United States Industrial Environmental Research EPA-600/2-79-118
Environmental Protection Laboratory June 1979
Agency Research Triangle Park NC 27711
Research and Development
Evaluation of
Hyperfiltration for
Separation of Toxic
Substances in Textile
Process Water
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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 nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
6. Scientific and Technical Assessment Reports (STAR)
7. Interagency Energy-Environment Research and Development
8. "Special" Reports
9. Miscellaneous Reports
This report has been assigned to the ENVIRONMENTAL PROTECTION TECH-
NOLOGY series. This series describes research performed to develop and dem-
onstrate instrumentation, equipment, and methodology to repair or prevent en-
vironmental degradation from point and non-point sources of pollution. This work
provides the new or improved technology required for the control and treatment
of pollution sources to meet environmental quality standards.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
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EPA-600/2-79-118
June 1979
Evaluation of Hyperfiltration
for Separation of Toxic Substances
in Textile Process Water
by
J.L Gaddis and H.G. Spencer
Clemson University
Department of Mechanical Engineering
Clemson, South Carolina 29631
Grant No. R805777
Program Element No. 1 LA760
EPA Project Officer: Max Samfield
Industrial Environmental Research Laboratory
Office of Energy, Minerals, and Industry
Research Triangle Park, NC 27711
Prepared for
U.S. ENVIRONMENTAL PROTECTION AGENCY
Office of Research and Development
Washington, DC 20460
-------
ABSTRACT
Three hyperfiltration membranes (cellulose acetate, poly ether/amide,
and dynamic zirconium oxide/polyacrylic acid) were used to separate textile
process water from scour and dye operations into permeate and concentrated
streams. Samples of feed, permeate, and concentrate from each run were
obtained and analyzed. Chemical analysis for organic and metal toxic
pollutants and bioassays for rat acute toxicity, fathead minnows and
Daphnia acute toxicity, microbial mutagenicity, and hamster ovary clone
cytotoxicity response were conducted.
Both the fathead minnows and Daphnia tests showed results in the active
range. The other bioassays did not. The results were consistent in
indicating a substantial reduction of toxicant in permeate samples from all
membranes and corresponding increases in toxicant in the residual concentrate
samples. Toxicant rejections of 55 to 100 percent were observed, and the
relative rejection by the three membranes was almost exclusively counter to
the relative rejection of salt. Mass balances of biological toxicant were
excellent, suggesting high confidence in the result.
Chemical analysis for organic compounds sensed 19 of the organic toxic
pollunts in low levels (300 mg/m^ and under). The results were difficult to
interpret for mass balance and membrane rejection of particular solutes.
Except for a few compounds, the data appears to suggest membrane separation.
An experiment set devised to enhance accuracy of analysis is recommended to
establish the rejections of pertinent substances.
Metal toxic pollutant concentrations were low. Analysis revealed only
three in high enough concentrations for reliable estimation of performance.
Other metals analyzed and the toxic metals results agree with the historical-
ly high rejection of metals (reference page 21).
This report was submitted in fulfillment of Grant R-805777 by Clemson
University under the sponsorship of the U.S. Environmental Protection Agency.
The report covers a period from January 1978 through October 1978 and work
was completed as of May 1979.
11
-------
CONTENTS
Abstract ii
Figures v
Tables vi
Symbols and Units vii
Acknowledgment viii
1. Introduction 1
2. Conclusion 3
3. Recommendations 5
4. Results and Discussion 6
Flows, Volumes, and Physical Parameters 6
Organic Solutes 11
Metals 17
Bioassay 22
Correlation of Rejection in Single-Solute
Solutions with Solute Solubility Parameter 29
5. Test Description 30
References 35
Appendix A Infrared Analysis 36
Appendix B Interpretation of Results 61
Appendix C Evaluation of Hyperfiltration Treated
Textile Wastewaters 68
1. Introduction 69
2. Summary 70
3. Sample Collection 78
4. Priority Pollutant Analysis 81
5. Bioassay Tests 99
6. Appendix CA: Priority Pollutant Analysis Fractions H5
7. Appendix CB: Raw Data from the Ames Mutagenicity Tests H8
8. Appendix CC: Raw Data for the CHO Cytotoxicity Tests 14°
9. Appendix CD: Characteristics of the 14 Wastewater
Samples and Reconstituted Water
10. Appendix CE: Characteristics of the Wastewater Samples
as a Function of Time and Mortality Data
Response
11. Appendix CF: Water Quality Analysis of the Wastewater
Samples as a Function of Test Solution
Concentrations and Raw Mortality Dose
Response for Daphnia Acute Toxicity Tests 206
111
-------
12. Appendix CG: Raw Data on Acute Oral Toxicity Study in
Rats 231
13. Conversion Factors and Metric Prefixes 260
Appendix D Dependence of Rejection Solubility Parameters 261
Appendix E Sampling Plan 270
IV
-------
FIGURES
Number
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Permeate Flow from Dynamic Membranes on Dye Waste
and Scour Waste
Permeate Flow Rates for Cast Membranes During Testing
Relation Between Feed and Permeate Concentration and
Membrane Rejection
Relative Concentrations of Toxicants and Arsenic
Relative Concentrations of Toxicants and Total Solids
Relative Concentrations of Toxicants and Copper
Correlation of Concentration Toxic to Fathead Minnows
with Concentration Toxic to Daphnids
Schematic of Fluid Acquisition and Operations
Page
7
8
10
25
26
27
28
31
-------
TABLES
Number
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
Table 7
Table 8
Table 9
Table 10
Table 11
Table 12
Table 13
Table 14
Table 15
Table 16
Experiment Results for pH, Solids, and Conductivity
Total Solids Balance and Recovery Data
Rejection by Membranes
Run 1 Cast Membranes on Dye Fluid
Run 2 Dynamic Membranes on Dye Fluid
Run 3 Cast Membranes on Scour Fluid
Run 4 Dynamic Membranes on Scour Fluid
Metal Analysis
Percent Rejection of Metals by Hyperfiltration:
Group I Results (Normal Confidence)
Percent Rejection of Metals by Hyperfiltration:
Group II Results (Reduced Confidence Level)
Lethal Concentration and Implied Toxicant
Concentrations
Rejection of Toxicity by Hyperfiltration
Mass Ratio of Toxicants
Operating Conditions Observed
Summary Log of Activities
Sample Disposition Log
6
9
11
12
13
14
16
18
20
21
23
24
25
30
33
34
VI
-------
SYMBOLS AND UNITS
Item Symbol (Unit)
Pressure p (N/m2)
Temperature T (°C)
Recovery R (no units)
Concentration c (g/m3)
Subscripts
Feed - f
Permeate - p
Concentrate - c
Units (S.I.) Multiply By To Get Unit
m 3.28 ft
°C (°K-273.16) 1.8 °F-32
MN/m2 1.44 x 10"1"2 psi
m3 " 264 gallon
m2 10.76 ft2
S (Siemens) 1.00 ohm'1 (mho)
£ (liter) is used generally rather than the S. I. unit dm3
Metric Prefixes
M denotes 106
k denotes 103
m denotes 10~3
u denotes 10~6
Vll
-------
ACKNOWLEDGMENT
The authors wish to acknowledge the participation of La France
Industries, a division of Riegel Textile Corporation, for allowing this
work to be performed on their premises. Dr. James E. Bostic, Jr. has
served as coordinator for Riegel. All of the La France personnel have
been extremely cooperative and helpful, but particular thanks are due to
Messrs. Perry Lockridge and Bill Williams in the dyehouse.
The authors also thank Dr. Max Samfield, EPA Project Leader for his
valuable guidance throughout the course of this work.
The chemical analysis and bioassay effort and report were coordinated
by Dr. Gary D. Rawlings, Monsanto Research Corporation. His contribution
was appreciated very much. The chemical analyses and bioassays were
performed by the Monsanto Research Corporation, EG and G Bionomics Marine
Research Laboratory and Litton Bionetics.
viii
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INTRODUCTION
The U. S. Environmental Protection Agency '(EPA) is implementing
limits on industrial plant discharge of Consent Decree Toxic Pollutants
and developing technologies for compliance with these limits. The
textile industry discharges large quantities of effluents with some
effluents containing detectable concentrations of several toxic
pollutants.1 Other chemicals not included in the Consent Decree such
as dyes which are toxic at concentrations as low as 100 g/nr* may be
present in the typical discharge.2 This report describes an investigation
of hyperfiltration as a technology for separating toxic materials occurring
in selected textile process effluents.
The purposes of the investigation were: 1) to determine the
effectiveness of representative commercial hyperfiltration membranes in
separating toxicity, as measured by EPA-approved short-term bioassay,
found in the untreated process effluents; 2) to compare the toxic
rejections of the membranes; 3) to obtain rejection coefficients of
the detectable solutes; and 4) to correlate toxicity with the presence
of detectable solutes, evaluating internal consistencies among both
the bioassay results and the chemical analysis results.
It was desired to obtain representative samples of untreated process
effluent and process them by hyperfiltration. Samples of feed, permeate,
and concentrate could be analyzed for specific chemicals and be subjected
to bioassay. The fluids selected were of a cotton scour and a cotton dye
process from a dye range.
Membranes selected have a reasonable expectation of industrial
applicability. Those selected were commercial cellulose acetate,
poly(ether/amide), and a dynamic membrane (zirconium oxide/poly
acrylic acid) prepared at Clemson University. The polyamides were
eliminated due to expected difficulties with plugging from the
industrial fluid, and other membranes were not considered to be
sufficiently commercial at the decision time.
A test program was designed for the fluids and membrane combinations
cited. The samples were analyzed by Monsanto Research Corporation or
designa ed subcontractor under separate contract to EPA. Analyses
•'•Rawlings, G.D. and Max Samfield," Source Assessment: Textile Plant
Wastewater Toxics Study Phase I," EPA 600/2-78-004h, March, 1978.
2"Dyes and the Environment," ADMI Report, Volume II, September, 1974.
-------
selected were organic toxic and metal toxic chemical analysis; rat
acute toxicity; Fathead minnow 96-hour acute toxicity; Daphnia 48-hour
acute toxicity; microbial mutagenicity response; and hamster ovary clone
cytotoxicity. In addition, measurements of total solids, electric
conductivity, pH, absorbance (410 nm), and infrared spectra were performed
at Clemson University.
-------
CONCLUSIONS
1. Hyperfiltration membranes have been shown to be effective in
producing a substantially less toxic (to aquatic organisms) permeate
while also producing a correspondingly more toxic concentrate when
operated on actual textile plant effluents.
2. While all membranes tested were effective, the relative
separation of toxicants was observed to be counter to the relative
salt separation. That is, the membrane having the best salt rejection
was not the best with regard to toxic material rejection.
3. The membranes exhibited high rejection, greater than 0.85,
of solute components detected by color, total solids, and conductivity
analyses.
4. All the metal toxic pollutants were detected, but only three
were present in concentrations sufficient to calculate reliable
rejection coefficients. These were high, the average values were:
above 0.89 for arsenic, 0.97 for copper, and 1.00 for zinc. This result
coupled with prior experience of generally high rejection of metal ions
found in textile process effluents provides good evidence for high
rejection of toxic pollutant metals in these effluents.
5. Only 19 organic toxic pollutants were detected, also at low
concentrations. Because of the analytical difficulties associated with
low concentration and difficulty in controlling concentrations of volatile
organic solutes at elevated temperatures during the experiments reliable
rejection coefficients were not obtained for the organic toxic pollutants.
However, using decreased solute concentration in the permeate and/or
increased solute concentration in the concentrate as indication of rejection,
most solutes were rejected in these process effluents, i.e., 43 of 51
comparisons showed positive rejection.
6. Because so few rejection coefficients were evaluated no cause/
effect correlations between toxic response and specific toxic pollutants
were apparent. Correlations between aquatic organism toxicity and
concentrations of copper and arsenic appear strong. However, the metal
concentrations were likely too low to account for the toxicity.
7. Toxicant concentrations implied by the aquatic organism toxicity
assays permitted calculation of reasonable toxicant mass balances. The
toxicant concentrations were substantially proportional to the total solids
concentrations.
-------
8. It should be noted the correlation coefficient relating the
toxicant concentrations implied by the two aquatic organisms Fathead
minnows and Daphnia was high, 0.94, suggesting that for these two
discharge streams, a measurement of either individual assay would
have produced parrallel data.
9. Rat toxicity and bacterial mutagenicity tests produced no
response. Concentrates were cytotoxic, but no cytotoxicity was observed
in feeds and permeates. Cytotoxicants were probably concentrated (rejected)
-------
RECOMMENDATIONS
1. The observed significant separation of toxicity provides a
basis for recommending hyperfiltration be considered further as a
technology for toxic control of industrial effluents.
2. Continued research to quantify the applicability of this
technology is recommended. Specifically, the analytical and concen-
tration control difficulties experienced in this field experiment
suggest well controlled, repeatable, zero recovery laboratory experiments
using a few selected solutes to determine accurate rejection coefficients.
The solutes should be selected to provide a breadth of properties sufficient
to test models for the prediction of rejections of all the toxic pollutants.
In addition, experiments using process effluents spiked with known
quantities of selected solutes should be completed to permit the quantita-
tive analysis of membrane performance under conditions approaching the
field experiments conducted in this investigation.
3. Research to identify the process effluent components responsible
for the toxicity to aquatic organisms is recommended.
-------
RESULTS AND DISCUSSION
FLOWS, VOLUMES, and PHYSICAL PARAMETERS
Experiments were carried out using three hyperfiltration membranes,
poly(ether/amide) composite (PEA), asymmetric cellulose acetate (CA), and
zirconium oxide/poly(acrylic acid) dynamic membrane (DM), using two types
of process effluents, cotton scour and dye wash. The permeate flow rates
of the three membranes during the course of the experiments are presented
in Figures 1 and 2.
Total solids, electric conductivity, absorbance, and pH of the samples
are shown in Table 1. The general level of solids shows the effect of
membrane separations and is in agreement with the concentrate levels as
well. Infrared spectra obtained from sample residuals are included in
Appendix A.
TABLE 1.
Sample
Number
Run
Run
Run
Run
1
2
#3
3
4
5
6
#4
7
8
9
#1
10
11
12
13
#2
14
15
16
Experiment Results
Description
Plant
Apparatus
Sc-1,
Sc-1,
Sc-1,
Sc-1,
Sc-2,
Sc-2,
Sc-2,
Dye-1,
Dye-1 ,
Dye-1 ,
Dye-1,
Dye- 2,
Dye-2,
Dye-2,
feed
permeate , PEA
permeate CA
concentrate
feed
permeate , DM
concentrate, DM
feed
peameate, PEA
permeate , CA
concentrate
feed
permeate, DM
concentrate
pH
6.
7.
9.
7.
7.
9.
10.
9.
9.
6.
6.
6.
7.
7.
8.
8.
for
pH , Solids and
Conductivity
Conductivity Total Solids Absorbance
(ps/cm) (g/m3) (410 nm)
6
2
7
2
7
8
4
3
4
5
9
7
6
5
2
4
106
157
710
25
24
3830
957
280
2870
271 (228)a
20
22
1800
929
106
3230
15
43
730
105
32
6020
870
205
3840
462 (391) a
15
45
2670
760
60
2160
0
0
0
0
0
0
0
0
0
0
0
0
0
2
0
7
*
.055
.050
*
•
.050
.03
.01
.15
.1 (0.08)
•
*
.65
.0
•
.8
aln Run 1, the feed was concentrated by an estimated 18 percent before the
feed sample was obtained. The estimated actual feed conductivity, solids,
and absorbance values are respectively shown in parentheses.
-------
LU
Q.
110
100
90
80
70
60
50
40
30
20;
10
00
e *
A-- -A DATA/ SCOUR (RUN 4, T = 77c, 16//MIN/ 5,9 lIN/M2)
3,14 FT2 ACTIVE AREA
A—
DATA, DYE (RUN 2. T = 70c, 16JC/MIN, 4,5 FWM2)
O
0 5 10 15 20 25 30 35 40 45 : 50 55 60 ^ 65 70 75 80
TIME * HOURS
FIGURE 1 PERMEATE FLOW FROM DYNAMIC MEMBRANE ON DYE WASTE AND SCOUR WASTE
-------
= 2,8 x 10
03
.
UJ--2
UJ
D-
4.0
3.0
2.0
1.0
FLOW * 15 //MIN
END RUN 3
END RUN 1
TEMPORARY INTERRUPT RUN 1
O PEA
•Q CA.
A PEA
OCA
} DYE/ RUN 1
I SCOUR/ RUN 3
10 ! 20 • 30 ,40 50 60 70 80 90
TIME/ MINUTES
FIGURE 2 PERMEATE FLOW RATES FOR CAST MEMBRANES DURING TESTING
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The volumes of permeate, feed, and concentrate have been refined as
described in Appendix B., based on the total solids measurements. In general,
the refined volumes agree well with direct observations forming a reasonable
consensus. The volumes observed have been modified to the recovery (volume
of permeate/volume of feed) shown in Table 2. These recoveries indicate
the best combined agreement with final (solute) mass to initial mass ratio,
rejection performance indicated by total solids analysis, and original
volume estimates. The recovery ranges from 0.73 to 0.89 for the four tests,
averaging 0.83. An overall mass ratio of total solids as shown in Table 2
is excellent except in run 2 where 26 percent of the original mass is not
accounted for.
TABLE 2. Total Solids Balance and Recovery Data
Run i ! ! 1
Fluid Dye Dye Scour Scour
Membrane Cast Dynamic Cast Dynamic
Recovery3
Overall 0.863 0.730 0.890 0.820
Cellulose acetate 0.379 - 0.418
Poly(ether/amide) 0.484 - 0.472
Mass ratio
(final/initial) , 0.99 0.74 0.99 0.99
aSee Appendix B for details of the calculation of recovery.
^To calculate mass ration, use solids data from Table 1.
mass in PEA permeate = 0.484 x 15 =7.26
mass in CA permeate = 0.379 x 45 =17.05
mass in Concentrate = (1 - .484 - .379) x 2670) = 365.8
Total, mass at end of run = 390.1 g/irr of feed
Mass in feed = 1 x 391 = 391 g
390
Mass ratio = —- =0.99
.391
In Run 2, a leak of 7 percent of feed during the run must be accounted for,
depressing the mass at the final condition.
An effort to refine the calculation of rejection to include individual
toxic components was made but was considered not appropriate for the
analytical results obtained. The accuracy estimates given by Monsanto
Research Corporation are -100 percent for organics and -20 percent for the
metal analysis. The calculated rejections are, therefore, not highly
accurate estimates. A simple, yet reasonably accurate, estimate of
rejection based on permeate and feed concentrations was used. It can be
shown that such a calculation is only mildly dependent on the recovery and
therefore a single relation of rejection versus permeate to feed concen-
tration ratio was used for simplicity.
Figure 3 shows the proposed relation between rejection and permeate to
feed concentration ratio. It is based on a simple assumption of uniform
rejection, independent of concentration, and a volume recovery of 0.85. The
-------
100
(85% RECOVERY)
0 10 20 30 ' 40 50 60 70 80 90
PERMEATE TO FEED CONCENTRATION RATIO GDO§=)
FIGURE 5 RELATION BETWEEN FEED AND PERMEATE CONCENTRATION AND MEMBRANE REJECTION
lo
-------
effects of vapor loss, small leaks, and recoveries different from 0.85 are
estimated to be relatively minor. The use of Figure 3, or equivalent, is
used to obtain rejection from permeate and feed analysis data.
The data presented in Table 1 has been analyzed for rejection and
presented in Table 3. All membranes are effective in rejecting total solids
and ionic solutes. The lower rejection of the solutes in scour by the
dynamic membrane is probably due to its passage of ions at the pH % 10
operating condition in this fluid. All membranes were effective in removing
color as evidenced by the absorbances in Table 1. The cellulose acetate
permeate did not foam, while the others did produce some foam.
TABLE 3.
Membrane/Fluid
Cellulose acetate/dye
Cellulose acetate/scour
Poly (ether /amide) /dye
Poly (ether /amide) /scour
Dynamic/Dye
Dvnamic/S cour
Rejection by Membranes
Rejection based on
Run Number Solids
1 0.94
3 0.98
1 0.98
3 0.93
2 0.97
4 0.88
Conductivity
0.95
0.99
0.96
0.99
0.95
0.85
Organic Solutes
Chemical and bioassay tests were conducted under separate contract
to Monsanta Research Corporation (MRC). The complete test results as
obtained from MRC are appended to this report as Appendix C for convenience.
The data obtained thusly are described in detail in the following.
Tables 4, 5, 6, and 7 show the results obtained for toxic organic
solutes in the four runs. The concentrations of the feed sample, permeate
sample(s), and concentrate samples are shown followed by the mass ratio
calculated thereform. The calculation of mass ratio is illustrated by the
following example in Run 1, Bis(3-ethylhexyl) phthalate (see Table 4).
Volume data from Table 1
Concentration data from Table 4
mass in PEA permeate = 0.484 x 31 = 15.0
mass in GA permeate = 0.379 x 3 = 1.1
mass in concentrate = 0.137 x 51 = 7.0
end of run, total = 23.1
mass in feed =1x3.4=3.4
mass ratio =
3.4
The value 3.4 is 4 -5- 1.18 where 1.18 is the estimated concentration which
occurred in Run 1 before securing the feed sample. Only on Run 1 is this
factor appropriate. No effect of the solute mass in the leak during Run 2
is accounted for in Table 5 mass ratio data.
