOO
MOO
MOO
54
MOO
MOO
97
55
MOO
MOO
MOO
MOO
MOO
MOO
MOO
>100
MOO
MOO
MOO
MOO
76
MOO
MOO
MOO
MOO
MOO
Silicon
3.3
1.4
4.4
2.3
16
15
6.8
7.3
11
8.6
10
13
2.4
3.2
17
15
18
17
23
IS
6.0
6.9
15
14
11
6.9
5.7
4.8
13
11
14
15
Tin
0.05
0.02
0.018
0.041
0.0?
0.12
0.02
0.02
0.03
0.01
0.07
0.04
0.03
0.02
0.019
O.OSS
0.03
0.05
0.05
0.04
0.019
0.007
4.0
3.5
0.012
a
0.06
0.004
0.003
a
0.007
0.018
'Phosphorus
1.2
0.50
12
6.5
4.0
4.1
1.6
1.0
1.9
1.4
24
9.5
- 6.4
6.1
0.99
0.20
3.3
0.6
1.9
0.93
2.2
1.6
3.99
3.46
0.43
5.2
5.7
2.2
3.9
.0.66
1.6
5.0
Titanium
0.012
0.008
0.0039
a
0.020
0.012
0.008
0.0001
0.018
0.012
0.0028
0.0039
0.011
a
0.009
a
1.5
0.06
0.0035
0.0104
0.010
0.0013
0.0092
a
. 0.010
0.001
0.006
a
0.071
a
0.0005
O.OOO5
VanadiuB
0.060
0.59
0.058
0.030
0.29
0.40
0.03
0.03
0.019
0.021
0.014
0.013
' 0.030
0.022
0.03S
0.032
0.13
0.11
0.012
0.035
0.012
0.033
0.042
0.037
0.013
0.037
0.02
0.02
0.021
0.016
0.008
0.024
Metals concentration below instrument detection limit - see Table 18 for detection limits.
Secondary effluent sample only.
Secondary effluent samples inadvertently collected prior to settling pond.
C"
-------
TABLE 17 (continued)
en
Plant
code
T
U
V
W
X
Y -
Z
Y-001
C-001
JJ
KK
LL
KM-1
-2
-3
-4
NN
OO
PPb
Hetals concentration - waste, g/ra3
- secondary effluent, 5/m*
Aluminum
0.20
0.075
0.90
0.17
0.20
0.16
6.0
0.77
0.14
0.94
0.076
0.018
0.023
v a
0.57
6.3
0.29
2.7
1.8
0.38
0.26
0.22
0.17
0.44
3.7
0.48
0.50
1.0
1.3
0.24
0.15
0.052
Barium
0.013
0.008
0.01
0.006
0.014
0.013
0.33
0.15
0.028
0.005
0.13
0.029
a
a
0.015
0.011
0.070
0.048
0.042
0.060
0.051
0.008
0.004
0.010
0.005
0.020
0.011
0.002
0.001
0.018
0.016
0.004
Boron
0.73
0.34
0.032
0.28
3.4
0.49
0.44
0.14
0.28
0.31
1.4
2.3
0.12
0.36
0.045
0.68
0.11
0.23
0.24
1.0
0.85
0.043
0.091
1.1
1.0
1.1
1.1
0.043
0.800
1.1
1.1
0.017
Calcium
12
9.9
5.8
8.0
3.2
3.6
93
36
8.3
9.3
2.1
8.9
2.5
2.1
68
61
22
7.3
16.5
12.4
11.8
4.5
3.7
5.7
5.1
4.3
8.9
0.85
5.7
3.3
3.5
0.021
Cobalt
0.008
0.00.1
0.005
a
a
0.004
0.14
0.005
0.034
0.024
0.026
0.004
O.O09
0.006
0.32
0.27
0.019
0.012
0.010
0.004
0.009
O.O04
0.004
0.004
0.009
0.005
0.006
0.010
0.008
0.004
0.006
0.007
Iron
0.14
0.43
0.68
O.30
0.37
0.47
5.7
1.7
0.22
0.35
0.14
0.17
0.16
O.075
1.2
1.1
2.9
2.3
1.5
0.42
0.46
0.16
0.10
0.39
0.30
0.25
0.40
0.28
0.75
0.37
0.10
0.009
Magnesium
4.2
3.5
2.5
2.5
1.4
1.7
14
5.0
1.4
1.9
3.2
5.8
0.65
1.2
3.1
3.0
8.1
2.2
4.0
14.4
11.6
2.0
2.0
3.7
3.5
2.9
3.8
0.26
0.55
•1.3
0.88
0.0004
Manganese
0.18
0.17
0.076
0.016
0.063
0.072
0.89
0.38
O.O20
0.010
0.044
O.046
O.O14
0.010
0.12
O.O8
0.45
0.14
0.19
0.054
0.036
0.040
0.021
0.030
0.025
0.017
0.032
0.12
0.008
0.020
0.019
0.002
Molybdenum
a
a
a
a
a
a
a
0.021
0.008
0.012
a
a
a
a
0.031
0.042
0.011
0.026
0.024
0.010
0.015
0.015
O.016
0.001
0.010
0.004
0.004
0.005
0.006
0.001
0.012
0.001
Sodium
MOO
MOO
MOO
MOO
94
MOO
65
51
82
MOO
MOO
MOO
MOO
MOO
65
59
MOO
88
MOO
MOO
MOO
MOO
MOO
MOO
MOO
MOO
MOO
MOO
MOO
52
66
0.196
Silicon
15
6.6
3.7
3.0
4.5
4.5
20
10
7.9
8.4
9.8
8.2
8.8
6.2
7.8
7.7
12
9.8
7.8
31.9
27.8
5.7
7.1
7.7
5.9
7.6
7.7
2.3
1.6
5.2
6.3
0.059
Tin
0.042
0.028
0.028
a
0.013
0.005
0.030
0.030
0.006
0.002
0.001
0.002
a
a
0.068
0.058
0.040
0.057
0.042
0.077
0.064
0.033
0.031
0.028
0.021
0.009
0.003
0.052
0.051
0.053
0.036
0.002
Phosphorus
12
17
3.5
3.7
0.75
0.78
5.1
0.15
4.6
5.4
16
IS
1.1
0.5
11.7
6.8
2.7
3.5
2.3
6.3
6.4
18.8
28.8
1.9
0.78
1.9
2.1
48.8
46.8
4.6
0.66
0.08
Titanium
O.OO09
0.0021
0.006
0.003
0.003
a
0.035
0.012
0.011
0.001
0.001
a
0.002
a
0.020
0.008
O.O10
0.096
0.056
0.042
0.017
0.014
0.008
0.006
0.011
0.009
O.OO8
0.006
0.007
0.004
0.006
0.0001
Vanadium
0.045
0.037
0.023
0.019
0.019
0.013
0.14
0.036
0.022
O.O29
O.019
0.037
0.089
0.086
0.039
0.088
0.072
0.044
0.049
0.091
0.077
0.032
O.O86
0.036
0.06S
' O.O34
0.039
0.019
O.O25
0.020
0.020
0.006
°Metals concentration below instrument detection limit - see Table 18 for detection limits.
b
Secondary effluent sample only.
Secondary effluent samples inadvertently collected prior to settling pond.
-------
TABLE 18. LOWER DETECTION LIMIT OF METALS ANALYSIS SYSTEMS
Metals analyzed by ICAPa
Metal
Detection
limit, g/m3
Metal
Detection
limit, g/m3
Aluminum
Arsenic
Barium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
0.050
0.005
0.0002
0.0001
0.0005
0.0002
0.0002
0.0005
0.0002
0.005
0.001
0.001
Manganese
Molybdenum
Nickel
Phosphorus
Sodium
Silicon
Silver
Tin
Titanium
Vanadium
Zinc
0.005
0.0006
0.01
0.01
0.050
0.003
0.0050
0.001.;
0.001
0.002
0.025
Metals analyzed by atomic absorption
Antimony
Beryllium
Mercury
0.0005
0.0001
0.0005
Selenium
Thallium
0.005
0.005
1 g/m3 equals 1 mg/1.
AQUEOUS/ORGANIC/SOLID
0.02-m'AMPLE
DISSOLVED 02
pH, COLOR, ODOR
0 01 - m3 SAMPLE
AQUEOUS/ORGANIC/1
0.02-m3SAMPU
t^Q
EXTRACT ORGAN1CS WITH 1
METHYLENE CHLORIDE , I 1
t t
AOiiFnil<; ORGANIC I DRY AND WEIGH
AgutUUS MATERIAL | FOP TOTAL
1 1 SUSPENDED SOLIDS
DISCARD | FILTER
* i
INORGANIC
aEMENTS
"1
LEACHABLE
ANI'ONS
AC
_£
SOLID
' TRANSFE
CAREFULLY
ENTRAINMB
R 0.001m3
TO AVOID AIR
IT-BOD5.COD
0. 009 -m3 SAMPLE
I FILTRATE"!
\
EXTRACT ORGANICS
WITH METHYLENE CHLORID
1
f
AQUEOUS
1 DIVIDE
INTO 3 PARTS
i 1 t
IDIFY 1 LEAVE 1
i<2 I AS IS I
BASIFYl
E
*
ORGANIC 1
MATERIAL I
1
REMOVE
SOLVENT
WEIGH AND
DISCARD
Figure 16. EPA-recommended field handling scheme
for liquid/slurry samples (1).
54
-------
The 0.01 m3 portion for organic analysis is extracted in the
field with methylene chloride. The aqueous portion is discarded;
the methylene chloride extract is filtered and sent to the lab-
oratory for organic chemical analysis.
Because textile wastewaters from stable emulsions with methylene
chloride, the field handling procedure was modified, with the
approval of the EPA-Process Measurements Branch, IERL/RTP. It
was not feasible to conduct methylene chloride extractions in
the field because previous experience has shown that it requires
from 1 hr to 3 hr of concerted effort to break these emulsions.
Centrifugation was necessary occasionally. The modified field
handling scheme for textile wastewater is shown in Figure 17.
The fundamental difference between the two schemes is that the
field analysis of some of the common wastewater chemical species
is performed on unextracted water samples. Cyanide samples were
not extracted before analysis.
The filter obtained from the field was dried and the concentra-
tion of total suspended solids determined. Leachate analyses
were not performed because each textile plant sampled was meeting
its EPA effluent standard for TSS at the time samples were
collected. The filter paper was ashed by means of low tempera-
ture plasma, digested and analyzed for metals by spark source
mass spectrometry (SSMS) and conventional atomic absorption (AA).
Level 1 chemical analysis protocoal requires SSMS for metals,
analysis and AA for those metals not accurately detected by SSMS.
SSMS can be used for analysis of nonvolatile compounds, such as
inorganic solids and trace elements. The spark produces ions
from the sample by high voltage breakdown across two electrodes.
One electrode usually consists of or contains the sample mate-
rial. In the spark ion source, the spark is sustained between
ions from the rf spark source are accelerated through a potential
field and focused with dual collimating slits. The ion beam
passes into the mass spectrometer where the ions are separated.
Photographic plates are used to record the emissions spectra for
various mass fractions that correspond to specific elements or
compounds.
A detailed flow diagram for Level 1 organic analysis is shown in
Figure 18 (2). First,. 0.01 m3 ;of the sample is extracted with
methylene chloride. The aqueous phase is saved and the organic
phase separated for analysis.
The modification to the Level 1 organic analysis scheme, as
recommended by th^ EPA, was employed in addition to the basic
Level 1 chemical analysis procedure.
55
-------
FIELD HANDLING
AND ANALYSIS "
U1
CTi
LABORATORY <
ANALYSIS
r".
-
WASTEWATER
SAMPLE 0.02m3
1
DO, pH, COLOR
ODOR
10"2m3 1 I0"2m3 10~3m3
| 0.009m3
SHIP TO LAB FILTER
CAD Apr AMI rc
ANALYSIS
t
SHIP FILTER SHIP PROPERLY PRESERVED
TO LAB INDIVIDUAL SAMPLES TO THE
. . . LAB FOR ANAL Y5 IS Or •
ORGANICS METALS
WEIGHT AQUEOUS T°S C>
OtTERWINAi (ON nuacr
i »
.____. _ - _. -
SEE FIGURE 18
FOR DETAILS
-
IFTSS
ARE LOW
,
DRYmDDT"IGH METHYLENE CHLORIDE
EXTRACTION
^YES 1 • — i '
t^OLEACHATE _^^ SAMPLES FOR ORGANIC PHASE: AQUEC
^ [— •- SSM"S,AA REMOVE SOLVENT. METAL
-* ANALYSIS DRY, WEIGH BY SS
HCI LEACHATE
I
_ - . DISCARD
i nyu TFMP
PLASMA ASH
f *
SSMS AA
ANALYSIS ANALYSIS
SHIP TO LAB FOR
" BOD5 AND COO ANALYSIS
0-3 3
1
FIELD ANALYSIS OF
SELECTED WATER
PARAMETERS
'ANIDE
... - - ^~ __„ ...
1 1
' STANDARD
DRY AND WEIGH METHODS
1 ANALYSIS
HJS PHASE :
S ANALYSIS
MS AND AA
Figure 17. MRC-modifled Level 1 field sampling and analysis scheme.
-------
SAMPLE FROM FIELD
ttOlm3
EXTRACT WITH
METHYLENE
CHLORIDE
AQUEOUS PHASE
ORGANIC/EMULSION
BREAK EMULSION
RETAIN Ixllf'm3
T02xlO~*m3
EXTRACT FOR GC
ANALYSIS OF
ORGANICS
CONCENTRATE VIA
ROTOVAPTO
3xlO~4m3
CONCENTRATE VIA
K-DTO
JxloT'm3
DILUTE TO lxlo"5m3
AMBIENTEVAP.
5xlO"6m3
CONSTANT WT.
IR ANALYSIS
LIQUID CHROMATOGRAPHS USING 8 SOLVENT MIXTURES
riiii MI
IR IR IR IR IR IR IR IR
I I I I I III
LRMS LRMS LRMS LRMS LRMS LRMS LRMS LRMS
LRMS - LOW RESOLUTION MASS SPECTROMETER
Figure 18. Level 1 organic analysis scheme (2)
57
-------
Funds to perform the additional methods development required" for
the modification were supplied under another EPA contract.3
The principal feature of this modification is that the organic
extract is not evaporated to dryness prior to liquid chromato-
graphy (LC) and/or low resolution mass spectrometry (LRMS). Two
new sample handling steps are required:
• A quantitative assay procedure using GC/MS to complement
gravimetric analysis and give quantitative data on Cj-C\i
hydrocarbons.
• A solvent exchange step to transfer the sample from methy-
lene chloride extract to nonpolar solvent and allow lower
boiling material (less than C7) to pass through the chro-
matograph and subsequent steps.
The new flow diagram incorporating these changes is shown in
Figure 19.
Results of Inorganic Analyses
The Level 1 chemical analysis scheme is divided into inorganic
and organic analysis. Inorganic analysis includes: BOD5, COD,
TSS, TDS, metals analysis by spark source mass spectrometer
(SSMS) of filtered solids and the filtrate, field analysis of
selected species, and total nonvolatile organic concentration.
Table 19 shows the concentrations of the following parameters in
15 plant effluents: BOD5, COD, TSS, TDS, and total organic con-
centration. Data from the field analysis of effluent samples are
found in Table 20. Metals concentrations of the suspended solids
collected on the filter paper are listed in Table 21. Results of
SSMS analyses of the filtrate are given in Table 22. Concentra-
tions are reported as g/m3 (ppm) of textile effluent sample. All
elements for which values are not entered have concentrations
below the detection limit.
Results of Organic Analyses
As illustrated in Figures 17 and 19, there are four points with-
in the Level 1 chemical analysis scheme where organic analysis
takes place. At each plant one portion of the sample was fil-
tered and a part of this volume was extracted with methylene
chloride. The aqueous phase was used for metals analysis by
SSMS, and the organic phase was dried and weighed' to determine
the concentration of total methylene chloride extractable
organics. Results of these analyses are shown in Table 23.
aContract No. 68-02-1411, Task 19, "Analysis Support of Textile
Environmental Assessments."
58
-------
YES
ALIQUOT
FORIR
ALIQUOT
FORLC
SOLVENT EXCHANGE
10-6m3 HEXANE
PLUS SILICA GEL
I
4
EACH FRACTION:
METHYLENE CHLORIDE
EXTRACT PREPARED AS USUAL
1X10-4 m3 JO 2X10-3 m3
TOTAL VOLUME
CONCENTRATE AS NECESSARY
-------
TABLE 19. SELECTED PARAMETERS OF TEXTILE EFFLUENT
SAMPLES FROM THE INORGANIC SEGMENT OF
LEVEL 1 CHEMICAL ANALYSIS PROTOCOL
Plant
code
A
B
C
E
F
G
K
L
N
S
T
U
V
W
X
5-day Chemical
biochemical oxygen
oxygen a demand
demand, g/m3 g/m3
168
b
25
<5
69
42
<5
13
36
59
32
24
<5
84
15
1652
99
396
78
276
502
131
234
286
1035
414
748
128
837
,258
Total
suspended
solids,
g/m3
234
7
24
10
10
5
14
42
13
349
44
111
b
217
1.3
Total
dissolved Total organic
solids, concentration,
g/m3 g/m3
1,725
1,681
2,924
13,120
2,006
276
1,256
725
1,352
692
660
1,331
b .
1,648 '
437
63.7
3.18
28.2
3.60
16.0
27.2
2.73
18.3
9.24
5.40
17.8
14.6
b
15.0
13.5
7i g/m3 equals 1 mg/1.
Analysis not performed.
Portions (1 to 2 x 10~6 m3) of each sample were then analyzed by
gas chromatography (GC) to determine the concentration of C7 to
Ci2 hydrocarbons. The Following GC columsn were used: a) 1.8 m
x 3.2 mm stainless steel column packed with 10% VC, W98 on 80 to
100S, and b) 1.8 m x 3.2 mm stainless steel column packed with
10% SP-2100. Each column was held at 50°C for 4 min, then pro-
grammed at 16°C/min to 250°C and held at 250°C for 4 min.
Results of these analyses indicated that in 13 of the 14 samples
the C7 to C12 hydrocarbon concentrations were below the threshold
detection limit of 1.0 g/m3 (ppm). The secondary effluent
sample from plant X contained a total concentration of about
3.0 g/m3 of C7 to Ci2 hydrocarbons.
60
-------
TABLE 20. FIELD ANALYSIS OF SELECTED SECONDARY WASTEWATER PARAMETERS ON ILTERED,
UNEXTRACTED SAMPLES AS PER LEVEL 1 ANALYSIS PROTOCOL (2).
Hater parameters, g/m30
Plant
code
A
B
c
E
F
G
K
L
N
S
T
U
V
H
X
Color,
APHA
2,000
90
1,920
0
80
500
270
370
90
240
350
2,480
500
1,900
>10
Specific .
conductivity Nitrate
1,500
1,2OO
2,400
310
1,900
155
875
555
990
640
460
770
360
1,250
285
1.9
0.002
23.3
79.2
0
1.32
4.4
13.5
5.5
4.4
0.8
0.8
0.88
12.3
0.033
Hydrogen
Nitrite sulfide
0.06
<0.005
4.64
0.016
0.043
0.076
0.056
0.864
0.003
0.033
0.04
<0.005
0.264
0.145
0.44
4
0.20
5
0.1
0.1
<2
2
3
0.1
0.3
6
3.5
0.5
0.1
0.01
Sulfate
8.5
368
40
12
10
56
>1
460
640
ISO
100
0
. 57
0
1
Methyl
orange
acidity
0
0
0
0
0
0
0
0
20
0
0
0
0
0
0
Dissolved
oxygen
5.5
7
6
8.5
5
8
5
4
9
7
8
9
9
5
7.2
Ammonia
12.8
2.5
3.4
3.4
1.54
72.5
3.1
0.5
12.8
72.5
13.6
5.44
2.5
0.38
0.05
o-phosphate
1.0
7.3
1.08
2.0
0.56
4
2.5
0.88
11.2
72
6.4
2.96
1.7
0.075
0
Total
alkalinity
100
S-*
8.3
35
2.4
30
710
30
0 '
130
300
120
0.4
950
140
Chromium
0.18
0.004
0.031
0.004
0.004
0.003
0.004
0.03
1.8
0
0
0.014
0.003
0.003
0.039
pa
7.3
7.5
10
7.5
7.4
7.5
7.2
5.8
7.0
7.8
7.4
7.3
7.1
8.1
7.2
1 g/m3 equals 1 mg/1
Units pfflhos at 25°C.
-------
TABLE 21. LEVEL 1 SPARK SOURCE MASS SPECTROMETER METALS
ANALYSIS OF THE SUSPENDED SOLIDS COLLECTED ON
THE FILTER PAPER DURING FIELD FILTRATION
Plant A
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
ytterbium
Thullium
Erbium
Holmium
Dysprosium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium ,
ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.04
<0.04
3.0
42
<0.04
-
<0.08
<0.03
<0.11
<0.04
<0.03
<0.03
<0.01
Plant B
0.05
0.2
.
-
/•
0.003C
<0.02
Plant C
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium.
Erbium
Holmium
Dysprosium
0.7
0.6
0.07,,
a
0.07,
0.01C
0.1
0.07
0.07
Concentration : rag/m3
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Concentration:
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
<0.03
<0.04
<0.04
<0.07
<0.09
<0.05
1.3
0.91
15
0.33
1.6
13 .
-
2.5
1.1
mer/m3
0.1
0.02
0.01
1.8
0.01
_0
Concentration: mg/m*
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
0.07
0.07
0.1
0.07
57
0.03
0.03
0.1
0.2b
V
0.07
Detection limit: 0.01 ma/m3
Ruthenium
Molybdenum
Niobium
Zirconium
yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
iron
Manganese
Chromium
Detection
Ruthenium
Molybdenum
Niobium
Zirconium
yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
17
0.03
1.9
0.08
6.0
1.0
3.0
0.25
3.4
0.03
0.13
300
172
26
2.0
582
11
56
limit: 0.61
0.04
0.1
0.08
0.02
1.8
0.3
0.07
13
0.3
0.5
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
g/m*
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
318
9.5
0.07
1,220
265
15
1,987
1,590
450
874
702
4,636
28 a
a
«~
_*
<0.04
1.3
•
0.02
0.3
30
11
0.3
24
120
340
25
130
240
6.8
-"
_
_'
0.04
Detection limit: 0.05mg/mJ
Ruthenium
Molybdenum
Niobium
Zirconium
yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
0.1
0.2
0.03
1.6
2.8
1.1
0.07
29
4
0.9
0.07
20
0.9
0.8
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
(
1.1
0.8
0.1
83
330
43
110
700
1,100
530
160
2,800
33 „
_a
_a
a
"a
0.3
continued
Not reported. Internal standard. Instrument source.
62
-------
TABLE 21 (continued)
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
•Dysprosium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
Terbium
Gadolinium
0.07 Europium
0.09 samarium
Neodymium
a Praseodymium .
Cerium
Lanthanum
Barium
Cesium
b Iodine
0.09" Tellurium
Antimony
-------
TABLE 21 (continued)
Plant L
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
ytterbium
Thullium
Erbium
Holmium
Dysprosium
O.OS
9
c*
J>
0.001C
<0.1
Concentration: rag/a*
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tim
Indium
Cadmium
Silver
0.3
0.05
O.OS
0.2
0.1
23
0.2
O.OS
Palladium Palladium
Plant N
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
ytterbium
Thullium
Erbium
Holmium
Dysprosium
0.04
4.0
*a
_b
0.04 ,
0.003C
<0.08
<0.08
Plant S
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
ytterbium
Thullium
Erbium
Holmium
Dysprosium
0.08
0.21
2.4
7.1
<0.02,
M
k
u
<0.11r
0.55C
<0.13
<0.02
<0.07
<0.02
<0.04
0.01
0.01
Rhodium
Concentration: mg/m*
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
' 0.08
<0.04
0.1
0.1
0.1
0.5
0.1
14
I
0.4
0.2
0.3.