11
-------
Table 4
Run 1 Cast Membranes on Dye Fluid
(values in mg/m )
Compound
Bis (2-ethylhexyl)
phthalate
Dimethyl phthalate
Di-n-butyl phthalate
Butylbenzyl phthalate
Diethyl phthalate
Acenaphthene
Anthracene
to Fluoranthene
Pyrene
Naphthalene
Phenanthrene
Phenol
Chloroform
Toluene
Trichloroethylene
Benzene
Chlorobenzene
Ethylbenzene
Methylene chloride
Triphenyl phosphine
Triphenyl phosphine oxide
«-Terepineol
Permeate
Poly
Feed Ether/Amide
CTHF 10 CTHF 11
4
55
1
3
0.6
0.8
0.2
19
10
2
5
5
.de 5
30
31
45
0.8
1
0.8
0.7
31
11
0.6
0.4
45
2
5
20
Permeate
Cellulose
Acetate Concentrate
CTHF 12 CTHF 13
3 51
290
6
7
7
3
0.4 1
4
24
1
4 4
7 10
10 30
30 50
Mass Ratio
End/Start
6.8
1.2
1.3
OO
0.5
0.7
0.0
3.6
1.0
1.7
00
0.3
5.7
1.1
2.3
1.1
Comments
mixed rejections,
positive rejection,
positive rejection,
concentrated
membrane may be source
not detected
rejected and concentrated
rejected and concentrated
not detected
not detected
sorbed or vaporized
not detected
not rejected,
concentrated
mixed rej ected, not
concentrated
not rejected
membrane source
rejected, perhaps
vaporized
not detected
not detected
membrane possibly source
container source
container source
slight rejection
-------
Table 5
Run 2 Dynamic Membrane on Dye Fluid
(yalues in mg/nrl
Compound
Bis(2-ethylhexyl)
phthalate
Dimethyl phthalate
Di-n-butyl phthalate
Butylbenzyl phthalate
Diethyl phthalate
Acenaphthene
Anthracene
Fluoranthene
Pyrene
Napthalene
Phenathrene
Phenol
Chloroform
Toluene
Trichloroethylene
Benzene
Chlorobenzene
Ethylbenzene
Methylene chloride
Triphenyl phosphine
Triphenyl phosphine
oxide
Oi Terepineol
2-Mercepto
benzthiazole
l-Cyano-2-
benzyloxyethane
Benzothizole
Feed Permeate Concentrate
CTHF 14 CTHF 15 CTHF 16 Mass Ratio Comments
170
1
3
0.7
0.1
0.8
0.2
96
0.6
0.6
5
10
10
50
40
60
200
4
1
0.05
0.1
0.4
1
3
10
10
5
30
10
3
10
200
100
250
0.905 rejected, concentrated
0.02 rejected, not concentrated
1.0 not rejected, not concentrated
not detected
CO
0.0 sorbed
0.1 rejected, not concentrated
0.0 sorbed
not detected
0.0 sorbed
not detected
0.0 detected
0.0 vaporized
0.94 slight rejection
1.22 negative rejection
not detected
not detected
not detected
0.6 slight rejection, not concentrated
1.0 container source
0.86 container source
0.07 sorbed
1.9 slight rejection
0.57 rejected
0.34 rejected
-------
Table 6
Run 3 Cast Membranes on Scour Fluid
(values in mg/m3)
Comments
Bis(2-ethylhexyl)
phthalate
Dimethyl phthalate
Di-n-butyl phthalate
Butylbenzyl phthalate
Diethyl phthalate
Acenaphthene
Anthracene
Fluoranthene
Pyrene
Naphthalene
Phenanthrene
Phenol
Chloroform
Toluene
Trichloroethylene
Benzene
Chlorobenzene
Ethylbenzene
Methylene chloride
Triphenyl phosphine
Feed
CTHF 3
Permeate Permeate
Poly Cellulose
Ether/Amide Acetate
CTHF 4 CTHF 5
3 3
9
7
2
0.4
1
0.8
0.5
18
0.8
0.3
5
2
18
15
1
6
0.5
3
22
29
0.4
1
0.7
5
2
13
41
5
6
21
15
00
0.98
30
2.4
OO
OO
oo
1.3
00
Concentrate
CTHF 6 Mass Ratio Comments
.30 mildly rejected, but not
concentrated
oo permeate possibly contami-
nated by previous run
(CTHF 11)
0.1 rejected, but not concen-
trated
not detected
not detected
0.05 rejected, but not concen-
trated
0.0 rejected, but not concen-
trated
0.0 rejected, but not concen-
trated
0.0 rejected, but not concen-
trated
not detected
concentrated
poor rejection
concentrated
concentrated
concentrated
concentrated
concentrated
not rejected, but concen-
trated
container source
-------
Table 6 (continued)
Comments
Triphenyl phosphine
oxide
ot-Terepineol
2-Atercapts
benzothiazole
l-Cyano-2-
benzyloxyethane
Benzothiazole
Laurie Acid
Myristic Acid
Palmitic Acid
Permeate Permeate
Poly Cellulose
Feed Ether/Amide Acetate Concentrate
CTHF 3 CTHF 4 CTHF 5 CTHF 6 Mass Ratio Comments
5
10
10
30
400
10
30
20
5
2
10
0.5
600
3000
1000
1000
1.76
1.42
0.96
00
2.4
0.9
00
00
container source
not rejected
not rejected
membrane source
rej ected, concentrated
rejected, concentrated
concentrated
concentrated
-------
Table 7
Run 4 Dynamic Membrane on Scour Fluid
(value? in mg/m3!
Feed
CTHF 7
9
Compound
Bis(2-ethylhexyl)
phthalate
Dimethyl phthalate
Di-n-butyl phthalate
Butylbenzyl phthalate
Diethyl phthalate
Acenaphthene
Anthracene
Fluoranthene
Pyrene
Naphthalene
Phenanthrene
Phenol
Chloroform ' 34
Toluene 0.8
Trichloroethylene
Benzene
Chlorobenzene
Ethylbenzene
Methylene chloride 4
Triphenyl phosphine
Triphenyl phosphine oxide 2
#-Terepineol 25
2-Mercapto-benzothiazole 10
Benzothiazole 40
Laurie acid
Palmitic acid
Stearic acid
Permeate
CTHF 8
Concentrate
CTHF 9
0.7
2
2
5
5
30
0.5
0.7
2
5
100
100
400
200
Mass Ratio Comments
0.0
0.0
0.0
0.0
O*
0.0
0.84
tOr
aa
1.11
09
11.6
0
0
0.76
00
00
00
sorbed
not detected
sorbed
not detected
not detected
sorbed
not detected
not detected
not detected
not detected
sorbed
source possibly in residual of
previous fluid
vaporized
rejected mildly, possibly vaporized
not detected
not detected
negative rejection
container .source
container source
sorbed
sorbed
rejected, concentrated
concentrated
concentrated
concentrated
-------
A value of one in mass ratio indicates a consistent total solute mass.
Values greater or less than one imply that the mass is estimated to have
increased or decreased. Increases in mass imply a source of solute either
from carryover from a previous run or desorption from the membrane or equip-
ment. Since care was taken to use only stainless steel and teflon in the
system, and the membranes were flushed reasonably well the latter source
was as small as was practical. The plastic (polyethylene) covers on the
tanks could have served as sources for phthalates when the condensing vapors
dripped into the tank. The possibility of carryover from the previous run
are acknowledged in the comments on the tables.
Many of the solutes subject to analysis expected in the concentrate were
not detected there. This is especially true of the base neutral compounds
in the dynamic membrane tests (Table 5 and 7). These compounds are not
highly volatile, but may have been sorbed into the apparatus or rendered not
extractable for analysis. The more volatile compounds chloroform and benzene
probably vaporized. Toluene may have been sourced from the cellulose
acetate and poly(ether/amide) membranes and as such the rejection may be
masked.
The number and level of concentration of toxic organic compounds was
low in all runs. Because of this and the analytical inaccuracy (-100 percent)
the calculation of rejection is not meaningful.
However, if either decreased permeate concentrations or increased
concentrations of concentrate can be used to signal positive rejection,
forty-three of fifty-one show positive indication and eight indicate
corroborating data for low rejection. Chloroform, toluene, trichloro-
ehtylene, and methylene chloride all show a somewhat consistent trend to
low rejection. The evidence for rejection is mixed for phenol and
di-n-Butyl phthalate. The remainder of compounds have at least some
evidence in each set of data to indicate positive rejection. These
observations are actually stronger than is actually substantiated by
the data, but represent the trends which are apparent.
A few additional organic compounds detected without the use of
standards are identified in Appendix C. Those most prominent are the acid
complement to certain detergents (lauric acid, myristic acid, palmitic
acid) which were noted almost exclusively in the concentrated samples.
Benzothiazole was detected in three runs and was rejected effectively.
Metals
Metal analyses for toxic pollutants and other metals were performed
by Monsanto Research Corporation. Analysis for arsenic was performed
by conventional atomic absorption, the others were analyzed in neat and
digested samples. The neat analysis results were suspected of showing an
effect due to organic loading. The digested samples do not show such
effects. Raw analysis for the digested samples has been corrected for
metals in dilution water and reagent acid which were added during digestion.
The results, as corrected are shown in Table 8. Very low levels of most
toxic metals are notable.
17
-------
Plant Apparatus
Table 8 Metal Analysis
Concentration in Streams (mg/m^)
Permeate, Permeate, Permeate,
Feed PEA CA Concentrate Feed DM Concentrate
Metal
Aluminum
Antimony
Arsenic
Barium
Beryllium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Molybdenum
Nickel
Phosphorus
Silicon
Silver
Sodium
Strontium
Tin
Titanium
Vanadium
Zinc
CTHF-1 CTHF-2
106
<0
<1
98
-
553
—
13,300
206
-
36
965
51
5,060
256
-
-
-
13,150
-
57,800
130
-
-
19
216
95
<0
<1
78
-
308
-
16,100
355
2
178
269
82
6,680
146
-
79
1,320
17,800
-
103,000
153
-
-
30
202
CTHF-3 CTHF-4 CTHF-5 CTHF-6 CTHF-7 CTHF-8 CTHF-9
794
38
19
82
-
47,200
6
15,900
306
15
72
445
263
9,260
356
16
61
3,926
17,300
11
378,000
142
64
35
69
106
77
23
<1
-
-
11,900
6
608
310
-
6
212
112
194
20
-
-
95
2,200
-
11,720
-
-
1
5
-
164
70
1
8
-
8,900
5
892
286
0
14
119
223
320
22
28
29
526
2,250
11
23,400
-
-
15
9
8,180
3,690
364
160
578
-
81,000
38
113,700
775
65
738
2,800
602
71,950
2,600
118
405
33,200
29,600
83
1,672,000
1,040
520
75
480
3,120
1,270
103
35
118
—
56,000
9
15,200
350
11
74
332
262
6,154
716
21
127
4,830
20,600
31
610,000
138
68
21
55
46
1,260
79
5
6
-
31,000
11
1,078
390
13
48
212
276
362
22
55
137
1,100
7,200
47
242,000
—
-
5
29
-
4,890
308
-14
348
-
81,000
110
63,500
555
45
622
1,900
762
28,750
2,860
190
393
23,600
22,000
169
1,544,000
560
260
59
230
6,146
-------
Table 8 (continued)
Permeate , Permeate ,
Feed PEA CA Concentrate
Metal
Aluminum
Antimony
Arsenic
Barium
Beryllium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Molybdenum
Nickel
Phosphorus
Silicon
Silver
Sodium
Strontium
Tin
Titanium
Vanadium
Zinc
CTHF-10 CTHF-11 CTHF-12 CTHF-13
431
120
35
62
-
2,500
12
16,300
55
19
358
328
362
11,350
276
56
145
7,200
23,400
51
185,000
144
60
17
86
6,780
165
65
15
-
-
430
11
618
350
9
26
192
266
94
12
39
127
375
120
37
11,600
-
-
5
15
—
116
54
<1
-
34
1,160
39
252
186
37
32
55
263
66
36
60
-
366
3,010
—
7,190
-
60
1
-
—
10,900
208
221
398
-
3,950
30
113,100
575
61
3,040
950
542
81,800
1,860
112
405
49,400
39,200
85
894,000
1,000
200
35
530
4,946
Feed
CTHF-14
1,090
124
2
194
-
772
10
25,500
415
13
10,600
300
362
12,950
696
66
111
49,400
11,600
39
528,000
258
50
9
96
6,190
Concentrate
CTHF-15 CTHF-16
640
98
<1
-
-
683
25
532
346
13
82
105
333
200
10
124
149
3,870
12,900
53
90,000
-
12
11
21
-
2,900
248
9
478
_
1,232
60
70,100
835
59
35,400
1,240
982
41,150
396
358
525
140,800
19,800
95
1,247,000
718
120
27
290
12,390
-------
The rejection of metals in the proces water is difficult to estimate
in most cases due to the low concentration levels. As has already been
mentioned, digestion of metal samples was performed, with the result that
metal addition from nitric acid and distilled water occurred. In many cases,
the metal addition was of the same magnitude as the total concentration in
the feed sample. Thus the correction applied was as large as was the metal
inclusion. Since membranes have shown an excellent rejection3 for metals
regardless of form (ionic, complexed, etc.) the anticipated level in a
permeate is at least an order of magnitude lower than the feed concentration.
In such a case the permeate analysis is subject to very large errors due to
ordinary uncertainty. For this reason the results for rejection have been
separated into three groups.
Group I (results shown in Table 9) contains the :data for which the feed
and permeate level is /sufficiently high to provide a normal estimate of
rejection. The criterion used is that the feed content is at least five
times that amount added during digestion of the sample.
TABLE 9. Percent Rejection of Metals by Hyperfiltration: Group I Results
(Normal Confidence)
Metal Toxic
Pollutants
Arsenic
Copper
Zinc
Other Metals
Aluminum
Barium
Boron
Calcium
Iron
Magnesium
Manganese
Phosphorus
Silicon
Sodium
Strontium
Tin
Titanium
Vanadium
Poly(ether/amide)
Scour Dye
98
96
100
88
98
70
99
97
98
94
98
100
100
>99
97
75
97
100
100
92
98
99
98
97
100
97
100
100
92
Cellulose Acetate
Scour Dye
98
90
96
92
98
87
98
97
95
94
97
100
100
74
94
>98
96
100
100
72
>99
99
95
97
95
98
±00
<0
100
Dynamic Membrane
Scour Dye
94
1
97
64
97
97
98
89
81
77
100
100
65
>69
>99
100
60
100
32
99
99
98
96
0
92
100
88
90
Omission from this table implies a low value of feed concentration.
text for details...
See
3Brandon, C. A., J. J. Porter, and D. K. Todd, "Hyperfiltration for
Renovation of Composite Wastewater at Eight Textile Finishing Plants," Final
Report, EPA Grant 802973.
20
-------
Group II (results shown in Table 10) contains the data for which the
feed has less than five times but more than twice the amount added during
digestion of samples. Rejections thus obtained are subject to greater
uncertainty than normal and the values should be treated as an indication
of rejection.
TABLE 10. Percent Rejection of Metals by Hyperfiltration: Group II Results
(Reduced Confidence Level)
Toxic Metals
Copper
Lead
Other Metals
Aluminum
Cobalt
Iron
Titanium
Poly(ether/amide)
Scour Dye
96
75
96*
70*
>99*
97*
44
78
70
60
85
Cellulose Acetate
Scour Dye
91
32
90*
87*
74*
96*
46
77
<0
92
97
Dynamic Membrane
Scour Dye
55
1*
55
88
99*
25
60*
80*
Omission from this table implies a near absence in feed. See text for
details.
*Values marked are higher confidence data from Table 9.
Group III contains the data having feed solute mass less than twice
that added in digestion. For these data, the uncertainty in feed and
product is such that the respective values of concentration may overlap
resulting in about as many negative as positive calculated rejections.
These data are not presented in rejection form because they are not
considered to be meaningful.
In all the data of Tables 9 and 10 the curve of rejection as
dependent on permeate and feed concentration ratio has been employed from
Figure 3.
According to the foregoing criteria, some metals were present in such
low concentration that the analysis cannot be expected to provide even an
indication of the rejection. These metals are Antimony, Beryllium, Cadmium,
Chromium, Nickel, and Silver from the toxic pollutant list. In some runs
zinc and lead also were below the concentration criterion. Arsenic was
present in low levels (20 mg/m3) but was analyzed without digestion such
that analysis is expected to be accurate. Some copper and zinc levels
were high enough to> qualify for normal rejection assessment. These
appear to be the only toxic metals present in the process water and occur
only in the dye effluent.
Despite the limited data for rejection of metals obtained in this
effort, membranes have historically shown excellent rejection for metals.
This trend is corroborated by the data in Table 9. Three unusually low or
21
-------
negative rejection data are shown in the "other metal" list: aluminum on
the scour with the dynamic membrane, silicon on the dye with the dynamic
membrane, and tin on the dye with the cellulose acetate membrane. In each
of these cases reference to the concentrate data of Table 8 shows that the
element was concentrated. Therefore, it is considered that some anomaly
of analysis is involved and that probably the rejections are not as low as
indicated .
Bioassays
The values of LCso (or ECso) obtained from each sample may be
heuristically related to the concentration of an unknown substance. The
concentration of that unknown substance which produces 50% mortality is
expected to be a reasonably- repeatable value, say C*. When a volume of
fluid contains LCso of a sample and (1-LCso) of diluent water the
concentration of unknown substances is C*. Also one can use this fact
to determine the concentration (C) from OLCso - C*. Therefore, the
concentration (C) of toxic substance is inversely proportional to the
value of
Obviously ±he foregoing statement applies to the simplest, single
toxicant solution. However, if the membrane is not highly selective in
rejection for a multicomponent mixture a very similar result would obtain
for comparison .of toxic effects of feed and concentrate, etc. Therefore,
the data for "LCso have been used to calculate relative values for the implied
concentration of toxic substance to enable the calculation of membrane
rejection. The bioassay tests results for LC50 are presented together with
the implied concentration of toxicant in Table 11. The values for
concentration of toxicant are simply 100 divided by its respective LCso
value. The information in Table 11 is organized in the order of actual
test sequence which is different from the sample numbering sequence.
Values of implied concentration from Table 11 are used to calculate
the rejection again using Figure 3 as a basis. All rejections of toxicant
concentration are substantial as shown in Table 12. The toxic level of
each concentrate was 5 to 11 times higher than that of the feed, providing
consistent evidence of membrane separation. A mass ratio of the implied
concentration of supposed toxicant is presented in Table 13. Mass ratio is
the combined mass of solute in permeate and concentrate divided by the mass
of solute in the feed. A sample calculation is provided in Appendix B. The
results are reasonably consistent (mass ratio * 1), ranging from 0.65 to 1.55.
The rejections shown in Table 12 are of considerable interest. As
already mentioned the bioassay results are consistent in showing reduced
toxic effect in permeates and corroborating increased toxic effects in the
concentrate. The rejection of material toxic to the daphnids is uniformly
lower than that of toxic to Fathead minnows. The toxicant rejections are
opposite to the rejection of inorganic salts. That is, the dynamic membrane
produces superior separation to the cellulose acetate which is superior to
the poly (ether /amide) on toxic substances. By contrast the inorganic (salt)
rejection exactly counters the ordering. Simply stated this only means the
22
-------
Table 11
Lethal Concentration and Implied Toxicant Concentrations
96 Hour Minnows
48 Hour Daphnia
CJ
50
Sample LC_
Fluid and Type No. % Solution
Run 1
Dye-feed 10 9.7
Dye-PEA permeate 11 82
Dye-CA permeate 12 >100
Dye-concentrate 13 1.6
Run 2
Dye-feed 14 25
Dye-DM permeate 15 NAT*
Dye-concentrate 16 5.3
Run 3
Scour-feed
Scour-PEA permeate
Scour-CA permeate
Scour-concentrate
Run 4
Scour-feed 7 13
Scour-DM permeate 8 NAT*
Scour-concentrate 9 2.0
*NAT - no acute toxicity
**by Implied Concentrations in headings
Implied
Concentration
No Units**
8.5***
1.2
<1.
62.
4.
0.
19.
3
4
5
6
16
28
>100
1.5
6.
3.6
<1.
67.
Implied Concentration =
100
LC
50
7.7
0.0
50.
LC50
% Solution
33.5
60 to 100
60 to 100
4.1
49
80
17
26
53
42
5.1
25
>100
9.9
Implied
Concentration
No Units**
2.5***
1 to 1.7
1 to 1.7
24.
2.0
1.2
5.9
3.8
1.9
2.4
20.
4.
1.
***Values lowered due to concentration of sample removed for feed
-------
Scour Fluid
Membrane
Dynamic ZrO/PAA
Cellulose Acetate
Poly(ether/amide)
Dye Fluid
Membrane
Dynamic ZrO/PAA
Cellulose Acetate
Poly(ether/amide)
Data in this table are
TABLE 12. Rejection of Toxicity by Hyperfiltration
Daphnia Toxicant
>88
55
68
Daphnia Toxicant
60
62 to 82
62 to 82
obtained from the procedure
% Rejection
Fathead Minnow Toxicant
100
>92
60
Rejection
Minnow Toxicant
100
96
95
Fathead
C.R. =
Implied concentration of permeate (from Table 11)
Implied concentration of feed (from Table 11)
C. R. is the concentration ratio used as abscissa for Figure 3. The
reiection is read as the ordinate of Fioure 3.
TABLE 13. Mass Ratio3 of Toxicants
Run 1 Cast Membranes on Dye Fluid
Toxicant to
Fathead minnows
Daphnids
Run 2 Dynamic Membrane on Dye Fluid
Toxicant to
Fathead minnows
Daphnids
Run 3 Cast Membranes on Scour Fluid
Toxicant to
Fathead minnows
Daphnids
Run 4 Dynamic Membrane on Scour Fluid
Toxicant to
Fathead minnows
Daphnids
Mass Ratio (final/initial)
0.94
1.315
Mass Ratio (final/initial)
1.28
1.23
Mass Ratio (final/initial)
1.55
1.08
Mass Hatio ffinal/initial)
1.17
0.65
A mass ratio calculation example is shown in Appendix B.
membranes developed to achieve high salt rejection for desalination
applications do not necessarily have proportional rejections of toxic
(presumably non-electrolytic) compounds.
In an attempt to determine cause and effect, the toxicant concentration
profile from Table 11 may be compared with measured concentrations of
substances. Three of the best fit profiles are shown in Figures 4 through
6. The relative toxicant concentrations are shown for the Daphnia and
Fathead minnows as compared with total solids, arsenic, and copper in the
succeeding figures. None of the organic toxic pollutants has a concentration
pattern remotely similar to the bioassay results. Arsenic and total solids
shown patterns resembling the bioassay results, whilte copper fails badly
24
-------
FATHEAD
MINNOW
TOXICANT
DAPHNIA
TOXICANTV
SCOUR FLUID SAMPLE NUMBER
10
11 12 13 14
DYE FLUID SAMPLE NUMBER
15
FIGURE
RELATIVE CONCENTRATIONS OF TOXICANTS AND ARSENIC
25
-------
a:
FATHEAD
HINNOW
TOXICANT
567
SCOUR FLUID SAMPLE NUMBER
11 12 13 14
DYE FLUID SAMPLE NUMBER
15
16
FIGURE 5 RELATIVE CONCENTRATIONS OF TOXICANTS AND TOTAL SOLIDS
26
-------
FATHEAD
MINNOW
TOXICANT
DAPHNIA
TOXICANT-v
5 6 7
SCOUR FLUID SAMPLE NUMBER
99 A
15
16
DYE FLUID SAMPLE NUMBER
FIGURE 6 RELATIVE CONCENTRATIONS OF TOXICANTS AND COPPER
27
-------
SCOUR DYE
o ° 48 HOUR
• • 24 HOUR
20 30 40 50 60 70
CONCENTRATION G™) TO DAPHNIDS
LC50
90 100
FIGURE 7 CORRELATION OF CONCENTRATION TOXIC TO FATHEAD MINNOWS WITH
CONCENTRATION TOXIC TO DAPHNIDS
28
-------
for the samples 14 and 16. The run in which samples 14 and 16 were taken
contained a much larger copper content than any of the other runs and yet
did not show proportionally high toxic effects. The presence of a dye
containing complexed copper could account for this result. It is doubtful that
the low values of arsenic could be toxic. Therefore, no simple cause/effect
can be determined; and, further it is likely that one or more of the gross,
non-analyzed compounds served as toxicant assuming its separation reasonably
paralleled that of the metals or total solids.
A correlation of toxic concentration to Fathead minnows and to Daphnids
may be investigated. A plot of implied toxicant concentration for minnows
versus the concentration for daphnids is shown in Figure 7. Alternatively,
Figure 7 may be viewed simply as a plot of reciprocal LC5Q data. There is
a high correlation coefficient of 0.94 suggesting that for this fluid a
measurement of either individual bioassay would have supplied essentially
the same information. A plot of concentration at which no effects were
observed (reciprocal ECO) is similar but shows a far greater range and
scatter. The Daphnia were more sensitive than the minnows to the test
fluid, judged by nine pf fourteen values in Table 11.
Rat toxicity and bacterial mutagencity tests produced no effective
response. The concentrates from each run produced responses at about 90
percent dilution suggesting that the feed may also have been marginally
cytotoxic. Neither the feed nor permeates produced position cytotoxicity
results. Appendix C contains the detailed results.
Correlation of Rejection in Single-Solute Solutions with Solute Solubility
Parameter
Hyperfiltration rejection of organic nonelectrolytes in single-solute
has often been correlated with the molecular weight of the solute although
for low molecular weight compounds the correlation is sometimes poor,
especially for cellulose acetate membranes. The dependence of rejection on
solute solubility parameter has been demonstrated using published
hyperfiltration results. Appendix D describes the results of this
correlation.
If this correlation is satisfactory, or can be developed into a
reliable model, it would greatly reduce the experimental work required to
characterize the effectiveness of a membrane to reject toxic pollutants.
Rejections of a few solutes could be determined for a solution-membrane
system and the rejection of other solutes estimated.
29
-------
TEST DESCRIPTION
Fluid samples were obtained at the overflow of the first washer on the
Kttsters dye range at the La France Industries plant (see Figure 8). The
effluent was collected in a plastic pail fitted with a 40-meter rubber hose
connected to a 80£/min centrifugal type transfer pump. The pail and entire
hose had been previously used extensively with the fluids from the range.
Non-stainless steel parts of the pump hardware were replaced with stainless
steel. The pump was all stainless steel with ceramic seals. The fluid was
passed through a one-micron polypropylene cartridge filter. New filters were
used for the bleach) (scour) acquisition. The fluid line was purged before
each new fluid acquisition.
All fluid lines and wetted parts in the test system were Teflon, stain-
less steel or ceramic except one line having a rubber tube joining two steel
tubes in a non-flowing channel used as a connection to a suction pressure
protection device. The feed and permeate tanks were cleaned with a
commercial cleaner used to clean becks at La France. Following this the
tanks and the skid-mounted pump station were flushed thoroughly for one-half
hour in 1 M NaOH and rinsed with plant water, until no pH elevation was
present. The tanks were covered with new polyethylene film to assist in
vapor and volatile retention and to prevent entrance of the airborne lint.
Pressure, temperature, and flow to the membrane were controlled at the
skid mounted pump station. Conditions were maintained during the runs at the
values shown in Table 14. The range of pressure and temperature shown in
Table 14 was selected in the dynamic membrane tests to allow stable operation
at a rate to achieve a reasonable time to acquire samples. All values are
approximate and varied slightly from the conditions listed. The other
membranes were operated at conditions determined in concert with the
manufacturer.
TABLE 14. Operating Conditions Observed
Fluid
Dye
Scour
Dye
Scour
Membrane
PEA-CA
PEA-CA
Dynamic
Dvnamic
Temperature
(°C)
40
40
70
77
Outlet Flow
a/min)
16
16
16
16
Inlet Pressure
(MN/m3 )
2.8 (400 psi)
2.8 (400 psi)
4.5 (650 psi)
5.9 (850 psi)
The dye test fluids are the wash water obtained while using a dye pad
formulation for direct dyeing cotton. These dye pad formulations contain
30
-------
CLOTH FLOW
DYE
I PAD
JET WASHER
COLLECTION
PAIL
CARTRIDGE FILTER
SKID
MOUNTED
PUMP STATION
L,
APPROXIMATELY
40 METERS
FEED TANK
-fiB-
FEED
OR
CONCENTRATE
SAMPLE
MODULE 1 H MODULE 2
L
PERMEAT
BARREL
1,
PERMEATE
: $ SIPHON
PERMEATE
FANK
L
PERMEATE DRAIN
FIGURE 8 SCHEMATIC OF FLUID ACQUISITION AND OPERATIONS
-------
a thickener, dispersing-wetting agents, and the direct dyes. Typical tests
fluids have pH 6 to 8, conductivity 200-1,000 yS cm'1 and total solids
400-2,800 g/m3.
The scour test fluids are the washer effluents taken while the scour
pad contains hydrogen peroxide, sodium carbonate, and a disper sing-wetting
agent. The pH is typically 8 to 10. The fluid also contains size, motes,
and other materials washed from the cloth and usually dyes and auxiliary
chemicals remaining in the washers from the previous dyeing operation.
Table 15 shows the sequence of runs and events which apply to the test
operation. The operation was marred by taking a delayed feed sample (about
15 percent concentrated) on the first run and by the failure of the solder
joint on the scour run with the dynamic membrane. The module was readily
repaired but some contamination could have occurred in reconstitution of the
feed sample with a small gear-type, plastic transfer pump or in the materials
used for the repair itself.
Table 16 shows the time at which the various samples were collected,
shipped, and received. All samples were refrigerated as soon as praxrtical
after collection.
All samples were collected according to the sampling plan which is
included as Appendix E. Samples were withdrawn through stainless steel tubes;
±he use of plasticized tubing was avoided. All collection barrels were
stainless steel.
32
-------
Table 15
Summary Log of Activities
Date Time Activity
6/01/78 1430 Obtain "clear" water sample from range: 280fc at 60°C
drawn through 1 micron polypropylene filter. Previous
dye formula was 9127.
6/01/78 1450 Operate membranes (PEA and CA) at 300 psi. 53°C feed
cooled at 46°C by heat exchanger.
6/01/78 1500 Stop, apparatus blank run. Take sample half from
permeate, half from concentrate.
6/01/78 1545 Drain all tanks.
6/01/78 1600 Obtain dye batch: dye formula 9204. 654X,. Allow to
cool overnight.
6/02/78 1114 Start unit. Discard first liter of product.
6/02/78 1140 Stop unit at 15.6% recovery. Obtain slightly concen-
trated feed sample.
6/02/78 1215 Resume operation.
6/02/78 1312 Stop unit, obtain samples at 90% recovery. Drain
tanks.
Obtain batch and feed sample from dye formula 1211,
partly unfiltered batch. Install dynamic membrane
0.3 m2. Start operation at 4MN/m2 (580 psi) with poor
rejection of color.
Return permeate in clean glass bottle to feed.
Permeate has cleared.
6/11/78 2215 Stop small leak from plumbing.
6/12/78 0730 Stop operation, obtain samples. Approximate 80%
recovery.
6/12/78 0830 New polypropylene feed filter installed. PEA and CA
membranes connected after flushing. Scour feed batch
obtained of 429£. Sample taken.
6/12/78 1107 Start run on scour.
6/12/78 1213 Stop run at 83% recovery. Obtain samples.
6/12/78 2000 Obtain scour batch for dynamic membrane; 465£.
6/13/78 1000 Connect dynamic membrane, gather feed sample, start
operation.
6/14/78 0400 Module failure - soldered joint failed.
6/14/78 0800 Repair module.
6/14/78 1130 Return fluid to feed using 2m vinyl hose and plastic
gear pump. Restart test.
6/17/78 2300 Stop test at 80% recovery, obtain samples.
6/18/78 0100 Obtain plant water blank sample.
6/09/78 1100
6/09/78 2230
33
-------
TABLE 16. Sample Disposition Log
OJ
*>
Sample
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Date Shipped
Date - Hour Taken
6/18 -
6/01 -
6/12 -
6/12 -
6/12 -
6/12 -
6/12 -
6/17 -
6/ -
6/02 -
6/02 -
6/02 -
6/02 -
6/09 -
6/12 -
6/12 -
0100
1500
0830
1300
1300
1300
1000
2300
2300
1200
1300
1300
1300
1200
0730
0730
Chemical
6/19
6/05
6/13
6/13
6/13
6/13
6/13
6/19
6/19
6/05
6/05
6/05
6/05
6/13
6/13
6/13
Fish
_
-
6/14
6/20
6/20
6/14
6/14
6/20
6/20
6/08
6/08
6/08
6/08
6/14
6/14
6/14
Rat
_
-
6/19
6/19
6/19
6/19
6/19
6/19
6/19
6/19
6/19
6/19
6/19
6/19
6/19
6/19
Date Received
Chemical
6/20
6/06
6/14
6/14
6/14
6/19a
6/14
6/20
6/20
6/06
6/06
6/06
6/06
6/14
6/14
6/14
Fish
_
-
6/15
6/21
6/21
6/15
6/15
6/21
6/21
6/09
6/09
6/09
6/09
6/15
6/15
6/15
Rat
_
-
6/20
6/20
6/20
6/20
6/20
6/20
6/20
6/20
6/20
6/20
6/20
6/20
6/20
6/20
aNote length of time between date shipped and date received.
-------
REFERENCES
1. G. D. Rawlings and Max Samfield, "Source Assessment: Textile Plant
Wastewater Toxics Study Phase I, EPA 600/2-78-004h, March, 1978.
2. "Dyes and the Environment," ADMI Report, Volume II, September, 1974.
3. C. A. Brandon, J. J. Porter, and D. K. Todd, "Hyperfiltration for
Renovation of Composite Wastewater at Eight Textile Finishing Plants,"
Final Report, EPA Grant 802973.
35
-------
APPENDIX A
Infrared Spectra of Sample and Process Chemical Residues
Infrared spectra were obtained of the evaporation residues of the
hyperfiltration solutions, i.e., feed, permeate, and concentrate; the scour
chemicals; and the auxiliary dye bath chemicals. A measured volume of each
solution was evaporated to apparent dryness in an oven at ca. 105°C. The
larger residues were scraped from the evaporating dishes and stored in vials.
The permeate residues were quite small and firmly attached to the evaporating
dishes, so they were softened with a drop or two of water and the slurry
scraped into a mortar and the water evaporated again by placing the mortar
in the oven. The spectra were obtained with a Perkin-Elmer 317 infrared
spectrophotometer using the KBr pellet technique. In the case of permeate
samples, the KBr was added to the mortar and ground to a fine power to
incorporate the residue in the pellet.
Table Al identifies the samples and describes the appearance of the
residues. A film like material was observed in some residues, presumably
composed of the high molecular weight thickener and/or size removed by the
scour. This observation is identified by the film notation. Table A2
identifies the process chemicals, other than dyes.
TABLE Al. Hyperfiltration Samples and Residues Characteristics
Total
CTHF Solids Absorbance
No. Identification (mg/m^) 410 m
1 Plant Water 15,000 0
2 Apparatus Water 43,000 .005
3 Scour-1, feed 730,000 .050
4 Scour-1, PEA permeate 105,000 0
5 Scour-1, CA permeate 32,000 0
6 Scour-1, Concentrate 6,020,000 .50
7 Scour-2, feed 870,000 .03
8 Scout-2, EM permeate 205,000 .01
9 Scour-2, concentrate 3,840,000 .15
10 Dye-1, feed 462,000 .10
11 Dye-1, PEA permeate 15,000 0
12 Dye-1, CA permeate 45,000 0
13 Dye-1, concentrate 2,670,000 .65
14 Dye-2, feed 76,000 2.0
15 Dye-2, DM permeate 60,000 0
16 Dve-2. concentrate 2.160.000 7.75
Description
of
-Residue
Not determined
Brown powder
Light yellow powder.
Colorless deposite
Colorless deposite-
Light brown, film
Cream powder
Colorless powder
Light brown, film
Green-brown particles
Colorless deposite
Colorless deposite
Dark brown, film
Dark red, powder and
film
Slightly pink powder
Dark red, fii™
36
-------
TABLE A2. Process Chemicals
Identification
Sodium carbonate
Hydrogen peroxide
Size
Thickener
Dispersing
Wetting Agent-1
Dispersing
Wetting Agent-2
Description
Colorless solution
Colorless solution
Colorless powder
Yellow powder
Brown-or ange
solution
Yellow solution
Occurrence in Process
Scour bath
Scour bath
May wash off cloth in scour
Dye bath
Dye bath
Dye bath and scour
The infrared spectra are presented in Figures Al - A22.
The hyperfiltration solutions are multicomponent and the infrared spectra
of their residues are complicated. Little information about the relative
passage of the components through the hyperfliters is obvious. The spectra
have been analyzed using two simple methods. First, the relative absorbances,
AA, of the strongest three peaks are compared for each hyperfiltration
experiment. Selectivity of the membranes with respect to the ir-active
components is indicated if the relative absorbance of the peaks differ in
the feed and permeate and/or feed and concentrate. The results of this
analysis are provided in Table A3. The comparison of Ag.o/A?.! for the scour
experiments and Ag. ]/A7 _ 0 for the dye experiments indicates membrane
selectivity of the ir-active components. The observed appearance and
disappearance of other peaks also indicated selectivity.
TABLE A3 . Relative Absorbance of Strong Infrared
CTHF
No.
10
11
12
13
14
15
16
3
4
5
6
7
8
9
Identification
Dye-1, feed
Dye-1, PEA permeate
Dye-1, CA permeate
Dye-1, concentrate
Dye-2, feed
Dye-2, DM permeate
Dye-2, concentrate
Scour-1, feed
Scour-1, PEA permeate
Scour-1, CA permeate
Scour-1, concentrate
Scour-2 , feed
Scour-2, DM permeate
Relative
A9.0/A7.1
1.8
1.6
(essentially
2.8
3.3
1.0
4.9
1.5
2,6
(essentially
1.7
WA7.0
1.5
.8
1.4
Absorbance
A6.2/A7.1
0.90
1.0
KBr spectrum)
1.0
A6.2/A7.1
1.6
.78
2.0
A6.2/A7.0
.84
.72
KBr spectrum)
.81
A6.2/A7.0
.71
.4
1.2
Maxima
Comments
shift in 7.1 peak
shift in 7 . 0 peak
37
-------
4000 3000
7 8 9 10 11
WAVELENGTH (MICRONS)
FIGURE Al CTHF-2 RESIDUE
-------
4000 3000
100
2000
1000 900
800
700
U)
VD
7 8 9 10 11
WAVELENGTH (MICRONS)
12
13
14
15
. FIGURE. A2 SCOUR-L FEED RESIDUE/ CTHF~3
-------
4000 3000
.... 1111 •
100
900
800
700
3 4 5 6 7 8 9 10 11
WAVELENGTH (MICRONS)
14 15
FIGURE A3 SCOUR-L PEA PERMEATE RESIDUE/ CTHF-4
-------
4000 3000
100
1000 900
800
700
7 8 9 10
WAVELENGTH (MICRONS)
11
12
13
14 15
FIGURE AJ| SCOUR-L CA PERMEATE RESIDUE/ CTHF~5
-------
4000 3000
900
800
700
7 8 9 10 11
WAVELENGTH (MICRONS)
12
13
14 15
FIGURE A5 SCOUR-L CONCENTRATE RESIDUE/ CTHF-6
-------
4000 3000
.. -ii 111 i i I i
100
2000
1500
CM
1000 900
800
700
11100
7 8 9 10
WAVELENGTH (MICRONS)
11
12 13 14 15
FIGURE A6 SCOUR-2, FEED RESIDUE/ CJHF-7
-------
4000 3000
2000
1000 900
800
700
0
7 8 9 10
WAVELENGTH (MICRONS)
15
FIGURE A/ SCOUR-2, DM PERMEATE/ CTHF-8
-------
4000 3000
100
1000 900
800
700
7 8 9 10 11
WAVELENGTH (MICRONS)
12
13
14
15
FIGURE A8 SCOUR-2/ CONCENTRATE, CTHF~9
-------
WAVELENGTH (MICR
700
FIGURE A9 DYE-L FEED, CTHF-10
-------
1000 900
i i
800
700
7 8 9 10 11
WAVELENGTH (MICRONS)
FIGURE AlO DYE-L PEA PERMEATE, CTHF-11
-------
4000 3000
800
700
7 8 9 10 11
WAVELENGTH (MICRONS)
12 13 14 15
FIGURE All DYE-L CA PERMEATE/ CTHF-12
-------
4000 3000
100
1000 900
800
700
0
7 8 9 10 11
WAVELENGTH (MICRONS)
12
13
14
15
FIGURE A12 DYE-L CONCENTRATE, CTHF-13
-------
4000 3000
2000
1000 900
800
700
7 8 9 10 11
WAVELENGTH (MICRONS)
12
13
14
15
FIGURE Al3 DYE-2, FEED/ CTHF-J4
-------
4000 3000
1000 900
800
700
Ul
5 ' 6
7 8 9 10 11
WAVELENGTH (MICRONS)
12
13 14 15
FIGURE AW DYE-2, DM PERMEATE/ CTHF~15
-------
4000 3000
700
4
7 8 9 10
WAVELENGTH (MICRONS)
11 12 13 14
FIGURE A]5 DYE-2, CONCENTRATE/ CTHF-16
-------
WAVELENGTH^ (MICRONS)
12
13 14
100
15
IOC
4000 3000
2000
1500
1200
CM'1
1000 900
800
700
FIGURE Al6 N*2C03 RESIDUE
-------
4000 3000
800
700
7 8 9 10 11
WAVELENGTH (MICRONS)
12 13 14 15
FIGURE A17 NaHC03 POWDER
-------
Ul
100
--80
Z60
£
§4ft|
20
I WAVELENGTH (MICRONS)
3 ' 4 5 6 7 8 9 10 11 12
i i i i i i i i I _i i < i i i i i i I
13
14
15
4000 3000
2000
1500
1200
CM"'
1000 900
800
700
FIGURE A18 SIZE/ POWDER
-------
100
6
WAVELENGTH (MICRONS)
11
12
13
14 15
i I .1
4000 3000
2000
1500
1200
CM1
1000 900
800
700
FIGURE A19 THICKNER/ POWDER
-------
4000 3000
900
800
700
100
0
7 8 9 10 11
WAVELENGTH (MICRONS)
12
13
14
15
FIGURE A20 DISPERSING-WETTING AGENT-L RESIDUE
-------
4000 3000
2000
1500
1000 900
800
700
100
U1
00
7 8 9 10 11
WAVELENGTH (MICRONS)
12
13
14
15
FIGURE A£L DISPERSING-WETTING AGENT-2/ RESIDUE
-------
4000 3000
900
800
700
0
7 8 9 10
WAVELENGTH (MICRONS)
11
12
13
14
15
FIGURE A22
PELLET
-------
A second analysis was attempted. The absorption peaks in the spectra of
the hyperf iltration solution residues were compared with three selected peaks
in the spectra of the process chemicals. If absorption peaks were found to
match the peaks of all those selected for a process chemical it is listed as
present. If more matching peaks are present than are absent for a process
chemical, it is listed as possibly present.
Table A4 lists evidence for the presence of the process chemicals in the
hyper filtration solutions residues. Hydrogen peroxide is not expected to
occur in the residue even if present in the solution. The chemicals
evaluated are carbonate and bicarbonate, size, thickener, dyes, and two
dispersing/wetting agents. Color, indicating dye presence in residue, is
denoted by/ C. Visual evidence, e.g., film formation in the. residue
indicating the presence of either or both the high molecular weight size and
thickener, is indicated by R. If the formulation indicates the chemical's
presence, F is used. Presence indicated by the ir matching-peaks analysis
is ir and (ir) , representing the presence and possibly present categories.
TABLE A4. Presence of Components in the Hyperf iltration Solution Residues
CTHF No.
10
11
12
13
14
15
16
3
4
5
6
7
8
9
Carbonate/
Bicarbonate
(ir)
F,ir
ir
F,ir
ir
ir
Size
R
R
R,(ir)
ir
ir
R,(ir)
Components
Thickener
F,ir
(ir)
*
R,ir
F,ir,R
R,ir
(ir)
R,(ir)
(ir)
R,ir
DW-1
(ir)
(ir)
F
(ir)
DW-2
F,ir
ir
ir
F
F,ir
ir
(ir)
Dyes
F,C
C
F,C
*A component in the KBr has a sharp peak at 7.3 ym, this peak is observed.
C - color in residue indicates dyes.
R - observation in residue of a film, indicating presence of either size
or thickener, or both.
F in both formulations, ir-presence. (ir)-presence possible.
The cellulose acetate membrane appears to reject most of the ir-active
components of both the scour and the dye feeds. The poly(ether/amide) and
the dynamic membranes show selectivity with respect to ir-active components.
The dyes and the high molecular weight thickener and size appear to be
highly rejected. Rejection of the dispersing/wetting agents may not be as
effective.
60
-------
APPENDIX B
Interpretation of Results
The experiment involved concentrating an initial volume while producing
a permeate volume. Therefore, it is necessary to appropriately interpret the
results to estimate the values of the rejection. In the following discussion
M symbolizes the mass of fluid in the concentrate flow, m symbolizes a mass
flow rate, C symbolizes the mass concentration of solute, t symbolizes time,
and y symbolizes the rejected fraction of solute. Only values for the mean
value of rejection may be calculated. Subscripts are used as follows: "c"
pertains to the concentrate, "f" to the feed, "p" to the permeate, "e" to
evaporation, and "1" to leak.
The initial mass, Mf, is depleted in general by evaporation, leaks (if
applicable), and permeation. The following equation is expected to apply for
two membranes.