•*
0.08
Concentration: mg/m3
Terbium
Gadolinium
Duropium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
0.04
0.05
0.09
1.0
0.36
3.4
4.0
49
0.99
0.38
188
31 h
_U
1.1
0.25
Detection
Ruthenium
Molybdenum
Niobium
zirconium
yttrium
Strontium '
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
Detection
Ruthenium
Molybdenum
Niobium
Zirconium
yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
Detection
Ruthenium
Molybdenum
Niobium
Zirconium
yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
Haiti '6.65 maV3
0.1
0.3
1.0
0.3
0.4
1,900
31
3.6
0.05
380
2.2
13
limit: 6.04 n
0.2
1
0.6
0.4
0.8
0.08
0.1
0.04
0.8
0.04
150
12
2.9
1.7
600
12
680
limit: 0.01
6.5
0.1
1.6
0.33
6.8
11
14
<0.22
7.0
0.16
0.56
13
306
16
0.44
2,588
56
26
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Hitrogen
Carbon
Boron
Beryllium
Li'thium
i
io/m^
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
mg/m3
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
1.0
3.2
<0.1
1,200
95
37
360
1,200
4,600
440
650
5,000
33 a
_
0.3
0.4
5.6
<0.2
280
44
3.8
1,240
2,000
2,000
200
110
520
76 a
a
^fl
-'
0.2
6.1
40
0.14
4,353
859
129
1,882
4,824
1,106
2,000
1,035
694
56 a
a
_*
d
3
<0.05
0.58
(continued)
aNot reported. Internal standard. CInstrument source.
64
-------
TABLE 21 (continued)
Plant T
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
S3
-*
_b
.
Plant U
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
Plant
Uranium
Thor ium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.02
<0.04
<0.05
7.4
<0.02
~a
_b
<0.07
<0.23C
<0.09
<0.06
<0.04
W
0.5
<0.2
14
<0.1
-*
_
<0.2
0.006
<0.3
<0.2
<0.1
Concentration: mg/m'
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
0.4
150
0.7
1.1,
-
Concentration: mg/m3
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
0.02
0.02
0.02
0.01
0.62
0.38
12
0.01
0.28
1.7
W
_D
0.44
0.23
Concentration: mg/m3
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
0.1
0.5
0.2
<0.9
1.5
1.3
8.0
2.3
230
0.1
0.07
0.4
1.3b
0.3
0.07
Detection limit i 0.01 mg/m3
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
Detection
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
Detection
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
0.2
0.9
0.2
1.6
1.1
1.8
98
29
1.6
0.2
320
4.9
5.1
limit: <0.01
5.2
2.2
0.16
6.4
3.3
5.5
0.09
<0.13
0.18
60
29
11
0.20
560
12
23
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
mg/m3
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
0.4
7.3
1,500
330
•11
140
1,300
1,040
550
530
2,700
890 .
9
_*
0.7
0'.49
18
0.1
756
402
12
2,680
597
378
1,220
116
1,950
96
-
-*
_ ™
_a
0.40
•
limit: 0.05 mg/m 3
0.5
0.2
1.9
1.1
19
3.1
2.5
13
2.3
66
23
17
8.0
4,500
370
10
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phorphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
15
160
0.4
15,000
1,500
290
330
1,700
2,600
6,700
3,700
3,300
390
_a
_a
_a
"a
0.2
13
Not reported. Internal standard. Instrument source.
(continued)
65
-------
TABLE 21 (continued)
Plant X Concentration: mg/m3
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Indium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.09
5.0
<0.06.
_a
0.06,
0.08
<0.1
<0.09
-------
TABLE 22. LEVEL 1 SPARK SOURCE MASS SPECTROMETERS METALS
ANALYSIS OF FILTERED SECONDARD EFFLUENT
; (detection limit of 0.001 g/m3)
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.003
<0.003
<0.006
0.38
<0.004,
_a
<0.008.
0.008C
<0.010
<0.002
<0.007
<0.002
<0.007
<0.003
<0.003
<0.005
0.17
<0.002
_a
^
.
-
<0.007
0.10C
<0.009
<0.006
<0.002
<0.003
<0.005
<0.005
<0.010
<0.25
<0.007,
O
<0.013
<0.045
<0..017
<0.003
<0. Oil
<0.003
<0.007
<0.002
<0.003
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
.Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Plant A
<0.003
<0.002
<0.002
<0.004
0.009
0.002
0.004
0.51
0.003
0.011
0.095
o.iob
D
0.005
0.001
Plant B,
<0.003
<0.002
<0.002
<0.003
<0.002
70.003
70.002
0.20
0.001
0.004
0.019
0.017
_D
0.006
0.001
Plant C,
<0.002
<0.004
<0.003
<0.006
<0.005
0.004
<0.003
<0.004
1.1
0.001
0.024
<0.002
0.62
0.082
_b
0.030
10
. q/m3
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Garmanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
g/m'
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
g/m 3
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
0.043
0.031
0.003
0.70
0.19
0.47
0.003
<0.015
13
0.14
1.0
0.021
6.1
0.48
1.4
£0.007
0.004
0.002
0.22
0.25
0.26
0.006
0.28
1.2
0.16
0.027
0.008
2.4
0.41
0.24
0.011
0.004
0.009
0.001
1.1
0.38
13
0.027
<0.025
0.004
1.2
0.63
0.45
0.018
2.1
0.18
0.053
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
2.5
0.087
<0.006
170
8.8
11
130
6
18
6.4
10
180
1-1 a
a
-a
<0.003
0.12
0.005
0.079
<0.003
21
79
2.1
300
45
8.0
0.96
14
190
3.7
—
a
a
_a
<0.003
0.032
4.4
0.71
<0.017
41
7.7
2.3
340
20
27
0.99
1.8
370
12 a
a
3
3
-fl
<0.005
0.044
Not reported. Internal standard. cInstrument source.
(continued)
67
-------
TABLE 22 (continued)
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.002
0.063
<0.002,
Q
_
<0.003r
0.001C
<0.003
<0.002
<0.002
<0.003
<0.004
<0.007
0.033
<0.005,
fl
k
u
<0.009_
<0.003C
<0.011
<0.002
<0.007
<0.002
<0.005
<0.002
0.10
Terbium
Gadolinium
Europium
Samarium
Neodymium •
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony f
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Plant E,
<0.002
0.007
0.004
0.022
0.019
0.29
0.011
0.13
0.006
_D
0.004
0.001
Plant F,
<0.003
<0.002
< 0.004
<0.004
0.002
0.003
0.002
0.16
0.002
<0.002
0.19
0.004
_b
0.004
Plant
0.004
0.004
0.23
0.001
0.007
•
1.2
0.032
_t>
0.002
0.003
q/m1
Ruthenium
Molybdenum
Niobium
Zirconium
yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
q/m 3
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
G, g/m"
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
0.004
0.009
0.094
0.071
0.92
0.002
0.035
0.76
0.10
0.038
0.001
0.86
0.035
0.043
0.005
0.011
0:28
0.054
0.16
0:006
<0.017
0.001
1.5
0.49
0.024
O.t)21
2.7
0.13
0.015
0.006
0.011
0.089
0.017
0.036
0.001
0.014
0.84
0.11
0.038
0.13
1.6
0.17
0.018
'
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
;
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
0.018
0.13
36
29
1.3
290 *
9.2
2.9
0.39
2.1
70
13
o
_
A
_a
0.014
0.022
0.10
<0.011
27
10
0.83
56
38
48
4.6
15
490
2.9 ,
_a
A
_d
~a
J»*^
<0.003
0.004
0.12
0.11
16
2.0
1.1
43
16
8.7
2.2
1.4
48
3.2
a
_
_a
0.32
aNot reported. Internal standard. CInstrument source.
(continued)
68
-------
TABLE 22 (continued)
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.002
<0.002
<0.004
0.017
a
_
<0.005r
<0.015C
<0.006
<0.004
<0.003
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Plant K,
<0.002
<0.002
<0.002
<0.002
<0.002
0.19
23
0.048
0.011
_D
0.003
~i753
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
0.006
0.005
0.19
0.027
2.9
0.32
0.010
0.001
0.77
0.11
0.014
0.012
0.72
0.008
0.090
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
'Boron
Beryllium
Lithium
0.015
0.018
<0.002
24
26
650
105
6.8
120
0.71
13
120
87 _a
-
-
-
<0.002
0.005
Plant L, g/m3
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
ytterbium
Thullium
Erbium
Holmium
Dysprosium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.002
0.14
.
-
<0.003.
0.003C
<0.004
<0.003
0.001
<0.004
<0.004
<0.008
0.95
<0.002
_a
-
<0.010
'0.011
<0.012
<0.002
<0.008
<0.003
<0.004
<0.002
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine .
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
<0.007
0.011
0.005
0.37
0.007
0.005
0.30
0.046
_b
0.003
0.001
Plant N,
<0.003
<0.003
<0.005
<0.004
0.002
0.005
0.008
1.3
0.001
0.12
0.12
0.008
_a
0.004
0.002
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
g/m 3
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
0.015
0.005
0.026
0.001
0.99
0.20
0.51
0.002
0.031
2.4
0.54
0.25
0.033
4.5
0.27
0.26
0.030
0.001
0.054
0.017
2.1
0.51
0.19
0.063
0.40
0.005
0.002
580
0.11
0.39
0.46
80
27
44
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogne
Carbon
Boron
Beryllium
Lithium
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
0.23
0.24
110
5.3
2.1
330
10
15
0.44
4.6
78
2.5
d
a
a
_a
2.1
0.033
0.089
<0.004
570
58
1.1
1,400
110
54
110
42
150
41 a
O
fl
_™
_a
<0.004
0.033
Not reported. Internal standard. Instrument source.
(continued)
69
-------
TABLE 22 (continued)
Plant S. q/m3
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
Utanium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.002
<0.002
0.012
0.085
<0.005
<0.012
<0.004
<0.003
<0.002
<0.003
0.002
0.042
< 0.002
<0.002
<0.002
<0.002
<0.002
<0.002
0.006
^
<0.002
<0.002
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
<0.002
<0.004
<0.005
<0.005
<0.007
"0.097
0.002
0.017
0.84
0.024
_b
0.008
0.001
Plant T,
0.002
0.002
0.006
0.022
0.001
0.002
0.009
0.005
_b
0.001
0.002
Plant U,
<0.026
<0.081
"0.16
0.002
0.076
0.19
0.003
_D
0.003
Ruthenium
Molybdenum
Niobium
Zirconium
Ytttium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
• Cobalt
Iron
Manganese
Chromium
g/m3
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
'
g/m 3
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
0.021
0.002
0.016
0.027
0.81
13
0.001
0.064
0.29
0.28
0.005
1.0
0.10
0.016
0.005
0.006
0.002
0.030
0.13
0.13
0.005
<0.003
0.29
0.040
0.045
0.002
0.57
0.059
0.058
0.007
0.002
0.001
0.32
0.043
0.55
0.018
0.14
16
0.099
0.058
0.10
0.12
0.53 '
0.005
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
0.006
0.010
<0.002
11
73
2.5
24
15
18
18
2.8
95
A
a
-
<0.002
0.011
0.014
0.019
5.2
36
0.51
0.70
0.79
1.9
3.3
1.4
40
0.95,
a
-
0.008
0.002
0.024
0.003
180
37
170
16
6.4
14
0.24
11
83
2.4
a
_
"a
0.023
Not reported. Internal standard. Instrument source.
(continued)
70
-------
TABLE 22 (continued)
Plant V, g/m3
Uranium
Thorium j
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
0.003
0.089
-
<0.002
<0.002
<0.001
<0.002
Terbium
Gadolinium
Europium
Samarium
Neodymium
Praseodymium
Cerium
Lanthanum
Barium
Cesium
Iodine
Tellurium
Antimony
Tin
Indium
Cadmium
Silver
Palladium
Rhodium
<0.002
<0.002
"0.005
0.003
0.36
<0.003
0.010
0.009
—D
0.011
Ruthenium
Molybdenum
Niobium
Zirconium
Yttrium
Strontium
Rubidium
Bromine
Selenium
Arsenic
Germanium
Gallium
Zinc
Copper
Nickel
Cobalt
Iron
Manganese
Chromium
0.005
0.002
0.001
0.19
0.022
1.1
0.012
3.1
2.0
0.010
0.073
4.7
0.31
0.066
Vanadium
Titanium
Scandium
Calcium
Potassium
Chlorine
Sulphur
Phosphorus
Silicon
Aluminum
Magnesium
Sodium
Fluorine
Oxygen
Nitrogen
Carbon
Boron
Beryllium
Lithium
0.012
0.12
43
2.0
1.0
45
4.2
9.5
4.0
7.4
42
1.4
-
-
-
-
0.041
Plant W, g/m3
Uranium
Thorium
Bismuth
Lead
Thallium
Mercury
Gold
Platinum
Iridium
Osmium
Rhenium
Tungsten
Tantalum
Hafnium
Lutecium
Ytterbium
Thullium
Erbium
Holmium
Dysprosium
<0.003
<0.003
<0.005
0.17
<0.002
-
_
<0.007r
<0.003C
<0.009
<0.001
<0.006
-------
TABLE 23. CONCENTRATION OF METHYLENE CHLORIDE EXTRACTABLE
ORGANICS IN FILTERED SECONDARY EFFLUENTS
Plant
Organic ,
concentration,
g/m3 '
Plant
Organic
concentration,
g/m3 .
A
B
C
E
F
G
K
L
63.7
3.18
28.2
3.60
16.0
27.2
2.73
18.3
N
S
T
U
V
W
X
,9.24
5.40
17.8
14.6
a
15.0
13.5
(
Analysis not performed.
The methylene chloride extract then went through a solvent ex-
change step to transfer the sample to a nonpolar solvent. This
extract was then passed through a liquid chromatography column
that divided the sample into eight fractions. Each ;of the eight
fractions was analyzed by an infrared (IR) spectrophotometer and
then by a low resolution mass spectrometer (LRMS) ." .The classes
of organic compounds and their relative intensities ifound in
each fraction are presented in Tables 24 and 25. \
t
i
IR analyses indicated the presence of aliphatic hydrocarbons, C=O
esters and acids, aromatics, phthalate esters, and fatty acid
groups. LRMS analyses identified the following major classes of
compounds: paraffinic/olefinic, alkyl benzenes> alcoholic ethers,
di-n-octyl phthalate, bis(hydroxy-t-butyl phenol) propane, tri-t-
butyl benzene, alkyl phenols, dichloroaniline, toluene-sulfonyl
groups, vinyl stearate, and azo compounds.
72
-------
TABLE 24. LEVEL 1 INFRARED ANALYSIS OF THE ORGANIC EXTRACTS
Fraction No.
Interpretation
Plant A
Before extraction
1
2
3
4
5
6
7
8
Bonded OH, aliphatic CH, C=O ester and acid, ketone or aldehyde, conjugated OC, possible aromatic OC, ether groups (CRj) >
where n = >4
All aliphatic hydrocarbons
Aliphatic and aromatic hydrocarbons, aromatic OC
Aliphatic OH, ester or aldehyde OO, conjugated OC, various CH2 groups, or aromatic substitution bonds
Similar to fraction 3
Similar to fraction 3
Bonded OH; aliphatic C-H; acid, ketone, or aldehyde OOi conjugated and aromatic OC; possible phthalate ester; ether group or Si-O
Bonded OH, aliphatic CH, ester or aldehyde C-O, water in material, ether or Si-O groups
Bonded OH and 1,630 cm'1 absorption-water, aliphatic CH trace of C-O, SiO2> poorly defined organic
Plant B
00
Before extraction
1
2
3
4
5
6
7
8
Bonded OH, aliphatic CH, ether, (CHaJi,, OO, or OC?
Aliphatic CN, trace aromatic CH, Si-CHjt?), methylene CH-j groups >4
Very strong background adsorption—only aliphatic CH visible
Bonded OH, aliphatic CH, ester C-O, acid, aldehyde or ketone C-O (CHa) or >4.
Bonded OH, trace aromatic CH, aliphatic CH, ester OO acid, aldehyde or ketone OO, various CHj groups.
Similar to 4, but stronger bonded OH, less ester OO, more acid, aldehyde or ketone OO, various CH2 groups complex spectrum.
Bonded OH, aliphatic CH, some ester OO, acid, aldehyde or ketone OO, ether groups, may contain glycol ether type compounds.
Bonded OH, aliphatic CH, ester C=O, acid, aldehyde or ketone OO, strong ether group, glycol ether type of compound.
Strong bonded OH, weak C-H (aliphatic) trace OO, nonconjugated OC, SiOa present.
Plant E
Before extraction
1
2
3
4
5
6
6 (repeat)
7
7 (repeat)
8
Aliphatic CH, diffuse OO region, carboxylate ion, ether group complex spectrum
Aliphatic hydrocarbons, no indication of number of CHa groups.
Poor spectrum—aliphatic CH, COj, and water vapor in spectrum.
Poorly defined spectrum—aliphatic CH, numerous ill-defined bonds. No OO or OC.
Aliphatic CH, ester C=O, strong background adsorption.
Aliphatic CH, ester=O, moot likely aliphatic ester, possibly acetate—may be single compound.
Spectrum too strong—bonded OH, aliphatic CH, ester OO, acid, ketone or aldehyde C-O, large portion of compound in Ho. 5 plus
additional carbonyl compounds.
Mixture of several comps. Ester OO, and acid, ketone or aldehyde OO. Ether group.
Bonded OH, aliphatic CH, acid, ketone or aldehyde C-O, spectrum too strong for good identification.
Bonded OH, aliphatic CH, acid, aldehyde or ketone OO, evidence of both acid and acid salt (carboxylate ion) CH(CR3>2 group
possible, no long chain (CH2) groups.
Bonded OH—evidence of water (3,350 cm"1 and 1,635 cm'1) SK>2 present aliphatic CH.
(continued)
-------
TABLE 24 (continued)
Fraction No.
; Interpretation
Plant P
Before extraction
1
2
3
4
5
6
7
8
Bonded OH, aliphatic CH, diffuse C=0, and C=C regions Si-O possible, trace CH2Cl2.
Long chain aliphatic hydrocarbons. Unknown 1,265 cm"1 bond. Si-(CH3>-?
Aliphatic CH, ester C=O, conjugated C=C, trace 1,265 cm"1.
Weak bonded OH, aliphatic CH, aromatic CH-?, ester OO, conjugated OC, progression of OS substitution bonds, series of CHj bond.
Similar to No. 3, series of ill-defined bonds below 1,400 cm'1.
Bonded OH, trace aromatic CH, aliphatic CH, ester or aldehyde C=O, conjugated C=C, series of CH2 groups aromatic substitution bonds-?
Bonded OH, aliphatic CH, ester OO, nonconjugated OC or amides, ether or Sio groups, secondary amide possible, CHj groups n >4 .
Similar to No. 6.
Strong bonded OH, aliphatic CH, weak ester or aldehyde OO, HjO present (1,640 cm"1) SiO2 present.
Plant G
Before extraction
1
2
3
4
5
6
7
8
Aromatic and aliphatic CH, residual CHjCla in spectrum.
Aliphatic hydrocarbon — chain length >Ci», possible C(GH3>3 group.
Aliphatic CH, ester C=O, phthalate bonds, various chain lengths of CH2.
Aliphatic CH, ester C=O, some OC, various CHz groupings.
Bonded O-H, aliphatic CH, ester OO, some acid, aldehyde or ketone OO, OC, possible fatty acid group, various CH2 groups.
Identical to No. 4.
Considerable bonded OH, aliphatic CH, ester OO conjugated OC, Si-O or ether group.
Similar to No. 6.
Considerable O-H, aliphatic C-H, ester OO, SiO or ether groups.
Plant K
Before extraction
1
2
3
4.
5
6
7
8
Bonded OH, aliphatic CH, diffuse C=O region Si (013) group? Diffuse spectrum.
Appears to contain water, aliphatic C-H, Si-CH3, mainly hydrocarbon compounds.
Aliphatic hydrocarbons, silicones.
Mainly silicone type materials.
Bonded OH, some aromatic CH, aliphatic CH, ester OO, acid, ketone or aldehyde OO, carboxylate ion, conjugated OC, some silicone
material, (CH2) where n >4.
Strong background adsorption, aliphatic CH, ester OO various (CH2) • groups, some silicone adsorption.
Bonded OH, aliphatic CH, trace aromatic CH, ester OO, aromatic OC, silicone adsorption.
Similar to No. 6, not as strong a spectrum.
Strong OH adsorption, very weak C-H, trace of OO (may be HzO background) SU>2 adsorption, low organic content.
(continued)
-------
TABLE 24 (continued)
Fraction No.
Interpretation
Plant 2.
Before extraction
1
2
3
4
5
6
7
8
Bonded OK, aliphatic CH, OO, aromatic or conjugated OC, ether group. Complex spectrum CSN, SiOj-?
Aliphatic CH, trace OO, OC, CHj group >4, possible 01(0)3)2 group—ma inly aliphatic hydrocarbons.
Aliphatic hydrocarbons — branched chain, trace of OO, OC no long (CHj) groups.
Bonded OH, aliphatic CH, ester OO, conjugated OC strong ether group, possible glycol ether.
Bonded OH, aliphatic CH, ester OO, nitrite (CSN) group, strong ether group, various straight and branched CHj groups.
Bonded OH, some aromatic CH, aliphatic CH, CBN nitrite group, ester OO, conjugated or aromatic OC, ether group, very complex
mixture spec t rums.
similar to No. S but less CHN, more bonded OH, ester OO, conjugated or aromatic OC, some aromatic C-H, aliphatic CH ether
grouping — complex spectra .
Some bonded OH, aliphatic CH, trace CEN, ester OO, carboxylate ion, possible C-C1 group.
Contain water and SiOj, plus some of materials found in No. 7. Carboxylate ion.
Plant JJ
Before extraction
1
2
3
4
5
6
7
a
Bonded OR, aliphatic CB, diffuse OO, OC region, silicons (CHjU ether?
Long chain aliphatic hydrocarbon.
Aliphatic CH, ester OO, possible (0X0)3)2 group, spectrum not very distinct.
Meak bonded OH, aliphatic CD, ester OO, conjugated OC, 01(0)3)2 group, various CHj groups.
Weak bonded OH, aliphatic CH, ester OO, possible fatty acid groups.
Bonded OH, aliphatic CH, medium ester OO,-nonconjugated OC, spectrum not very distinct.
Bonded OH, aliphatic CH, ester OO, acid, aldehyde or ketone OO, conjugated OC, aliphatic ketone or ester group, possible ether
group, various CH2 chain lengths.
Bonded OH, aliphatic CH, ester OO, conjugated OC, possible fatty acid gropus, ether group.
Strong OH, weak aliphatic CH, ketone, acid or aldehyde OO, ester OO, strong OC, ether group.
Plant S
Before extraction
1
2
3
'
4
5
6
7
8
Bonded OH, aliphatic CH, ester OO, (0)2) groups, broad diffuse spectrum, ether groups possible.
Aliphatic hydrocarbon, chain length diffuse spectrum.
(continued)
-------
TABLE 24 (continued)
Fraction No.
Interpretation
Plant T
Before extraction
1
2
3
4
5
6
Before extraction
1
2
3
4
5
6
7
8
Bonded OH, aliphatic CH, OO, OC, ether of SiO groups, (0)2) groups. Very diffuse spectrum.
Bonded OH, aliphatic CH, ester OO, some OC, CHj groups >&,.
Bonded OH, aliphatic CH, medium ester OO, possible fatty acid groups, no definite CH2 groupings.
Bonded OH, aliphatic CH, strong ester OP, nonconjugated OC, ether or SiO groups. Various chain lengths of (CH2)n-
Bonded OH, medium aliphatic CH, medium ester OO, nonconjugated OC, ether or SiO group, various (CH2)n groups, possible acid
salts.
Similar to No. 3.
Bonded OH, aliphatic CH, medium ester OO, series of 5 unknown bonds medium intensity 1,510 cm"1 to 1,610 cm"1, ether or SiO group,
various (CH2) groups.
Similar to No. 6, but weaker OH, OCO, conjugated OC, fatty acid groups? ether or SiO group. (C»2)n groups.
Very strong bonded OH group, medium aliphatic CH, weak ester OO, strong nonconjugated OC, possible fatty acid group, ether or
SiO group, no CH2
Plant 5"
Trace bonded OH, aliphatic CH, acid, ketone or aldehyde OO, silicones, some (CB2> groups.
Aliphatic hydrocarbon, conjugated or aromatic OC, weak, (CH2> —no >4.
Aliphatic hydrocarbon, conjugated OC (alkene?) , diffuse 0)2 groups.
Aliphatic C-H, ester OO, conjugated or aromatic OC, possible unsaturated ester-fumazate, maleate, etc.
Bonded OH, trace sec N-H or oxcetone OO, aliphatic C-H possible trace CHN, ester OO, conjugated OC, unsaturated ester group.
Complex spectra below 1,500 cm"1.
Aliphatic C-H, ester OO, diffuse spectra below 1,100 cm"1.