— =-m_m_- ^
Integration yields an expression for the mass at any time
M(t) = Mf - Q^ (me + n^ + mpl + mp2)dt (B2)
When t becomes the elapsed time for the experiment, the corresponding M value
becomes MC/ the concentrate mass. Separate observations of M(t), measured
as fluid depth during the experiment, allow an estimate of the value of me
(evaporation rate). The absolute measurement of Mf and other values is un-
certain due to ignorance of the volume of pumps, fittings, modules.
Corrections may be applied to promote the integrity of the volume estimate
.based on relative values of concentrate and feed data provided by analysis.
The volumes recorded during operation of the test procedure are shown in
Table Bl. The initial volumes and concentrate volumes were obtained by
measuring the lev.el in the tank (top of tank to fluid level) . To the value
thus obtained was added 20 dm3 to account for the internal volume of pipes,
etc. The permeate volumes were obtained by measurement of fluid level in the
containers and by integration of the permeate rates observed. The leak in
Run 2 was measured in terms of its duration and rate. The vaporized volume
is simply the volume required to close the fluid balance. Runs 1 and 3 show
the permeate volume as the sum of two numbers which are, respectively, the
PEA permeate and CA permeate. All values in Table Bl are subject to errors
61
-------
in observation through at least the following mechanisms: (1) Poor approxi-
mation in system hold up volume, (2) tanks not exactly level, (3) ordinary
measurement of length uncertainty, and (4) difficulty with foaming fluid
level sensing. Therefore the use of total solids measurements to ijnprove the
volume estimates has been employed. The following describes the methodology
of calculating the values of solute in the leak fluid and the determination
of the permeate volume fraction from concentration data. An equation for the
concentration of a particular solute may be written based on a differential
mass balance, using y to symbolize rejection:
- Y2)C
(B3)
Evaporation has been deleted from this equation by assuming that the solute
is non-volatile. Expanding d(MC) to CdM + MdC and substitution from equation
(Bl) for dM/dt gives
meC.
TABLE
Initial Volume (dm3)
Concentrate Volume (dm3)
Permeate Volume (dm3)
Leak Volume (dm3)
Vaporized Volume (dm3)
„ feed-concentrate
Bl. Recorded Volumes
Run 1 Run 2
593 371
90 60
282 + 221 214
0 29
0 68
n QAa r> QTQ
Run 3
429
73
188 + 168
0
0
n HOA
Run 4
465
65
300
0
100
n o<;n
feed
Division by MC renders the variables separated if y is independent of C
Y m
+ m
M
dt
(B4)
Substitution of equation (B2) for M allows straightforward.integration, which
must be done numerically except for special cases. One important special
case has no leak (ra^ = 0) , a neglibible evaporation rate, and constant
permeate rates. Equation (B4) after substitution of equation (B2) for M
yields
,'m
y2m
Y m + y m
- mpl
t -
V*
dt =
6.2
-------
Ylmpl +
Cc
Cf M - M - M2 (B5)
In equation (B5) M.^ and M are the permeate total mass associated with mem-
branes 1 and 2, respectively. Equation (B5) holds for one membrane as well
and is also valid for non-constant flow rates if the rates are proportional.
A global mass balance equation may be written as
- MfCf - McCc + MlEpl + M2S2 (B6)
The overbar designates mixed average permeate. Division by Cf , substitution
of equation (B5) for Cc/Cf and substitution of MC = Mf - Mj_ - M2 leads to
C . C R (1 - yJ + Rod - Y2)
pi r>2 • 1 _ 1 _ £ _ £_
Rl Cj- + R2 C^ = 1 - (1-R> R (B7)
where R^ = M^/M^, R2 = M2/Mf ' and R = Ri + Ro- R is commonly called the
recovery.
Equation (B5) may be used to calculate the average rejection of the two
membranes based on the recovery and chemical analysis of concentrate and
feed. Equation (B7) may be used to calculate the rejection based on analysis
of permeate and feed. An auxiliary equation permits the calculation of
1 - n = Si CBS)
1 - Y2 Cp2
rejection for either membrane by itself. Ideally at the recovery value
observed the solute mass balance (B6) is satisfied and the calculated
rejections from (B5) and (B7) are identical. This never happens precisely
due to experimental uncertainty. In the interpretation herein calculations
of mass balance in terms of ratio -of the right side to left side of equation
(B6) , rejection based on permeate and feed data have been made. These
calculations have been made at various levels of recovery near the actual
recovery noted in testing. The results for total solids determination
have been employed for this exercise. Table B2 shows a typical result for
the fourth run (scour fluid with dynamic membrane) . Using volume observa-
tions the best estimate of recovery (vapor + permeate) /feed was 0.86 while
the recovery which yields essentially unity for the mass balance ratio (right
side divided by left side of equation (B6) ) was 0.82 (less than 4% different).
In this case the use of the best mass balance yields good agreement between
the rejection estimates and the recovery of 0.82 is adopted as the best
estimate of actual volume recovery. The values of rejection calculated from
permeate and feed data differ by less than 1 percent, while the rejection
calculated using concentrate and feed data differ by over 10 percent. When
63
-------
differences occur in the rejection estimated from concentrate and feed
compared to the rejection estimated from permeate and feed, the value of
rejection should be estimated on the basis of permeate and feed data due to
the reduced sensitivity to experimental uncertainty.
TABLE B2. Effect of Recovery on Mass Balance and Rejection
Recovery with
Based on Volumes Best Mass Balance
Recovery 0.860 0.820
Mass Balance Ratio (final initial) 0.862 1.0
Rejection (using concentrate) 0.783 0.866
Rejection (using permeate) 0.882 (K87J5
In each of the four runs a calculation similar to that described for run
4 was made. In Runs 1 and 3, an improved recovery estimate using total
solids data was found to agree reasonably well with the preliminary volume
estimates. In Run 2 the total solids data do not allow a reasonable change
in the preliminary estimate and which forces a solute mass balance. The best
interpretations for the exact recovery are presented in Table 2 of the main
text of the report. As shown in the table, 26 percent of the initial mass
in Run 2 could not be accounted for in the sum of permeate and concentrate.
Recheck of solids analysis shows no change and the conductivity data tend to
collaborate the preliminary volume estimates so that no substantial improve-
ment of the recovery estimate can be made. Run 2 was complicated by a leak
and evaporation which tend to increase the difficulty in interpretation.
A computer program was prepared for detailed analysis of the specific
solute analytical data. Upon receipt of data and execution of the program
it was apparent that such an exercise would not be meaningful. For example,
in Tables 4 through 7 few results for mass balance ratio were near unity.
Therefore for the metals, a simple rejection calculation was adopted based
on use of permeate and feed data. The concentrate and feed comparison is
much more sensitive; i.e., errors in analysis or recovery estimated are
amplified in rejection estimate. Only in cases where the permeate and feed
data may not yield clearly defined results due to low concentrations will the
concentrate analysis be important. In such a case, the presence of concen-
trated material in the concentrate stream indicates rejection has occurred.
For interpretation of organic solute data, depletion in the permeate
together with enrichment in the concentrate yield confidence in estimating a
substantial rejection of solute. Many solutes have such results and others
have conflicting indications. Those which conflict by having one indication
of rejection and another of no or negative rejection will be in violation of
the mass balance. The more likely erroneous datum may be selected from such
logic. The relatively unsatisfying statements "probably rejected", "probably
not rejected", or "mixed indications" are really all the information that can
be gleaned.
Following are a group of comments pertinent to the interpretation of the
64
-------
bioassays. A single toxicant is presumed to have a lethal fraction versus
concentration curve F(C) as illustrated in the sketch below.
1-
C5Q C
The curve shows no lethal effect below some minimum concentration rising to
complete mortality at a higher concentration. The value of C = C^Q will
produce lethal effects in half the subjects. If two lethal species are pre-
sent the lethal fraction F may be determined on the basis of the individual
components. Let F-^(C^) be the lethal distribution curve for specie 1 and
F2(C2) that for specie 2. Those dying from exposure to specie 1 will be
F (C ) of those not dying from specie 2. Those dying from specie 2 will be
F2(C2^ of tnose not dying from specie 1. If PX and P2 represent the frac-
tions killed by -specie 1 and 2 respectively,
Pi = (l-P2)F1(Ci)
P2 = (l-p1)F2(C2)
without synergistic effects, the total fraction killed is the sum of these,
or F is
P1^C1) ~ F2(C2} F2 - Vci> [FI(C!) -F2(C2)12
F = __ _ + _ - + -
1 - F (C2) 1 - F^C^) [l-F2(C2)][l-Fi(Cj)]
If the relative amounts of specie 1 and 2 are the same, or if F± and F2 are
identical functions, then F(C1+C2) wil 1 behave exactly as a single component.
Similar results are expected from situations with three or more components.
In many but not all hyperf iltration systems the rejections of individual
toxicants will not be largely different from each other so that the relative
concentrations of substances will be preserved, it is not unexpected then
for mixtures of toxicants to behave as a single toxicant (even with
synergistic effects) .
The action of hyperf iltration on a single toxicant is expected to
produce a dilute and concentrated stream. Their permeate stream has volume
R at concentration Cp whereas the feed stream has unit volume at concen-
tration G£. The concentrate will have volume (1-R) and the concentrate
concentration, Cc, according to: mass balance information will be
1 - R(C/C,)
When each stream is subjected to bioassay a set of dilutions is determined
which produce effects on half the population. These dilutions are 6p, 6f ,
and 6C for the permeate, feed, and concentrate, respectively. The dilution
6 is the fraction of sample in a unit of total fluid, so that 6 = LCso- One
expects the single solute to produce a medium effect at a concentration, CC
independent of permeate or feed or concentrate source
65
-------
'50
5pCp =
Solving for each individual concentration value
CP =
'50
= C50
(BIO)
Cf '
C50
The concentration of toxicant is seen to be inversely proportional to the
value of 6 (6 = LCsg)• Substitution of each value from equations (BIO) into
relation (B9) yields
^f_ = 1 - R(6f/«p)
1 - R
(Bll)
Equation (Bll) is a kind of mass balance for the toxicant. Written as a
ratio of solute mass in permeate and concentrate to solute mass in feed the
mass balance ratio is
M.B.R. = 6,(R/6 + [l-R]/6 )
f p p
(B12)
Equation (Bll) is useful in predicting the value of 6C knowing 6p, 6f, and R
while the ratio in equation (B12) is useful in evaluating the internal con-
sistency of the data for all parameters. As noted in the report the mass
balance ratios were - 60 percent of unity, which is felt to be very reason-
able for biological assay data.
Example for calculation of mass balance ratio for Run 1 with Fathead minnows.
feed
PEA permeate
CA permeant
concentrate
(Table 11)
9.7
82
>100
1.6
Concentration
(Table 11)
10.
1.2
62.
Volumes
1
0.379
0.484
0.137
mass of toxicant in concentrate 0.137 x 62 = 8.494
in PEA permeate 0.379 x 1.2 = 0.4548
in CA permeate 0.484 x (<1) = 0.48
Total end of run = 9.43
66
-------
in feed 1 x 10 =10
9.4
mass balance ratio = ^Q = 0.94
this value is shown as the first entry in Table 13.
67
-------
APPENDIX C
EVALUATION OF HYPERFILTRATION
TREATED TEXTILE WASTEWATERS
by
G. D. Raw!ings
Monsanto Research Corporation
1515 Nicholas Road
Dayton, Ohio 45407
Contract No. 68-02-1874
ROAP No. 21AXM-071
Program Element No. 1AB015
1 November 1978
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
68
-------
SECTION 1
INTRODUCTION
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. versus Train) requiring EPA to accelerate develop-
ment of effluent standards for 21 industrial point sources in-
cluding textile-manufacturing. Among other requirements, the
Court's mandate focused federal water pollution control efforts
on potentially toxic and hazardous chemical compounds. The con-
sent decree required that "65 classes" of chemical compounds be
analyzed in wastewater samples. Recognizing the difficulty of
analyzing for all chemical species present in each category of
compounds, EPA developed a surrogate list of 129 specific com-
pounds representative of the classes of compounds listed in the
consent decree. These compounds are referred to as "priority
pollutants."
The consent decree obligates EPA to identify which priority pollu-
tants are present in industrial wastewaters and to determine the
ability of various wastewater treatment technologies to remove
priority pollutants. It is the second item above to which this
project is directed. Under EPA Grant No. R805777, Clemson Uni-
versity is evaluating the ability of a hyperfiltration unit to
treat textile manufacturing wastewaters. Samples of two waste-
water feeds and hyperfiltration permeates and concentrates using
three types of membranes were sent to Monsanto Research Corpora-
tion (MRC) for priority pollutant analysis and bioassay testing.
The following bioassay tests were performed to evaluate the
reduction in toxicity by hyperfiltration of wastewater: Ames
mutagenicity and cytotoxicity tests, and fathead minnow, daphnia,
and 14-day rat acute toxicity tests.
This report discusses the analytical and bioassay procedures used
by MRC and its subcontractors and the results of the analyses.
69
-------
SECTION 2
SUMMARY
Under EPA Grant No. R805777, researchers J. L. Gaddis and
H. G. Spencer at Clemson University are evaluating the effective-
ness of hyperfiltration to cleanup various textile plant waste-
waters for discharge and possible recycle of chemical feedstocks.
The skid-mounted hyperfiltration unit was field tested on two
types of wastewater (scour bath and dye waste) at a textile woven
fabric finishing plant. Three types of hyperfiltration membranes
were tested: polyether amide (PEA), cellulose acetate (CA), and
dual-layer hydrous zirconium oxide (ZrO)-polyacrylate (PAA) dy-
namic membrane.
A total of 16 wastewater samples consisting of the hyperfiltration
feed, permeate, and concentrate were sent to MRC for priority
pollutant analysis and bioassay testing. The sample coding sys-
tem and corresponding description of the sample collected is
shown in Table 1. MRC performed the priority pollutant analyses,
Ames mutagenicity test, and cytotoxicity test (using Chinese
hamster ovary - CHO cells). Fathead minnow and daphnia acute
toxicity tests were performed for MRC by BG&G Bionomics Marine
Research Laboratory. The 14-day rat acute toxicity tests were
performed for MRC by Litton Bionetics.
Results of the analysis of 16 wastewater samples and a reagent
blank for the presence of the 114 organic priority pollutants are
shown in Table 2. Analysis of the data indicates no organic
priority pollutants are introduced due to the sample workup pro-
cedures or analysis contamination at MRC. Samples CTHF-1 and
CTHF-2 were samples of the textile plant intake water and hyper-
filtration unit rinse. Analyses of these samples indicate that
possibly chloroform and toluene are introduced from these two
sources.
In addition to the organic priority pollutant species, several
other organic compounds were detected in the wastewater samples.
Triphenyl phosphine and triphenyl phosphine oxide were detected
in all wastewater samples (except CTHF-6) and in the reagent
blank sample. These compounds probably result from glass clean-
ing detergents and were introduced from the sample containers and
laboratory glassware. Other organic compounds detected include:
70
-------
TABLE Cl. SAMPLE CODING SCHEME AND DESCRIPTION
OF SAMPLE COLLECTED
Sample Description
CTHF-1 Plant water
CTHF-2 Apparatus water
CTHF-3 Scour-1, feed for PEA and CA hyperfiltration
CTHF-4 Scour-1, permeate from PEA hyperfiltration
CTHF-5 Scour-1, permeate from CA hyperfiltration
CTHF-6 Scour-1, concentrate from PEA and CA hyperfiltration
CTHF-7 Scour-2, feed for DM hyperfiltration
CTHF-8 Scour-2, permeate from DM hyperfiltration
CTHF-9 Scour-2, concentrate from DM hyperfiltration
CTHF-10 Dye-1, feed for PEA and CA hyperfiltration
CTHF-11 Dye-1, permeate from PEA hyperfiltration
CTHF-12 Dye-1, permeate from CA hyperfiltration
CTHF-13 Dye-1, concentrate from PEA and CA hyperfiltration
CTHF-14 Dye-2, feed for DM hyperfiltration
CTHF-15 Dye-2, permeate from DM hyperfiltration
CTHF-16 Dye-2, concentrate from DM hyperfiltration
a-terepineol, 2-mercaptobenzthiazole, l-cyano-2-benzyloxy ethane,
benzthiazole, lauric acid, myristic acid, palmitic acid, and
stearic acid. Results of the priority pollutant metals analysis
for the 16 wastewater samples are shown in Table 3. Three pri-
ority pollutant metals (mercury, selenium, and thallium) were not
analyzed in this program because previous research indicated the
absence of these metals in textile plant wastewaters.
Because of the metals analytical technique used, 16 other trace
metals were analyzed in the samples: aluminum, barium, boron,
calcium, cobalt, iron, magnesium, manganese, molybdenum, phos-
phorus, silicon, sodium, strontium, tin, titanium, and vanadium.
Results of the phenol (total) and cyanide (total) analyses are
also shown in Table 3.
Fourteen of the sixteen wastewater samples (excluding CTHF-1 and
CTHF-2) were subjected to a battery of bioassay tests to determine
the reduction in toxicity by application of hyperfiltration to
various wastewaters. MRC performed the Ames mutagenicity test
and CHO cytotoxicity test on the samples. MRC directed Clemson
University to ship samples to EG&G Bionomics Marine Research
Laboratory, Wareham, Massachusetts, for freshwater static acute
toxicity tests using fathead minnows (Pimephales promelas) and
daphnids (Daphnia magna). Samples were likewise sent to Litton
Bionetics, Kensington, Maryland, for 14-day rat acute toxicity
testing.
71
-------
TABLE C2. ORGANIC PRIORITY POLLUTANT SPECIES DETECTED IN SPECIFIC WASTEWATER STREAMS
-4
NJ
Blank
Organic compound water CTHF-1
Bis ( 2 -ethylhexyl) phthalate 1.1
Dimethyl phthalate
Di-n- butyl phthalate 0.4
Butylbenzyl phthalate
Diethyl phthalate 0.3
Ac enaphthene
Anthracene
F luor anthene
Pyrene
Naphthalane
Phenanthrene
Phenol
Chloroform 58
Toluene 3
Tr ichloroe thyl ene
Benzene
Chlorobenzene
Ethylbenzene
Methylene chloride 6
Concentration in stream
CTHF-2 CTHF-3
9 9
18
4
7
2
0.4
1
0.9
31 18
22 0.8
0.3
2
34 5
CTHF-4
3
9
1
0.8
0.5
2
18
15
1
6
CTHF-5
3
3
22
29
0.4
1
0.7
5
CTHF-6 CTHF-7 CTHF-8
-9
3
7
2
13
34
41 0.8 0.7
5 2
6 2
21
15 4 5
(continued)
Note.—Blanks indicate compound is below detection limits.
-------
TABLE C2 (continued)
Concentration in stream
Organic compound
Bis (2-ethylhexyl) phthalate
Dimethyl phthalate
Di-n-butyl phthalate
Butylbenzyl phthalate
Diethyl phthalate
Acenaphthene
Anthracene
Fluor an thene
Pyrene
Naphthalene
Phenanthrene
Phenol
Chloroform
Toluene
Trichloroethylene
Benzene
Chlorobenzene
Ethylbenzene
Methylene chloride
CTHF-9 CTHF-10
4
55
1
3
0.6
0.8
1 0.2
19
0.5 10
0.7 2
2 5
CTHF-11
31
45
0.8
1
0.8
0.7
31
11
0.6
0.4
45
CTHF-12 CTHF-13
3 51
290
6
7
7
3
0.4 1
4
24
1
4 14
CTHF-14
2
170
1
3
0.7
0.1
0.8
0.2
96
0.6
0.6
5
CTHF-15
1
4
1
0.05
0.1
0.4
1
3
CTHF-16
4
1
1
3
Note.—Blanks indicate compound is below detection limits.
-------
TABLE C3.
CONCENTRATION OF PRIORITY POLLUTANT METALS, PHENOL,
AND CYANIDE DETECTED IN SPECIFIC WASTEWATER STREAMS
Species
Priority pollutant
metal species :
Antimony
Arsenic
Beryllium
Cadmium
Chromium
Copper *
Lead
Nickel
Silver
Zinc
Other species :
Phenol (total)
Cyanide (total)
Detection
limit
10
2
0.04
2
4
4
22
36
5
1
1
1
Concentration in stream
CTHF-1 CTHF-2 CTHF-3 CTHF-4
12
<1
-
5
540
54
168
-
-
630
-
1
<7
30
<1
-
9
840
200
240
154
24
616
33
4
100
19
-
15
640
90
380
132
42
520
6
<4
90
<1
-
15
720
26
250
70
26
360
12
72
CTHF-5
132
1
-
14
620
32
340
100
42
8,600
16
30
CTHF-6 CTHF-7 CTHF-8
436
160
-
48
1,260
760
760
480
114
3,540
_a
a
170
35
-
16
760
94
400
200
62
460
4
<4
146
5
-
20
800
414
210
78
248
<1
<7
(continued)
Sample arrived at MRC 4 days after sample collection and at room temperature; therefore, no
analysis was performed due to poor sample integrity.
-------
TABLE C3 (continued)
en
Concentration in stream
Species
Priority pollutant
metal species :
Antimony
Arsenic
Beryllium
Cadmium
Chromium
Copper
Lead
Nickel
Silver
Zinc
Other species :
Phenol (total)
Cyanide (total)
CTHF-9
380
M.4
-
120
1,040
644
920
468
200
6,560
13
62
CTHF-10 CTHF-11 CTHF-12
192
35
-
22
540
480
520
220
82
7,200
19
<1
132
15
-
20
760
46
404
200
68
360
r
20
<1
116
<1
34
48
520
50
380
62
20
140
18
<1
CTHF-13
280
221
_
40
1,000
3,060
700
480
116
5,360
64
8
CTHF-14 CTHF-15
196
2
-
20
900
10,600
520
186
70
6,600
12
<4
160
<1
-
34
680
100
450
220
84
188
3
<4
CTHF-16
320
9
_
70
1,320
35,400
1,140
600
126
12,800
26
20
-------
Results of the bioassays are shown in Table 4. None of the
samples were mutagenic in the Ames test in the range of sample
concentrations tested - 10 to 1,000 y£/plate. Two samples
(CTHF-6 and 13) indicated- acute toxicity to CHO cells. Sample
CTHF-16 exhibited acute toxicity but in a sample concentration
higher than that tested. Analysis of the fathead minnow and
daphnia acute toxicity data indicated the four permeate samples
(CTHF-5, 8, 12 and 15) produced no or very little mortality. The
most toxic samples were the concentrates (CTHF-6, 9, 13, and 16).