Complex spectra, bonded OH, NH or OO overtone, aliphatic C-H, ester OO, acid, aldehyde or ketone OO (weak), ether group,
complex bond pattern below 1,500 cm*1.
Bonded OH, aliphatic C-H, ester OO, conjugated OC, possible ether group, miscellaneous (CH2> groups.
Bonded OH strong, aliphatic CH, strong OC, ether group.
SK>2, possible glycol ethers.
Plant V
Before extraction
1
2
3
4
5
6
7
8
Trace bonded OH, aliphatic CH, ester OO, acid, aldehyde or ketone OO, silicone adsorption, some (CH2) groups.
Aliphatic CH, (CH2) where n >4, hydrocarbons plus possible 81-013.
Aliphatic and aromatic CH, some ester OO, silicones.
Poor spectrum—low organic content? aliphatic CH strong background adsorption.
Bonded OH, aromatic and aliphatic CH, ester OO, conjugated OC, silicones.
Identical to No. 4.
Aliphatic CH, ester OO, silicones, CH2 various groups ester stronger, silicones weaker than in No. 4 or No. 5.
Bonded OH, aliphatic CH, ester OO, conjugated OC ether group—possible glycol ether-type compounds.
Strong bonded OH, weak aliphatic CH, ester OO, strong OC, some ether under S1O2 adsorption.
(continued)
-------
TABLE 24 (continued)
Fraction No. ( ^ ^ Interpretation
Plant W
Before extraction Bonded OH, aliphatic CH, ester OO, acid, ketone or aldehyde OO. (CH2) -n where n >4, diffuse spectrum 1,300 cm'1 to 900 cm*1.
1 Bonded OH, aliphatic CH, ester OO, possible ether group (CHj) where n >4% Not typical fraction 1.
2 Aliphatic CH groups, ester OO, weak spectrum.
3 Aliphatic ester compounds, ester OO, no aromatic CH.
4 Trace OH, aliphatic CH, ester OP, weak diffuse spectrum.
5 Weak diffuse spectrum, poor background, OH (water?), aliphatic CH, weak ester OO, nonconjugated OC.
6 Bonded OH, aliphatic CH, ester OO, long chain CH2 groups, unsaturated acid, aldehyde or ketone, conjugated OC, possible amide
groups.
7 poor spectrum. Bonded OH, aliphatic CH, numerous OO types, OC, amide possible, very diffuse below 1,400 cm'1.
8 Weak spectrum, bonded OH—likely HaO, very weak aliphatic CH, low organic content.
Plant X
Before extraction Bonded OH, aliphatic CH, OO, ether or SiO, (CHj) , numerous broad diffuse bonds.
n
1 Aliphatic hydrocarbons, possible Si (0)3).
2 Aliphatic CH, ester OO, conjugated OC, hydrocarbon.
3 Bonded OH, aliphatic ch, ester OO, phthalate plus other types of ester materials.
4 Similar to fraction No. 3.
S Bonded OR, trace aromatic CH, aliphatic CH, ester OO, acid, aldehyde or ketone OO, conjugated OC, various straight and
branched CHj chains.
6 Bonded OH, aliphatic CH, ester OO, conjugated and aromatic OC, ether group, long CRj chains.
7 Very similar to No. 6.
8 Strong OH adsorption, similar to No. 6 and No. 7 but more OH and presence of SiOj.
-------
TABLE 25. LEVEL 1 LOW RESOLUTION MASS SPECTROMETER
ANALYSIS OF ORGANIC FRACTIONS
Plant
and
organic
fraction
1
2
3
4
5
6
7
e
Fraction
•eight.
aq
J>
16. 1
5.1
J>
_b
10.2
_b
21.0
cataooriee preaent
Relative.
intanaity
_b
100
100
100
100
J>
_b
100
10
10
10
J>
10
100
Category
_b
Aliphatic*0
Aroaatice
Allphetlca
Aroaatice
_b
J>
JUiphatio
Aloabola/ethere
Pbeoole
Eatara
_b
Alipbatica
AlcoholA/etnera
Bubcateooriea preeant
aalatlve
intaaalt
_b
100
100
100
100
_b
J>
100
10
10
10
_b
10
100
» , Epacifie TitT"'r'ifla
riant A
_b
Paraffinic/oletiaic (or cyelic-paraffinic. etc.l. •*
AUcyl benaenaa (91, 105, 114, 133 iona pr*aant.°>*
Parafrinlc/olafinle (or cycUc-paraffiaic, *tc.).d'*
lUtyl banaanaa (91, 105, 114, 133 iona pruant).a>*
_b
"b
Parafflnic/olafinlc (or crcUc-vuaffinie, atc.).4>e
Alcoholic athara (45 iona to 89 Iona) ."•' f
Ita (by4ro)cy-t-bi)tyl phanyl) Dtopana (Ci|H)202) m 340.
Di-n-ootrl phthalata (C2tI|aOi,) W390.
_b
Mnffiaio/olafiritc (or oyolic-paralfinic, atc.).4><
Aloobolic^athara (45 iona to 89 iona) ^*®
Btbarai m-n-octyl phthalata (C2%B]«Oi,) MV390.
othar unknoiro ooipounJa praaant
_b
Mo aaaaaa abcva 49B.
101368(100), 369(45), 353(20)
1.345(100), 396(35), 411(30)
_b
_b
Ho aaaaaa abova 414.
_b
No auaaa abon 368.
10Q,2$4(lflOl 126OfM 1971111
Awl f *3* \XWJJ , AADI4UI, H I \iyj
Plant B
1
1
3
4
5
6
7
8
1
2
3
4
5
6
7
8
_b
10.3
J>
3.6
J>
13.9
_b
15.0
9.4
0.3
1.9
3.3
1.4
18.6
51.7
42.7
J>
100
100
_b
10
J>
1
ion
XWI
_b
10
10
10
100
100
100
1
noo
10
100
10
10
10
1
10/100
100
1
10/100
10
100
10
100
10
100
J>
Aliphatice
Eatare
_b
Aliphatic
_b
Aliphatic*
PtMnflla
rnenoie
_b
Allphatica
AloohoUtathere
Eaten
Aliphatica
Aliphatica
Eaters
Aliphatica
Aroaatice
Eatara
Aliphatica
Aroaatica
latara
Aliphatica
Aroaatice
Phaaola
Eatara
Aroaatlca
Phenols
Eatara
Aliphatic
Phanola
Eatara
Aliphatica
Eatara
_b
100
100
J>
10
J>
i
1M^
1W
J>
10
10
10
100
100
100
1
100
10
100
10
10
10
1
10
100
100
1
10
100
10
100
10
100
10
100
_b
BaM aa Plant A, fraction 2.
Saa» aa aatan. Plant A, fraction 6.
.b
Saa» a* Plant A, rraction 2.
.b
Sax* aa Plant A, fraction 2.
a tnyuuAj1 • u~Mui.yl pnanon propana 1(^2383202) MI39O.
_b
Saaa aa Plant A, fraction 2. *
Alcoboltaaanhra- (4S't»aa to 89 iona).
Saw aa aaUra, Plant A, fraction 6.
Plant f
Prfcaarily paxa«inic.d'e
8aa» a Plant A, fraction 2.
saaia aa avtara. Plant A, fraction 6.
AliphaUca.d>e
AUqrl bauanaat Irl-t-bQtyl bantaM (Ci6H30) MJ246.
Stmt aa aatara. Plant A, fraction «.
Sana aa Plant A, fraction 2.
Trl-t-bntyl banaana (Cjaljo) M>246.
Sana aa aatara. Plant A, fraction 6.
Baa* aa Plant A, fraction. 2.
Alkyl banaanaai Trl-t-butyl banana (CijHjo) Mn46.
Alkjl phanola (135, 107, 121, 148 iona) .*
BiaOiydroxr-t-butyl pbuiyl] propana (023113202) MM390.
Saaa aa aatara. Plant A, fraction 6.
Sam aa Plant A, fraction 2.
Saaa aa aatara. Plant A, fraction 6.
J>
No aaaaaa abova 354.
_b
Mo aaaaaa abon 483.
10,279(100), 294(28), 280(12)
100,341(100), 356(36), 342(27)
100i381{100), 396(15), 382(11)
100,410(100), 151(37), 411(31)
100i429(100), 444(23), 445(15)
_b
HD aaaaaa aboia 437.
J>
Mo aaaaaa abova 414.
1001294(100), 127(20), 128(12)
NO aaaaaa abova 446.
Mo aaaaaa abova 354.
K> aaaaaa abora 354.
NO aaaaaa abova 381.
No aaaaaa abova 340.
10,239(100), 240(20), 254(25)
Ho aaaaaa abova 354.
100,45(100), 42(90) 12 coapounda)
100,59(100) (4 coapounda)
Ho aaaaaa abova 354.
No aaaaaa above 354.
Ralativa intenaity 6f tha aaaa-to-charga ratio Untenaity ralativa to ooMinant ion) .
Organic w«lght of fraction balo« oravixatric thraanold of 0.1 aoi tharafora, no analyala vaa parfonaad.
^Oanarally all iona up through 498 preaent in allphatic-typa patt«m, homvar, all naoa >100 ar« abnormally atrono for
No aolacular night range datarainatlon poaaibla.
No conpoaition dataraination poaaibla.
Molecular weight.
typical aliphatica.
(continued)
78
-------
TABLE 25 (continued)
plant
and
organic
fraction
1
2
3
4
S
6
\
a
i
2
3
4
5
6
7
8
Fraction
Might,
31.8
7.2
2.3
0.9
0.3
29.0
13.9
15.5
• v
12.7
2.2
8.6
8.6
9.7
55.1
11.7
18.1
Cataooriaa creaant Sobaateonriea prevent
wlativa
intanaltv
100
1
100
100
1
100
1
10
1
100
100
100
1
1
100
100
1
1
100
10
1
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100/10
100
100
100
Category
Aliphatica
Aliphatica
later.
Aliphatica
latara
Aliphatica
Batere
Aliphatica
Phaoola
Eatara
Aliphatica
Aloohole/ethera
Phanola
Batara
Aliphatica
Alcohole/ethera
Phenola
tatara
Aliphatic.
Aloohole/ethere
Satara
Aliphatica
Aliphatica
Arowtica
Phanola
Satara
Aliphatica
Arowtic.
Phanola
Batara
Aliphatica
Phanola
Eatara
Phanola
Batara
Aliphatica
Phanola
Setare
Aliphatica
Satara
telatlva
intanaity Specific namniMiirte
100
1
100
loo
i
100
1
10
1
100
100
100
1
1
100
100
1
1
100
10
1
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Plant C
Saw aa Plant F, Fraction 2.
Swa aa plant A, Fraction 2.
Swa aa eatara. Plant A, Fraction 6.
Saw aa Plant A, Fraction 2.
Swa aa aatara. Plant A, Fraction 6.
Swa aa Plant A, Fraction 2.
saw aa aatara. Plant A, Fraction 6.
Sana aa Plant A, Fraction 2.
Swa aa phanola. Plant A, Fraction 6.
Sana aa eatara. Plant A, Fraction 6.
Saw aa Plant A, Fraction 2.
Aloohollo ether. Kith 41, 43, 45 iono
and 55, 57, 59 iona.»«*
Saw aa phanola, Plant A, Fraction 6.
Saw aa aatara. Plant A, Fraction 6.
Saw aa Plant A, Fraction 2.
Saw aa Fraction 6.
Saw aa phanola. Plant A, Fraction 6.
saw aa eater. , plant A, Fraction 6.
Swa aa Plant A, Fraction 2.
Saw aa Fraction 6.
save aa eatara. Plant A, Fraction 6.
Plant L
Saw aa plant F, Fraction 1.
Saw aa Plant F, Fraction 1.
saw aa Plant A, Fraction 3.
Alkyl phanola (135, 107, 121, 149 iona).d><
swa aa aatara. Plant A, Fraction 6.
Sew aa Plant F, Fraction 1.
Saw aa Plant F, Fraction 3.
Di-t-eutyl phenol (CitH2*°) NI206.
Swa aa eatara. Plant A, Fraction" 6.
Saw w Plant F, Fraction 1. A ,
Alkyl phanol (135, 107, 121, 149 ion.).
Bwa u eatar.. Plant A, Fraction 6.
Alkyl phanola 1135, 107, in, 149 ion,) .d>*
Phthalata, probably 4i-c» alkyl but «ith a nev
eeriaa** ion added. (223, '237, 251, 265, 279).
saw aa Plant A, Fraction 2. H
Alkyl phanola (135, 107, 121, 144 lone).
Sia (hydiiuy~l~l)ut.yl pbenyl) propane (C2)H3202) "^
Saw ea Plant A, Fraction 6.
Saw aa Plant A, Fraction 2.
Saw aa aatara. Plant A, Fraction 6.
Other unknown coround.. preaent
-
no auuuaa above 410.
Mo oasaea above 378.
Bo uaaen above 381.
•0 anna., abova 325.
BO .oases abova 354.
1,69(100), 41(87), 43(78)
Ho naasea above 354.
10,69(100), 41(80), 43(78)
Bo wsaea abova 354.
>•
u
Bo wana abova 367.
No news abova 367.
Bo aoaaes above 354.
10.69(100), 41(80), 43(78)
Bo wswa above 429.
Ho aueseo above 429.
10.69(100), 41(80), 43(78)
Ho •aaoea ebove 340.
No massea above 381.
100,69(100), 41(80), 43(78)
MO. ItlSS
1,200(100), 201(86)
10,280(100), 279(98)
No aaaaea above 279.
Itceeiun iodide from infrared
plataa.
Relative intensity of th« mass-to-charge ratio (intenaity relative to dominant, ion).
No molecular weight rang* dattrmlnation poisibl*.
eHo ccopoilcton d«t-nination possible.
(continued)
79
-------
TABLE 25 (continued)
Plant
and Fraction cateoorlea preaent
orgajiic weight, Relative.
fraction »g intanaity Category
1 44.5 100
10
2 1.0 100
100
100
3 3.9 100
100
10
4 2.1 100
10
5 1.0 100
10
100
6 20.5 100
10
10
10/10
10
7 8.9 100
10/1
1
8 . 16.6 100
10
100
Uiphatica
Eatera
Uiphatica
Aromatica
Eatara
Uiphatica
Aromatica
Eatera
Uiphatica
Eatera
Uiphatica
Phenola
Eatera
Uiphatica
Aromatica
Aminea
Phenol.
Eatara
Uiphatica
Phenola
Eatera
Uiphatica
Phenol.
Eatara
Subcategoriea preaent l
Relative
intenaity Specific compounde
100
10
100
100
100
100
100
10
100
10
100
10
100
100
10
10
10
10
100
10
1
1
loo
10
100
Plant II
Sam* aa Plant A, Fraction 2.
Sam* aa eetera. Plant A, Fraction 6.
Sam* aa Plant A, Fraction 2* A
Alky! banian* (91, 105, 119 ions).
Sam* aa eatera, Plant A, Fraction 6.
Sam* aa Plant A, Fraction 2.
Sam* aa plant A, Fraction 3.
Sam* aa eatara, Plant A, Fraction 6
Sam* aa Plant A, Fraction 2.
Sam* aa eatara. Plant A, Fraction 6.
Sam* aa plant A, Fraction 2.
Sam* aa Plant F, Fraction 5.
Sam* aa aatara. Plant A, Fraction 6.
Sam* aa Plant A, Fraction 2.
t-autyl dichlorobanaan* (CijHvjCl,) MK202
Dichloroanlllne (C6HjNClj) Ml 161
Sam* aa phenols, plant F, Fraction 5.
Sam* aa phenol e, plant A, Fraction 6.
Phthalete. probably di-C» alkyl but with a new aeriea
of lone added i 223, 237, 251, 26! , 279 (CII,H),O»>
Hf 390 (probably) .
Sam* aa Plant A, Fraction 2.
Sam* aa phenola. Plant F, Fraction 5.
Sam* aa phenola. Plant A, Fraction 6.
Sam* aa eatera, Plant A, Fraction 6.
Sam* aa Plant A, Fraction 2.
Sam* aa phenol., plant F, Fraction 5.
Sam* aa aatara. Plant A, Fraction 6.
Other unknown compounda preaent
No aaaaea above 428.
No maaaea above 394.
No maaaea above 498.
101368(100), 369(30), 353(20)
10,395(100), 396(30)
1:410(100), 411(30)
No maaaea above 499.
10:368(100), 369(30), 353(20)
10,395(100), 396(30)
10:410(100), 411(30)
Mo maaaea above 779.
No maaaea above 490.
1,155
No maaaea above 340.
No maaaea above 279.
l:CeaiuD iodide.
Plant S
1 12.3 100
2 4.7 100
100
) 19.7 100
10
1
4 U.I 10
100/10
10
10
S 6.6 100
100
6 26.8 1
100
10
10
7 6.5 1
10
100
8 13.6 100
10
Uiphatica
Uiphatica
Aromatica
Uiphatica
Aromatica
Eatera
Aliphatic.
Aromatica
Phenol.
Phenola
Uiphatica
Aromatica
Uiphatica
Aromatica
Phenol.
Eatera
Uiphatica
Aromatica
Eatera
Allphatics
Eatere
100
100
100
100
10
1
10
100
10
10
100
100
1
100
10
10
1
100
100
10
1
Sam* aa Plant A, Fraction 2.
Sam* aa Plant Fraction .
Sam* aa plant Fraction . '
Sam* aa Plant Fraction .
same aa plant Fraction .
Sam* aa Plant Fraction .
Sam* aa Plant A, Fraction 2.
lolnene-atilfonyl -group (91, 155 iona).
Beat identity la p-toluene aulronamlda.
Sam* aa Plant F, Fraction 3.
Di-t-butyl phenol (C]i,H2jO) HN206.
Sam* aa Plant A, Fraction 2.
loluene-aulfonyl group (91, 155 iona).
Beat identity p-toluene eulfonamid*.
Sama ai Plant A, Fraction 2. .
Toluene-aulfonyl group 191, 155 Iona).
Same as phenola, Plent A, Fraction 6.
Sam* aa eatara, plant A, Fraction 6.
Sam* as Plant A, Fraction 2.
Sane as aromatica. Plant 'A, Fraction 4.
Sam* aa eatera. Plant A, Fraction 6.
Sana aa Plant A, Fraction 2.
Sam* aa eetere, plant A, Fraction 6.
•ethyl eatera (T4, 87 lone)."'8
Ho maaaea abov* 446.
No maaaea abova 446.
No maaaea above 452.
No maaaes above 477 .
1,381(100), 382(27), 396(17)
No maaaes above 354.
100:98(100), 97(75)
No Bmaaea above 446.
10:90(100), 91(68), 106(58)
No maaaee above 446.
10:90(100), 91(68), 106(56)
No maaaea above 354.
dRelative intensity of the .use-to-charge ratio (intensity relative to dominant ion).
Ho molecular weight rang* determination possible.
Ho coropoBition determination possible.
(continued)
80
-------
TABLE 25 (continued)
Plant
and Fraction Categories present
organic weight. Relative
fraction mg intensity Category
Relative
intensitv
Subcategorles present
Specific compounds
Other unknown compounds present
Plant T
1 18.5 100
1
2 2.6 100
100
10
3 6.7 100/100
10
10
4 12.9 100
10
10
1
5 6.9 10
100
6 17.4 100
100/100
100
7 29.2 100
100
100
t
8 17.1 10
10
Aliphatics
Esters
Allphatica
Aromatic.
Esters
Aliphatics
AZOMtiCS
Esters
Aliphatics
AroMtics
Phenols
Esters
Aliphatics
Esters
Aliphatic.
Phenols
Esters
Aliphatics
Phenols
Esters
Aliphatics
Esters
100
1
100
100
10
100
100
10
10
100
10
10
1
10
100
100
100
100
100
100
100
100
10
10
SaM as Plant A, Fraction 2.
SSM as esters. Plant A, Fraction 6.
Same as Plant A. Fraction 2.
SSM as Plant A, Fraction 2.
Same as esters, plant A, Fraction 6.
SaM as Plant A, Fraction 2.
SaM aa Plant F, Fraction 1.
SaM as Plant F, Fraction 3.
Sams as eaters. Plant A, Fraction 6.
SaM aa Plant F, Fraction 1.
SaM as Plant F, Fraction 1.
Di-t-butvl phenol (CitHilO) M 206.
SaM as Plant A, Fraction 6.
Sams aa Plant A, Fraction 2.
Same as esters. Plant A, Fraction 6.
SaM aa Plant F, Fraction 1.
SaM as Plant F, Fraction 3.
SaM as phenols. Plant A, Fraction 6.
SaM as ester., plant A, Fraction 6.
SaM as Plant A, Fraction 2.
SaM as phenols, plant F, Fraction 5.
Phthalate, probably dl-Cj alXyl but with a new serin
of ions added! 223, 237, 151, 265, 279 ions '
(C^fcHjsOO Ms* 390.
Same as Plant A, Fraction 2.
SaM as estere, plant A, Fraction 6.
•o masses abovo 362.
Ho masses above 312.
10,69(100), 41 (BO), 43(78)
N» nasoes above 486.
1,314(100). 315(25)
10,410(100), 411(32)
Ho Baues above 396.
10,69(100), 41(80), 43(78)
•o masses above 396.
•o masses above 340.
100,99
•o Banes above 279.
1
t
VD .V.U.MI above 336.
100.117(100), 59(44)
10.69(100), 41(80), 43(78)
Also »«ny organo-ailiccfi ioosi
e.g., 207. 221. etc.
Plant 0
1 59.1 100
2 8.1 100
100
3 11.0 100
4 4.4 100
100/10
10
5 5.3 100
100
10
6 21.9 100
100
10
7 11.5 100
100
10/100
6 23.6 100
10
Aliphatics
Aliphatics
ArcMtlcs
Alipttatlcs
Aliphatics
Esters
Am
Aliphatics
Esters
Amines
Aliphatics
Esters
Amines
Aliphatics
Phenols
Esters
Aliphatics
Esters
100
100
100
100
100
100
10
100
100
10
100
100
10
100
100
10
100
100
10
SaM as Plant F, Fraction 1.
SaM as Plant A, Fraction 2.
Same as plant A, Fraction 2.
Same as plant A, Fraction 2.
SSM as Plant A, Fraction 2.
SBM as esters. Plant A, Fraction 6.
vinyl stearate (CjjHjsOj) MK310
(Ci6Hi20SJ2> HH248.
Same as plant A, Fraction 2.
Some as esters. Plant A. Fraction 6.
Halogenated aminesi chloroanlline (C6a6cl) mil 27.
SaM as plant A, Fraction 2.
SaM as esters, Plant A, Fraction 6, and vinyl
stearate (C20"3B°2) HH310.
Chloroanlline (C6H6C1) Ml 127.
Same as Plant A, Fraction 2.
Same as phenols. Plant A. Fraction 6.
Some as esters. Plant A, Fraction 6.
vinyl ctearate tCjo«)o°2> MH 31°
SaM as Plant A. Fraction 2.
Same as esters. Plant A, Fraction 6.
9o auaea abovo 404.
No nasaes above 410.
HD nasaes above 396 in aliphatic-
type pattern i all •.asses above
100 are abnormally strong for
typical aliphatic...
No oasaes above 411.
10.410(100). 411(30)
Ho Basses above 444.
10.429(100), 444(20)
Ho masses above 496.
No masses above 495.
Similar to unusual pattern in
Fraction 3 through mass 495.
No masses abovo 340.
100 i 155
'Relative intensity of the nass-to-chargn ratio (intensity relative to doainant ion).
No molecular weight range determination possible.
e(to composition determination possible.
10,254, 127 (diatomic iodine or
probably napthyl iodide) <'
l:Cesium iodide.
(continued)
81
-------
TABLE 25 (continued)
Plant
and Fraction Catagorlea preaent
Subcategorlea present
organic weight. Relative, Relative
fraction tag intenalty Category intensity Specific conpounda
otbar unknown coatpounda praaant
Plant V
1 15.3 100
10
2 4.9 100
10
3 3.8 100
1
100
4 S.S 10
10
100
5 11. a 10
100
6 3.9 100.
100
100
7 22.6 10
100
I
100
8 19.9 100
1 20.9 100/100
1
2 3.4 100
10
100
3 13.1 10
1
4 8.5 100
10
5 ' 5.7 100
10
10
6 4.2 100
100/10
10
7 14.1 100
10
Aliphatice
Bstera
Oroano-ailicon apeciee
Eaten
Aliphatic.
Aromatlca
(stare
JUiphatiei
Oroano-eilicon speclea
Eatare
Phenol e
Batara
Ulphatlce
Phenol a
Esters
Uiphatica
Aleonols/ethere
Esters
Eatara
Aliffcatlca
Katara
Uiphatics
ATOBAtiCa
Eatara
Uiphatica
Catara
Altfhatlca
Eatara
Aliphatlca
Phanol
Eatara
Uiphatica
Phanola
Batera
Uiphatica
Batara
100
10
100
10
100
1
100
10
10
100
10
100
100
100
100
10
100
100
100
100
100
1
100
10
100
10
1
100
10
100
10
10
100
100
10
10
100
10
Saae as plant p, Fraction 1.
Sane aa aatara, Plant A, Fraction 6.
73 (dominant) , 147, 207, 221, 355 lone.*1''
Sasie as aatara. Plant A, Fraction 6.
SUM as plant A, Fraction 2.
See* as Plant A, Fraction 2.
Saaw aa aatara. Plant A. Fraction 6.
Sake aa plant F. Fraction 1.
73 (dominant). 147 lone."'8
SaM aa aatara. Plant A, Fraction 6.
Saao aa phanola. Plant A, Fraction 6.
Baae aa aatara. Plant A, Fraction 6.
Sas* aa Plant F, Fraction 1.
Saxe aa phanola, Plant A, Fraction 6.
Baa* aa eatara. Plant A, Fraction 6.
SIM aa Plant P, Fraction 1.
Pattern Indlcataa aloobolic e there, 41, 43, 49 (do»-
lnant>«ut 55, 97, 59 (ddatlnant) Ion duatara.11'*
Baata mm wtara. Plant A, Fraction 6.
B«aa aa aatara. Plant A, Fraction 6.
plant V
n-p«raf.lna.d>t
SMa aa plant A, Fraotloa 2.
SBM aa aatars. Plant A, Fraction 6.
Ua» a* plant A, Fraction 2.
Ua» aa plant A, Fraction 2.
Sajaa aa mtara. Plant A, fraction 8.
Basta aa Plant A, Fraction a.
S«BV aa aatara, Plant A, Fraction 6.
SIOM aa Plant A, Fraction 2.
0aM aa aatara. Plant A» Fraction 6.
Saiaa aa Plant A, Fraction 2.
Soa aa phenol. Plant A, Fraction 6.
Sana aa aatara. Plant A, Fraction 6.
Saaw aa plant A. Fraction 2.
Saaa aa plant F, Fraction 5.
SaaM aa phanola. Plant A, Fraction 6.
8a*» aa aatara. Plant A, Fraction 6.
6as* aa Plant A, Fraction 2.
Saaa aa aatara. Plant A, Fraction 6.
Ho Baaaaa abcva 451.
Do aaaaaa abova 491.
m amaaaa abova 296.
10169(100), 41(80), 43(78)
Ho nataai abova 477.
100,69(100), 41(80), 43(78)
Ho aaaaaa abova 340.
10t69(100), 41(80), 43178)
»o amaaaa abova 253.
10(156(100), 155(35)
(Poaaibly hlpyrloyl or phanyl
cyclohaxadiana , MH156 aach)
»o aaaaaa abova 373.
ao auaaa abora 279.
BO m**mmm abova 424.
1,368(100), 369(45), 353(16)
So a*aaaa abova 409.
1001368(100), 169(45), 353(18)
•0 auaaa above 485.
1001366(100), 169(45), 353(18)
no acaaaa above 495.
10,69(100), 41(60), 43(78)
100,368(100), 169(45), 353(18)
Bo naaaaa abova 480.
100i368(100), 369(45), 353(18)
Me. Baaaaa abova 451.
No naaaaa abova 495.
10,69(100), 41(80), 43(78)
100 Uiphatica
> aa Plant A, Fraction 2.
100i384(100], 383(98), 368(90),
382(66)
Ho a*aaes abova 499.
lOiCaalun iodida or nathyl lodloa.
100,69(100), 41(80), 43(78)
10,383(100), 368(96), 382(72),
369(64)
8Relativ« intensity of the oaas-to-charge ratio (zntenalty relative to doainant ion).
No nolecular weiqht range determination posaibla.
Ho conpoaition detarnination posalble.
82
-------
SECTION 7
BIOASSAY OF SECONDARY EFFLUENTS
The primary objective of the entire wastewater toxicity study is
to determine the level of toxicity removal from secondary waste-
water achieved by the tertiary treatment technologies selected
for the ATMI/EPA BATEA study. To this end, the purpose of this
Phase I screening study was to provide chemical and toxicological
base-line data on secondary effluents from the 23 textile plants
and to select plants for the toxicity removal study (Phase II).
Bioassays used were selected by EPA and included tests for
assessment of both health and ecological effects (3). Health
effects tests estimated the potential mutagenicity, potential or
presumptive carcinogenicity, and potential toxicity of the
secondary effluent wastewater samples to mammalian organisms.
Ecological effects tests focused on the potential toxicity of
samples to vertebrates (fish), invertebrates (daphnids and
shrimp), and plants (algae) in freshwater, marine, and terres-
trial ecosystems.
Biological testing, as well as chemical and physical parameters,
must be considered when assessing the potential impact of indus-
trial or municipal/industrial wastewaters on the aquatic environ-
ment. Biological testing involves determination of toxicity for
samples of treated effluents. In a toxicity test, aquatic
organisms'will integrate the synergistic and antagonistic effects
of all the effluent components over the duration of exposure.
Although toxicity tests with aquatic organisms can be conducted
by applying wastewater samples directly to the test organisms, or
by injection or feeding, most tests are conducted by exposing the
test organisms to test solutions containing various concentra-
tions of effluent samples. One or more controls are used to
provide a measure of test acceptability by giving some indication
of test organism health and the suitability of dilution water,
test conditions, handling procedures, etc. A control test is an
exposure of the organisms to dilution water with no effluent
sample added. Bioassay tests are exposures of test organisms to
dilution water with effluent samples added. Generally the most
important data obtained from a toxicity test are the percentages
of test organisms that are affected in a specified way by each
concentration of wastewater sample added. The result derived
83
-------
from these data is a measure of the toxicity of the effluent
sample to the test organisms under the test conditions.
Acute toxicity tests are used to determine the level of toxic
agent that produces an adverse effect on a specified percentage
of test organisms in a short period of time. The most common
acute toxicity test is the acute mortality test. Experimentally,
50% effect is the most reproducible measure of the toxicity of a
toxic agent to a group of test organisms, and 96 hr is often a
convenient, reasonably useful exposure duration. The 96-hr
median lethal concentration (96-hr LCsg) is most often used with
fish and macroinvertebrates. Thus the acute mortality test is a
statistical estimate of the LCs0, which is the concentration of
toxicant in dilution water that is lethal to 50% of the test
organisms during continuous exposure for a specified period of
time. However, the 48-hr median effective concentration (48-hr
EC50), based on immobilization, is most often used .with daphnids.
The terms median lethal concentration (LCsg) and median effective
concentration (ECso) are, consistent with the widely used terms
median lethal dose (LDs0) and median effective dose (EDsg)/
respectively. "Concentration" refers to the amount of toxicant
per unit volume of test solution; "dose" refers to the measured
amount of toxicant given to the test organism.
A total of 8 biological systems' were used for wastewater toxicity
evaluation, utilizing 21 different tester organisms. Specific
tests used and the purpose of each bioassay are summarized in
Table 26. The tests, testing conditions, and toxicity results
for 23 secondary effluent samples are described in this section.
Under guidance of appropriate EPA Technical Advisors, four of the
eight bioassays were performed at commercial laboratories exper-
ienced with the bioassays. The remaining four bioassays were
performed by the EPA Technical1 Advisor, as shown in Figure 20.
Bioassay results were sent to MRC and are included in the fol-
lowing sections.
MICROBIOLOGICAL MUTAGENICITY
Introduction
The purpose of the mutagenicity bioassay was to determine if a
chemical mutagen was present in secondary effluents. Nine dif-
ferent bacteria strains and one yeast strain were used in the
test because of their individual sensitivities to various classes
of chemical compounds. Secondary effluent samples were shipped
to Stanford Research Institute (SRI) for mutagenicity testing by
in vitro microbiological assays with Salmonella typhimurium
(TA1535, TA1537, TA98, and TA100), Escherichia coli WP2, repair-
deficient and proficient strains of Bacillus subtilis H17 and
M45, and E. coli Pol A, and the yeast Saccharomyces cerevisiae
84
-------
TABLE 26. BIOASSAY TESTS USED TO EVALUATE THE TOXICITY OF SECONDARY EFFLUENTS
Bioassay test system
Indicator organisms
Purpose or teat
Microbial rautagenicity
CO
Ul
Cytotoxicity
Freshwater static bioassay
Freshwater algal assay
Marine static bioassay
Marine algal assay
Range finding acute toxicity
Terrestrial ecology
Salmonella typhimurium (Ames test) .
(Strains TA1535, TA1537, TA98, TA100)
Eeahepiohi-a aoli
(Strains WP2, W3110, p3478)
Bacillus aubtilie
(Strains H17 and M4S)
Saccharomycea cerevieial
(Strain D3)
Rabbit alveolar macrophage (RAM)
(viability and ATP determinations)
Chinese hamster ovary cells
Pimephales promelua
(fathead minnow)
Daphnia pulex
(daphnid)
Selenaetrum oaprioornutum
Cyprinodon variegatue
(sheepshead minnow)
Palaemonetee pugio
(grass shrimp)
Skeletonema costatum
Rats (Charles River CD strain)
Soil microorganisms
To determine if a chemical mutagen (possibly a
carcinogen) is present. These microbial strains
were selected because of their sensitivity to
various classes of chemical compounds.
To measure metabolic impairment and death in
mammalian cells. These primary cell cultures have
some degree of metabolic repair capability.
To detect potential toxicity to organisms in aquatic
environments.
To detect potential toxicity to aquatic plants.
To detect potential toxicity to organisms in a
marine environment.
To detect potential toxicity to marine plants.
To detect potential toxicity to whole animals.
To determine potential effects on soil ecosystems.
-------
BIOASSAYAND
TESTING LABORATORY
MUTAGENICITY
SRI ANDMRC
CYTOTOXICITY
NORTHROP AND MRC
FATHEAD MINNOW
AND DAPHNIA
EPA
FRESHWATER ALGAE
EPA
SHEEPSHEAD MINNOW
AND GRASS SHRIMP
BIONOMICS
MARINE ALGAE
EPA
14 - DAY RAT
ACUTETOXICITYTFST
LITTON BIONETICS
EPA TASK OFFICER
SOIL MICROCOSM
EPA
MRC PROJECT LEADER
SAMPLE COLLECTION
MRC
EPA TECHNICAL
ADVISOR
HERL - RTP
M. WATERS
ERL - NEWTOWN
W. HORNING
ERL - GULF BREEZE
J. WALSH
HERL-CINC
J. STARA
EPA-CORVALLIS
B. LIGHTHART
Figure 20.
Laboratories and EPA technical advisors involved
in biotesting of effluent samples.
86
-------
D3. An Aroclor 1254-stimulated, rat-liver homogenate metabolic
activation system was included in each procedure.
The assay procedure with S. typhimupium. has proven to be 85% to 90%
accurate in detecting mutagens, and it has about the same accur-
acy in identifying chemicals that are not carcinogenic (6). The
assay procedure with S. cerevisiae is about 50% accurate in detect-
ing carcinogens as agents that increase mitotic recombination.
The E. ooli WP2 assay and the microbial sensitivity assay are two
additional methods of detecting mutagens. The combination of
these assay procedures significantly enhances the probability of
detecting potentially hazardous substances.
To date the most sensitive assay for deoxyribonucleic acid (DNA)
damage is the induction of mutations in bacteria. The Ames test,
the most highly developed of the bacterial mutagenesis tests,
used mutant strains of S. typhimuriim which were specially selected
because of their abilities to detect specific types of mutations.
For example, the TA1535 strain was designed to detect mutations
due to base-pair substitutions. This strain responded particu-
larly well to alkylating agents. Similarly, the TA1537 and
TA1538 strains were used to detect frameshift mutations. Tester
strains also included mutations which greatly increase their
overall sensitivity to mutagens. One of these was responsibly
for loss of the DNA excision repair system, while the other was
responsible for loss of the lipopolysaccharide barrier that coats
the surface of the bacteria, thereby enhancing the penetration of
large molecules.
Mutant Salmonella tester strains lack the ability to synthesize
histidine and are therefore unable to grow unless histidine is
supplied. These bacteria are cultured in media containing mini-
mal levels of histidine to sustain growth. Under- these condi-
tions only microscopic colonies of bacteria develop during the
course of the test. However, if a mutagen is added to the medium,
a reversion occurs in a certain number of the bacteria, restoring
their ability to synthesize histidine. This reversion (back-
"mutation) is evidenced by the appearance of visible colonies in
the histidine-limited agar, thus indicating the presence of a
chemical mutagen.
Many compounds are not directly acting mutagen but are converted
to active forms by normal body metabolism. A special microsomal
preparation (usually liver) is added in the Salmonella tests to
simulate in vivo metabolic actions. In practice, the substance
is tested with and without this microsomal preparation to deter-
mine whether it requires metabolic transformation or is, itself,
mutagenically active.
(6) McCann, J., E. Choi, E. Yamasaki, and B. N. Ames. Detection
of Carcinogens as Mutagens in the Salmonella/Microsome Test:
Assay of 300 Chemicals. Proceedings of the National Academy
of Science, 72:5135-5139, 1975.
87
-------
The testing procedure used for each type of microbe and the bio-
assay results of each test are described below.
Bioassay Procedures
Each secondary effluent sample was shipped and stored in the lab-
oratory at 4°C. Preliminary experiments on the first two samples
received indicated microbial contamination when an aliquot of the
sample was incubated on a culture medium. Therefore, each sample
was filtered before it was tested in any microbial system. Nal-
gene filters (0.45 ym) were used. Approximately 50, x 10~6 m3 of
each sample was filtered; the remaining 200 x 10""6 m3 was stored
for possible future testing.
Four strains of S. typhimurium (TA1535, TA1537, TA98, and TA100)
were obtained from Dr. Bruce Ames of the University of California
at Berkeley and stored in 10% sterile glycerol at -80°C. New
stock cultures were prepared every two months from single colony
reisolates that were checked for their genotypic characteristics
and for presence of the plasmid. For each experiment, an inocu-
lum from the stock cultures was grown overnight at 37°C in
nutrient broth (Oxoid, CM67). After stationary overnight growth,
the cultures were shaken for 3 hr to 4 hr to ensure optimal
growth.
j
The metabolic activation mixture-used for each experiment con-
sisted of
• 1.0 x 10~6 m3 of S-9 rat liver fraction
• 0.2 x 10~6 m3 of magnesium chloride (0.4 M) and
potassium chloride (1.65 M)
• 0.05 x 10~6 m3 of glucose-6-phosphate (1 M)
• 0.4 x 10~6 m3 of nicotine adenine dinucleotide phosphate
• 5.0 x 10"6 m3 of sodium phosphate (0.2 M, pH 7.4)
• 3.35 x 10~6 m3 of water
For each experiment, the following solutions, listed in the order
of addition, were added to a sterile 13 mm x 100 mm test tube
placed in a 43°C heating block:
• 2.00 x 10~6 m3 of 0.6% agar (containing 0.05 mM
histidine and 0.05 mM biotin)
• 0.05 x 10~6 m3 of indicator organisms
• 0.50 x 10~6 m3 of metabolic activation mixture
• 0.05 x 10~6 m3 of the secondary effluent sample
Dimethylsulfoxide (DMSO) was added to each sample to improve the
water solubility of organic compounds. The resulting mixture was
gently stirred and poured onto minimal agar plates. These plates
88
-------
consisted of 15 kg of agar, 50 kg of glucose, 0.2 kg of magnesium
sulfate (MgBOi+'T H20) , 2 kg of citric acid monohydrate, 10 kg of
potassium orthophosphite, and 3.5 kg of sodium ammonium phos-
phate, per cubic meter. After the top agar had set, the plates
were incubated at 37°C for 2 days. Then the number of revertant
colonies was counted. Each sample was run in duplicate.
All samples were run with both positive and negative controls,
with and without metabolic activation at five concentrations.
Positive controls were run using various combinations of
2-anthramine, 9-aminoacridine, B-propiolacetone, sodium azide,
n-methyl-n'-nitro-n-nitrosoguanidine, daunomycin, and
2-(2-furyl)-3-(5-nitro-2-furyl) acrylamide (AF2). Controls were
also run on DMSO.
The S. aerevisiae tester strain was stored at -80°C. For each
experiment, the tester strain was inoculated in 1% tryptone and
0.5% yeast extract and grown overnight at 37°C with aeration.
The in vitro yeast mitotic recombination assay in suspension was
conducted as follows. The overnight culture was centrifuged, and
cells were resuspended at a concentration of 108 cells per 10~6 m3
in a 67-mM phosphate buffer (pH 7.4). The following solutions
were added to each sterile test tube:
• 1.30 x 10~6 m3 of S. cerevisiae
• 0.50 x 10~6 m3 of either the metabolic activation
mixture or buffer
• 0.20 x 10~6 m3 of the secondary effluent sample
Because many organic chemicals are not appreciably water soluble,
DMSO was added as the solvent for the secondary effluent sample.
Several doses of the chemical [up to 5%, weight-to-volume ratio
(w/v) or volume-to-volume ratio (v/v)] were tested in each experi-
ment, and appropriate controls were included.
The suspension mixture was incubated at 30°C for 4 hr on a roller
drum. The sample was diluted serially in a sterile physiological
saline, and volumes of 0.2 x 10~6 m3 of the 10~5 and 10~3 dilu-
tions were spread on tryptone-yeast agar plates; five plates were
used for the 10~3 dilution and three plates for the 10~5 dilution.
These plates were incubated for 2 days at 30°C, followed by
2 days at 4°C to enhance the development of the red pigment.
Plates of the 10"3 dilution were scanned with a dissection micro-
scope at 10 X magnification, and the number of red colonies or
red sectors (mitotic recombinants) was recorded._ The surviving
fraction of organisms was determined from the number of colonies
appearing on the plates of the 10"5 dilution.
The number of mitotic recombinants was calculated per 105 survi-
vors. A positive response in this assay was indicated by a
89
-------
dose-related increase of more than threefold in the absolute
number of mitotic recombinants per 10~6 m3 as well as in the
relative number of mitotic recombinants per 105 survivors.
Positive, negative, and reagent controls were run at four con-
centrations with each test, with and without metabolic activation.
Positive controls were performed using 1,2,3,4-diepoxybutane.
A procedure similar to the Ames Salmonella assay was used to
measure the reversion of E. aoli WP2 to tryptophan independence.
However, the minimal agar was supplemented with 1.25 g of Oxoid
nutrient broth (CM67) per 10~3 in3 to provide each plate with the
trace of tryptophan required for enhancement of any mutagenic
effect of the test chemical. No additional tryptophan was added
to the top agar. The positive controls used for the Ames test
were also used for the E. coli WP2 test.
As an alternative to reversion of the mutated tryptophan gene;
WP2 may undergo a forward suppressor mutation in a tryptophan
transfer ribonucleic acid (RNA) gene to obtain tryptophan
independence. The test did not distinguish experimentally
between true revertants and suppressor mutants (although the
latter tend to form smaller colonies).
E. aoli. strains W3110 and p3478 and B. subtilis strains H17 and
M45 were stored in the laboratory at -80°C. Inoculums from the
frozen stocks were grown overnight at 37°C with shaking in a
nutrient broth. The broth contained 1% of tryptone and 0.5%
yeast extract, and was supplemented with 5 vq of thymine per
10~6 m3. To 2 x 10~5 m3 of top agar containing 0.6% agar was
added 0.1 x 10~6 m3 of the test culture. This suspension was
mixed and poured onto plates containing nutrient broth and 2%
agar.
After the soft agar solidified, a sterile filter disc impregnated
with a secondary effluent sample was placed in the center of the
plate. Plates were incubated at 37°C for 16 hr, and the width
of the zone of toxicity or inhibition of growth was measured.
Several concentrations of chemical were tested to accurately
detect differences in the zones of growth inhibition, because
higher initial concentrations lead to steep concentration gradi-
ents that may reduce the difference in growth inhibition of the
test strains.
The positive control for this assay was 2 mg of l-phenyl-3,3-
dimethyltriazene placed on the disc. Larger zones of inhibition
were observed in the DNA repair-deficient strains (p3478 and M45).
The negative control was 20 yg of chloramphenicol. Equal zones
of inhibition were observed in all four strains since the toxic-
ity of chloramphenicol did not depend on a mechanism that leads
to DNA damage.
90
-------
Results
SRI tested secondary effluent wastewater samples from 22 of the
23 basic textile plants for bacterial mutagenicity using 10
tester microorganisms and 3 different assay systems. The second-
ary effluent sample from Plant E was lost in shipment.
The voluminous amount of mutagenicity raw data generated by SRI
are given in Reference 7 and a summary of the results is given
in the following paragraphs.
All 22 of the samples were tested twice in the standard Ames
Salmonella microsome procedure using four test strains: TA1535,
TA1537, TA98, and TA100. A metabolic activation mixture was
included in each experiment. Each sample was tested to a maximum
dose of 1 x 10~6 m3 of sample per plate (the maximum amount pos-
sible in 2 x 10~6 m3 of top agar). The second experiment was a
confirming experiment. None of the samples caused an increase in
the number of histidine-independent revertants above the normal
background, thus no chemical mutagen was detected.
Twenty-two samples were tested in the E. ooli WP2 strain. None
of the samples caused an increase in the number of tryptophan-
independent revertants above the normal background. Twenty-two
samples were tested in the S. cerevisiae D3 suspension assay,
with and without a metabolic activation system. The maximum con-
centration tested was 50% v/v. None of these samples caused an
increase in the number of mitotic recombinants above the normal
background.
The samples were also tested in the microbial inhibition assay
using DNA repair-deficient and -proficient strains of E. ooli
and B. subtilis. The maximum dose tested was 20 x 10~9 m3.
Each sample was applied to a filter disc on the plate. None of
the samples was toxic to any of the strains of the organisms
used.
All 22 secondary effluent samples were tested in three assay
systems. In each assay, the maximum possible dose was tested.
None of the samples gave a mutagenic or toxic response in any
strain in any assay or experiment.
(7) Poole, D. C. and V. F. Simmon. Final Report of in Vitro
Microbiological Studies of Twenty-two Wastewater Effluent
Samples. Contract 68-01-2458, U.S. Environmental Protection
Agency, Biomedical Research Branch, Research Triangle Park,
North Carolina, November 1977. Ill pp.
91
-------
MRC Ames Testing
To provide backup duplicate results to the Ames tests performed
by SRI, MRC performed the Ames test on eight randomly selected
secondary effluent samples from Plants D, H, J, M, P, R, Y, and
Z.
i
Biotest procedures used by MRG were the same as those used by SRI.
Final test results indicated no positive responses on filtered,
unconcentrated effluent samples. Samples from Plants D, P, and
R were rerun on selected strains with no indication of a positive
response.
i
CYTOTOXICITY ASSAY
Introduction '>
I
Cytotoxicity (cell toxicity) assays were performed to measure
quantitatively any cellular metabolic impairment and death
resulting from exposure in vitro to secondary effluent samples.
Primary cell cultures, such as the rabbit alveolar macrophage
(RAM) used in this study, exhibit many of the metabolic and func-
tional attributes of the in vitro' state. These cells can there-
fore combat, to some degree, the effects of chemical mutagens on
mammalian cells.
Recently this system has been applied in evaluating the relative
cellular toxicity of hazardous metallic salts and industrial air
particulates (8). As compared to conventional whole animal tests
for acute toxicity, these cytotoxicity assays are more rapid,
less costly, and require less sample. These tests provide useful
information about the relative toxicity of unknown samples. How-
ever, it should be understood that because the assays employ
isolated cells and not intact animals, they can provide only
preliminary information about the ultimate human health hazards
of toxic chemicals.
Two tests were used to measure the toxic effects of secondary
effluent samples on rabbit alvaolar macrophage: viability and
adenosine triphosphate (ATP) production. Viability refers to the
ability of cells to survive, and it was measured by the trypan
blue dye exclusion method. Living (viable) cells do not absorb
trypan blue dye. Therefore, the measure of cell mortality is the
number of blue (dead) cells counted after exposure to the sample.