Data from the 14-day rat test indicated that no rat deaths or
sample related physical effects occurred due to a single maximum
dosage. Therefore, no samples were subjected to the quantita-
tive bioassay.
76
-------
TABLE C4. SUMMARY OF BIOASSAY TEST RESULTS
Sample
CTHF-3
CTHF-4
CTHF-5
CTFH-6
CTHF-7
CTHF-8
CTHF-9
CTHF-10
CTHF-11
CTHF-12
CTHF-13
CTHF-14
CTHF-15
CTHF-16
Microbial Cy totoxicity ,
mutagenicity ECsg (% waste-
response water solution)
Negative
Negative
Negative
_d
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
NAT8
NAT
NAT
9
NAT
NAT
NAT
NAT
NAT
NAT
10
NAT
NAT
>20
Daphnia acute toxicity, ICso
(% wastewater solution)
26
53
42
5.1
25
(20
(45
(35
(4.
(20
to
to
to
2 to
to
34, "
62)
51)
6.2)
31)
Fathead minnow acute toxicity, Rat acute toxicity, LD50
LC>;n (% wastewater solution) (g-sample/kg body weight)
16
28
1.5
13
(13
(24
(1.
(7.
>100
9.9
33.5
4.1
49
80
17
(8.
(27.
>60
>60
(3.
(41
(71
(12
3 to
6 to
12)
50.4)
10
33) >10
>10
2.2) >10
8 to 22) >10
NAT
5 to
5 to
(21 to
<100
4 to
to
to
to
4.9)
58)
90)
23)
1.6
25
5.3
(1.
(21
(4.
NAT
2 to
to
NAT
1 to
>10
2.8) >10
12) >10
100) >10
>10
2.0) >10
39) >10
>10
6.8) >10
No acute toxicity.
Values in parentheses are 95% confidence intervals.
Only 30% mortality occurred in 100% solution of wastewater.
CTHF-6 could not be readily filter sterilized, therefore the Ames test could not be performed.
>60 <100 means
value is greater than 60% but less than 100%.
-------
SECTION 3
SAMPLE COLLECTION
Hyperfiltration is a separation process involving the filtering
of aqueous solutions by membranes capable of removing not only
suspended particles but also substantial fractions of dissolved
impurities, including organic and inorganic material. The pro-
cess is illustrated schematically in Figure 1. Application of
high pressure to the feed solution causes purified permeate water
to pass through the membrane. Remaining feed water becomes a
concentrated solution of suspended solids and higher molecular
weight compounds.
PRESSURE VESSEL
FEED
CONCENTRATE
\\l\\\\\\\
PERMEATE
Figure el. Schematic diagram of a hyperfiltration module.
In the Clemson University study, EPA Grant No. 805777, two waste-
water streams were used as feed: 1) scour bath wastewater, and
2) wastewater from dying operations. In addition, three hyper-
filtration membranes were tested on each wastewater: 1) polyether
amide (PEA) membrane, 2) cellulose acetate (CA) hyperfilter, and
3) a dynamic membrane (DM) of a dual-layer hydrous Zr(IV) oxide-
polyacrylate. The polyether amide and cellulose acetate membranes
were tested in series, resulting in two permeate samples and one
concentrate sample per feed tested. The resulting sample coding
system and volume of sample collected in the test program are
shown in Table 5.
78
-------
TABLE C5. COLLECTION SAMPLES FOR BIOASSAY
TESTS AND CHEMICAL ANALYSES
Sample
Description
Volume,
gal
CTHF-1 Plant water 5
CTHF-2 Apparatus water 5
CTHF-3 Scour-1, feed for PEA and CA hyperfiltration 25
CTHF-4 Scour-1, permeate from PEA hyperfiltration 25
CTHF-5 Scour-1, permeate from CA hyperfiltration 25
CTHF-6 Scour-1, concentrate from PEA and CA hyper-
filtration 10
CTHF-7 Scour-2, feed for DM hyperfiltration 25
CTHF-8 Scour-2, permeate from DM hyperfiltration 25
CTHF-9 Scour-2, concentrate from DM hyperfiltration 10
CTHF-10 Dye-1, feed for PEA and CA hyperfiltration 25
CTHF-11 Dye-1, permeate from PEA hyperfiltration 25
CTHF-12 Dye-1, permeate from CA hyperfiltration 25
CTHF-13 Dye-1, concentrate from PEA and CA hyper-
filtration 10
CTHF-14 Dye-2, feed for DM hyperfiltration 25
CTHF-15 Dye-2, permeate from DM hyperfiltration 25
CTHF-16 Dye-2, concentrate from DM hyperfiltration 10
Concentrate samples will be 2 gal to 5 gal, containing equivalent
solids to the feed sample.
Sample CTHF-1 was the textile plant intake water. The hyperfil-
tration unit was cleaned of residual materials using a sequence
of washes. A detergent wash followed by a caustic wash removed
residual greases, waxes, and organic materials. A nitric acid
wash followed to remove trace metals from the stainless steel
surfaces. The unit was then rinsed with plant intake water. The
unit was finally operated for several hours with plant water to
indicate whether materials were evolved within the plumbing.
Sample CTHF-2 was a sample of this water.
Samples generated in the testing program were analyzed for the
129 priority pollutants and subjected to five bioassay tests.
The priority pollutant analysis scheme is divided into the fol-
lowing fractions for sampling purposes: volatile organics, non-
volatile organics, metals, cyanide (total), and phenol (total).
Three separate samples were required for bioassay testing:
1) microbical mutagenicity (Ames test) and cytotoxicity, 2) 14-
day rat acute toxicity test, and 3) freshwater static acute
toxicity test with fathead minnows and daphnids. Samples for
priority pollutant analysis, Ames test, and cytotoxicity tests
were sent by Clemson University directly to MRC for analysis. To
79
-------
expedite sampling delivery and insure sample integrity, MRC
directed Clemson University to ship the remaining samples for
bioassay testing directly to the testing laboratories.
Table 6 shows the sample fractions collected, volume, and con-
tainers used for each stream sampled. Note that the plant intake
water (CTHF-1) and hyperfiltration rinse water (CTHF-2) were not
subjected to bioassay testing.
TABLE C6. BIOASSAY TESTS AND CHEMICAL ANALYSES,
TEST-SAMPLE CONTAINERS, AND TESTS
DESIGNATED FOR COLLECTION SAMPLES
- '
Test
Ho.
B.I
B.2
B.3
Description
Microbial mutagenicity
(Ames) and cytotoxicity
(hamster ovary cells)
Acute toxicity (rat)
Freshwater static bio-
Sample
volume
500 mi
500 m£
20 gala
Container
Amber glass, Teflon-
lined caps
Glass, Teflon-lined caps
5 gallon, plastic
Required for
collection
samples
(CTHP)
3 to 16
3 to 16
3 to 16
assay (Daphnia and
fathead minnows)
cubitainers
C.I
C.2
C.3
C.4
C.5
C.6
Volatile solutes
nonvolatile solutes
Metals
Cyanide
Phenols
Pesticides
2 x 40 mi
2x1 gal
500 m£
500 raft
500 raft
Aaber glass vials,
Teflon- lined septa
Amber glass, Teflon-
lined caps
Plastic bottles
Plastic bottles
Amber glass
1 to 16
1 to 16
1 to 16
1 to 16
1 to 16
(use part of test sample C.2)
Concentrate samples will be 2 gal
feed sample.
to 5 gal, containiag equivalent solids to the
80
-------
SECTION 4
PRIORITY POLLUTANT ANALYSIS
ANALYTICAL PROCEDURE
Analyses of the 16 wastewater samples for the 129 priority pollu-
tants were performed by MRC in accordance with the analytical
methodology recommended by EPA (1). It is important to realize
that the purpose of EPA's analytical scheme is to screen samples
to determine which of the 129 chemical species are present and to
estimate their general concentration range. Currently, the
recommended analytical protocol is in the developmental stage and
requires further verification and validation. Analytical results
must be considered as reliable estimates of which priority pollu-
tants were present, with concentrations accurate to within a fac-
tor of two.
Of the 129 priority pollutants, two species were not determined
in this project: 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 potential health hazard involved in prepar-
ing standard solutions from the pure compound (1). Asbestos was
eliminated, as recommended by the EPA Project Officer.
Priority pollutants are divided into the following fractions for
analysis purposes: volatile organics, base/neutral organics,
acid organics, pesticides, polychlorinated biphenyls (PCB),
metals, phenol (total), and cyanide (total) (1).
A brief discussion of the analysis procedures used and sample
analysis results are given in the following three subsections.
(1) Sampling and Analysis Procedures for Screening of Industrial
Effluents for Priority Pollutants. Draft final report, U.S.
Environmental Protection Agency, Cincinnati, Ohio, April
1977. 145 pp.
81
-------
Volatile Organics
The recommended method for volatile organic analysis was designed
by EPA to determine those chemical species which were amenable to
the Bellar purge and trap method (1). Appendix A lists those
priority pollutants classified as volatile organics.
Two hermatically sealed 40-m£ glass vials collected from each of
the 16 samples were composited in the laboratory for one analy-
sis. Two composited solutions were used, one for analysis and
one as a backup sample. Figure 2 is a simplified diagram of the
analytical scheme for volatile organics analysis.
An internal standard of 1,4-dichlorobutane was added to 5 mi of
the composited sample and the sample sparged with helium onto a
Tenax GOsilica-packed sample tube. Two tubes were prepared, one
for analysis and one duplicate. Tenax tubes were then sealed in
glass under a nitrogen atmosphere 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.
For compounds with boiling points below room temperature, cryo-
genic trapping at -40°C (liquid nitrogen cooling) was found to
give better reproducibility of retention time than using the sug-
gested temperature of 30°C. After desorption, the GC column
temperature was raised 8°C/min to 170°C.
SEAITOM
IN CUSS
UHBKNITROWN
W>STOM»T-18°C
Figure 2. Analytical scheme for volatile organics analysis
82
-------
Qualitative identification of a compound was made using three
criteria listed in the protocol (1): 1) retention time must co-
incide with known retention times, 2) three characteristic masses
must elute simultaneously, and 3) intensities of the character-
istic masses must stand in the known proper proportions. Quanti-
tation of volatile organics were made using response ratios of
the 1,4-dichlorobutane internal standard.
Nonvolatile Organics
Nonvolatile organics are divided into three groups for analysis:
base/neutral fraction, acid fraction, and pesticides and poly-
chlorinated biphenyls (PCB). A list of compounds that are classi-
fied as nonvolatile organics is given in Appendix A.
The analytical procedure is described in Reference 1. Figure 3
depicts the sample processing scheme for the base/neutral and
acid fractions. The sample solution, 2 £, was made alkaline (pH
greater than 11) with sodium hydroxide, and then extracted three
times with methylene chloride. The wastewater samples formed
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 fun-
nel. Separation was also enhanced by slowly dripping the emulsion
onto the wall of a slightly tilted flask.
METHYLENE CHLORIDE
EXTRACTION
BASES & NEUTRALS 1 ACIDS (PHENOLS) , UNEXTRACTABLES
BOTTOM LAYER TOP LAYER I
DRIED ON CHANGE pH < 2
ANHY. SODIUM SULFATE W/HYDROCHLORIC ACID
(
CONCB
IN K-C
TO!
!
GC/
IDENTIFIC
QUANTI
»
METHYLENE CHLORIDE
, EXTRACTION
YTRATE 1 -T- '
EVAP. ACIDS 1 AQUEOUS
J* I J
DRIED ON ANHY. SAVE 25 mi
SODIUM SULFATE IISCARD REMAINDER
MS 1
ATION& 1
i"llul" CONCFNTRATFD
IN K-D EVAP.
TOlmJ
1
GC/MS
IDENnFICAHON &
QUANTITATION
Figure 3.
Sample processing scheme for
nonvolatile organics analysis.
83
-------
Extracts were dried on a column of anhydrous sodium sulfate, con-
centrated to 1.0 ml in a Kuderna-Danish (K-D) evaporator with a
Snyder column, spiked with deuterated anthracene, sealed in sep-
tum 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 1.0 £ sample was used for analysis of the pesticides
and PCB (Aroclor® fluids). The basic sample processing scheme is
shown in Figure 4. 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 necessary, acetonitrile
partitioning and a Florisil® chromatography column were used for
further cleanup of the sample. All samples went through aceto-
nitrile partitioning cleanup, only.
AOWEOUS must
WSCARDED
I
we»HC PHASE
CC/EC SOKTN
NO
PARTITION
tC/fCRBOKEN
(WBTMER CIEAMUP ?»
aoBisn. COLUMN
CH80MATOCRAPHY
NO
GCftC KSCREEN
ffUBTWB CLEANUP ?)
res
SttlCIC ACID
CHROMATOC«APHY
CC/tCOUA*nTATION
ON HRST CQLWWI
GC/K VfBUFICATION
ON StCOW) COLUMN
Figvire 4. Sample processing scheme for
pesticide and PCB analysis.
84
-------
Confirmation of identity and guantitation were made using two
different GC columns: SP-2550 and Dexil 410. Compound verifica-
tion was made with the MS when the concentration was greater than
10 yg/fe- Concentrations of potential pesticides ranged from
0-1 yg/& to 10 yg/Jl; therefore, MS verification was not possible
in this study. Pesticide species identified only by GC below
10 yg/Jl were reported only if they met the following two criteria:
1) the retention time window between standards and unknown peaks
correlated within ±3 s, and 2) concentrations calculated from
both GC columns had to agree within ±20%. Unknown peaks not meet-
ing 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, measured as the total metal.
Sixteen metal samples were collected and shipped in low-density
polyethylene plastic bottles. Due to U.S. Department of Trans-
portation regulations regarding air freight of hazardous materials,
the samples were not acidified in the field. Upon arrival at MRC,
5 ma of redistilled nitric acid (HNOs) were added to each sample
and the sample allowed to stand for 24 hr before processing.
Each metals sample was beaker digested to reduce sample matrix
effects with HNOs for about 6 hr or until the solution became
clear. The digested solution was then taken up to 100 ml with
distilled deionized water and stored in low-density polyethylene
plastic bottles.
The following nine priority pollutant metals were analyzed on the
Jarrell-Ash Plasma Atomcomp, Model 975 with inductively coupled
argon plasma excitation (ICAP) at Monsanto Company's Physical
Sciences Center in St. Louis: antimony, beryllium, cadmium,
chromium, copper, lead, nickel, silver, and zinc. ICAP is an
optical emission spectroscopy analytical system for simultaneous
multi-element determination of trace metals. In this device, a
stream of inert gas (argon) is first ionized and then a concen-
tric, which is a source of a high frequency (HF) field, accel-
erates the electrons until they acquire sufficient energy to
excite and ionize atoms. The elements of the wastewater samples
introduced into this plasma are immediately raised to a higher
energy state from which they decay with ultraviolet (uv), visible,
and infrared (ir) emissions.
Of the remaining four priority pollutant metals, only arsenic
was measured in the 16 samples because previous research indi-
cated that mercury, selenium, and thallium were not in textile
(2) Rawlings, G. D. Source Assessment: Textile Plant Waste-
water Toxics Study - Phase I. EPA-600/2-78-004h, U.S.
Environmental Protection Agency, Research Triangle Park,
North Carolina, March 1978. 166 pp.
85
-------
wastewaters (2). Arsenic was measured by conventional atomic
absorption techniques in accordance with Referances 3 and 4.
In addition to the 16 samples, 5 other samples were included as
part of the quality assurance program. A certified U.S. National
Bureau of Standards trace metal in water standard (No. 1643) was
included with the set of samples. Two trace metal standards pre-
pared by MRC were included: one concentrated standard and one
5 mt/i. dilute standard. A separate standard was added for silver
and nickel quality assurance testing. Finally, one of the real
samples was split and submitted as a blind repeat.
Since ICAP simultaneously analyzes for 25 trace elements, the
results of the nonpriority pollutant metals is also reported.
Cyanide (Total)
Total cyanide was analyzed according to the procedure in Refer-
ence 1. Two standard solutions were prepared and sent with the
samples along with two blind repeats of the standards.
Phenol (Total)
In addition to specific phenolic compounds and phenol measured by
GC-MS in the acid fraction, total phenol was also measured by
typical wet chemistry techniques (1, 3, 4) .
Phenol samples were preserved in the field by adding 1.0 g
maintaining the pH to less than 4 with HaPOij and storing the
sample at 4°C. Recent research has indicated this preservation
technique is adequate for at least 8 days (5) . All phenolic
samples collected in this study were analyzed within 5 days of
collection.
RESULTS OF CHEMICAL ANALYSIS
Organic Species
Results of the analysis of 16 wastewater samples for the pres-
ence of the 114 organic priority pollutant species are shown in
(3) Manual of Methods for Chemical Analysis of Water and Wastes.
EPA-625/6-76-003a (PB 259 973) U.S. Envionmental Protection
Agency, Cincinnati, Ohio, 1976. 317 pp.
(4) Standard Methods for the Examination of Water and Wastewater,
Fourteenth Edition. American Public Health Association,
Washington, D.C., 1976. 874 pp.
(5) Carter, M. J., and M. T. Huston. Preservation of Phenolic
Compounds in Wastewaters. Environmental Science and Tech-
nology, 12(3):309-313, 1978.
86
-------
Table 7. A reagent blank using' organic free water was included
with the samples and worked up and analyzed like all the samples.
Results of this analyses are shown in the second column of Table 7
Analysis of the data indicates no organic priority pollutants are
introduced due to the sample workup reagents or analysis contami-
nation at MRC. Samples CTHF-1 and CTHF-2 were samples of the
textile plant intake water and hyperfiltration unit rinse. Analy-
ses of these samples indicate that possibly chloroform and toluene
are introduced from these two sources. The remaining organic
priority pollutant species in Table 7 are present in the waste-
water samples.
Samples with the largest number of organic species are the four
feed streams (CTHF-3, 7, 10, and 14). Species found in the con-
centrate and not found in the feed are due to the concentrating
mechanism of the hyperfiltration unit and the species are now
above detection limits. The detection limit for the 114 organic
priority pollutants are shown in Table 8.
In addition to the organic priority pollutant species, several
other organic compounds were detected in the samples, Table 9.
These compounds were identified by their characteristic fragmen-
tation pattern in the mass spectrometer based on their principle
ion and corresponding mirror ions. Qualitative concentration
values were determined based on the peak heights of known concen-
trations of priority pollutants which elute the gas chromatograph
in adjacent retention time windows.
Analysis of the data indicate that triphenyl phosphine and its
oxide are probably a result of glass cleaning detergents and was
introduced from the sample containers and laboratory glassware.
Metals
Results of ICAP and atomic absorption analyses of the 16 digested
metals samples are shown in Tables 10 and 11. Table 10 shows the
priority pollutant metals, while Table 11 shows the other metals
simultaneously measured by ICAP. Note that the second column in
both tables shows the detection limit for each metal.
Results of the trace metals quality assurance program are pre-
sented in Table 12.
Phenol (Total) and Cyanide (Total)
Results of the phenol (total) and cyanide (total) analyses were
also presented in Table 10. Sample CTHF-6 arrived at MRC four
days after sample collection and at room temperature. Therefore,
total phenol and cyanide were not measured due to poor sample
integrity.
87
-------
TABLE C7. ORGANIC PRIORITY POLLUTANT SPECIES DETECTED IN SPECIFIC WASTEWATER STREAMS
(yg/fc)
oo
oo
Blank
Organic compound water CTHF-1 CTHF-2
Bis ( 2-ethylhexyl) phthalate 1.1 9
Dimethyl phthalate 18
Di-n-butyl phthalate 0.4
Butylbenzyl phthalate
Diethyl phthalate 0.3
Acenaphthene
Anthracene
F luor anthene
Pyrene
Naphthalane
Phenanthrene
Phenol 0.9
Chloroform 58 31
Toluene 3 22
Trichloroethylene
Benzene 2
Chlorobenzene
Ethylbenzene
Methylene chloride 6 34
Concentration in stream
CTHF-3
9
4
7
2
0.4
1
18
0.8
0.3
5
CTHF-4
3
9
1
0.8
0.5
2
18
15
1
6
CTHF-5
3
3
22
29
0.4
1
0.7
5
CTHF-6 CTHF-7 CTHF-8
9
3
7
2
13
34
41 0.8 0.7
5 2
6 2
21
15 4 5
(continued)
Note.—Blanks indicate compound is below detection limits.
-------
TABLE C7. (continued)
00
Concentration in stream
Organic compound
Bis (2-ethylhexyl)phthalate
Dimethyl phthalate
Di-n-butyl phthalate
Butylbenzyl phthalate
Diethyl phthalate
Acenaphthene
Anthracene
F luor anthene
Pyrene
Naphthalene
Phenanthrene
Phenol
Chloroform
Toluene
Trichloroethylene
Benzene
Chlorobenzene
Ethylbenzene
Methylene chloride
CTHF-9 CTHF-10
4
55
1
...
3
0.6
0.8
1 0.2
19
0.5 10
0.7 2
2 5
CTHF-11
31
45
0.8
1
0.8
0.7
31
11
0.6
0.4
45
CTHF-12 CTHF-13
3 51
290
6
7
7
3
0.4 1
4
24
1
-
4 14
CTHF-14
2
170
1
3
0.7
0.1
0.8
0.2
96
0.6
0.6
5
CTHF-15
1
4
1
0.05
0.1
0.4
1
3
CTHF-16
4
1
1
3
Note.—Blanks indicate compound is below detection limits.