Adenosine triphosphate (ATP) is a coenzyme in mammalian cells
that plays an important role in energy metabolism. Living cells
synthesize ATP. Therefore, another measure of cell mortality 'or
(8) Waters, M. D., D. E. Gardner, C. Aranyi, and D. L. Coffin.
Metal Toxicity of Rabbit Alveolar Macrophages in Vitro.
Environmental Research, 9(l):32-47, 1975.
92
-------
inhibition was the quantity of ATP produced by a cell culture
exposed to the secondary effluent sample compared to the amount
produced by an identical culture not exposed to the solution.
Both of these methods were used to evaluate secondary effluent
toxicity to RAM. The measure of toxicity was expressed as EC2Q
or ECso; i.e., the concentration of secondary effluent that
inhibits RAM metabolism by 20% or 50% over a specified time
period (20 hr).
The following section discusses the test procedure used to evalu-
ate secondary effluent toxicity and the bioassay results.
Bioassay Procedure
Rabbit alveolar macrophage primary cell cultures were obtained
from New Zealand white rabbits of both sexes. Each of the 23
secondary effluent samples was filtered through a 0.45-ym filter.
There was no concentrating of the samples prior to testing. Each
sample was prepared in five dilutions: 6, 20, 60, 200, and
600 x 10~6 m3 sample per 10"3 m3 solution. To each concentration,
fetal calf serum (heat inactivated) was added to give a final
serum concentration of 10%. Antibiotics were added to give 100
units per 10~6 m3 penicillin, and 100 \ig combined streptomycin
and kanamycin per 10~6 m3. The pH of each concentration was
recorded, but no adjustments were made.
Samples were then added to Falcon cluster dishes, 1.5 x 10~6 m3
per well. A volume of 0.5 x 10~6 m3 of complete IX Medium 199
(with 10% fetal calf serum and antibiotics) containing approxi-
mately 2 x 106 rabbit alveolar macrophages per 10~6 m3 was added
to each well and gently mixed. Dishes were then incubated for
20 hr at 37°C in a humidified atmosphere of 5% C02 in air on a
Belico rocker platform.
At the end of the 20-hr exposure, the medium containing
unattached cells was removed from each well and transferred to a
separate test tube. A volume of 10~6 m3 of 0.25% trypsin was
added to each well and left until the cells were removed from the
dish. This was then combined with the original pouroff and mixed
to inactivate the trypsin.
Cell counts, viabilities, and ATP determinations were then .
performed.
Viability was determined by trypan blue dye exclusion. Filtered
samples were counted in the Cytograf; unfiltered samples, because
of the particulate matter present, were counted in a hemocytometer,
For hemocytometer counts, 1 part of 0.4% trypan blue was added to
5 parts cell suspension and counted after a 5-min exposure. For
Cytograf counts, dilutions, usually fourfold, were made with cold
93
-------
0.85% saline to yield a suspension of no more than 2 x 105 cells/
10~6 m3. Trypan blue was diluted (immediately before use) with
0.85% saline to a final concentration of 0.01% and added to an
equal volume of cell suspension. Simultaneous determinations of
cell viability and cell numbers per 10~6 m3 of cell suspension
were made. The numbers of viable cells were expressed as a per-
centage of the -number of cells in control cultures; viability was
expressed as the concentration of secondary effluent which
inhibited 20% and 50% of the test cells (EC2o and EC50).
ATP was determined according to a procedure supplied with the
Du Pont Model 760 Luminescence Biometer. Dimethyl sulfoxide
(0.4 x 10~6 m3) was used to extract ATP from a 0.1 x 10~6 m3
aliquot of trypsinized cell suspension containing 0.3 to 0.4 x 105
cells. After 2 min at room temperature, 2.5 x 10~6 m3 of cold
0.1 M morpholinopropane sulfonic acid (MOPS) at pH 7.4 was added
to buffer the extracted sample. The tube containing the buffered
sample was then placed in an ice bath. Aliquots of 10~8 m3 were
injected into the luminescence meter's reaction cuvette contain-
ing 0.7 mM luciferin (crystalline)/ 100 units luciferase (puri-
fied and-stabilized) , and 0.01 M magnesium sulfate in a total
volume of 10~7 m3 of 0.01 M MOPS buffer, pH 7.4 at 25°C. Light
emitted from the reaction cuvette was measured photometrically in
the luminescence meter and was proportional to the ATP concentra-
tion of the sample. ATP values were expressed per 106 cells and
as a percent of the control cells.
Nutrient agar plates were streaked with 0.5 x 10~6 m3 from each
sample (unfiltered) and antibiotic sensitivity discs were added
for a 24-hr incubation period. This was done to ascertain what
antibiotics were capable of suppressing growth of any bacteria
present in the samples. Antibiotics present in the culture
medium were found to be capable of inhibiting bacterial growth so
that the samples could be tested unfiltered.
Results
All determinations were performed in duplicate or triplicate.
The voluminous amount of cytotoxicity raw data generated by
Northrop are given in Reference 9 and a summary of the results
are given below.
Because cell viability could be considered a binomial response,
the arc-sine transformation was employed in the regression
(9) Campbell, J. A., H. F. Stack, and P. R. Williams. Cyto-
toxicity Screening of Twenty-three Textile Mill Effluent
Water Samples Utilizing the Rabbit Alveolar Macrophage
Assay. Contract 68-02-2566, U.S. Environmental Protection
Agency, Biomedical Research Branch, Research Triangle Park,
North Carolina, December 1977. 86 pp.
94
-------
analysis. Samples listed as nondeterminable were such high
extrapolations that they could not be considered significant
estimates.
Table 27 shows the estimated EC20 and £€50 concentrations for the
filtered effluents. The EC values are the concentrations
expected to cause a decrease in viability and ATP by 20% and 50%,
respectively. Control values were routinely 92% to 100%.
TABLE 27. ESTIMATED EC2o AND EC50 VALUES FOR CYTOTOXICITY
SCREENING OF FILTERED SECONDARY EFFLUENT SAMPLES
Percent effluent
Viability ATP3
Plant EC2° ECso EC2° EC5°
A
Bb
a" 16.8 6.1 33.5
cc d d
D
E
F 9.4
G
H
J
K
L 4.0 35.1
Plant
M
N
P
R
S
T
0
V
W
X
y
z
Percent effluent
Viability ATP3
EC£0 ECso EC20
13.3 3.8
d d d
2.5
13.7
4.8
ECso
12.8
d
Note.—Blanks indicate data not determinable.
Adenosine triphosphate. pH equals 9.1 not adjusted before testing.
C d
pH adjusted to 7.2. Test not performed.
Samples from Plant N and C caused the greatest response by viabil-
ity and ATP determinations. The response of Sample C was largely
due to its high (9.1) pH. When testing was repeated with the pH
adjusted to 7.2, there was much less response.
Because the antibiotic sensitivity testing showed that samples
could be tested without prior filtration, five samples were
retested without prior filtration. Much of the solid material
removed by filtration appeared to be biological material (e.g.,
algae) by microscopic observation. In each instance, the
unfiltered sample caused greater response than the filtered one.
MRC Clonal Assay
MRC performed clonal assay acute toxicity tests using Chinese
hamster ovary cells (CHO-K1) on selected secondary effluent
95
-------
samples. The purpose of these tests was to evaluate the response
of another test system to complex environmental samples. MRC has
developed this in vitro clonal assay for measuring acute toxicity
of compounds using CHO-K1 cells. This test is a modification of
a clonal assay described by Malcolm (10). Preliminary studies
indicate that the sensitivity of the CHO-Kl clonal assay appears
to be two orders of magnitude greater than that of the WI-38
assay.
Secondary effluent samples from eight textile plants (Plants D, H,
J, M, P, R, Y, and Z) were selected at random and analyzed accord-
ing to the following procedure.
Test Procedure—
Effluent samples for this test were filtered through 0.45-ym and
0.22-pm filters. Samples were run at 5 to 7 concentration levels
using approximately 300 to 500 cells that had been plated on the
previous day. After incubation at 37°C for 6 days to 7 days, the
media and sample were removed and the cells were fixed, stained,
and counted. Results are reported as experimental versus con-
trols or percent survival. A detailed test procedure is given in
Table 28.
Results—
Screening tests were first performed on the eight samples to
determine whether any of them were toxic and in what concentra-
tion range. Results showed that secondary effluent samples from
Plants P, R, Y, and Z had no acute toxicity to the CHO-Kl cells,
but samples from Plants D, H, J, and M did exhibit toxicity.
The latter four samples were therefore rerun using narrower sam-
ple concentrations.
Test results are presented in Figure 21. The range bar associ-
ated with each data point corresponds to the standard deviation
of that value.
Graphical interpolation of Figure 21 yields the following LC5o
values (in percent of secondary effluent sample) for Plants D, H,
J, and M: 2.4%, 13.3%, 18.5%, and 3.0%, respectively. These
values indicate that the secondary effluent from Plants D and M
contain chemical species more acutely toxic to CHO-Kl cells than
do samples from Plants H and J.
(10) Malcolm, A. R., B. H. Pringle, and H. W. Fisher. Chemical
Toxicity Studies with Cultured Mammalian Cells. In: Bio-
assay Techniques and Environmental-Chemistry, G. E. Glass,
ed. Ann Arbor Science Publishers, Inc., Ann Arbor,
Michigan, 1974. pp. 217-230.
96
-------
TABLE 28. CHO-Kl CLONAL CYTOTOXICITY TEST
Cell line: Chinese hamster ovary epithelial cells ATCC No. CCL 61
Medium: F-12 GIBCO No. H-17 10.8 x 103 g/m3
Sodium hydrogen carbonate
10% Fetal calf serum, virus, mycoplasma screened
GIBCO No. 629
Incubation: 37°C, 5% C02, Saturated humidity
Samples: 6 Controls (blank)
5 to 7 Concentrations of test compound in triplicate
5 Concentrations of a positive toxic control in triplicate
Test procedure .
To stock CHO-Kl, add 5 x 10~6 m3 0.25% trypsin at 37°C for 5 min.
Shake cells and add to centrifuge tube.
Add 5 x 10~6 m3 media to flask, shake, and add to centrifuge tube.
Centrifuge 5 min at 1,200 g, pour off liquid, retaining cells.
Add 10 x 10~6 m3 medium, shake, centrifuge 5 min, pour off medium.
Add 10 x 10~6 m3 medium, shake.
Make hemocytometer count of trypsinized cells.
Dilute so that 5 x 10~6 m3 media contain 300 to 500 cells.
Add 5 x 10~6 m3 media and cells to T-25 flasks.
Incubate 12 hr to 18 hr to allow attachment using normal media.
Replace 5 x 10"^ m3 of media and sample.
Incubate 6 days to 7 days total.
Fix with 10% formaldehyde/0.5% sodium chloride/4% methanol for 30 min.
Stain with crystal violet (0.04% for 15 min).
Count clonal colonies of remaining cells macroscopically using Fisher Count-
All Model 600.
Score with respect to experimental vs. controls as percent survival.
97
-------
20 40 60 80 100 120 140 160 180 200
SAMPLE CONCENTRATION, 10~3m3/m3
Figure 21. Results of CHO-Kl clonal assay.
FRESHWATER ECOLOGY TOXICITY
Algal Assay Bottle Test
Algal assay was performed to estimate the potential toxicity of
secondary effluents on aquatic plants. The algal assay was based
on the principle that growth is limited by the essential nutri-
ents that are in shortest supply. The test was designed to quan-
tify the biological response (algal growth) to changes in concen-
trations of nutrients, and to determine whether various effluents
were stimulatory or inhibitory to algae. These measurements were
made by adding a selected test alga to the effluent and deter-
mining its growth response.
The freshwater algae testing series was performed at the Environ-
mental Research Laboratory (ERL, Corvallis, Oregon) under the
direction of W. E. Miller. Each effluent sample (0.01 m3 from
each of the 23 plants) as apportioned in three autoclavable,
0.004-m3 plastic bottles, was packed in ice and flown air freight
to Corvallis, Oregon. Most samples arrived at the laboratory with-
in 36 hr from the time they were shipped. The following paragraphs
summarize the sample handling and analysis procedures used at ERL,
An EPA manual (3) gives a detailed description of the procedure.
98
-------
Each 0.01-m3 textile effluent sample was thoroughly composited in
the laboratory in a 0.02-m3 cubitainer, then redistributed into
bottles. A 0.001-m3 aliquot was taken for, soil microcosm studies.
Samples were stored in the dark at 5°C.
Well water from the Western Fish Toxicology Station was used for
dilution water and for control samples. The water was filtered
with a 0.45-wm porosity membrane filter and pad to remove any
particulate matter before mixing with raw effluent. Dilutions,
Table 29, were made in a 0.02-m3 cubitainer for a total of
0.016 m3 of diluted effluent.
TABLE 29. DILUTIONS USED FOR FRESHWATER ALGAL TESTS
"Effluent volume,Well water volume,
Dilution, % x IP"1* m3 x IP"3 m3
-
1
Total
1
2
5
10
20
volume used
3.2
8.0
16.0
32.0
59.2
15.68
15.20
14.40
12.80
58.08
Each dilution of 0.016 m3 was divided into four parts of 0.004 m3
each for four different treatments: One part was autoclaved and
filtered, one was autoclaved only, one was filtered and auto-
claved, and one was filtered only.
Diluted samples were autoclaved in polypropylene bottles (washed
with 10% hydrochloric acid and autoclaved before use) .at a pres-
sure of 108 kPa at 10 min/10~3 m3. Diluted samples were filtered
with a 0.45-pm porosity membrane filter and pad. During and
•"after treatment, diluted samples were stored in the dark at 5°C.
The~pH values were taken on raw effluent and on diluted effluent
before and after treatment to determine changes in pH that can
affect growth. Each secondary effluent sample was analyzed by
MRC for the following nutrient indicators: o-phosphate, ammonia,
nitrite, nitrate, total Kjeldahl nitrogen, total phosphorus, and
total organic carbon, Table 30 (4, 5).
The algal assay bottle test procedure used Selenastrum capri-
"oorriutum Printz as the test alga and was used to assess algal
growth response to 23 secondary effluent wastewater samples.
Growth response was measured as net biomass produced (gram dry
weight per cubic meter of sample), i.e., the total biomass in
the sample minus the biomass produced in a control sample using
well water. Algal response was expressed as percent stimulation
or inhibition at the 20% wastewater concentration as compared
to the control sample. Tests were performed on two types of
99
-------
TABLE 30. NUTRIENT ANALYSIS OF SECONDARY EFFLUENT SAMPLES
Plant
code
A
B
C
D
E
F
G
H
J
K
L
M
N
P
R
S
T
U
V
W
X
Y
Z
^*^^^MI^^*BW^^BH^
Nitrite
0.06
0
4.64
7.5
<0.02
0.04
<0.02
16.8
11.9
<0.02
0.86
0.27
<0.02
7.3
1.82
<0.02
0.04
<0.02
0.07
0.145
0.44
8.5
<0.02
•taH*^^^^«^^^^^^«^^BA
Nitrate
1.9
0.002
23.3
0.08
40
0
1.3
0.69
0.24
0.16
13.5
0.30
5.5
0.08
0.015
0.23
0.80
0.8
1.0
12.3
0.033
0.05
0.94
MM^^^MV^^m^BM^MIta^M
Ammonia
12.8
2.5
3.4
0.20
1.13
1:54
0.40
0.02
1.25
0.40
0.5
0.84
12.8
0.20
2.6
1.65
13.6
5.44
1.26
0.38
0.65
1.9
0.85
Nutrient,
Total
Kjehldahl
nitrogen
21.3
4.2
1.85
4.5
6.75
14.8
12
5.5
2.3
5.25
1.9
6.15
17.0
2.9
19.8
40
30.2
11.3
6.3
7.4
5.25
5.75
4.75
g/m3
«»
o -Phosphate
1.0
7.3
1.08
1.6
1.11
0.56
5.02
0.06
0.28
0.82
0.88
0.55
11.2
0.02
0.06
3.1
6.4
2.96
0.11
0.075
0
9.9
0.40
^^^^^^^^^MIAMWI^AWV^H^^H
Total
phosphorus
0:39
6.0
0.50
2.48
1.19
9.7
6.29
0.39
1.68
1.05
1.98
3.03
2.05
3.12
1.82
5.23
14.9
2.43
0.76
0.50
4.87
14.3
0.54
Total
organic
carbon
29.0
219
36
31.8
57.5
84
150
19.1
42.9
•84
90.4
54
244
261
142
199
94
41.8
40
Note.—Blanks indicate analysis,not performed.
pretreated samples: 1) filtered followed by autoclaving, and
2) filtered only. Results are given in Table 31.
TABLE 31. RESULTS OF FRESHWATER ALGAE BIOASSAY TESTS
14-Day growth response in 20* secondary effluent
aa compared to control well water
Filtered only
Plant
A
B
C
D
E
f
C
H
J
X
L
M
N
p
R
S
T
U
V
H
X
Y
Z
Percent
inhibition
53
0
0
0
95«
0
0
92
0
0
81
0
95»
0
95"
0
0
0
0
95
0
0 i
84
Percent
stimulation
0
83
187
100
0
598
390
0
76
57
0
149
0
38
0
382
1.911
377
232
0
163
261
0
Piltered-autoclaved
Percent
inhibition
33
0
0
0
95«
0
0
95
0
0
0
0
95«
5
93b
0
0
0
D
95
0
0
17
Percent ,
•timulation
0
44
146
125
0
866
578
0
217
243
98
291
0
0
0
365
2,362
639
503
0
348
365
0
.°95« growth inhibition in 2t solution of secondary effluent.
^Sample inadvertently collected prior to the lettling pond.
Five distinct growth patterns were discerned in the initial
screening; they are described in the following sample subsets,
100
-------
Two samples (Plants E and N) failed to support growth of the test
alga in all waste concentration levels (2%, 5%, 10%, and 20%).
This response has been attributed to toxicity since nitrogen (N)
and phosphorus (P) concentrations (2% level) were adequate to sup-
port up to 8.0 g dry wt/m3 of S. eaprieornu-tum.
Seven samples (Plants B, D, J, K, P, X, and Z) supported similar
growth at all dilutions ranging from 5.5 to 16.0 g dry wt/m3 of
S. oapricornutum, depending on nutrient bioavailability within
waste samples. Failure of the growth response to increase at a
rate proportional to the analyzed incremental concentration of
nitrogen and phosphorus in these samples suggests that some con-
stituent other than nitrogen and phosphorus is limiting the maxi-
mum yield. However, the suboptimal yield obtained is considered
to be stimulatory for the support of test alga.
Four samples (Plants H, L, R, and W) supported growth at the 2%
and 5% levels but inhibited growth at the 10% and 20%
concentrations.
Seven samples (Plants C, F, G, M, S, V, and Y) supported increas-
ing growth of test alga with similar increase in waste concentra-
tion, proportional to the bioavailable (but not chemically
analyzed) inorganic nitrogen content. The growth thus obtained
indicates that these wastes are highly stimulatory for the sup-
port of test alga.
Two samples (Plants T and U) were extremely stimulatory, 124.5
and 29.1 g dry wt/m3, respectively, for these wastes at the 20%
concentration level. Growth obtained in these samples was
directly proportional to their chemical nitrogen and phosphorus
content, indicating its complete availability for support of
algal growth.
Using test results from the filtered-and-autoclaved and filtered-
only samples, ERL was able to rank the secondary effluent samples
in terms of inhibition and stimulation; results are shown in
Table 32. In those samples categorized as nontoxic, the primary
limiting factor regulating growth response of the test alga was
bioavailability and utilization of the total soluble inorganic
nitrogen (TSIN equals nitrite, nitrate, and ammonia). Shiroyama,
Miller, and Greene (11) demonstrated that maximum yield for
Selenastrum capriaornutum Printz is predictable provided the
TSIN is known, other essential nutrients are in adequate supply,
and toxicants are absent. Under these conditions 0.001 g/ra3 of
TSIN can yield 0.038 g/m3 dry weight of the alga. Based on this
(11) Shiroyama, T., W. E. Miller, and J. C. Greene. The Efforts
of Nitrogen and Phosphorus on the Growth of Selenastrum
Caprioornutum Printz. EPA-606/3-75-034, U.S. Environmental
Protection Agency, Corvallis, Oregon, March 1975.
pp. 132-142.
101
-------
TABLE 32. RELATIVE RANKING OF TEXTILE PLANTS BY
TOXIC AND STIMULATORY EFFECTS OF
SECONDARY WASTEWATER ON S. CAPRICORNUTUM
Filtered and autoc laved
Toxic Plant
rating rank
Host toxic E, N
W,aH
Least toxic R. P, L, B, Z, A
Nontoxic O
C
Y
K
J
H
X
S
F
V
G
U
T
samples Filtered only
Stimulatory Toxic Plant
rating rating rank
Nonstimu!
P
H
Least at.
Latory Host
jnulatory Leas
Nontc
Host stimulatory
toxic E, N
w a
R," H
toxic A, Z, L
xic P
j
B
K
M
D
v
Y
C
u-
G
n
V
T
samples
Stimulatory
1 rating
Nonstimulatory
•
•
H
Least stimulatory
Host stimulatory
Sample inadvertently collected prior to the setting pond.
information a linear regression between biomass produced in the
textile waste samples and that predicted from the TSIN content
of the test water showed a correlation coefficient in the
filtered followed by autoclaving and filtered only samples of
0.980 and 0.989, respectively, resulting in dry weight yield
per unit concentration of TSIN relationship of 0.910 and 0.997,
respectively. These relationships indicate that the complexity
of the textile water samples under the two pretreated conditions
does not affect the algal from utilizing the essential nutrients
in obtaining its maximum yield.
Acute Static Bioassays with Freshwater Fish and Daphnia
The acute static bioassay technique with freshwater animals pro-
vided an easy measure of toxicity and was recommended by EPA for
the wastewater assessment (3). Fathead minnow (Pimephales
promelus) and Daphnia pulex were the selected test animals
because they are a readily available, hardy species, and they
can be conveniently and economically maintained in a laboratory
(3).
Primary objections to the following procedure are that the recom-
mended dilution water may not closely simulate receiving water
characteristics, and the fathead minnow may not be representative
of the most sensitive species in a given geographical area. How-
ever, the procedure does adequately serve to develop relative
toxicity data for the purpose of ranking industries based on the
toxicity of their effluents.
102
-------
This series of tests was performed at the EPA Fish Toxicology
Station (Newtown, Ohio) under the direction of Mr. W. Horning.
Each of the three 0.02-m3 glass bottles of effluent from 23
plants was packed in ice and shipped by air freight.
The fathead minnow test utilized 16 x 10~3 m3, wide mouthed jars.
A control dilution and effluent dilutions of 100%, 60%, 36%,
21.6%, 13%, 7.8%, and 4.7% effluent were set up. A total volume
of 0.015 m3 was used in each test jar. Ten fish were randomly
distributed to each jar. Duration of the test was 96 hr, and
temperature ranged from 20.5°C to 21.6°C. Aeration was not used
during the test. At the end of the test, the fish length and
weight were determined. Length ranged from 28 mm to 44 mm, with
an average length of 33 mm. Weight ranged from 0.18 g to 0.80 g,
with an average weight of 0.29 g.
Dissolved oxygen, pH, specific conductivity, temperature, and
turbidity were determined in each test jar at the beginning of
the test and every 24 hr through the end of the test. At the
end of each 24-hr period, the number of fish surviving was
recorded, and dead fish were removed.
The same test procedures and dilutions of effluent used for the
fathead minnow test with D. pulex. The D. pulex were from a
laboratory culture maintained at the Newtown Fish Toxicology
Station. Estimated LC5o and ECso values and 95% confidence
limits were reported where possible and are shown in Table 33.
Data were evaluated with Probit Analysis whenever possible, with
Moving Average Angle where Probit was not applicable.
TABLE 33. ACUTE TOXICITY DATA FOR FATHEAD
MINNOW AND DAPHNIA PULEX
Fathead minnow,
percent effluent concentration
Plant
code
A
B
C
0
E
P
G
H
J
K
L
M
N
Pn
R9
S
T
U
V
H
X
Y
Z
96-Hr
LCso
19.0
NAlC
46. 5
NAT
NAT
MAT
64.7
d
NAT
NAT
23.5
NAT
48.8
NAT
16.5
NAT
46.5
NAT
36.0
55.2
NAT
NAT
NAT
95% Confidence limit
15.5
37.6
57.0
IB. 3
38. 6
12.4
37.6
27.4
45.2
to
to
to
to
to
to
to
to
to
24.0
57.5
74. B
28.7
61.8
21.7
57.5
43.9
70.7
Statistical
analysis
MA"
HA
MA
MA
P
P
MA
MA
MA
baphnia pul«x,
percent effluent concentration
48-Hr
ECso 95t Confidence limit
9.0
NAT
41.0
NAT
7.8
81.7
62.4
e
NAT
NAT
28.0
60.0
f
NAT
8.0
NEAR
NAT
12.1
9.4
6.3
NAT
NAT
42.6
6.
32.
5.
66.
54.
40.
6,
a.
7.
3.
30.
,8 to
.4 to
.6 to
.7 to
.3 to
.7 to
.1 to
,7 to
.1 to
.7 to
,8 to
11.6
50.2
9.8
101.6
72.7
89.0
8.0
16.3
12.2
8.4
64.1
Statistical
analysis
Pb
MA
P
P
MA
MA
MA
MA
P
P
P
P
Moving average angle. Probit. cNo acute toxicity.
e40l Dead at 100% concentration. 100% Dead at all dilutions.
'sample inadvertently collected prior to the settling pond.
No statistical analysis; heavy solids concentration bbscured the analysis! the sample did not
appear to be acutely toxic.
103
-------
Generally, the toxicity of textile mill effluent samples was
exerted within the first 24 hr for both the fathead minnow and
D. pulex. The relative sensitivity of the animals, cannot be dif-
ferentiated on the basis of these samples. In 5 out of the 15
samples, D. pulex appeared to be somewhat more sensitive than the
fathead minnow. In one instance, the effluent was not acutely
toxic to D. pulex but was acutely toxic to fathead! minnow, with
an estimated LC50 value of 46.5% effluent dilution. It should
be noted that 8 samples were acutely toxic to fathead minnow and
10 were acutely toxic to D. pulex. Thus, it is desirable to use,
when feasible, more than 1 organism to evaluate the toxicity of
an effluent.
Reactions of test organisms to each effluent sample are briefly
described in Appendix D.
MARINE ECOLOGY TOXICITY
Bioassay with Unicellular Marine Algae \
t
Unicellular algae are important constituents of marine ecosystems.
They are comprised of a variety of species that have different
growth rates, photosynthetic rates, nutrient requirements, and
other functions that regulate species composition and diversity
in the community in relation to environmental parameters. The
algal community, through photosynthesis, produces most of the
food and oxygen used in the marine ecosystem. '
i
It is well known that algal species and communities are sensitive
to environmental changes. Species may be either inhibited or
stimulated by pollutants. In a community, a pollutant may affect
some species but not others, thereby causing changes in species
diversity and composition. This can be followed by changes in
composition of the animal community and altered routes of flow of
energy and materials. Often, the altered ecosystem is undesira-
ble from the human standpoint. On this basis, a bioassay program
designed to study effects of suspected pollutants should include
research on unicellular algae.
Marine algae tests were performed on 15 textile effluent samples
at ERL (Sabine Island, Gulf Breeze, Florida) under the direction
of Dr. J. Walsh. Fifteen wastewater samples were subjected to
this testing series instead of 23 samples because this bioassay
was integrated into the program after sampling began. EPA test
procedures (3) used for this analysis were modified as follows:
• Continuous lighting was used instead of a 14-hr-light,
10-hr-dark cycle.
• Salinity was 30 parts per thousand (ppt) instead of
10 ppt.
• Wastes were not sterilized.
104
-------
• Optical density of cultures was determined on days 3, 4,
and 5.
• Final biomass was not estimated.
• Only the 96-hr EC59 is reported.
The relationship between optical density as a measure of popula-
tion density and cell counts was determined by spot checks. In
all cases, it was found that optical density and cell counts
using a hemocytometer were closely correlated.
In order to estimate stimulation of growth, the ratio of popula-
tion density in treated samples to population density in controls
(T/C) was calculated, and the highest value for each waste is
reported.
Effects of the textile wastes on population density of
Skeletonema oostatum are given in Table 34. There was a wide
distribution of toxicity. Wastes L and N were by far the most
toxic, whereas B, S, U, and X were not toxic.
TABLE 34. RESULTS OF MARINE ALGAE ACUTE TOXICITY TESTS
Stimulation
Percent growth Percent
Inhibition, ECsg, over control, secondary
Plant percent secondary effluent T/C effluent^
B
C
E
F
G
K
L
N
P
S
T
U
V
W
X
_b
90
10 to 50
85
59
77
1.7
2.3
9.0
_b
70L
_b
94
50
_b
200
130
0
230
130 '
180
110
120
110
410
310
180
120
230
170
100
80
0
10
10
10 to 30
0.5
0.1 to 0.5
0.1
90
35
70
10 to 70
30
95
Percent of secondary effluent solution corresponding to highest
stimulation growth rate.
'inhibition less than 50% in 100% secondary effluent.
"Small volume of sample received, not enough to complete tests.
105
-------
Most wastes stimulated growth. Although there were not enough
data for proper statistical analysis, wastes with T/C ratios of
1.1 to 1.3 were not significantly different from the controls.
The importance of growth stimulation by waste must not be under-
estimated. Of the 15 samples, 8 definitely stimulated growth.
Five wastes caused population densities two to four times those
of the controls.
Another way to look at this is to estimate the concentrations
that doubled the population dens-ity (T/C equals 2) :
Plant Percent waste
T
S
F
W
B
10
30
30
100
Fewer than 10% of the wastes from Plants U, X, and K causes an
increase of 50% in population density (T/C equals 1.5).
Growth stimulation could have a substantial impact on natural
bodies of water by causing 1) eutrophication and/or 2) changes in
relative numbers of important phytoplankton.
Marine Animal Series
Marine animal bioassay testing was performed to ascertain the
concentration of secondary wastewater sample that was acutely
toxic to juvenile sheepshead minnows and to grass shrimp. Al-
though none of the textile plants discharge directly into a
marine environment, this biotest was performed to provide general
information about the toxicity of textile plant wastewaters and
to evaluate this new bioassay testing procedure. Since this
testing series was integrated into the program after sampling
began, only 14 textile plant wastewater samples were subjected
to this test.
The EPA static bioassay procedure incorporates several methods
(5, 12) and is the simplest, most economic marine animal assay
test available. Juvenile sheepshead minnows (Cyprinodon
variegatus) and adult grass shrimp (Palaemonetes pugio or P.
vulgaris) were used as test animals because they adapt easily to
a wide range of salinity and temperature in static bioassays.
(12) Sprague, J. B. The ABC's of Pollutant Bioassay Using Fish.
In: Biological Methods for the Assessment of Water Quality,
J. Cairns, Jr., and K. L. Dickson, eds. ASTM Special Tech-
nical Publication 528, American Society for Testing and
Materials, Philadelphia, Pennsylvania, 1973. pp. 6-30.
i
106
-------
Additionally, various phases of their life cycles may be studied
due to their short life span.
There are two objections to this procedure: Receiving water
characteristics are not closely simulated, and the test organism
may not be representative of the most sensitive species in a
given geographical area. However, the method is adequate for
ranking industries according to their effluent toxicity.
Tests on 14 textile plant wastewater samples were performed at
EG&G-Bionomics Marine Research Laboratory (BMRL; Pensacola,
Florida) under the direction of Dr. R. Parrish. Dr. J. Walsh was
the EPA Technical Advisor associated with the marine ecology
biotests.
A total of 0.02 m3 of secondary effluent sample was collected
from each of the 14 plants for sheepshead minnow and grass shrimp
acute toxicity analysis. Samples were shipped via air freight in
glass bottles, packed in ice, and were stored at BMRL in a room
with the temperature controlled at 15 ± 1°C until testing. A
description of the samples as they arrived at BMRL is given in
Table 35.
TABLE 35. PHYSICAL DESCRIPTION OF EFFLUENT SAMPLES AS
THEY ARRIVED AT BMRL FOR SHEEPSHEAD MINNOW
AND GRASS SHRIMP ACUTE TOXICITY ANALYSIS
Plant
A
B
C
E
F
G
K
L
N
S
T
U
W
X
Chlorinated; pH
particulate.
Nonchlorinated;
Nonchlorinated ;
particulate.
Nonchlorinated ;
Nonchlorinated;
Chlorinated; pH
Chlorinated; pH
Nonchlorinatedi
Nonchlorinated i
particulate.
Nonchlorinated ;
particulate.
Nonchlorinated ;
particulate.
Chlorinated; pH
particulate.
Nonchlorinated;
particulate.
Chlorinated; pH
particulate.
Description
6.2) cloudy, gray with considerable amount of fine
pH 7.6; clear, light yellow.
pH 10.2) clear, blue black with moderate amount of
pH 6.8; clear with small amount of particulate.
pH 7.5; clear, light salmon.
6.0; light olive with fair amount of particulate.
7.8; clear, light gray with small amount of particulate
pH 7.3.*
pH 3.7; clear, light gray with moderate amount of
pH 7.7; clear, light champagne with small amount of
pK 7.4) clear, blue green with moderate amount of
9.0; clear, dark amber with moderate amount of
pH 8.0; cloudy, orange with moderate amount of
7.1; clear, light gold with moderate amount of
Incomplete description
remained.
because subsample was lost and no other material
107
-------
Grass shrimp, 15 mm to 26 mm rostrum-telson length, were col-
lected from Big Lagoon, near BMRL. Shrimp were held in flowing
water and acclimated to a salinity of 10 ppt for a minimum of
2 days before testing. Mortality was less than 3% during the
acclimation.
Methods for the 96-hr, static tests followed EPA test procedures
(3) as modified by the EPA Technical Advisor (ERL—Gulf Breeze).
Sheepshead minnow and grass shrimp tests were conducted in
0.004-m3 uncovered glass jars which contained 0/003 m3 of test
solution. Five fish or shrimp were tested per jar, and all test
concentrations and controls were duplicated, except as noted.
There was no aeration; temperature was maintained at 20 ± 1°C
during the tests.
Test concentrations were prepared by mixing appropriate volumes
of effluent and dilution water directly in test containers. Dilu-
tion water was glass-distilled water adjusted to 10 ppt salinity
with Rila Marine Mix (Rila Products, Teaneck,'NJ). Batches of
dilution water were aged for at least 24 hr with aeration and
then filtered (5 ym) before mixing test concentrations. Salinity
of the effluents was also adjusted to 10 ppt with Rila Marine Mix
before preparing the test concentrations. Control gars received
0.003 rn3 of 10-ppt dilution water, but no effluent.
Range-finding tests (48-hr) were conducted with all effluents to
determine appropriate test concentrations for 96-hr definitive
tests. Range-finding tests in which no mortality had occurred
after 48 hr of exposure to 100% effluent were extended to 96 hr
of exposure. If mortality remained less than or equal to 50%, no
further tests were conducted with the effluent. In range-finding
tests, only five fish or.shrimp were tested, and test concentra-
tions were not duplicated.
In cases where mortality was greater than 50% during range-
finding tests, 96-hr definitive tests were conducted with five
fish or shrimp per jar, and all test concentrations and controls
were duplicated.
Based on the test results, 24-hr, 48-hr, and 96-hr LC5o's were
graphically interpolated. Two points, representing death in
concentrations that were lethal to more than one-half and less
than one-half of the fish at the specified times, were plotted
on semilogarithmic coordinate paper (test concentrations on the
logarithmic axis and corresponding percentages of dead fish on
the arithmetic axis, Figure 22}. The concentration at which a
straight line drawn between the two points crossed the 50% mortal-
ity line was the estimated LCsg.
108
-------
s
0 10 » 30 40 50 M 70
MORTALITY, %
90 in
Figure 22. Example of graphically interpolated 24-hr, 48-hr, 96-hr,
LCso's for juvenile sheepshead minnow (Cyprinodon
variegatus) exposed to textile effluent W.
Sheepshead Minnow Results—
Five of the 14 textile effluents caused greater than 50% mortal-
ity of minnows after 96-hr of exposure to effluent concentrations
less than or equal to 100% in static, artificial seawater
(Table 36). The most toxic effluent (lowest LC50 value) was from
Plant W, with a graphically interpolated 96-hr LC$Q of 37.5%
(solution containing 37.5% effluent sample). Plant N was less
toxic, with a 96-hr LC50 of 47.5%. The 96-hr LC50's for efflu-
ents from Plants A, C, and T were 62.0%, 69.5%, and 68.0%,
respectively. Four effluents, from Plants E, G, L, and S, caused
less than 50% mortality when sheepshead minnows were exposed to
100% effluent for 96-hr. Therefore, no LC50's were determined
for these effluents and values are reported as no acute toxicity.
Five effluents, from Plants B, F, K, U, and X, caused no deaths
when minnows were exposed to 100% effluent for 96-hr, and 96-hr
LC50's are reported as no acute toxicity effluent.
TABLE 36. ACUTE TOXICITY OF 14 TEXTILE EFFLUENTS TO
JUVENILE SHEEPSHEAD MINNOWS (C. VARIEGATUS)
LC50 , percent secondary effluent
Plant
A
B
C
E
F
G
K
L
N
S
T
U
W
X
24-Hr 48-Hr
75.0 69.5
72.0
75.0 75.0
42.0 41.0
96-Hr
62.0
69.5
47.5
68.0
37.5
Note.—Blanks indicate no acute toxicity.'
109
-------
Measured concentrations of dissolved oxygen (DO) in most tests
remained greater than 40% of saturation after 96 hr of testing'.
However, DO was low in test concentrations of Plant T effluent
(particularly in 100% effluent) early in the test. Apparently,
low DO concentrations in this effluent were due more to the
nature of the effluent than to the oxygen demand of test animals.
Grass Shrimp Results—
Five of the 14 textile effluents caused more than 50% mortality
of grass shrimp after 96 hr of exposure to effluent concentra-
tions less than or equal to 100% in static, artificial seawater
(Table 37). The most toxic effluent was from Plant C, with a
graphically interpolated 96-hr LC5Q of 12.8%. Plant W and A
effluents were less toxic, with 96-hr LC50's of 19.6% and 21.2%;
and the 96-hr LC50's for effluents from Plants N and T were 26.3%
and 34.5%, respectively. Three effluents, from Plants E, G,
and S, caused less than 50% mortality when grass shrimp were
exposed to 100% effluent for 96 hr. Therefore, no LC50's were
determined, and values are reported as no acute toxicity. Six
effluents, from Plants B, F, K, L, U, and X, caused no deaths
when grass shrimp were exposed to 100% effluent for 96 hr, and
96-hr LC5o's are reported as no acute toxicity.
TABLE 37. ACUTE TOXICITY OF 14 TEXTILE EFFLUENTS
TO GRASS SHRIMP (P. pugio)
Plant
percent secondary effluent
24-Hr 48-Hr 96-Hr
A
B
C
E
F
G
K
L
N
S
T
U
W
X
37.6 24.0 21.
17.7 15.5 12.
43.5 37.4 26.
34.
37.5 24.8 19.
2
8
3
5
6
Note.—Blanks indicate no acute toxicity.
110
-------
Measured concentration^ of dissolved oxygen (DO) in most tests
remained greater than 40% of saturation after 96 hr of testing.
However, DO was low in test concentrations of Plant T effluent
(particularly in 100% effluent) early in the test. Apparently,
low DO concentrations in this effluent were due more to the
nature of the effluent than to the oxygen demand of test animals.
RANGE-FINDING ACUTE TOXICITY 14-DAY RAT TEST
The major objective of any biological testing procedure is the
identification of toxicological problems at minimal cost. There-
fore, a two-step approach was used to evalute the acute in vivo
toxicity of samples containing unknown compounds. The first ap-
proach is based on the quantal (all-or-none) response; the second
is based on the quantitative (graded) response. Normally, the
quantal test is used to determine whether or not the quantitative
assay is necessary.
The Quantal Test
Five male and five female young adult rats (weighing approxi-
mately 250 g each) were purchased from the supplier and condi-
tioned at the laboratory for a minumum of 5 days. A single 10~5-
m3/kg dose of undiluted sample was administered by gavage to each
animal. Immediately following administration of the test sub-
stance and at frequent intervals during the first day, observa-
tions were recorded on all toxic signs or pharmacological effects
as described in Table 38 (3). The frequency and severity of the
signs were scored. Particular attention was paid to time of
onset and disappearance of signs. Daily observations were made
on all animals through a 14-day observation period. Effluent
samples which produced harmful effects in vivo and did not result
in deaths were further investigated. At termination of the
observation period, all surviving animals were killed and
necropsies were performed. Similarly, necropsies were performed
on all animals that died during the course of the study.
If mortality did not occur in the quantal study, no further work
was done on the test substance, and the LDso was reported as
greater than 10 g/kg.
The Quantitative Assay
If a single animal in the quantal study died in the 14-day obser-
vation period, a quantitative study was performed. Eighty
animals equally divided by sex were maintained for 7 days in
quarantine to determine good health in the study population.
From these, 40 animals then were randomly divided into 4 groups
of 5 male and 5 female animals per group. The test substance,
treated as in the quantal test, was administered in graded dos-
ages according to the following schedule: 3.0, 1.0, 0.3, and
0.1 g/kg. Dosage was related to the numbers of animals that
died and to the severities and types of signs. Observations,
111
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TABLE 38. PHYSICAL EXAMINATIONS IN ACUTE TOXICITY TESTS IN RODENTS (3)
Organ system
Observation and examination
Common signs of toxicity
to
Central nervous system
and somatomotor
Autonomic nervous system
Respiratory
Cardiovascular
Gastrointestinal
Genitourinary
Skin and fur
Mucous membranes
Eye
Others
Behavior
Movements
Reactivity to various stimuli
Cerebral and spinal reflexes
Muscle tone
Pupil size
Secretion
Nostrils
Character and rate of breathing
Palpataion of cardiac region
Events
Abdominal shape
Feces consistency and color
Vulva, mammary glands
Penis
Perineal region
Color, turgor, integrity
Conjunctiva, mouth
Eyeball
Transparency
Rectal or pay skin temperature
Injection site
General condition
Change in attitude to observer, unusual
vocalization, restlessness, sedation.
Twitch, tremor, ataxia, catatonia, paralysis,
convulsion, forced movements.
Irritability, passivity, anaesthesia,
hyperanaesthesia.
Sluggishness, absence.
Rigidity, flaccidity.
Myosis, mydriasis.
Salivation, lacrimation.
Discharge.
Bradypnoea, dyspnoea, Cheyne-Stokes breathing,
Kussmaul breathing.
Thrill, bradycardia, arrhythmia, stronger or
weaker beat.
Diarrhea, constipation.
Flatulence, contraction.
Unformed, black or clay colored.
Swelling.
Prolapse.
Soiled.
Reddening, flaccid skinfold, eruptions,
piloerection.
Discharge, congestion, hemorrhage cyanosis,
jaundice.—
Exophthalmus, nystagmus.
Opacities.
Subnormal, increased.
Swelling.
Abnormal posture, emaciation.
-------
duration of study, and necropsies were carried out as indicated
above. The LDs0 was calculated by the method described in Refer-
ence 3.
The range-finding tests were conducted at Litton Bionetics under
the direction of Dr. R. Beliles. Dr. J. Stara served as the EPA
Technical Advisor.
Actual experimental design parameters used in this study were as
follows. Young adult rats of the Charles River CD strain (CRL:
COBS CD (SD) BR) were obtained from the Charles River Breeding
Laboratories, Inc., Portage, Michigan. Animals were individually
housed in wire bottom cages in temperature-controlled quarters
under artificial illumination controlled to provide a 12-hr light
cycle. Water and Purina Laboratory Chow were provided ad libitum
with the exception of the night before treatment when food was
removed from cages.
Effluent samples were kept refrigerated until used. A single
undiluted dose of 10~5 mvkg of test material was administered by
gastric intubation to five rats of each sex. If any rats died at
this dose, an LD50 value was to be determined by giving addi-
tional doses of the test material.
The rats were observed frequently on the day of treatment and
daily thereafter. Animals were weighed on the day of treatment,
and on days 7 and 14 following treatment. All surviving animals
were killed 14 days after treatment and necropsies were performed.
In summary, no rat deaths occurred following the oral administra-
tion of 10~5 m3 of effluent sample per kilogram of rat body
weight for any of the 23 textile plant effluent samples. Resid-
ual effects of treatment were not evident from necropsy findings.
Twelve samples (from Plants A, C, E, G, K, M, T, U, V, W, X,
and Z) showed no effects that could be related to treatment.
Reduced activity was observed immediately after dosing with some
of the effluent samples (those from Plants D, H, J, P, and Y).
Rats treated with sample from Plant R showed signs of eye irrita-
tion which were related to treatment. Effluent samples were col-
lected between the aeration lagoon and the settling pond at
Plant R. Reduced body weights or weight gains were noted after
administration of five samples (those from Plants B, F, L, N,
and S) .
A more detailed description of the reactions of the test rats to
the effluent samples is given in Appendix E.
SOIL MICROCOSM TEST
Decomposition of dead organic matter by microorganisms is essen-
tial for maintaining ecological balance. It is becoming
increasingly apparent that as more and more toxic materials from
anthropogenic sources are introduced into the soil, they will
113
-------
progressively inhibit these organisms, ultimately creating a
critical imbalance.
Soil decomposition is a complex matrix of biological, physical,
and chemical processes, dependent on a myriad of variables. Any
test designed to detect and ultimately predict the effects of
toxic materials on soils must control as many variables as prac-
ticable. In addition, disposal of toxic test substances must be
simple, convenient, and practical. To meet these needs, a soil/
litter microcosm test contained in a "life support" system was
designed, constructed, and used to determine the soil response to
toxic materials (3).
A soil microcosm is, by design, a miniature model of the natural
system—in this case, the site of decomposition in the upper
(50 mm) soil layers. In the test microcosm, soil and litter are
separately homogenized and layered in an array of airtight con-
tainers in which carbon dioxide generation is monitored with time.
Carbon dioxide generation is an accepted measure of soil respira-
tion and, in this case, is assumed to be a measure of the micro-
bial activity in the microcosm during the decomposition of
organic matter.
Carbon dioxide generation rates were measured from1replicates of
each waste solution after 2 wk to 3 wk of incubation. For data
analysis, rates were compared with three sets of waste solutions
and controls using a linear analysis of variance. Treatments
showing statistically significant differences in the stimulation
of respiration indicated inhibition or stimulation of respiration.
The soil microcosm test was performed at EPA—Corvallis Environ-
mental Research Laboratory, Corvallis, Oregon under the direction
of Dr. B. Lighthart. The soil microcosm test used 0.001 m3 of
the 0.01-m3 freshwater algae samples. The bioassay procedure and
apparatus were those described by the EPA (3). A brief descrip-
tion of the materials and methods used is given in Appendix F.
Instead of reporting all data on carbon dioxide (C02) generation
rates, only the mean qualities of C02 produced over the 3-wk
incubation period for replicate samples were reported and are
presented in Table 39. Samples were analyzed in three batches
with one set of replicate controls (using organic-free water) run
per batch. The quantity of CO2 produced each day over the test
period was plotted on graph paper and normalized with the control
sample. The slope of the resulting curve is the normaalized
relative C02 rate change (Table 39). If the wastewater sample
stimulated the microorganisms, the slope of the curve was posi-
tive; if the sample was inhibitory or toxic to the microorganisms,
the slope was negative. Of the 23 secondary effluent wastewater
samples tested, 15 inhibited the production of C02 and 8 stimu-
lated the production of C02.
114
-------
TABLE 39. RESULTS OF SOIL MICROCOSM TESTS
ON SECONDARY WASTEWATER SAMPLES
Plant
Run code
1 D
H
J
M
Pr
RC
Y
Z
Control
2 E
G
K
L
S'
V
W
Control
i 3 A
B
C
F
N
T
U
X
Control
Mean total Normalized
COa prbduced, relative COa.
m3 rate change
221.
220.
211.
229.
238.
253.
204.
222.
246.
275.
302.
291.
285.
288.
275.
305.
298.
246.
270.
285.
236.
269.
276.
277.
274.
254.
9
2
6
9
7
3
3
2
8
9
0
9
5
4
3
5
7
7
4
7
8
7
6
3
6
2
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
io-6
io-6
lO"6
ID'6
io-6
io-6
IO-6
ID'6
io-6
io-6
io-6
10"6
io-6
io-6
ID'6
10~6
10~6
10~6
10~6
10~6
10"6
IO"6
10~6
10~6
10~6
10~6
-0
-0
-0
-0
0
-0
-0
-0
-0
0
-0
-0
-0
-0
0
-0
0
-0
-0
0
0
0
0
.099
.083
.163
.059
.022
.062
.172
.112
.048
.017
.004
.020
.017
.066
.031
.032
.020
.005
.039
.059
.020
.055
.047
F-valueb
789
242
442
224
11
10
611
465
245
13
33
312
12
247
12
0
55
119
6
25
62
55
59
.1
.9
.5
.6
.4
.4
.14
.4
.15
.2
.4
.0
.5
Note.—Blanks indicate information not applicable.
aNegative sign indicates inhibition of C02 generation rate
compared to a control sample; positive sign indicates C02
stimulation.