-------
TABLE C8. MINIMUM DETERMINABLE CONCENTRATIONS
(ygA)
Compound
Detec-
tion
limit
Compound
Detec-
tion
limit
Acids:
2-Chlorophenol 0.09
Phenol 0.07
2,4-Dichlorophenol 0.1
2-Nitrophenol 0.4
p-Chloro-m-cresol 0.1
2,4,6-Trichlorophenol 0.2
2,4-Dimethylphenol 0.1
2,4-Dinitrophenol 2.0
4,6-Dinitro-O-cresol 40.0
4-Nitrophenol 0.9
Pentachlorophenol 0.4
Volatiles:
Chloromethane 0.2
Dichlorodifluoromethane 0.2
Bromomethane 0.2
Vinyl chloride 0.4
Chloroethane 0.5
Methylene chloride 0.4
Trichlorofluoromethane 2.0
1,1-Dichloroethylene 2.0
1,1-Dichloroethane 3.0
£rons-l,2-Dichloroethylene 2.0
Chloroform 5.0
1,2-Dichloroethane 2.0
1,1,1-Trichloroethane 2.0
Carbon tetrachloride 4.0
Bromodichloromethane 0.9
Bis-chloromethyl ether 1.0
1,2-Dichloropropane 0.7
Trans-1,3-dichlorproopene 0.4
Trichloroethylene 0.5
Dibromochloromethane 0.3
Cis-1., 3-dichloropropene 0.5
1,1,2-Trichloroethane 0.7
Benzene 0.2
2-Chloroethylvinyl ether 1.0
Bromoform 0.6
1,1,2,2-Tetrachloroethene 0.9
1,1,2,2-Tetrachloroethane 0.6
Toluene 0.1
Chlorobenzene 0.2
Ethylbenzene 0.2
Direct injeatables:
Acrolein 200
Acrylonitrile 100
Base neutrals:
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
Pluorene
2,6-Dinitrotoluene
1,2-Diphenylhydrazine
2,4-Dinitrotoluene
N-nitrosodiphenylamine
Hexachlorobenzene
4-Bromophenyl phenyl ether
Phenanthrene
Anthracene
Dimethyl phthalate
Diethylphthalate
Fluoranthene
Pyrene
Di-n-butyl phthalate
Benzidine
Butyl benzyl phthalate
Chrysene
Bis(2-ethylhexyl)phthalate
Benzo (a) anthracene -.„.-;
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Indeno(1,2,3-c,d)pyrene
Dibenzo(a,h)anthracene
Benzo(g,h,i)perylene
N-nitrosodimethylamine
N-nitrosodi-n-propylamine
4-Chlorophenyl phenyl ether
3,3'-Dichlorobenzidine
All pesticide and PCB's
0
0.02
0.04
0.1
0.05
0.06
0.08
0.09
0.007
0.07
0.2
0.08
0.06
0.02
0.02
0.04
0.06
0.02
2
0.02
0.02
0.07
0.05
0.1
0.01
0.01
0.03
0.03
0.02
0.01
0.02 .
0.02
0.03
0.02
0.04
0.02
0.02
0.02
0.02
0.02
0.02
0.01
0.8
0.2
0.03
1.0
1.0
90
-------
TABLE C9. OTHER ORGANIC COMPOUNDS DETECTED IN THE 16 SAMPLES
<£>
Blank Approximate concentration in stream
Compound water CTHF-1 CTHF-2 CTHF-3 CTHF-4 CTHF-5 CTHF-6 CTHF-7 CTHF-8
Triphenyl phosphine 5 50.5 0.52 5
Triphenyl phosphine
oxide 5 50 10 5 10 10 2 3.0
cx-Terepineol 40 10 30 25
2-Mercapto
benzthiazole 10 20 0.5 10
l-Cyano-2-benzyloxy
ethane 5
Benzthiazole 30 2 600 40 5
Laurie acid 4*00 3,000
Myristic acid 1,000
Palmitic acid 1,000
Stearic acid
Approximate concentration in stream
Compound CTHF-9 CTHF-10 CTHF-11 CTHF-12 CTHF-13 CTHF-14 CTHF-15 CTHF-16
Triphenyl phosphine 5 2 7 10 10 10 10
Triphenyl phosphine
oxide 55 5 10 30 10 10 5
a-Terepineol 30 20 30 50 50 5
2-Mercapto
benzthiazole 40 30 200
1-Cyano-2-benzyloxy
ethane 60 10 100
Benzthiazole 100 200 250
Laurie acid 100
Myristic acid
Palmitic acid 400
Stearic acid 200
Note.—Blanks indicate compound is below detection limits.
-------
TABLE CIO.
NJ
PRIORITY POLLUTANT METALS ANALYSIS OF
DIGESTED SAMPLES, PHENOL, AND CYANIDE
Species
Priority pollutant
metal species:
Antimony
Arsenic
Beryllium
Cadmium
Chromium
Copper
Lead
Nickel
Silver
Zinc
Other species:
Phenol (total)
Cyanide (total)
Detection
limit
10
2
0.04
2
4
4
22
36
5
1
1
1
Concentration in stream
CTHF-1 CTHF-2 CTHF-3 CTHF-4
12
<1
-
5
540
54
168
-
-
630
<1
<7
30
<1
-
9
840
200
240
154
24
616
33
4
100
19
-
15
640
90
380
132
42
520
6
<4
90
<1
-
15
720
26
250
70
26
360
12
72
CTHF-5
132
1
-
14
620
32
340
100
42
8,600
16
30
CTHF-6 CTHF-7 CTHF-8
436
160
-
48
1,260
760
760
480
114
3,540
a
-
170
35
-
16
760
94
400
200
62
460
4
<4
146
5
-
20
800
68
414
210
78
248
<1
<7
(continued)
Sample arrived at MRC 4 days after sample collection and at room temperature; therefore, no
analysis was performed due to poor sample integrity.
-------
TABLE C1CK (continued)
u>
Concentration in stream
Species
Priority pollutant
metal species:
Antimony
Arsenic
Beryllium
Cadmium
Chromium
Copper
Lead
Nickel
Silver
Zinc
Other species :
Phenol (total)
Cyanide (total)
CTHF-9
380
^14
-
120
1,040
644
920
468
200
6,560
13
62
CTHF-10 ClUF-ll CTHF-12
192
35
-
22
540
480
520
220
82
7,200
19
<1
132
15
-
20
760
46
404
200
68
360
20
<1
116
<1
34
48
520
50
380
62
20
140
18
<1
CTHF-13
280
221
-
40
1,000
3,060
700
480
116
5,360
64
8
CTHF-14 CTHF-15
196
2
-
20
900
10,600
520
186
70
6,600
12
<4
160
<1
-
34
680
100
450
220
84
188
3
<4
CTHF-16
320
9
-
70
1,320
35,400
1,140
600
126
12,800
26
20
-------
TABLE Cll. CONCENTRATION OF OTHER METALS DETECTED BY
ICAP ANALYSIS OP SPECIFIC WASTEWATER STREAMS
(vg/*)
Metal
Aluminum
Barium
Boron
Calcium
Cobalt
Iron
Magnesium
Manganese
Molybdenum
Phosphorus
Silicon
Sodium
Strontium
Tin
Titanium
Vanadium
Detection
limit
12
0.2
1
0.04
6
2
0.1
0,5
10
70
15
26
0.2
15
1
2
Concentration
CTHF-1
202
100
610
13,720
-
1,040
5,100
260
28
260
13,500
59,200
132
-
6
28
CTHF-2
204
80
396
16,600
10
370
6,720
150
30
1,720
18,200
104,000
160
-
7
40
CTHF-3
800
84
47,200
16,300
22
520
9,300
360
52
4,260
17,600
379,600
144
64
42
78
CTHF-4
180
-
11,900
1,050
-
300
240
24
36
460
2,600
13,500
-
«•>
8
14
in stream
CTHF-5
260
10
8,900
1,280
8
194
360
26
64
860
2,600
24,800
-
-
22
18
CTHF-6
3,800
580
81,000
114,200
72
2,900
72,000
2,600
160
33,600
30,000
1,674,000
1,040
520
82
480
CTHF-7
1,470
120
56,000
15,600
18
420
6,200
720
60
5,200
21,000
612,000
140
68
28
64
CTHF-8
1,360
8
31,000
1,520
20
300
408
26
94
1,470
7,600
244,000
—
—
12
38
(continued)
-------
TABiJT ell (continued)
Ul
Concentration
Metal
Aluminum
Barium
Boron
Calcium
Cobalt
Iron
Magnesium
Manganese
Molybdenum
Phosphorus
Silicon
Sodium
Strontium
Tin
Titanium
Vanadium
CTHF-9
5,000
350
81,000
64,000
52
2,000
28,800
2,860
232
24 , 000
22 , 400
1,546,000
560
260
66
240
CTHF-10
540
64
2,600
16,800
26
428
11,400
280
98
7,600
23,800
187,200
146
60
24
96
CTHF-11
268
-
500
1,060
16
280
140
16
78
740
500
13,400
-
-
12
24
CTHF-12
212
-
1,220
640
44
130
106
40
96
700
3,360
8,600
-
60
8
10
in stream
CTHF-13
11,000
400
4,040
113,600
68
1,050
81,800
1,860
154
49,800
39,600
896,000
1,000
200
42
540
CTHF-14
1,200
196
860
26,000
20
400
13,000
700
108
46,800
12,000
530,000
260
50
16
106
CTHF-15
740
-
740
920
20
180
240
14
160
4,200
13,300
92,000
-
12
18
30
CTHF-16
3,000
480
1,320
70,600
66
1,340
41,200
400
400
141,200
20,200
1,249,000
720
120
34
300
-------
TABLE C12. TRACE METALS QUALITY ASSURANCE RESULTS
1643
BBC Concentrate
As/Mi dilute
MRC dilute
Sample Sample Sample
Standard, value. Percent Standard, value. Percent Standard, value. Percent
M9./4 M9/t error V8/1 V9/JI error Mg/l lig/fc. error
Sample
Standard, value. Percent
pg/t ug/t error
Arsenic
Bariun b
Beryllium
Boron .
cad«liu«b
Calcium .
Chromium
copper
Magnesium
Himgawse
Hicfcel"
Pnosphorus
77
19
27,000d
15
17
16
75
20H
7,000d
29
105
49
66.6 14
SilV
Sodium
Strontium
Tin
Titanium
10.
212
50
«5
15.4
16.4
7.0
26,900
16.9
19.0
IS. 5
73.0
8.5
6,140
27.3
95.2
66.1
11,500
244
87,2
£1.6
14
14
12
0.3
13
12
16
3
57
12
8
9
35
15
15
74
5
30,000
4,000
10,000
5,000
30,000
50,000
20,000
33,140 10
4,089
10,040
5,055
29,230
50,870
20,790
2
0.4
1
3
2
40
1,500
200
500
250
1,500
2,500
1,000
39 3
1,421 5
205
502
250 236 5.6
250 269 7.6
2
0.1
253 1
1,473 2
2,523 0.9
1,001 0.1
10,000
10,36D
500
514
2.8
Alanum*
antimony
Arsenic
Bar ion
Beryllitaa
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
lead
Magnesium
Manganese
Molybdenum
Bickel
Phosphorus
Silicon
Silver
Sodium
Strontium
Tin
Titanium
Vanadium
Zinc
1,360
146
a
-
31,000
2O
1,520
800
20
68
300
414
408
26
94
210
1.470
7.600
78
244.000
-
-
12
38
248
280
-
-
31,800
28
1.880
820
22
90
320
500
440
26
122
260
1,800
8,080
100
262,400
12
42
16
42
320
6
31
_
—
3
33
21
2
10
28
6
19
7
0
26
21
20
6
25
7
-
-
29
10
25
3.080
320
480
-
1,320
70
7O,600
1,320
66
35,400
1,340
1,140
41,200
400
400
600
141,200
20,200
126
1,249,000
720
120
34
300
12,800
J.OOO
388
480
-
1,360
70'
• 70, BOO
1,350
74
35,400
1,360
1,250
41,600
406
416
650
143,000
23,400
134
1,245,200
720
136
54
300
21,600
0
17
0
-
3
0
0.3
2
11
0
1
9
1
1
4
a
i
15
-6
0.3
0
12
45
0
51
130
88
3
O.3
140
12
680
740
7
28
145
286
75
6
54
82
500
550
32
3,380
3
4
7
n
350
US
92
2
0.6
57
13
511
370
12
25
92
240
50
5
51
114
480
490
51
1.440
2
..
•" 7
14
690
Slacks indicate no metal standard included.
Priority pollutant metal.
CBariun is not certified because of the large difference betwen its initial concentration
(39 ng/g), corresponding to the amount added to the water, and the stabilized concentra-
tion (18 ng/g). Hovever, this stabilized concentration has remained constant in the
test-bottles for over four months.
4
SHI 1643 approximates the elemental composition of fresh water - 27 ug/g calcium,
10 »g/g sodium, 7 ng/g magnesium, and 2 ug/g potassium.
6«etal standard concentration is below detection limit.
96
-------
For quality assurance standard solutions in two concentration
ranges of phenol and cyanide were prepared according to Refer-
ence 4 procedures. A blind repeat of each standard was also
included. Results of the analyses are shown in Table 13.
97
-------
TABLE CIS. QUALITY CONTROL SAMPLES FOR PHENOL (TOTAL)
AND CYANIDE (TOTAL)
StandardSample
value, value, Percent
Compound mg/1, mg/A error
Phenol (total) 10 11 10
10 15 50
300 300 0
300 270 10
Cyanide (total) 10 4 60
10 4 60
300 384 28
300 348 16
98
-------
SECTION 5
BIOASSAY TESTS
MICROBIAL MUTAGENICITY (AMES TEST)
Test Procedure
The purpose of the mutagenicity bioassay (Ames test) was to
determine if a chemical mutagen (possibly a carcinogen) was pres-
ent in the 14 samples tested. The plant intake water (CTHF-1)
and hyperfiltration rinse water (CTHF-2) were not tested in any
of the bioassays.
To date the most sensitive assay for deoxyribonucleic acid (DNA)
damage is the induction of mutations in bacteria. The Ames test,
the most developed of the bacterial mutagenesis tests, used
mutant strains of Salmonella typhimur-Lum which were specially
selected because of their abilities to detect specific types of
mutations. The assay procedure with S. typhimurium has proven to
be 85% to 90% accurate in detecting mutagens, and it has about
the same accuracy in identifying chemicals that are not carcino-
genic (6). For example, the TA1535 strain was designed to detect
mutations due to base-pair substitutions. This strain responded
particularly 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 respon-
sible 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 penetra-
tion of large molecules.
Mutant S. typhimuTium tester strains lack the ability to synthe-
size histidine and are therefore unable to grow unless histidine
is supplied. These bacteria are cultured in media containing
minimal levels of histidine to sustain growth. Under these con-
ditions 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
(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.
99
-------
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 to the S. typhimuvium tests
to simulate in vivo metabolic actions. In practice, the sub-
stance is tested with and without this microsomal preparation to
determine whether it requires metabolic transformation or is,
itself, mutagenically active.
The 14 samples were tested following the procedure of Ames des-
cribed in Reference 7. An outline of the procedure used by MRC
is given in Table 14. MRC currently purchases the S-9 fraction
from Litton Bionetics. In addition to the details of the pro-
cedure the following steps were taken. All samples were stored
at 4°C until analyzed. Each sample was filter sterilized then
passed through three filters (1.2 ym, 0.45 ym, and 0.22 ym) in
series. Each filtrate was then tested in duplicate, with and
without microsome addition. Dose response tests were conducted
by adding the following amounts of sample per plate, 10 y£, 50 y£,
100 yH, 500 uAf and 1,000 y£. Several samples were retested be-
cause of poor growth or the possibility of an increased response.
Results of the Ames Tests
None of the 14 samples were mutagenic in the Ames S. typhimurium
mutagenicity test under the conditions tested. The criteria used
to evaluate the samples were that the sample must increase the
number of revertants by a factor of two over the spontaneous
revertants of the controls and exhibit a dose response with the
increase plate dosage. Toxicity was observed by a sparse lawn
at the highest concentrations (1,000 y£/plate) for samples
CTHF-3, 4, 5, 9, 14, and 15. Sample CTHF-6 could not readily
be filter sterilized and when streaked on nutrient agar showed
bacterial contamination. Therefore, this sample was not tested
by the Ames test due to the quantity of sample needed. Table 15
summarizes the data. The actual test data including positive
controls and background controls are listed in Appendix B.
(7) Ames, B. N., J. McCann, and E. Yamasaki. Methods for the
Detection of Carcinogens and Mutagens with the Salmonella/
Mammalian-Microsome Mutagenicity Test. Mutagenicity Re-
search, 31:347-364, 1975.
100
-------
TABLE C14. PROCEDURE FOR AMES MUTAGENICITY TEST
Tester strains:
TA98, TA100, TA1535, TA1537, TA1538, TA92, HisG46
Strains are stored frozen in nutrient broth at
-80°C
Strains are routinely checked for their histidine
requirements, rfa~ deletion, urvB" deletion,
plasmids, and spontaneous revertants.
A. S-9 Mix Preparation;
Reagent
MgCl2/KCl
G-6-P
Na Phosphate
NADP
Sterile H20
S-9 fraction (thawed,
kept on ice)
Stock solution mJl/S-9 mixture Storage
0.4M/1.65M
0.5M
0.2M (pH 7.4)
0.1M
0.2 mJl
100 y£
5.0 m£
0.4 m£
3.3 mi,
1.0 mZ
refrigerate
frozen
refrigerate
frozen
frozen
B
Preparation of Top and Bottom Agar;
1. Top Agar - 0.6% Agar (6 g/3.)
0.5% NaCl (5
Autoclaved, stored in sterile bottles (50 m& and 100 mJl
aliquots)
Top is melted in autoclave or steam bath before use.
0.5 mM L-Histidine • HC1/0.5 mM Biotin solution is added
to the melted top agar just before each test.
2. Bottom Agar
g/i
MgSO^ • 7H2O 0.2
Citric Acid H2O 2.0
K2HPOit (anhydrous) 10.0
NaNH^HPOij • 4H2O 3.5
Agar 15.0
Glucose 20.0
C. Preparation of Test Substance;
A compound is dissolved in sterile H2O, DMSO, ethanol or p-
dioxane. Up to 100 \iH ethanol or p-dioxane can be used per
plate.
D. Assay;
1. 2 m£ molten top agar (45°C) (with histidine and biotin)
2. 0.1 mA bacteria
3. 1-100 u& test compound
4. 0.5 mJl S-9 mix (when added)
Plates are incubated at 37°C, in dark, 48 hr. Revertants are
scored for test compounds and controls.
101
-------
TABLE C15. RESULTS OF AMES MICROBIAL
MUTAGENICITY TESTS
Sample
number
CTHF-3
CTHF-4
CTHF-5
CTHF-7
CTHF-8
CTHF-9
CTHF-10
CTHF-11
CTHF-12
CTHF-13
CTHF-14
CTHF-15
CTHF-16
Result
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Amount tested
per plate, yJl
10
10
10
10
10
10
10
10
10
10
10
10
10
to
to
to
to
to
to
to
to
to
to
to
to
to
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
1,000
CYTOTOXICITY TEST
Test Procedure
MRC performed clonal assay acute toxicity tests on 14 samples
using Chinese hamster ovary cells (CHO-Kl). The purpose of cy-
totoxicity tests was to measure metabolic impairment and death in
mammalian cells due to exposure to the wastewater samples. These
primary cell cultures have some degree of metabolic repair
capability.
Samples were tested according to the procedure described in
References 8 and 9 and shown in Table 16.
In general, the test procedure involves*trysinizing stock cul-
tures of CHO-Kl cells and counting. A cell dilution was made
with media to concentration of 60 cells/mi. Five milliliters
cells) of this dilute cell suspension were added to a T-25
(8) 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.
(9) Wininger, M. T., F. A. Kulik, and W. D. Ross. In Vitvo
Clonal;Cytotoxicity Assay Using Chinese Hamster Ovary Cells
(CHO-Kl) for Testing Environmental Chemicals. In Vitro.
14;381, 1978.
102
-------
TABLE C16. PROCEDURE FOR CHO-K1 CLONAL CYTOTOXICITY TEST
The details of the CHO-K1 clonal toxicity test are as follows:
Cell line: Chinese hamster ovary epithelial cells ATCC
#CCL 61
Medium: F-12 GIBCO #H-17 10.8 g/£
NaHC03 1.18 g/A
10% fetal calf serum, virus, mycoplasma
screened GIBCO #629
Incubation: 37°C, 5% C02, saturated humidity
Samples: 6 controls (blank)
5-7 concentrations of test compound in triplicate
Test Procedure:
• To stock CHO-K1, add 5 mfc 0.25% Trypsin at 37°C for 5 min
to 10 min
• Shake cells and add to centrifuge tube
• Add 5 mJl media to flask, shake and add to centrifuge tube
• Centrifuge 5 min at 500 g, pour off liquid, retaining
cells
• Add 10 mi, medium, shake, centrifuge 5 min, pour off medium
• Add 10 mA medium, shake
• Make hemocytometer count of trypsinized cells
• Dilute so that 5 m£ media contains 300 to 500 cells
• Add 5 mJl media and cells to T-25 flasks
• Incubate 12 to 18 hours to allow attachment using normal
media
• Replace with 5 m£ of premixed media and sample
• Incubate 6 to 7 days total
• Fix with 10% formaldehyde/0.5% NaCl/4% methanol for
30 min
• Stain with crystal violet (0.04% for 15 min)
• Count colonies of remaining cells macroscopically using
Fisher Count-All Model 600
• Score with respect to experimental versus controls at
% survival
flask. Cultures were incubated for 18 hr to permit cell attach-
ment using normal media. All wastewater samples were stored at
4°C and filter sterilized through a series of 1.2 ym, 0.45 ym,
and 0.22 ym filters. Filtrates were then applied to the plates
18 hr after seeding the plates. These plates were incubated at
37°C in a carbon dioxide incubator for 5 days.
At the end of this time period, the cells were fixed with 10%
formaldehyde, 0.5% sodium chloride, and 4% methanol for 30 min
103
-------
followed by staining with 0.04% crystal violet for 5 min. Stained
colonies were then counted. The percent colony formation was
calculated by comparing the control plates with the wastewater
sample containing plates. EC50 (effective concentration at which
50% of the cells show metabolic impairment) determinations were
calculated from dose response curves.
The concentrations used for these samples were 0.2 y£, 2.0 y&,
10 y£, 50 y£, 100 yJl, 150 yJl, and 200 y& of sample per milliliter
of media. All samples were tested in triplicate at each concen-
tration and retests were performed on those samples that exhibited
toxicity.
Cytotoxicity Results
Results of the CHO cytotoxicity test are shown in Table 17. The
raw data collected in the tests are given in Appendix C.