Results are significant at a 95% confidence level for
F > 5.99 and at a 99% confidence level for F > 13.7.
Samples inadvertently collected prior to the settling
pond.
The final column in Table 39 as reported by the EPA Technical
Advisor is a measure of the statistical significance of the data.
A.standard "Student t" test was employed using the F-table and
all the individual C02 generation rate data. Based on those data,
if the value of F is greater than 5.99, then the probability of a
Type I error is 5%. If the value of F is greater than 13.7, then
the probability of a Type I error is 1%. A Type I error means
that one rejects the hypothesis when it is in fact true.
From the data in Table 39, all secondary effluent samples have a
statistically significant effect at the 5% confidence level.
Data for Plant A have a greater than 5% probability of a Type I
error. Of the 23 samples, 17 have a. statistically significant
effect at the 1% confidence'level. '
115
-------
SECTION 8
DISCUSSION OF RESULTS
PLANT RANKING BY RELATIVE WASTEWATER TOXICITY ;
v
The primary objective of the Phase I screening study was to rank
textile plants according to the toxicity of their secondary
wastewater and to select plants for detailed toxicity evaluation
in Phase II. To accomplish this objective, members.of the EPA
Bioassay Subcommittee met to evaluate the bioassay data. Members
of the Subcommittee are illustrated as EPA Technical Advisors in
Figure 20 (Section 7). A summary of all the bioassay results is
given in Table 5 (Section 3).
Data evaluation began with ranking of the plants in each set of
bioassays. Results are discussed in the following sections.
Freshwater Ecology Series
Results from these tests showed sufficient variation to permit
relative ranking of the toxicity of effluent samples. A com-
posite ranking based on the responses of fathead minnows and
Daphnia is shown in Table 40. No general rule can be made
concerning the relative response between fathead minnows and
Daphnia. For example, Plant E's effluent was significantly
toxic to Daphnia but not toxic at all to fathead minnows; at
Plant T, the reverse was true.
i1
TABLE 40. RELATIVE TOXICITY RANKING BY BIOASSAY TEST
Relative
toxicity
C_v_It_
fraahwatar
acology (fathaad
Cytotoxicity •innov and
(RAM) _ Daphnia}
coapo»it« .
Marine ecology
(ah««pahaad minnows, Clonal
graaa •hrinp, and toxicity
m«rin« alqfc«) _ (CHO-Kl)
NOIt tOXlc
C,»
l,»,T
Inunxdlat*
Toxicity
C,V
A,F,L,T,M,» E
U
T
A,C,t,«
N,B,J
L*ut toxic M P,H,M
Nontoxic B,E,G,«,U,D, B,X,8,X,D,J,P,T
B,J,P,R,V,
Not an«lyio
-------
At the time of the evaluation, freshwater algae results were not
available, but as seen in Table 32 (Section 7), ranking of plants
by algal inhibition is similar to that for fathead minnow and
Daphnia. Because the focus of the program is toxicity removal
by treatment technologies and not ecological impacts, algal
stimulation effects were not considered in the ranking.
Marine Ecology Series
Based on toxicity data for sheepshead minnows, grass shrimp, and
marine algae, ranking of effluents by toxicity was accomplished
and is shown in Table 40. In all samples, grass shrimp were
more sensitive than sheepshead minnows. Also, the fathead min-
nows were more sensitive in the majority of the samples than
sheepshead minnows. No general correlation was seen between the
response of Daphnia and grass shrimp.
Cytotoxicity
Rabbit alveolar macrophage tests indicated that none of the
samples was highly toxic. Two samples, N and C, were moderately
toxic and the following seven samples were slightly toxic: L,
W, T, X, A, F, and J.
Only eight samples were tested by MRC using the clonal toxicity
test: D, H, J, M, P, R, Y, and Z. Of the eight samples, four
showed significant toxicity: D, M, H, and J.
Mutagenicity
None of the 23 effluent samples produced a positive response in
any of the bacterial tester strains. The Bioassay Subcommittee
expressed concern that the detection limits for this bioassay
series were not sensitive enough to detect the presence of sig-
nificant concentrations (0.001 to 0.1 g/m3) of chemical
mutagens.
Rat Acute Toxicity Tests
No acute toxicity was observed from the maximum dose (10~5 m3/kg)
of rat body weight) ingested by the rats. However, six effluent
samples showed potential body weight effects: F, N, C, L, S,
and B. The subcommittee expressed concern about the detection
limits of this test also.
Plant Ranking
Based on all of the above analyses, the subcommittee ranked the
23 textile plants in descending order of secondary effluent
toxicity, and results are shown in Table 41.
117
-------
TABLE 41. PRIORITIZATION OF TEXTILE PLANTS BY
TOXICITY OF SECONDARY EFFLUENT
Toxicity ranking Plant
Most toxic N,A
W
C,T
V,L
Least toxic S,P
Nontoxic B,D,E,F,G,H,J,K
M,U,X,Y,Z
From the above list, the subcommittee recommended that the fol-
lowing nine textile plants be tested to determine the removal of
toxicity achieved by the tertiary treatment technologies being
tested in the ATMI/EPA Grant Study: N, A, L, T, C, P, S, W, and
V. (Plant R was also recommended for study under Phase II
because its secondary effluent samples were inadvertently col-
lected prior to the settling pond.) In addition, they recom-
mended that the freshwater ecology series be used to measure
reduction in wastewater toxicity by the treatment technologies.
The marine ecology series was not selected because none of the
textile plants discharge wastewater into a marine environment.
PROGRAM OUTLINE FOR PHASE II STUDY
The objective of the second part of the textile wastewater
toxicity study is to determine reduction in priority pollutant
concentrations and in acute toxicity as a result of applying
the ATMI/EPA BATEA tertiary treatment technologies to the
secondary effluent at the 10 textile plants.
Pilot plants are scheduled to be at each (10) textile plant for
from 6 wk to 8 wk. For the first 4 wk, seven tertiary treatment
systems will be tested to determine which one provides the best
removal of criteria pollutants. A treatment system consists of
one or more of the six tertiary treatment technologies. From
the data collected, the "best" system will be identified. This
system will then be set up and operated at steady-state condi-
tions for a final period of 2 wk.
For toxicity and priority pollutant analyses at each plant,
24-hr composited samples will be collected during the 2 wk of
steady-state operations from the one system identified as the
"best available technology." Since the tertiary treatment
system will be composed of several of the six treatment tech-
nologies, samples will be collected before and after each unit
operation in the system, resulting in approximately four samples.
118
-------
In order to evaluate the reduction in toxicity and priority
pollutant concentrations, 24-hr composited secondary effluent
samples will also be collected. Due to hydraulic retention time
through the pilot plant, secondary effluent sampling will lead
the tertiary treatment sampling by the appropriate time for the
tertiary treatment system selected.
A 24-hr composited sample of the intake water to the textile
plant will be collected at each of the 10 plants to understand
the presence of priority pollutants in wastewater samples.
Either continuous or grab samples will be collected depending
upon the sampling conditions around the intake water facilities.
Samples will be collected for volatile organics, nonvolatile
organics, and metals analyses. Therefore, a total of approxi-
mately 6 samples will be collected at each of the 10 plants as
illustrated in Table 42.
TABLE 42. SAMPLE SCHEDULE AT EACH OF THE 10 TEXTILE PLANTS
Sample site
No. of
samples
Analyze for
Plant intake water
Secondary effluent
Best tertiary
treatment system
1
a
129 priority pollutants
129 priority pollutants
and freshwater ecology
series
129 priority pollutants
and freshwater ecology
series
The freshwater ecology series consists of bioassay tests on the
following three test organisms: fathead minnows, Daphnia, and
freshwater algae. Five sample fractions are collected for
priority pollutant analysis: volatile organics, nonvolatile
organics, metals, cyanide, and phenol. Criteria pollutant
analyses will not be performed since these analyses will be
routinely performed under the ATMI/EPA Grant Study.
119
-------
REFERENCES
1. Draft Final Report: Sampling and Analysis Procedures for
Screening of Industrial Effluents for Priority Pollutants.
U.S. Environmental Protection Agency, Cincinnatij Ohio,
March 1977. 145 pp.
i
2. Hamersma, J. W., S. L. Reynolds, and R. F. Maddalone. IERL-
RTP Procedures Manual: Level 1 Environmental Assessment.
EPA-600/2-76-160a (PB 257 850), U.S. Environmental Protection
Agency, Research Triangle Park, North Carolina, June 1976.
147 pp. I
3. Duke, K. M., M. E. Dav'is, and A. J. Dennis. IERL-RTP Pro-
cedures Manual: Level 1 Environmental Asssessment Biological
Tests for Pilot Studies. EPA-600/7-77-043 (PB 268 484), U.S.
Environmental Protection Agency, Research Triangle Park,
North Carolina, April 1977. 114 pp.
4. Manual of Methods for Chemical Analysis of Water and Wastes.
EPA-625/6-76-003a (PB 259 973), U.S. Environmental Protection
Agency, Cincinnati, Ohio, 1976. 317 pp.
5. Standard Methods for the Examination of Water and Waste-
water, Fourteenth Edition. American Public Health Associa-
tion, Washington, D.C., 1976. 874 pp.
6. McCann, J., E. Choi, E. Yamasaki, and B. N. Ames. Detection
of Carcinogens as Mutagens in the Salmonella/Microsome Test:
Assay of 300 Chemicals. Proceedings of the National Academy
of Science, 72:5135-5139, 1975.
7. Poole, D. C. and V. F. Simmon. Final Report of in Vitro
Microbiological Studies of Twenty-two Wastewater Effluent
Samples. Contract 68-01-2458, U.S. Environmental Protection
Agency, Biomedical Research Branch, Research Triangle Park,
North Carolina, November 1977. Ill pp.
8. Waters, M. D., D. E. Gardner, C. Aranyi, and D. L. Coffin.
Metal Toxicity of Rabbit Alveolar Macrophages i'n Vitro.
Environmental Research, 9(l):32-47, 1975.
120
-------
9. Campbell, J. A., H. F. Stack, and P. R. Williams. Cyto-
toxicity Screening of Twenty-three Textile Mill Effleunt
Water Samples Utilizing the Rabbit Alveolar Macrophage
Assay. Contract 68-02-2566, U.S. Environmental Protection
Agency, Biomedical Research Branch, Research Triangle Park,
North Carolina, December 1977. 86 pp.
10. Malcolm, A. R., B. H. Pringle, and H. W. Fisher. Chemical
Toxicity Studies with Cultured Mammalian Cells. In: Bio-
assay Techniques and Environmental Chemistry, G. E. Glass,
ed. Ann Arbor Science Publishers, Inc., Ann Arbor,
Michigan, 1974. pp. 217-230.
11. Shiroyama, T., W. E. Miller, and J. C. Greene. The Efforts
of Nitrogen and Phosphorus on the Growth of Selenastrum
Capriocornutum Printz. EPA-606/3-75-034, U.S. Environmental
Protection Agency, Corvallis, Oregon, March 1975.
pp. 132-142.
12. Sprague, J. B. The ABC's of Pollutant Bioassay Using Fish.
In: Biological Methods for the Assessment of Water Quality,
J. Cairns, Jr. and K. L. Dickson, eds. ASTM Special Tech-
nical Publication 528, American Society for Testing and
Materials, Philadelphia, Pennsylvania, 1973. pp. 6-30.
13. Standard for Metric Practice. ANSI/ASTM Designation:
E 380-76e, IEEE Std 268-1976, American Society for Testing
and Materials, Philadelphia, Pennsylvania, February 1976.
37 pp.
121
-------
APPENDIX A
RECOMMENDED LIST OF PRIORITY POLLUTANTS
TABLE A-l. RECOMMENDED LIST OF PRIORITY POLLUTANTS
Compound name
Acenaphthene
Acrolein
Acrylonitrile
Benzene
Benzidine
Carbon tetrachloride (tetrachloromethane)
Chlorinated benzenes (other than dichlorobenzenes)
Chlorobenzene
1,2,4-Trichlorobenzene
Hexachlorobenzene
Chlorinated ethanes (including 1,2-dichloroethane,
1,1,1-trichloroethane and hexachloroethane)
1,2-Dichloroethane
1,1,1-Trichloroethane
Hexachloroethane
1,1-Dichloroethane
1,1,2-Trichloroethane
^ 1,1,2,2-Tetrachloroethane
Chloroethane
Chloroalkyl ethers (chloromethyl, chloroethyl and
mixed ethers)
Bis(chloromethyl) ether
Bis(2-chloroethyl) ether
2-Chloroethyl vinyl ether (mixed)
Chlorinated naphthalene
2-Chloronaphthalene
(continued)
122
-------
TABLE A-l (continued).
Compound name
Chlorinated phenols (other than those listed elsewhere;
includes trichlorophenols and chlorinated cresols)
2,4,6-Trichlorophenol
p-Chloro-m-cresol (4-chloro-3-methylphenol)
Chloroform (trichloromethane)
2-Chlorophenol
Dichlorobenzenes
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Dichlorobenzidine
3,3'-Dichlorobenzidine
Dichloroethylenes (1,1-dichloroethylene and
1,2-dichloroethylene)
1,1-Dichloroethylene (vinylidine chloride)
1,2-Trans-dichloroethylene
2,4-Dichlorophenol
Dichloropropane and dichloropropene
1,2-Dichloropropane
1,3-Dichloropropylene
(ois and trans-l,3-dichloropropene)
2,4-Dimethylphenol
Dinitrotoluene
2,4-Dinitrotoluene
2,6-Dinitrotoluene
1,2-Diphenylhydrazine
Ethylbenzene
Fluoranthene
(continued)
123
-------
TABLE A-l (continued).
Compound name
Haloethers (other than those listed elsewhere)
4-Chlorophenyl phenyl ether
4-Bromophenyl phenyl ether
Bis (2-chloroisopropyl) ether
Bis (2-chloroethoxy) methane
i
Halomethanes (other than those listed elsewhere)
Methylene chloride (dichlorometharie)
Methyl chloride (chloromethane) i
Methyl bromide (bromomethane)
Bromoform (tribromomethane)
Dichlorobromomethane
Trichlorofluoromethane
Dichlorodifluoromethane
Chlorodibromomethane
Hexachlorobutadiene
Hexachlorocyclbpentadiene
": i
Isophorone (3,5,5-trimethyl-2-cyclohexen-l-one)
Naphthalene
Nitrobenzene
Nitrophenols (including 2,4-dinitrophenol
and dinitrocresol)
2-Nitrophenol
4-Nitrophenol
2,4-Dinitrophenol
4,6-Dinitro-o-cresol
Nitrosoamines
i
N-nitrosodimethylamine
N-nitrosodiphenylamine
N-nitroso-di-n-propylamine
Penta chlorophenol
Phenol
(continued)
124
-------
TABLE A-l (continued).
Compound name
Phthalate esters
Bis(2-ethylhexyl) phthalate
Butyl benzyl phthalate
Di-n-butyl phthalate
Diethyl phthalate
Dimethyl phthalate
Di-n-octyl phthalate
Polynuclear aromatic hydrocarbons
Benz(a)anthracene (1,2-benzanthracene)
Benzo(a)pyrene (3,4-benzopyrene)
3,4-Benzofluoranthene
Benzo(k)fluoranthene
(11,12-benzofluoranthene)
Chrysene
Acenaphthylene
Anthracene
Benzo(ghi)perylene (1,12-benzoperylene)
Fluorene
Phenanthrene
Dibenz(ah)anthracene
(1,2,5,6-dibenzanthracene)
Indeno(1,2,3-cd)pyrene
(2,3-o-phenylenepyrene)
Pyrene
Tetrachloroethylene
Toluene
Trichloroethylene
Vinyl chloride (chloroethylene)
Pesticides and metabolites
Aldrin
Dieldrin
Chlorodane (technical mixture and metabolites)
DDT and metabolites
4,4'-DDT
4,4'-DDE (p,p'-DDX)
4,4'-ODD (p,p'-TDE)
(continued)
125
-------
TABLE A-l (continued).
Compound name
Endosulfan and metabolites
a-Endosulfan
g-Endosulfan
Endosulfan sulfate
Endrin and metabolites
Endrin
Endrin aldehyde
Heptachlor and metabolites
Heptachlor
Heptachlor epoxide
Hexachlorocyclohexane
ct-BHC
e-BHC
X-BHC (lindane)
6-BHC
Polychlorinated biphenyls (PCB)
PCB-1242 (Arochlor 1242)
PCB-1254 (Arochlor 1254)
PCB-1221 (Arochlor 1221)
PCB-1232 (Arochlor 1232)
PCB-1248 (Arochlor 1248)
PCB-1260 (Arochlor 1260)
PCB-1016 (Arochlor 1016)
Toxaphene
Elements
Antimony (Total)
Arsenic (Total)
Asbestos (Fibrous)
Beryllium (Total)
Cadmium (Total)
Chromium (Total)
Copper (Total)
Cyanide (Total)
Lead (Total)
(continued)
126
-------
TABLE A-l (continued).
Compound name
Elements (continued)
Mercury (Total)
Nickel (Total)
Selenium (Total)
Silver (Total)
Thallium (Total)
Zinc (Total)
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)
127
-------
APPENDIX B
PRIORITY POLLUTANT ANALYSIS FRACTIONS
TABLE B-l. VOLATILE COMPOUNDS
1
Compound
Compound i
Chloromethane
Dichlorodifluoromethane
Bromomethane
Vinyl chloride
Chloroethane
Methylene chloride
Trichlorofluoromethane
1,1,-Dichloroethylene
1,1-Dichloroethane
trans-1,2,-dichloroethane
Chloroform
1,2-Dichloroethane
1,1,1-Trichloroethane
Carbon tetrachloride
Bromodichloromethane
Bis(chloromethyl) ether
1,2-Dichloropropane
trans-1,3-dichloropropene
Trichloroethylene
Dibromochloromethane
Cis-1,3-dichloropropene
1,1,2-Trichloroethane
Benzene
2-Chloroethyl vinyl ether
Bromoform
1,1,2,2-Tetrachloroethylene
1,1,2,2-Tetrachloroethane
Toluene
Chlorobenzene
Ethylbenzene
Acrolein
Acrylonitrile
128
-------
TABLE B-2. BASE NEUTRAL EXTRACTABLE COMPOUNDS
Compound
Compound
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachloroethane
1,2-Dichlorobenzene
Bis (2-chloroisopropyl) ether
Hexachlorobutadiene
1,2,4-Trichlorobenzene
Naphthalene
Bis(2-chloroethyl) ether
Hexachlorocyclopentadiene
Nitrobenzene
Bis (2-chloroethoxy) methane
2-Chloronaphthalene
Acenaphthylene
Acenaphthene
Isophorone
Fluorene
2,6-Dinitrotoluene
1,2-Diphenylhydrazine
2,4-Dinitrotoluene
N-nitrosodiphenylamine
Hexachlorobenzene
4-Bromophenyl phenyl ether
Phenanthrene
Anthracene
Diethyl phthalate
Dimethyl phthalate
Fluoranthene
Pyrene
Di-n-butyl phthalate
Benzidine
Butyl benzyl phthalate
Chrysene
Bis(2-ethylhexyl) phthalate
Benz(a)anthracene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Indeno(1,2,3-cd)pyrene
Dibenz(a,h)anthracene
Benzo(g,h,i)perylene
N-nitrosodimethylamine
N-nitroso-di-n-propylamine
4-Chlorophenyl phenyl ether
3,3 '.-Dichlorobenzidine
2,3,7,8-Tetrachlorodibenzo-
p-dioxina
Bis-(chloromethyl) ether
This compound was specifically listed in the consent decree.
Because of TCDD's extreme toxicity, EPA recommends that labora-
tories not acquire analytical standards for this compound.
TABLE B-3. ACID EXTRACTABLE COMPOUNDS
2-Chlorophenol
Phenol
2,4-Dichlorophenol
2-Nitrophenol
p-Chloro-m-cresol
2,4,6-Trichlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
4,6-Dinitro-o-cresol
4-Nitrophenol
Pentachlorophenol
129
-------
TABLE B-4. PESTICIDES AND PCB
Compound
B-Endosulfan
a-BHC
Y-BHC
g-BHC
Aldrin
Heptachlor
Heptachlor epoxide
a-Endosulfan
Dieldrin
4,4'-DDE
4,4'-ODD
4,4'-DDT
Endrin
Endosulfan sulfate
6-BHC
Chlordane
Toxaphene
PCB-1242 (Aroclor 1242)
PCB-1254 (Aroclor 1254)
PCB-1221 (Aroclor 1221)
PCB-1232 (Aroclor 1232)
PCB-1248 (Aroclor 1248)
PCB-1260 (Aroclor 1260)
PCB-1016 (Aroclor 1016)
TABLE B-5. METALS AND OTHER COMPOUNDS
Metals,
total Others
Antimony Asbestos
Arsenic Cyanide
Beryllium
Cadmium
Chromium
Copper
Lead
Mercury
Nickel
Selenium
Silver
Thallium
Zinc
130
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APPENDIX C
ADDRESSES OF PERSONS ASSOCIATED WITH THIS TEXTILE STUDY
1. Mr. James Campbell
Northrop Services
U.S. Environmental Protection Agency
Research Triangle Park, NC 27711
919-541-2997
2. Dr. Dale A. Denny, Chief
Chemical Processes Branch, MD-62
Industrial Environmental Research Lab-RTP
U.S. Environmental Protection Agency
Research Triangle Park, NC 27711
919-541-2547
3. Dr. James A. Dorsey, Chief
Industrial Processes Branch, MD-62
Industrial Environmental Research Lab-RTP
U.S. Environmental Protection Agency
Research Triangle Park, NC 27711
919-541-2557
4. Dr. James D. Gallup
Effluent Guidelines Division
Environmental Protection Agency
919B WH-552, Waterside Mall
401 M Street, S.W.
Washington, DC 20460
202-426-2554
5. Dr. W. M. Haynes
Monsanto Research Corporation
Station B, Box 8
Dayton, OH 45407
513-268-3411, x300
6. Dr. Roger D. Holm
Monsanto Research Corporation
Station B, Box 8
Dayton, OH 45407
513-268-3411, x354
131
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7. Mr. Bill Horning (Technical Advisor)
EPA Fish Toxicology Laboratory
Environmental Protection Agency
3411 Church Street .
Cincinnati, OH 45244
513-684-8601
8. Dr. Joellen Huisingh (Technical Advisor)
Health Effects Research Lab, MD-82
Environmental Protection Agency
Research Triangle Park, NC 27711
919-541-2948
9. Dr. Larry D. Johnson
Industrial Environmental Research Lab, MD-62
Environmental Protection Agency
Research Triangle Park, NC 27711
919-541-2557
10. Dr. Bruce Lighthart (Technical Advisor)
Environmental Research Laboratory
Environmental Protection Agency
200 S.W. 35th Street
Corvallis, OR 97330 *
503-757-4832
11. Mr. W. E. Miller
Corvallis Environmental Research Lab |
Environmental Protection Agency I
200 S.W. 35th Street
Corvallis, OR 97330
503-757-4775
12. Mr. O'Jay Niles
BATEA Grant Project Manager
American Textile Manufacturers Institute
Wachovia Center, Suite 2124
400 South Tryon Street
Charlotte, NC 28285
704-334-4734
13. Mr. Rod Parrish
EG&G, Bionomics Marine Research Laboratory
Route 6, Box 1002
Pensacola, FL 32507
904-453-4359
14. Dr. Gary D. Rawlings (Project Manager)
Monsanto Research Corporation
Station B, Box 8
Dayton, OH 45407
513-268-3411, x396
132
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15. Mr. W. D. Ross
Monsanto Research Corporation
Station B, Box 8
Dayton, OH 45407
513-268-3411, x362
16. Dr. Max Samfield (Project Officer)
Industrial Environmental Research Lab, MD-62
Environmental Protection Agency
Research Triangle Park, NC 27711
919-541-2547
17. Dr. A. D. Snyder
Monsanto Research Corporation
Station B, Box 8
Dayton, OH 45407
513-268-3411, x216
18. Dr. Jerry Stara
Health Effects Research Lab-Cine.
Environmental Research Center, Room 640
U.S. Environmental Protection Agency
26 West St. Clair
Cincinnati, OH 45268
513-684-7407
19. Dr. Gerald Walsh
Environmental Research Laboratory
Environmental Protection Agency
Sabine Island
Gulf Breeze, FL 32561
904-932-5311
,»
20. Dr. Michael D. Waters
Health Effects Research Lab, MD-82
Environmental Protection Agency
Research Triangle Park, NC 27711
919-541-2537
21. Dr. Robert J. Weir
Litton Bionetics
5516 Nicholson Road
Kensington, MD 20795
301-881-5600
133
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APPENDIX D
REACTION OF FATHEAD MINNOWS AND
DAPHNIA TO TEXTILE SECONDARY EFFLUENTS
Plant A
This sample was acutely toxic to both the fathead minnow and D.
pulex, resulting in estimated 96-hr LCso and 48-hr ECs0 values
of 19.0% and 9.0% effluent .dilution, respectively. The 2-day
delay from collection of the sample to the beginning of the test
probably did not affect sample toxicity.
Control fish ranged in length 30 mm to 39 mm, with the average
being 34.1 mm. Their weight ranged from 0.2 g to 0.6 g, with
the average being 0.37 g.
Plant B
This sample was not acutely toxic to either the fathead minnow
or D. pulex.
Control fish ranged in length from 25 mm to 51 mm, with the
average being 36.5 mm. Their weight ranged from 0.2 g to 1.25 g,
with the average being 0.56 g.
Plant C
This sample was acutely toxic to both fathead minnow and D.
pulex, resulting in estimated 96-hr LCso and 48-hr EC so values
of 46.5% and 41.0% effluent dilution, respectively. All fish
died in 100% effluent, and half of the fish died in 60% effluent
dilution within the first 1/2 hr. The remaining fish in the 60%
dilution were dead by the next morning.
Control fish ranged in length from 28 mm to 38 mm, with the
average being 33.5 mm. Their weight ranged from 0.2 g to 0.5 g,
with the average being 0.30 g.
Plant D
At the end of the test, fish from the control were measured and
weighed. Fish ranged in length from 32 mm to 48 mm, with an
average 38.6 mm. Fish ranged in weight from 0.15 g to 0.73 g,
with an average weight of 0.34 g. At the end of the test, two
134
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fish had died in the 60% dilution and one in the 4.7% dilution.
In the D. pulex test, no animals died during the 48-hr period.
Results indicate that this sample was not acutely toxic to fat-
head minnows or D. pulex.
Plant E
At the end of the test, fathead minnows from the control jar
were measured and weighed. Fish ranged in length from 21 mm to
37 mm, with an average length of 31 mm. Their weight ranged
from 0.1 g to 0.4 g, with the average being 0.29 g.
Data indicate that this textile mill sample was not acutely
toxic to fathead minnows over a 96-hr period. However, the
sample was acutely toxic to the D. pulex, with an estimated
48-hr EC50 of 7.8 % waste.
Plant F
This sample evidenced no acute 96-hr toxicity to fathead minnows.
However, the estimated 48-hr ECs0 value of 81.7% indicated the
sample was somewhat toxic to D. pulex.
Fish ranged in length from 28 mm to 38 mm, with the average
being 32.1 mm. Their weight ranged from 0.2 g to 0.5 g, with
the average being 0.28 g.
Plant G
This sample indicated acute toxicity to both fathead minnow and
D. pulex with estimated 96-hr LCso and 48-hr EC so values of
64.7% and 62.4% effluent dilution, respectively. In this in-
stance, toxicity was essentially the same for both species.
Fish from the control jar ranged in length from 26 mm to 35 mm,
with an average length of 29.6 mm. Their weight ranged from
0.15 g to 0.40 g, with the average being 0.26 g.
Plant H
The fathead minnow bioassay is inconclusive for this sample. It
appears that the fish were diseased. There was considerable
mortality in all of the jars including the control. This was
not a reliable test run.
The bioassay run using the D. pulex, however, was meaningful.
Mortality in the 100% effluent was 40%. In the other concentra-
tions, 10 D. pulex were alive at the end of the test. The 100%
concentration of effluent showed some indication of toxicity,
although an EC50 value could not be determined.
135
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Plant J
Fish from the control jar were weighed and measured at the end
of the test. Fish ranged in length from 32 mm to 44 mm, with an
average of 37 mm. They ranged in weight from 0.3 g to 0.75 g,
with an average weight of 0.46 g. At the end of the test, one
fish had died in the 100%, one in the 60%, two in the 7.8%, and
three in the 13% dilutions. Three of the D. pulex died in 100%
effluent; one died in the 60% dilution; and one died in the con-
trol jar. i
Based on the results from these test, this sample was not acute-
ly toxic to the fathead minnows or the D. pulex.
Plant K
This sample indicated no acute toxicity to fathead minnows and
little toxicity to D. pulex. Four D. pulex died in 100% effluent
during the 48-hr period of the test.
Control fish ranged in length from 28 mm to 35 mm, .with an aver-
age length of 30 mm. Their weight ranged from 0.15 g to 0.4 g,
with the average being 0.26 g.
Plant L ,
This sample was quite acutely toxic to both fathead minnows and
D. pulex, resulting in estimated 96-hr LCso and 48-hr ECso
values of 23.5% and 28.0% effluent dilution, respectively. The
two statistical procedures used could not be utilized to calcu-
late the toxicity values with 95% confidence limits for the D.
pulex.
Control fish ranged in length from 26 mm to 40 mm, with the
average being 30.6 mm. Their weight ranged from 0.2 g to 0.6 g,
with the average being 0.33 g.
Plant M
At the end of the test, fathead minnows in the control jar weigh-
ed an average 0.29 g, with a range of 0.15 g to 0.50 g. Average
length of the fish was 34 mm, with a range of 28 mm to 48 mm.
Data indicate that this sample was not acutely toxic to the fat-
head minnow, but it was acutely toxic to D. pulex. A 48-hr
ECso value of 60.0% effluent, with 95% confidence limits of
40.72% to 88.95%, was determined with the moving average angle
procedure.
136
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Plant N
This sample was acutely toxic to both fathead minnow and D.
pulex. The estimated 96-hr LCso value for the fish was 48.8%
effluent dilution. All fish in 100% effluent were dead within
19 hr after the beginning of the test. The fish showed evidence
of hemorrhaging aroung the mouth and tail. The pH of the 100%
effluent was 4.0 at the beginning of the test and only 4.5 24 hr
later. Thus, low pH was responsible for this mortality. The
sample was extremely toxic to D. pulex, with all animals being
killed within 24 hr in as low as 13% effluent dilution, and all
were dead in every dilution at the end of 48 hr. The 2-day de-
lay from the time of collection to the beginning of the test
probably did not affect the toxicity. The temperature of the
sample was 2°C when it reached the Newtown Fish Toxicology
Station. Biological degradation would be minimal under these
conditions.
Control fish ranged in length from 28 mm to 45 mm, with the
average being 36.2 mm. Their weight ranged from 0.2Jg to 1.0 g,
with the average being 0.55 g.
Plant P
At the end of the test (96 hr), there were nine fish surviving
in each test jar. None of the fish in the control jar died.
The sample did not indicate 96-hr acute toxicity.
At the end of the test there wer 10 Daphnia alive in each con-
tainer.
Results indicate that this sample was not acutely toxic to
either fathead minnows or D. pulex.
Plant R
Based on appearance, this sample, was both high in suspended
solids and highly turbid (149 units). Turbidity of previous
samples ranged from a low of 6 units to a high of 54 units.
Considerable sample agitation achieved only 2.8 g/m3 dissolved
oxygen in the 100% effluent at the beginning of the test.
At the end 6f the test, fish from the control jar averaged 0.28
g in weight, with a range of 0.15 g to 0.65 g. Average length
was 35.4 mm, with a range of 28 mm to 47 mm.
Mortality in both the fathead minnow bioassay test and the D.
pulex test usually was observed within the first 24 hr. BOD
appears to have been a contributing factor. Dissolved oxygen
dropped to low levels in the four high effluent volumes. How-
ever, after 24 hr, dissolved oxygen was still at a level (0.3
gYm3 to 1.8 g/m3) wherein fathead minnows can survive for a
137
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considerable period of time. In 18 hr there was a complete kill
of the fish in 100% and 60% effluent. In 36% and 21.6% effluent
there was 80% and 30% mortality, respectively, in 24 hr.
Probit analysis of the data at the end of the fathead bioassay
(96 hr) indicates an LC50 value of 16.45% effluent, with 95%
confidence limits of 12.39% to 21.72%. A 48-hr LCso value for
D. pulex could not be determined with probit analysis. The
moving average angle procedure indicated the 48-hr ECso value to
be 7.96% effluent, with 95% confidence limits of 6.14% to 7.96%.
Thus, the data show this particular textile mill effluent sample
to be acutely toxic to both fathead minnows and D. pulex. The
D. pulex were more sensitive than the fathead minnows.
Plant S
This sample exhibited no 96-hr acute toxicity to fathead minnows
and little toxicity to D. pulex. There were three, two, and one
D. pulex deaths in 100%, 60%, and 36% effluent dilution, respec-
tively.
Unlike all other samples, this sample contained a heavy, floccu-
lent, fibrous material that settled to the bottom of the test
containers (to a depth approximately 5.1 cm from the 100%
effluent container). Fish hid in this material throughout the
test and were not adversely affected.
Control fish ranged in length from 27 mm to 34 mm, with the
average being 29.6 mm. Their weight ranged from 0.20 g to 0.35
g, with the average being 0.25 g.
Plant T
This sample was acutely toxic to fathead minnow, with an esti-
mated 96-hr LCso value of 46.5% effluent dilution. All of the
fish were dead in 100% effluent within 18 hr after the test was
started. Slight evidence of toxicity to D. pulex was indicated
with three animals dying in 100% effluent and one in the 60%
dilution. The 2-day delay from the time of collection to the
beginning of the test probably did not affect the toxicity of
the sample.
Control fish ranged in length from 26 mm to 38 mm, with the
average being 31.3 mm. Their weight ranged from 0.2 g to 0.5 g,
with the average being 0.33 g.
Plant U
This sample was not acutely toxic in 96 hr to fathead minnow.
However, it was toxic to D. pulex, having an estimated 48-hr
EC50 value of 12.1% effluent dilution. The 2-day delay from
138
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sample collection to the beginning of the test probably did not
affect its toxicity.
The length of the control fish ranged from 29 mm to 46 mm, with
the average being 37.7 mm. Their weight ranged from 0.2 g to
1.0 g, with the average being 0.51 g.
Plant V
This sample indicated acute toxicity to both fathead minnows and
D. pulex. All fish died in 100% effluent within 1 hr; all died
in the 60% dilution within 2.5 hr. The estimated 96-hr LCs0 was
36% effluent dilution. D. pulex were much more sensitive with
an estimated EC50 value of 9.4% effluent.
Fathead minnows in the control jar at the end of the test ranged
in length from 27 mm to 38 mm, with an average length of 29 mm.
Their weight ranged from 0.1 g to 0.45 g, with an average weight
of 0.23 g.
Plant W
This sample was acutely toxic to fathead minnow and D. pulex,
resulting in estimated 96-hr LCso and 48-hr ECso values of 55.2%
and 6.3% effluent dilution, respectively. The sample was much
more toxic to D. pulex than to fathead minnow.
Control fish ranged in length from 28 mm to 41 mm, with the
average being 34 mm. Their weight ranged from 0.2 g to 0.7 g
with the average being 0.37 g.
Plant X
This sample was not acutely toxic to either fathead minnows or
D. pulex. In contrast with a 2-day delay for most samples, it
should be noted that there was a delay of 2 day from the time of
sample collection until the test was begun. It is possible that
the toxic components may have decomposed during the extra day of
storage, although this seems unlikely when compared to other
samples.
Control fish ranged in length from 28 mm to 42 mm, with the
average being 35.3 mm. Their weight ranged from 0.2 g to
0.85 g, with the average being 0.48 g.
Plant Y
The fathead minnow bioassay was not significant with this sample
because a disease caused the loss of 30% of the control fish.
Despite the disease problem, there was 70% to 90% survival of
the fish throughout the range of effluent dilutions. In the
139
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D. pulex test, only one animal died in 100% effluent and one in
the 21.6% dilution.
Based on the results from these tests, this sample was not
acutely toxic to fathead minnows or D. pulex.
Plant Z
At the end of the test, fathead minnows in the control jar
averaged 0.5 g in weight with a range of 0.2 g to 1.1 g. Aver-
age length of the fish was 35.8 mm, with a range of 28 mm to
50 mm.
Data indicate that this particular sample was not acutely toxic
to fathead minnow. However, the sample was acutely toxic to
D. pulex. A 48-hr ECso value of 42.57% with 95% confidence
limits of 30.79% to 64.07% was determined with probit analysis.
140
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APPENDIX E
REACTIONS OF RATS TO TEXTILE
PLANT SECONDARY EFFLUENT
Plant A
Although reduced activity was observed immediately after treat-
ment, there were no deaths. Necropsy showed mottled kidneys;
however, this has been observed frequently in untreated rats in
the Litton Bionetics laboratory and was not considered treatment
related. Mean body weights (grams) are tabulated below.
Day
Dose Sex 0 7 14
10~5 m3Ag M 240 312 340
F 240 250 266
Data did not suggest any treatment effect.
Plant B
There were no deaths following treatment. One male had soft
stools during several days of the observation period. Necropsy
findings were limited to kidney changes described above. Mean
body weights (grams) are tabulated below.
Day
Dose Sex 0 7 14
10~5 m3/kg M 249 248 300
F 202 233 241
Body weight loss of males between 0 and 7 was not normal.
Plant C
There were no deaths and no signs of toxicity following treat-
ment. Necropsy findings were limited to kidney changes pre-
viously described. Mean body weights (grams) are tabulated
below.
141
-------
Day
Dose Sex 0 7 14
10~5 m3Ag M 297 328 365
F 214 221 223
Data did not suggest any treatment-related effect.
Plant D
There were no deaths following treatment. Reduced activity
immediately after treatment among all male rats and a soft
stool in one male rat 1 day after treatment were observed.
Necropsy showed mottled kidneys; however, this was not con-
sidered treatment related. Mean body weights (grams) are
tabulated below.
Day
Dose Sex 0 7 14
10~5m3Ag M • 164 235 274
F 171 208 216
Body weight data did not suggest any adverse effect.
Plant E
There were no deaths following treatment. Signs of eye irrita-
tion appeared in 6 of 10 rats near the end of the 14-day observ-
ation period, but were not considered treatment related.
Necropsy findings, limited to changes in heart surface and
kidneys, were observed only among male rats; they also were
not considered treatment related. Mean body weights (grams)
are tabulated below.
Day
Dose Sex 0 7 14
10"5 m3/kg M 176 225 283
F 157 175 214
Body weight data did not suggest any treatment effect.
Plant F
There were no deaths and no signs of toxicity following treat-
ment. Necropsy findings were limited to kidney changes pre-
viously described. Mean body weights (grams) are tabulated
below.
142
-------
Day
Dose Sex 0 7 14
ID"5 m3Ag M 187 244 291
F 210 235 229
Decreased body weights were observed in four of five females
during the last 7 days of the observation period.
Plant G
There were no deaths following treatment. Signs of eye irrita-
tion were observed in a few rats near the end of the observation
period. Because of the time elapsing between treatment and the
onset of changes, no treatment relationship was judged to be
present. Necropsy findings were limited to heart and kidney
changes previously described. Mean body weights (grams) are
tabulated below.
Day '
Dose Sex 0 7 14
l
-------
I Day
Dose Sex 0 7
10~5 m3/kg M 201 275
F 187 221
Body weight data did not suggest any adverse effect.
Plant K :
i
There were no deaths following treatment. Signs were limited to
redness around the eyes of one female rat. Necropsy findings
were limited to mottled kidneys. Mean body weights (grams) are
tabulated below.
i
Day
Dose Sex 0 7 .14
10"5 inVkg M 209 277 313
I F 192 233 248
i
Body weight data did not suggest any treatment effect.
i
Plant L
There were no deaths and no signs of toxicity following treat-
ment. Necropsy findings were limited to kidney changes pre-
viously described among the male rats. Mean body weights
(grams) are tabulated below. i
Day
Dose Sex 0 7 14
10~5 m3/kg M 335 354 380
F 238 240 251
Weight gain among female'rats was less than normal.
i
Plant M I
i
I
There were no deaths andino signs of toxicity except for soft
stool in one male rat on day 11 after treatment. Necropsy find-
ings were limited to mottled kidneys. Mean body weights (grams)
are tabulated below. j
i
i . Day
Dose • Sex 0 7 14
10~5 n»3/kg ' M 262 320 332
i F 196 227 233
144
-------
Data did not suggest any treatment-related effect.
Plant N
There were no deaths and no signs of toxicity following treat-
ment. Except for kidney changes previously described, Necropsy
findings were limited to a distended cecum in one male rat.
Mean body weights (grams) are tabulated below.
Day
Dose Sex 0 7 14
l
-------
Plant S
There were no deaths following treatment. Redness around the
eye of one male rat seen near the end of the observation period
was not judged to be related to the treatment. Necropsy find-
ings were limited to kidney changes previously described. Mean
body weights (grams) are tabulated below.
Day
Dose Sex 0 7 14
10"5 mVkg M 312 341 364
F 237 239 248
Weight gain of females in the first seven days after dosing was
judged to be below normal.
Plant T
There were no deaths and no signs of toxicity following treat-
ment. Necropsy showed mottled kidneys in only one rat. Mean
body weights (grams) are tabulated below.
Day
Dose Sex 0 7 14
1(T5 m3Ag M 172 268 289
F 208 234 249
Data did not suggest any treatment-related effect.
Plant U
There were no deaths and no signs of toxicity following treat-
ment. Necropsy findings were limited to kidney changes pre-
viously described. Mean body weights (grams) are tabulated
below.
Day
Dose Sex 0 7 14
10~5 m3/kg M 360 403 435
F 206 231 239
Data did not suggest any treatment-related effect.
Plant V
There were no deaths following treatment. Signs suggestive of
eye irritation occurred in 4 of 10 treated rats. These appeared
4 to 5 days after administration of test material and were not
146
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judged to be related to treatment. Except for mottled kidneys,
necropsy signs were limited to rough appearance of heart
ventricles. Mean body weights (grams) are tabulated below.
Day
Dose Sex 0 7 14
10~5 m3/kg M 174 233 270
F 158 171 208
Body weight data did not suggest any adverse effect.
Plant W
There were no deaths and no signs of toxicity following treat-
ment. Necropsy findings were limited to kidney changes pre-
viously described among male rats. Mean body weights (grams)
are tabulated below.
Day I
Dose Sex 0 7 14
10~5 m3/kg M 350 378 394
F 226 234 244
Data did not suggest any treatment-related effect.
Plant X
There were no deaths and no signs of toxicity following treat-
ment. Necropsy findings were limited to kidney changes pre-
viously described. Mean body weights (grams) are tabulated
below.
Day
Dose Sex 0 7 14
10~5 m3Ag M 291 359 374
F 203 235 247
Data did not suggest any treatment-related effect.
Plant Y
Although reduced activity was observed immediately following
treatment, there were no deaths. One female rat developed an
ulceration on the ventral thorax 9 days after treatment, but
this was not considered treatment related. Mean body weights
(grams) are tabulated below.
147
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Day
Dose Sex 0 7 14
1CT5 m3/kg M 220 277 318
F 191 214 233
Body weight data did not suggest any treatment-related effect.
Plant Z
There were no deaths and.no signs of toxicity following treat-
ment. Except for mottled kidney changes, Necropsy findings
were limited to an unusual rough appearance of the right
ventricle in one male rat. Mean body weights (grams) are
tabulated below.
Day
Dose Sex 0 7 14
10"5 m3Ag M 249 311 339
F 198 228 236
Data did not suggest any adverse effect.
148
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APPENDIX P
PROTOCOL TO TEST EFFECTS OF WASTE MATERIALS ON MICROBIAL
RESPIRATION (CARBON DIOXIDE REDUCTION) IN A SIMPLE SOIL MICROCOSM
Materials:
• Homogenized soil from site in question
• 12 - 9.46 x lO'1* m3 Mason jars with airtight lids
• 12 - 3 x 10~5 m3 carbon dioxide trap bottles with air-
tight lids
• 20°C incubator
• Approximately 0.5 N NaOH solution ,
• Approximately 0.6 N HC1 solution
• 8 - bent glass rods
• 10 x 10~6 m3 burette or titrometric device
• Trizma (to prepare standard)
Methods:
(A) Soil preparation:
• Air dry soil and grind to pass a 1- to 2-mesh screen.
Ball milling or crushing may be required to break
larger particles.
(B) Microcosm preparation:
• Weigh 100 g (air dried) sieved and homogenized soil into
each of eight Mason jars. One set of four replicates
each for test and control treatments.
• Optional soil inoculum solution:
- Thoroughly mix approximately 200 g of fresh nondried
soil with 10~3 in3 water.
- Separate microorganisms from sediment by filtration
or light centrifugation.
• Moisten soil in each Mason jar from 60% to 80% of field
water holding capacity (FWHC) by uniformly pipetting
dropwise over the surface in each of four replicates,
25 x 10~6 m3 of either test or control solution, plus
5 x 10-6 m3 of inoculum and/or water to bring soil to
desired FWHC.
149
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GLOSSARY
acute toxicity: Toxic effects to an organism due to a short-
term exposure.
concentration: Amount of sample (or toxicant) by weight or
volume per unit volume of test solution.
criteria pollutants: Pollutant species identified by EPA -
Effluent Guidelines Division which require effluent stand-
ards and include BOD5 COD, TSS, chrome, phenol, color,
sulfide, and pH for the textile industry.
cytotoxicity: Toxicity to mammalian cells.
dose: Measured weight or volume of sample (or toxicant) fed to
test organism.
EC 59: Effective concentration at which 50% of the test organisms
reach the desired effect. The "effect", for example, can be
growth inhibition or stimulation.
gastrointubation: Insertion of a tube into the intestinal tract
to feed effluent sample to test animal.
gavage: Forced feeding of an animal through a tube.
hemocytometer: Microscope slide with square rulings used for
counting blood corpuscles or other cells.
in vitro: Describing a biological reaction which can be per-
formed outside the living organism, such as in a test tube.
in vivo: Describing a biological reaction which takes place
within the living organism.
LCso: Lethal concentration fifty - calculated concentration of
substance which is expected to cause death in 50% of the
test organism population, as determined from their exposure
to the substance.
LD50: Lethal dose fifty - calculated dose of chemical substance
which is expected to cause death in 50% of the test organ-
ism population, as determined from their exposure to the
substance.
150
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moving average angle: Iterative computer model designed to
estimate the median of a tolerance distribution using number
of test species used and number that dried due to exposure
to the sample.
necropsy: Sacrificing the test organism to perform an autopsy.
priority pollutants: The 129 chemical species identified by EPA
as a result of the consent decree.
probit analysis: Iterative computer model designed to calculate
LC5Q values from dose response tests using dosage values,
number of test species in control and those exposed to the
effluent sample, and probability values of response.
raw waste: Untreated wastewater as it leaves the textile plant
and enters the wastewater treatment facility.
secondarlr effluent: Textile wastewater treated by aerated
lagoons and clarified.
;em; i:
soil microcosm: Miniature model of the natural system; in this
case, the site of decomposition in the upper (5 i?m) soil
layer.
viability: Capacity of an organism to live and grow.
151
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CONVERSION FACTORS AND METRIC PREFIXES (13)
To convert from
Grams/meter3 (g/m3)
Kilogram (kg)
Meter (m)
Meter3 (m3)
Meter3 (m3)
CONVERSION FACTORS
to
Milligrams/liter
Pound-mass (avoirdupois)
Inch
Gallon (U.S. liquid)
Liter
Multiply by
1.0
2.205
3.937 x 101
2.642 x 102
1.0 x 103
METRIC PREFIXES
I Prefix Symbol Multiplication, factor
Kilo
Milli
Micro
k
m
103
io-3
10~6
Example
5 kg
5 mg
5 yg
5 x 103 grams
5 x 10"3 gram
5 x 10~3 gram
(13) Standard for Metric Practice. ANSI/ASTM Designation:
E 380-76e, IEEE Std 268-1976, American Society for Testing
and Materials, Philadelphia, Pennsylvania, February 1976.
37 pp.
152
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