TABLE C17. RESULTS OF THE CHO-Kl CYTOTOXICITY TEST
Sample
CTHF-3
CTHF-4
CTHF-5
CTHF-6
CTHF-7
CTHF-8
CTHF-9
CTHF-10
CTHF-11
CTHF-12
CTHF-13
CTHF-14
CTHF-15
CTHF-16
Concentration range
Results tested/ y£/m£ media
No acute toxicity
No acute toxicity
No acute toxicity
EC 50 = 90 y£/mJl media
=9% solution
No acute toxicity
No acute toxicity
No acute toxicity
No acute toxicity
No acute toxicity
No acute toxicity
EC50 = 100 yJl/mA media
=10% solution
No acute toxicity
No acute toxicity
EC50 > 200 yJl/mJl media
> 20% solution
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
2
2
2
2
2
2
2
2
2
2
2
2
2
2
to
to
to
to
to
to
to
to
to
to
to
to
to
to
200
200
200
200
200
200
200
200
200
200
200
200
200
200
At the concentration range tested (0.2 y£/m& to 200 y£/mJl) toxi-
city was exhibited with the CTHF-6, 13, and 16 samples, while all
104
-------
other samples were nontoxic. Toxicity is reported as EC50 (ef-
fective concentration at which 50% of the cells shown metabolic
impairment) . EC50 values for samples CTHF-6 and CTHF-13 are
90 y&/m5, and 100 u£/m&, respectively. Expressed in different
terms, a wastewater concentration of 9% for CTHF-6 and 13% for
CTHF-13 would metabolically impair 50% of the CHO cells. Sample
CTHF-16 showed toxic effects only at the highest concentration
of 200 yjl/m£, thus no EC50 could be calculated and is expressed
as EC50 >200 yJl/mA or EC50 >20%.
Cadmium chloride was used as a positive control and had an aver-
age EC50 of 0.15 u£/m& or 0.015% solution.
ACUTE STATIC BIOASSAYS WITH FATHEAD MINNOWS AND DAPHNIDS
Fourteen of the sixteen wastewater samples (excluding CTHF-1 and
2) were collected by Clemson University in 5-gal plastic con-
tainers for subsequent bioassay analysis with fathead minnows
(Pimephales promelas) and daphnids (Daphnia magna) . Clemson
University sent the samples directly to the laboratory designated
by MRC: EG&G Bionomics Marine Research Laboratory, Wareham,
Massachusetts. The following subsections describe the test pro-
cedures and materials used and results of the tests.
Fathead Minnow Test Procedures
Procedures used in this 96-hr, static acute toxicity test fol-
lowed those described in Reference 10. Wastewater samples were
delivered by Clemson University, Clemson, South Carolina, in
June 1978, in 5-gal. plastic containers. A characterization of
each sample is presented in Appendix D.
The fathead minnows (Pimephales promelas) used in the determina-
tions were obtained by EG&G from a commercial fish supplier in
Missouri. These fish were assigned a lot number and held in
500-5, fiberglass tanks. Well water characterized as having total
hardness and alkalinity, as calcium carbonate, ranged from 28 mg/£
to 44 mg/£ and 20 rng/S, to 30 mg/Jl, respectively (4) , a pH range
of 6.7 to 7.4, a temperature of 22 ± 1°C and a specific conduct-
ance range of 95 to 170 micromhos per centimeter flowed through
the tank at a minimum of 4 £/min. The specific conductance was
measured with a Model #33 YSI conductivity meter. Experimental
animals were maintained under these conditions for a minimum of
14 days. During this time period, all fish were fed a dry pel-
leted food, ad libitum, daily and ground liver weekly, except
during the 48 hours prior to testing. Mortality observed during
this 2-day period ranged from 0.16% to 0.80%.
(10) Peltier, W. Methods for Measuring the Acute Toxicity of
Effluents to Aquatic Organisms. EPA-600/4-78-012 (PB 276
690) U.S. Environmental Protection Agency, Cincinnati, Ohio,
January 1978. 63 pp.
105
-------
Toxicity tests were conducted in 19.6-A glass jars which con-
tained 15 H of test solution. The diluent water used was soft
water reconstituted according to recommended procedures (10).
A characterization of this water also appears in Appendix D.
Wastewater samples were mixed with diluent water to provide the
appropriate percentage concentrations. A control jar containing
the same dilution water and maintained under the same conditions
as test concentrations, but containing no wastewater sample, was
established. Test solution temperatures were controlled by a
system designed to maintain test temperatures at 22 ± 1°C. Test
solutions were not aerated, except where noted.
Ten fathead minnow with a mean and range (N=30) net weight of
0.53 g (0.21 g to 1.1 g) and a total length and range of 40 mm
(31 mm to 52 mm) were randomly distributed to each test jar
within 3 hours after the test solutions were mixed. This time
period was necessary to warm the solutions to 22 ± 1°C.
During toxicity determinations, pH and dissolved oxygen concen-
trations of test solutions were measured at 0-, 24-, 48-, 72-,
and 96-hr in the control, high, middle and low test concentra-
tions. Temperatures were also measured in the control jars at
the above-mentioned time intervals. Specific conductance, total
hardness and alkalinity were measured in the control, high, middle
and low test concentrations at 0-hr. The pH was measured with a
Model #175 Instrumentation Laboratory pH meter and combination
electrode and the temperature and dissolved oxygen (DO) with a
Model #57 YSI combination oxygen-temperature meter and probe.
Test concentrations and corresponding percentage mortality data
derived from each test were used to calculate 24-, 48-, 72-, and
96-hr median lethal concentrations (LC5o's) and 95% confidence
intervals by means of the moving average angle method (11). The
LCso is defined as the calculated nominal concentration of the
wastewater sample in diluent water which caused 50% mortality in
the fathead minnow population at the stated exposure interval.
Prior to analysis by this method,.nominal concentrations were
transformed to logarithms and corresponding percentage mortali-
ties to angles. Each group of three successive angles was then
averaged and the LCso was estimated by linear interpolation
between the successive concentrations whose average angles
bracketed 45°.
Daphnid Test Procedure
Daphnia magna (<24 hr old) used in this 48-hr acute toxicity
test were from laboratory stocks cultured at EG&G, Bionomics.
(11) Harris, E. K. Confidence Limits for the LD50 Using the
Moving Average Angle Method. Biometrics, 4(3):157-164, 1959.
106
-------
Deionized, reconstituted well water with a total hardness of
200 mg/Jl as CaC03, a pH of 8.1, a temperature of 22 ± l°c and a
dissolved oxygen (DO) concentration of greater than 60% of satu-
ration was used to culture these animals. A description of each
of the 14 samples is given in Appendix D.
Procedures used in this acute toxicity test were based on proto-
cols in Reference 12 except where stated otherwise.
Two independent tests involving two different series of sample
concentrations were performed in this study. A preliminary
(range-finding) test was performed to define a narrower range of
concentrations to be used in a subsequent definitive test. Mor-
tality data derived from the definitive test were used to calcu-
late a median lethan concentration (LC50) and its 95% confidence
limit utilizing the moving average method (13). The LCso is tne
calculated nominal concentration of the wastewater sample in dil-
uent water which produces 50% mortality in the daphnid population
at the stated times of exposure.
Static toxicity tests were conducted in 250-mA beakers which con-
tained 150 m& of test solution. Diluent water used in this study
had the same water quality characteristics as described in
Appendix D. For each test concentration, the appropriate amount
of the wastewater sample was introduced into the required volume
of diluent water to total 750 mZ and mixed with a magnetic stir-
rer. This solution was then divided into three 150-rnA aliquots
in triplicate beakers to provide replicate exposure treatments.
The remaining 300 m£ were used for 0-hr DO, pH, specific conduct-
ance, alkalinity, and total hardness determinations.
A control, consisting of the same dilution water and conditions,
but with no effluent, was established. All test vessels were
maintained at 22 ± 1°C and test solutions were not aerated during
the test. Five daphnids were randomly assigned to each test ves-
sel within 30 minutes after the compound was added for a total
of 15 daphnids per concentration.
During the tests, the dissolved oxygen concentration, pH, and
temperature of test solutions were monitored at the initiation
and termination of the toxicity test in the control, high, middle
and low test concentrations. Total hardness, specific conduct-
ance and alkalinity were monitored at the initiation of the study
in the control, high, middle and low test concentrations. DO and
(12) Methods for Acute Toxicity Tests with Fish, Macroinverte-
brates, and Amphibians. EPA-660/3-75-009 (PB 242 105) U.S.
Environmental Protection Agency, Duluth, Minnesota, March
1975. 61 pp.
(13) Personal communication with C. E. Stephan, U.S. Environmental
Protection Agency, Duluth, Minnesota, 1978.
107
-------
temperature were measured with a USI dissolved oxygen meter and
combination oxygen- temperature probe. pH was measured with an
Instrumentation Laboratory pH meter. The total hardness determi-
nations of diluent water were conducted according to Reference 4.
Salinity and specific conductance were determined with an American
Optics refractometer and a YSI conductivity bridge, respectively.
Results of Fathead Minnow and Daphnia Bioassay
Appendix E shows the raw data as reported by EG&G Bionomics for
each of the 14 fathead minnow bioassay samples. The first table
for each sample shows the pH, DO, specific conductance, total
hardness and alkalinity measurements made during the 96-hr test.
The second table associated with each sample shows the percent
mortality raw data used to calculate LCso values.
Appendix F shows the raw data as reported by EG&G Bionomics asso-
ciated with each sample for the daphnia acute toxicity tests.
The first table for each sample shows the water quality analyses
of each as a function of concentration tested. The second table
shows the raw mortality data used to calculate LCso values.
Results of the two static acute toxicity tests are shown in
Table 18. In addition to LCso values, the mortality data was
used to extrapolate the concentration of wastewater sample which
would produce no discernible effect on the fathead minnows or
daphnids. Analysis of the data indicate the four permeate sam-
ples (CTHR-5, 8, 12, and 15) produced no or very little mortality
to fathead minnows. The most toxic wastewater samples were the
concentrates (CTHF-6, 9, 13, and 16).
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 evaluate the acute in vivo
toxicity of samples containing unknown compounds. The first
approach was based on the quantal (all-or-none) response; the
second was based on the quantitative (graded) response. The
quantal test was used to determine whether or not the quantitative
assay was necessary. Fourteen wastewater samples were shipped
by Clemson University to the MRC designated laboratory: Litton
Bionetics, Kensington, Maryland. The following sections describe
the test procedures used and bioassay results.
The Quantal Test Procedure
Five male and five female young adult rats (weighing approximately
250 g each) were purchased by Litton Bionetics from the supplier
and conditioned at the laboratory for a minimum of 5 days. A
single 10 m£/kg dose of undiluted sample was administered by
gavage to each animal. Immediately following administration of
108
-------
TABLE CIS. RESULTS OF STATIC ACUTE TOXICITY TESTS TO FATHEAD MINNOWS AND DAPHNIDS
Fathead minnow acute toxicity
LC50 at
Sample
CTHF-3
CTHF-4
CTHF-5
CTHF-6
CTHF-7
CTHF-8
CTHF-9
CTHF-10
CTHF-11
CTHF-12
CTHF-13
CTHF-14
CTHF-15
CTHF-16
24-hr
a
>11 <24
28 (24 to 33)
NATC
>2.4 <5.3
16 (7.8 to 32)
NAT
>3.2 <5.3
17 (14 to 21)
>100
NAT
21 (1.6 to 27)
41 (35 to 48)
NAT
9.7 (6.3ito 14)
16
28
1.6
13
2.7
10
1.7
41
5.3
time intervals, %
48-hr
(13 to 21) b
(24 to 33)
NAT
(1.2 to 2.3)
(7.8 to 22)
NAT
(2.2 to 3.3)
(8.1 to 12)
>100
NAT
(1.2 to 2.1)
(35 to 48)
NAT
(4.1 to 6.8)
16
28
1.6
13
2.2
10
87
1.7
25
5.3
sample solution
72-hr
(13 to 21)
(24 to 33)
NAT
(1.2 to 2.3)
(7.8 to 22)
NAT
(1.6 to 2.9)
(8.1 to 12)
(20 to 100)
NAT
(1.2 to 2.1)
(21 to 29)
NAT
(4.1 to 6.8)
16
28
1.5
13
2.0
9.7
82
1.6
25
5.3
Daphnia acute toxicity
No discernible No discernible
effect LDso at time intervals, effect
concentration % sample solution concentration
96-hr
(13 to 21)
(24 to 33)
_d
(1.0 to 2.2)
(7.8 to 22)
NAT
(1.5 to 2.8)
(7.5 to 12)
(21 to 100)
NAT
(1.2 to 2.0)
(21 to 29)
NAT
(4.1 to 6.8)
at 96-hr, %
0.53
4.6
_e
0.24
<7.8
_e
<0.41
<4.6
36
_e
0.78
11
_e
<0.36
24-hr
59 (49 to 72)
>60 <100
60 (36 to 100)
10 (7.3 to 15)
>100
>100
13 (11 to 16)
>100
>100
>60 <100
11 (2.5 to 48)
>60 <100
>100
>100
48-hr at 48-hr, %
26
53
42
5.1
25
9.9
33.5
4.1
49
80
17
(20 to 34)
(45 to 62)
(35 to 51)
(4.2 to 6.2)
(20 to 31)
>100
(8.3 to 12)
(27.6 to 50.4)
>60 <100
>60 <100
(3.4 to 4.9)
(41 to 58)
(71 to 90)
(13 to 23)
<13
22
13
2.
60
4.
22
60
60
2.
<13
2.
8
1
8
8
>11 <24 = 24-hr LCsg value is greater than 11% but less than 24% solution of the wastewater.
Values in parentheses are 95% confidence interval.
CNo acute toxicity.
Only 30% mortality occurred in 100% solution of wastewater.
Not calculated due to insufficient mortality.
-------
the test substance and at frequent intervals during the first
day, observations were recorded on all toxic signs or pharmaco-
logical effects as described in Table 19 (14). 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.
Wastewater 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 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 LD50 was reported as
greater than 10 g/kg.
The Quantitative Assay Procedure
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 dosages according
to the following schedule: 3.0 g/kg, 1.0 g/kg, 0.3 g/kg, and
0.1 g/kg. Dosage was related to the numbers of animals that died
and to the severities and types of signs. Observations, duration
of study, and necropsies were carried out as indicated above.
The LD5o was calculated by the method described in Reference 14.
The range-finding tests were conducted at Litton Bionetics under
the direction of Dr. R. Beliles. Actual experimental design
parameters used in this study were as follows. Young adult rats
of the Charles River CD strain [CRLrCOBS CD (SD) BR] were ob-
tained 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 ex-
ception of the night before treatment when food was removed from
cages.
Wastewater samples were kept refrigerated until used. A single
undiluted dose of 10 m&/kg of test material was administered by
(14) Duke, K. M., M. E. Davis, and A. J. Dennis. IERL-RTP Proce-
dures Manual: Level I 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.
no
-------
TABLE C19. PHYSICAL EXAMINATIONS IN ACUTE TOXICITY TESTS IN RODENTS (3)
Organ system
Observation and examination
Common signs of toxicity
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.
-------
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,
Results of Rat Acute Toxicity Test
Final reports from Litton Bionetics by Dr. R. Beliles state that
no rats died as a result of single maximum dosages of the 14
wastewater samples. Necropsy results indicated no sample related
effects were observed. Therefore, no samples were subjected to
quantitative analysis.
112
-------
REFERENCES
1. Sampling and Analysis Procedures for Screening of Industrial
Effluents for Priority Pollutants. Draft final report, U.S.
Environmental Protection Agency, Cincinnati, Ohio, April
1977. 145 pp.
2. Rawlings, G. D. Source Assessment: Textile Plant Waste-
water Toxics Study - Phase I. EPA-600/2-78-004h, U.S.
Environmental Protection Agency, Research Triangle Park,
North Carolina, March 1978. 166 pp.
3. 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.
4. Standard Methods for the Examination of Water and Wastewater,
Fourteenth Edition. American Public Health Association,
Washington, D.C., 1976. 874 pp.
5. Carter, M. J., and M. T. Huston. Preservation of Phenolic
Compounds in Wastewaters. Environmental Science and Tech-
nology, 12(3):309-313, 1978.
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. Ames, B. N., J. McCann, and E. Yamasaki. Methods for the
Detection of Carcinogens and Mutagens with the Salmonella/
Mammalian-Microsome Mutagenicity Test. Mutagenicity Re-
search, 31:347-364, 1975.
8. 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.
9. Wininger, M. T., F. A. Kulik, and W. D. Ross. In Vitro
Clonal Cytotoxicity Assay Using Chinese Hamster Ovary Cells
(CHO-K1) for Testing Environmental Chemicals. In Vitro,
14:381, 1978.
113
-------
10. Peltier, W. Methods for Measuring the Acute Toxicity of
Effluents to Aquatic Organisms. EPA-600/4-78-012 (PB 276
690) U.S. Environmental Protection Agency, Cincinnati, Ohio,
January 1978. 63 pp.
11. Harris, E. K. Confidence Limits for the LD50 Using the
Moving Average Angle Method. Biometrics, 4(3):157-164, 1959.
12. Methods for Acute Toxicity Tests with Fish, Macroinverte-
brates, and Amphibians. EPA-660/3-75-009 (PB 242 105) U.S.
Environmental Protection Agency, Duluth, Minnesota, March
1975. 61 pp.
13. Personal communication with C. E. Stephan, U.S. Environmental
Protection Agency, Duluth, Minnesota, 1978.
14. Duke, K. M., M. E. Davis, and A. J. Dennis. IERL-RTP Pro-
cedures Manual: Level I 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.
15. 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.
114
-------
APPENDIX CA
PRIORITY POLLUTANT ANALYSIS FRACTIONS
TABLE CA1.. VOLATILE COMPOUNDS
Compound
Compound
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 (chloromethy1) ether
1,2-Dichloropropane
trans-1,3-dichloropropene
Trichloroethylene
Dibromochloromethane
Ci8 -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
115
-------
TABLE GA2 . 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 CAS. ACID EXTRACTABLE COMPOUNDS
2-Chlorophenol
Phenol
2,4-Dichlorophenol
2-Nitrophenol
p-Chloro-m-cresol
2,4,6-Trichlorophenol
2,4-DimethyIphenol
2,4-Dinitrophenol
4,6-Dinitro-o-cresol
4-Nitrophenol
Pentachlorophenol
116
-------
TABLE CA4. PESTICIDES AND PCB
Compound
B-Endosulfan
a-BHC
Y-BHC
8-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 CA5. METALS AND OTHER COMPOUNDS
Metals,
total Others
Antimony Asbestos
Arsenic Cyanide
Beryllium
Cadmium
Chromium
Copper
Lead
Mercury
Nickel
Selenium
Silver
Thallium
Zinc
117
-------
APPENDIX CB
RAW DATA FROM THE AMES MUTAGENICITY TESTS
AS DIRECTLY REPORTED BY MRC
118
-------
Sample CTHF- 5
Hssay Amount Revertants Per Plate
Test Condition Sample (yl/plate) TA98 TA100 TA1535 TA1537 // /
V3 50 57, ^ )*, I* '*/ " 3°'
With Activation Control v^-^v Jif -j *>
2-aminoanthracene 20 yg ?V7 ""a ^^ 9/
100
SCO
j 000 /$ , 10 AZI 2 (°
-> i i iT /9/7 / *
-------
Sample
- 3
Test Condition
Sample
With Activation
Control
2-aminoanthracene
H
to
O
Without Activation Control
2-nltrofluorene
Assay Amount
(Hi/Plate)
20 yg
10
SO
100
Soo
1000
2.0 ug
9.0 mg
TA98
Revertants Per Plate
TAlOb
YA1S55
If 9,
\U, 137
-------
Sample CT H p ~
Test Condition
Sample
Assay Amount
hi/Plate)
With Activation Control
2-aminoantnracene 20 yg
/£>
/ 000
TA98
, .35"
Revertants Per Plate
TA100
57
35-31
3S1,
TA1535
TAI537
//
/fe
9,
3?, 3f
li
Without Activation . Control 3 «2( 35
2-nitrofluorene 2.0 ug / /
Nafi02 9.0 mg _
100
Sec
I OCc
37
35"
33. 3-)
n,
/O
10
/•?,
to, /O
*, 9
'>> V
'* //
-------
Sample C~T tf F -
Test Condition
Sample
Assay Amount
(pi/Plate)
With Activation
Control
2 -ami noanthracene
20 yg
10
50
I DO
£GO
I O GO
TA98T
Revertants Per Plate
"TAlOO tAl53S
TA1537"
173
Without Activation . Control
2-nitrofluorene
NaMG,
2.0 ug
9.0 mg
-------
Sample CTtjF- <5~
Assay Amount ; Revertants Per Plate
Test Condition Sample (pi/plate) TA9§ WOO TA1535 TAT537
to
U)
Without Activation . Control %. If 3.1*}. ll^j 2H t S.(o l^>, Mo
~~~ i I *
2-nitrofluorene 2.0 yg
NaN02 9.0 mg
/O
50 24, 55"
I vis A, ^-1 J *
600 / 33 £ /I//
2-aminoanthracene 20 yg ///x
^^r J i^S i ^M
10 (<>(*, 12 2 ', o , /
^^ <-,)(*£ 2l*>, 2ol n, n !<*, I*
6O /*/»-» ' '
yoo ^/0,
5"^> ^> 70 'VS ^^ / 8"^ "/ ^ ' ^
/£>OD 5^, 4?/ ^°Vy /9o //, /-? ^/ /3 3^3^
' / f
-------
Sample
Revertants Per Plate
Test Condition Sample
With Activation Control
2-ami noanthracene
-
Without Activation . Control
2-nitrofluorene
NaNC2
-
(yl/plate) TA98
(o$, SI
20 yg nil
10 **£>, 6%
50 51, bf
loo *^ ^, '
ill, 13% &, 73 IL, if ^v,ot l(* "^' 3^
/347 /Of //x /7 ^i /6) 3S, 35^
/JTOx /3<9 /3, 4» ^/ /D 33' ^
^/^ //^/ 21,2-^ /'3, /^. /V, /3
5"69 — — //5-
— 35-3 - __
^/S,/77 /C, /^ /*, /y /(,, //
/W, /t/ /^/ -?o /^ 7 /y, /3
^^7, /74 /j, /r /^ 9 /^ /y
•/J,
too* ** j* it n ?. y 3, /s ^ /s
-------
Sample (-7H F ~
Assay Amount
Test Condition Sample (yl/plate) TA98
/ Q %0
With Activation Control to~'
2-aminoanthracene 20 yg % #96
10 *?. ^
IOO 35", 38
600 fot S~l
iooo tf, 35"
Without Activation Control 66^ ^ '1
2-nitrofluorene 2.0 pg 3^7
NaNC2 9.0 mg — •
/D 8%5"9
5"o 5^/ ^°
/oo 5"3, 5"5T
5"^o VJ?( 5^
Revertants Per Plate
TAlOO TA1535 TA1537 "*9 X53g'
«2/5 jZ^> 5^ /6/ -^<< -5/y •5*< -^ ^/ 77
J? v/ m H4,it '*'*' y^
ii~), in '*, /4; ' ' ^j
„», ^9 '% ^
a«, »/ 33, /« 3', /» *->,**
^ $
^ :^ i;/5 ^'-
^fTS 327 ^ /^ ^' ^ /3'
y . . . , a / 1
II.
-------
Sample
Test Condition
With Activation
Assay Amount
Sample (yl/plate)
Control
2-aminoanthracene 20 yg
ib
fo
JOV
600
TA98
6V, *0
ZZHo
5-), ^
$2, a
5*, S(o
S7, S<~
Revertants Per Plate
TA100 TA1535
m^t ZD£ H*, 21
•tfWb &K>
Ml, XW W, '*)
3o^ 35-8 **>, />
^/fc/ /(,» ^ *4
27^ ^^0 ^ /6
^. ^ + s\ . i » t **** /*i.
TA1537 1M/339
3t, I* 3^ ^
'•? 7o / *?0o
At, 30 57, 5-y
52^ T7 35, VJ
-•7 <" / fT <^ 7 3 •
3.3 f 13 1 /, ->*
22, '(* Vyx 3
,26/3 -2/,3
CTl
/coo
/£, 6t? ^^ a6/ 3"^
Without Activation Control ^ "X
2-nitrofluorene 2.0 yg
NaNOo 9.0 mg
2*, 27
/o
-7^ 6"? 36~3 V/o
/ r\ /-, Q-' t /•! •* n -^ Jie / '• —' . •'/
500 5-9, ^2 333,
30,
-------
Test Condition
Sample
Assay Amount
(yl/plate)
With Activation
Control
2-ami noanthracene
20 ug
ID
)00
500
NJ
Sample CjHF " 10
i
Revertants Per Plate
TA98
3110
-------
Sample CTt+F ' II
to
CO
Assav Amount
Test Condition Sample (yl/plate) TA98
'?/, 1H
With Activation Control '
2-aminoanthracene 20 ug 5O" 7
10 **' ^
So 63, 63
i^ *
l&D 6>q, bl
r
60V G> 7, $1
/OVO 66, 5"V
t/< %(
1 J *
Without Activation Control
£/ ^ T
2-nitrofluorene 2.0 pg <^V /?, ^
*
., , ^ x*V /*^ J al /
V 3 ?7
X, ^-O*' ^^ ^^,
^6^x ^^5 ^,^^
Atl, 22*3 3o; ^/
/7^x £/t
V
37
-------
Test Condition
Sample
Assay Amount
With Activation
Control
2-aminoanthracene
,20
Sample
TA98
Revertants Per Plate
TATS3F
(O
Without Activation Control
2-nitrofluorene
NaNO,
2.0 pg
9.0 mg
10
50
loo •
5-00
/DOO
52 7
n,
-------
Sample
Assay Amount Reyertants Per Plate
Test Condition Sample (pi/plate)" TA98 TAlbo' TftT555
With Activation Control
acene 20 yg ^O
70
*,
"/ '*
10
1(0
•7/ JV 2Z1, ^>Y I/, *" *2j l(o 3*/
ITn& -7 LJVQQ i%~7 317
2-aminoanthracene 20 yg $017 «Y.
55; ^^
Without Activation ControT V^/ 33»x J?W /^V ^
2-nitrofluorene 2.0 pg V^5" ^4>O
NaNG2 9.0 mg — ^ 5" 3
4,7,73 3«^w ^'/7 7///
5-0 7^,77 33^^^ «^/^^ /^/// ^^
100 73,*/ 3^ 34>4> ^^^ ^/^ % y
5(90 ^^ 7/ W% 33
-------
Sample
Test Condition
Sample
Assay Amount
(pi/plate)
With Activation
Control
2-ami noanthracene
20
TAW
Revertants Per Plate
TAlW
Without Activation Control
2-nitrofluorene
NaNO-,
2,0 ug
9.0 ing
10
I DO
£0
Hi,
it,
-------
Sample C Tftf - I 3
Assay Amount Reyertants Per Plate.., -rorr
Test Condition Sample (yl/plate) TA9& TAfbO YA1S35 TAIS37
Without Activation . Control
2-nitrofluorene 2.0 pg
NaN02 9.0 mg — — O
fO *?0, 7*
17,
With Activation Control "' ' ' ' ' ^^ -,-,
x-na-y U-ZQQ 127 *'7
2-aminoanthracene 20 yg OUf/
10 7V, 5to
t>3, 2o
~ ", 3./
/ 00 to*, /' ~~ '
f*° ^'77 *,,0 !«,«>
I 000
SO
100 It, 43 ***>' 3n
600 *,rr *** *** '*>'* *' 4 ^ '3
/OOO $1,71
-------
Sample CTftf -/3
Assay Amount Revertants Per Plate
Test Condition __ Sample _ (pi/plate) TA98 TAlOO TAl535 TA1S5T
With Activation Control
2~aminoanthracene 20
U)
U)
"7 c?
Without Activation Control ^/ '" ._,
2-ni trofluorene 2.0 wg 53-7 , _
NaN&? 9.0 mg - — -^ O
^ 3
6, O
LOO &st
7
1000
-------
Sampl
e CTH-F " /H
u>
Assav Amount Kevertaru
Test Condition Sample (yl/plate) TA98 TA100 ._
y/ 3/ 5^,60
With Activation Control -*QU 35S~?
2-aminoanthracene 20 yg o ° VT
,0 3^,33 **, *^
5-0 ,2*, 3*> 6V, 7?
/00 <#?, 5"3 k£^ 5"7
5"^^ 3^ V* *// 7:?
y£)^ V^,^ 7*, <^^
//, ;M
5^ /6X ^ ^f3y ^/
/oo *7^t ***,**
5 oo Mt*l *so}*-n
1000 ?i 7 33*. *SO
* r YZfe5 TA153/ ' *9/.53S
9X ^ //x/^ ^^^
330 ££* 5;?^
29,^ /?^^ ^'a<
,o,1 '*,'* ***'*'
it u n<« *7'**
' ,, G £ f/30
&,/* "' V ,'.
5~/ f ^ */ ^/
^ /^ 5/ 7
yo ^/ ^0/ /x V/ ^
/J^// _!_ ^47
/ W — ~~
/ ^- ^ / ^? t3, JO
l(o,l£ £Ll,o '
11,1 %» '*'*3
ft// /^^ ^/X
d ,* /^ /6 ^f
^9 ^^
3X 3 */ Y
-------
Sample CTliF- I <-{
Test Condition
Sample
Assay Amount
(yl/plate)
Revertants Per Plate
TA98
TA100
TA1535
TAT 537 Tfl/53&
With Activation
CO
01
Control
2-aminoanthracene
20 yg
10
So
|OO
5oo
iooo
13 IH
Without Activation
Control
2-nitrofluorene
NaKO,
2.0 ug
9.0 mg
-------
Sample C,THF ~
u>
Assay Amount Revertants Per Plate
Test Condition Sample (vl/plate) TA96 TA10& YA1S35
With Activation Control ' ,-si/j «^o
^S'^/V ^ ' ^
2-amlnoanthracene 20 wg 0* "/ ^
1 OO Cft 3J? ; 2 9 i ~7I <& '£*
1 v *-/ \s \J -JO /^\0'*' '
X /
5OO 95; ^^ ^^ /77 7; //
IOOO ^g /*?0 /9^ /O^ *L/
2-n1trofluorene 2.0 pg 3 (o 8 — —
NaN02 9. 0 mg — — / ^ ^
lo */ 7 - /^/ ^
SO 3H>f U 21$ t WO 1 ^, ^
» 00 /^ £0 ,206, ;?;# ^3, ^
5^0 , ft ;4, ^5-^ ^^^ 7, /$"
l°0o . 7. // *tq i** y, /3
TA1537 7# /^^
J i J '0 "^ £•. ^5 ^s^
mi i &\ &*• ^^ / ^^ +J
1 3. /3 /5>^5*
5"x /^ / -
/?, // ^ ^-«
^ /J /^ =2
/^ /4 5, 9 /^ '/
//, 7 /•*, /o
/•>/// ^ /o
xl xl . jf y/ 1 £~
CC«C'& tl. f O
$, *Z % /6
-------
Sample
- / 5
Test Condition
Sample
Assay Amount
(pi/plate)
TA98
Revertants Per Plate
TA1o6
YAlSJS
TA1537
With Activation
Control
2-aminoanthracene
20 yg
Ul
Without Activation
Control
2-nitrofluorene
NaN0
2.0 ug
9.0 mg
ID
50
I O O
5oo
I D CO
1>/ f
5O3
6,8
-------
00
Sample C T HF - / 1&
Assay Amount Revertants Per Plate
Test Condition Sample (pi/plate) TA98 TA100 TA1535
. f-.y r-£ £-£, ft If
With Activation Control '/ ' '
2-aminoanthracene 20 yg ^cTV3 IHfof Y
,0 39, 3
5O 5"^ ^/5" 35"x ^3 /4, /9
100 3^,5"^? 6'^/ ^ 3 / 5", /^
f r
5 O o c/ 5" 3-7 ^y 5"x 3 3 / 5", / V
1^0 -57 /fr V7, ^fc ^; 7
Without Activation . Control ( % . /
-------
Sample CJ ftp— I
Test Condition
Sample
Assay Amount
(ul/plate)
Revertants Per Plate
TA98
TA100
TA1535
TA1537
With Activation
CO
Control
2-aminoanthracene
20 yg
ID
50
loo
Boo
Jooo
Without Activation Control
2-nitrofluorene
NaN02
2.0 vg
9.0 mg
-------
APPENDIX CC
RAW DATA FOR THE CHO CYTOTOXICITY TESTS
AS DIRECTLY REPORTED BY MRC
140
-------
•YTOTOXICITY DATA FOR CADMIUM CHLORIDE STANDARD
;ELL LINE: CHO
6/5/78
PAGE REFJ 1207195
CONTROL
BACKGROUND)
UALUES
430
440
450
445
430
445
MEAN
VALUE:
440
STANDARD
DEVIATION
8
;t)NCENTRATIGN
CMG/ML)
REPLICATE
VALUES
MEAN
VALUE
STANDARD
DEVIATION
PERCENT
SURVIVAL
,001
,.0005
..0004
-0003
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
..0001
90
J.5
15
141
-------
MONSANTO COMPANY
N2 1214106
.UHJECT
i ' H .
>OB NO.
,» r
BY (SIGNATURE)
DATE
(OTOXICITY DATA FOR CADMIUN CHLORIDE
, L LINE: CHO
6/22/78
PAGE REFJ 1214103
CONTROL
nCKGROUND)
VALUES
421
369
374
524
449
473
iJCENTRATION
(MG/ML)
-001
. 0005
0004
• 0003
• '002
. "001
MEAN STANDARD
VALUE DEVIATION
435 60
REPLICATE MEAN STANDARD PERCENT
VALUES VALUE DEVIATION SURVIVAL
0 0 0 0
0
0
000 0
0
0
000 0
0
0
000 0
0
0
2 10 11 2
22
6
78 312 204 100
409
449
READ AND UNDERSTOOD BY DATE
14? =
-------
'•YTOTOXICITY DATA FOR CADMIUM CHLORIDE STANDARD
OELL LINE; CHO
7/6/78
PAGE REF: 1214115
CONTROL
•BACKGROUND)
VALUES
625
61:1.
614
607
MEAN
VALUE
617
STANDARD
DEVIATION
8
615
;ONCENTRAT1'ON
(MB/ML)
, 001
,0005
.0004
.0003
. 0002
REPLICATE MEAN
VALUES VALUE
0 0
0
0
0 0
0
0
7 8
7
9
25 21
13
25
573 575
642
511
647 638
615
653
STANDARD PERCENT
DEVIATION SURVIVAL
0 0
0 0
1 1
7 3
66 100
20 100
in:., f.i
143
-------
:YTOTOXICITY DATA FOR CADMIUM CHLORIDE STANDARD
:ELL LINE: CHO
7/11/78
PAGE REF: 1214126
CONTROL.
BACKGROUND)
VALUES
557
547
518
520
528
546
MEAN
VALUE
536
STANDARD
DEVIATION
16
ONCENTRAT10N REPLICATE MEAN
(MG/ML) VALUES VALUE
,001 0 0
0
0
.0005 0 0
0
0
. 0004 0 0
0
0
.0003 0 0
0
0
.0002 57 34
32
12
-0001 520 516
521
508
STANDARD PERCENT
DEVIATION SURVIVAL
0 0
0 0
0 0
0 0
23 6
7 100
-------
:YTOTOXICITY DATA FOR CTHF-S
:;ELL LINE.' CHO
7/6/78
PAGE REF: 1212115
CONTROL
BACKGROUND)
VALUES
625
611
A1A
607
627
615
MEAN
VALUE
617
STANDARD
DEVIATION
8
.ONCENTRATION
(UL/ML)
REPLICATE
VALUES
MEAN
STANDARD
DEVIATION
PERCENT-
SURVIVAL.
200
J. 00
lo
687
707
725
723
733
685
702
696
707
738
684
655
643
691
652
586
577
584
609
569
608
706
19
702
69
662
58
595
42
26
loo
100
100
100
100
94
100
145
-------
CYTOTOX1CITY DATA FOR CTFH-4
CELL LINE: CHO
7/6/78
PAGE REF: 1214115
CONTROL
(BACKGROUND)
VALUES
624
612
614
607
627
615
MEAN
VALUE
617
STANDARD
DEVIATION
8
CONCENTRATION
(UL/HL)
200
150
100
10
REPLICATE
VALUES
711
734
709
685
670
695
674
652
647
625
644
645
647
608
550
582
597
614
613
611
599
MEAN
VALUE
718
683
658
638
602 ,
598
608
STANDARD
DEVIATION
14
13
14
11
49
16
8
PERCENT
SURVIVAL
100
100
100
100
100
100
100
:ASIC
146
-------
"YTOTOXICITY DATA FOR CTHF-5
;;ELL LINE: CHO
7/6/78
PAGE REF: 1214115
CONTROL
BACKGROUND)
UALUES
625
6:1 J.
6:1.4
60 /
627
6.1.5
MEAN
VALUE
617
STANDARD
DEVIATION
8
.ONCLNTRATIQN
(UI.../ML)
REPLICATE
VALUES
MEAN
VALUE
STANDARD
DEVIATION
PERCENT
SURVIVAL
200
616
665
667
649
29
100
10
664
638
647
626
678
660
635
613
634
628
617
587
608
602.
630
614
602
590
650
655
627
611
613
602
13
26
12
21
15
12
100
100
100
100
100
100
147
-------
MONSANTO COMPANY
N2 1214107
SUBJECT
JOB NO.
YTOTOXICITY DATA FOR CTHF-6
ELL LINE: CHO
6/22/78
PAGE REF: 1214103
CONTROL
BACKGROUND)
VALUES
421
369
374
524
449
473
MEAN
VALUE
435
STANDARD
DEVIATION
60
Q8CEWTRATIQN
REPLICATE
VALUES
HEAH
VALUE
STANDARD
DEVIATION
PC8GENT
SURVIVE
200
150
100
50
10
4
4
20
5
10
122
166
170
396
424
378
465
469
453
466
387
256
245
363
212
153
399
46?
370
273
8
27
23
16
106
79
35
100
100
100
63
RKAO AH0 UMD*M«TOOO BY
DATE
-------
MONSANTO COMPANY
N2 1214149
'.UBJECT
liTOXICITY DATA FOR CTMF
LINE; cuo
HRLPARKD BY (SIGNATURE!
DATE
7/25/78
PAGE REF: 1214143
ONTROL
K6ROUND)
•MI. utrs
497
480
4S4
450
480
478
MEAN
VALUE
478
STANDARD
DEVIATION
i'.N'lRATION
•. UL./IU.)
REPLICATE
VALUES
MEAN
VALUE
STANDARD
DEVlAdUN
PERCENI
SURVIVAL
.1.1
.10
|50
4.!. 9
431
19
At;
4!.j4
'•• •,'",)
A 8
4'.r,
16
100
.1.00
149
READ AND UNDERSTOOD BY
DATE
-------
oToxieiTY DATA FOR CTHF--?
.1. LINE: CHO
7/6/78
PAGE RER 12.1.4116
CONTROL.
BACKGROUND)
VALUES
625
61 1.
<•> .I 4
607
627
6J5
MEAN
VALUE
617
STANDARD
DEVIATION
8
CONCENTRATION
(UI.../ML)
REPLICATE
VALUES
MEAN
VALUE
STANDARD
DEVIATION
PERCENT
SURVIVAL
200
'.SO
t 00
706
700
675
695
650
605
655
695
654
633
628
635
6.15
637
6:1.8
623
6.19
580
602
617
694
650
668
632
623
607
605
16
45
23
12
11
100
100
100
100
100
100
100
150
-------
•Y10TOXICITY DATA FOR C
;EI...L LINE; CHO
THF-8
7/6/78
PAGE;: REF: 1214116
CONTROL
BACKGROUND)
VALUES
625
6.1.1.
614
607
627
615
MEAN
VALUE
617
STANDARD
DEVIATION
8
'.(ihfLElNTRATIQN
liUL/ML)
150
100
REPLICATE
VALUES
677
678
618
604
581
575
631
650
615
640
619
601
594
60 A
605
631
606
624
606
626
633
MEAN STANDARD PERCENT
VALUE DEVIATION SURVIVAL
658 34 100
587 15 95
632 18 100
620 20 100
601 6 97
620 13 100
622 14 10°
'•': 1 C
151
-------
CYTOTOXICITY DATA FOR CTHF-9
:ELL LINE* CHO
7/6/78
PAGE REF: 121411*
CONTROL
iBACKGROUND)
625
611
614
607
627
615
(UL/ML)
200
150
100
50
10
739
70O
730
735
712
716
676
694
714
695
677
616
609
590
591
620
617
609
638
627
726
«*5
409
625
12
13
16
15
STANDARD
DEVIATION
8
PERCENT
SURVIVAL
100
100
100
100
100
100
100
iASIC
152
-------
MONSANTO COMPANY
N2 1214108
SUBJECT
.U-teLu.fc.vL
JOB NO.
PREPARED BY (SIGNATURE)
-!••"• '
DATE
CYTOTOXICITY DATA FOR CTHF-10
CELL LINE: CHO
6/22/78
PAGE REF: 1214103
CONTROL
C BACKGROUND) HEAN
VALUES VALUE
421 435
369
374
524
449
473
STANDARD
DEVIATION
60
CONCENTRATION REPLICATE MEAN STANDARD PERCENT
(1JL/HL) VALUES VALUE DEVIATION SURVIVA
200 291 228 66 52
234
159
150 495 385 168 100
192
468
100 492 483 10 100
484
473
50 486 510 25 100
536
507
10 491 481 22 100
497
456
2 94 100 9 23
110
96
.2 450 449 16 10°
46S
433
READ AND UNDER1TOC
153
>O B 1 DATE
_..,. -„
-------
GYTOTOXICITY DATA FOR CTHF-10
r:t:LL LINE: CHO
7/11/78
PAGE REF: .1214126
CONTROL
KACKGKOUNB > MEAN
VALUES VALUE
557 536
547
518
520
528
546
ONCENTRATION REPLICATE MEAN
(UL/ML) VALUES VALUE
200 596 588
603
565
150 558 552
543
554
1.00 550 538
532
531
50 518 525
544
514
10 500 511
521
511
2 524 519
518
515
. 2 532 530
528
530
STANDARD
DEVIATION
16
STANDARD PERCENT
DEVIATION SURVIVAL
20 tOO
8 100
11 100
16 100
11 100
5 100
2 100
:A3IC
154
-------
,-YTOTOXICITY DATA FOR CTHF-11
CELL LINE: CHO
7/11/78
PAGE REF: 1214126
CONTROL
(BACKGROUND)
VALUES
557
547
5.18
520
528
546
MEAN
VALUE
536
STANDARD
DEVIATION
16
NPFNTRATION
IXL*fc.li I i\n i **•*!>
(UL/ML)
200
150
100
50
10
")
* *
REPLICATE MEAN STANDARD PERCENT
VALUES VALUE DEVIATION SURVIVAL
566 564 18 100
580
545
549 531 16 100
526
518
476 489 18 91
482
510
535 515 23 100
490
520
524 548 23 100
570
550
514 510 19 100
527
490
515 522 7 100
523
528
155
-------
:YTOTOXICITY DATA FOR CTHF-12
CELL LINE; CHO
7/11/78
PAGE REF: 1214126
CONTROL
BACKGROUND)
VALUES
557
547
518
520
528
546
MEAN
VALUE
536
STANDARD
DEVIATION
16
,,'QNCENTRATION
(UL/ML)
200
150
.!. 00
50
10
REPLICATE
VALUES
530
518
545
550
560
516
542
572
524
513
528
552
456
517
468
483
490
535
520
485
528
MEAN
VALUE
531
542
546
531
480
503
511
STANDARD
DEVIATION
14
23
24
20
32
28
23
PERCENT
SURVIVAL
100
100
100
100
90
100
100
i A 81C
156
-------
CYTOTOXICITY DATA FOR CTHF-13
CELL LINE: CHQ-KI
6-12-78
PAGE REFJ 1207186
CONTROL
(BACKGROUND)
VALUES
430
445
430
440
450
445
MEAN
VALUE
440
STANDARD
DEVIATION
8
CONCENTRATION
(UL/ML)
200
150
100
50
2
BASIC
>
REPLICATE
VALUES
0
0
0
30
70
25
260
195
155
370
520
470
450
MEAN
VALUE
0
42
203
445
470
450
STANDARD
DEVIATION
25
53
106
0
0
PERCENT
SURVIVAL
0
46
100
100
100
157
-------
rum x i. c :i. i Y D A r A F o R c i H F • 13
ll I. .I.N::.: CHQ
7/11/78
PAGF REF: 1214126
(.'KGhi)LIND '>
UAI. lit. 8
MEAN
VALUE
STANDARD
DEVIATION
16
: UL./MI..)
REPLICATE
VALUES
94
170
".'> ") '")
440
451
383
540
521
541
570
574
531
528
530
525
514
515
500
507
435
514
MEAN
VALUE
162
425
534
558
528
510
502
STANDARD PERCENT
DEVIATION SURVIVAL
64 30
37 79
11 100
24 100
3 100
8 95
t
15 94
158
-------
MONSANTO COMPANY
N2 1214112
{SUBJECT
V ". *v
,. !•-- i iM.rrf.-is-.., •> .. ;> i H ;„ - ....
PREPARED 8Y (SIGNATl
rTOTOXICITY DATA FOR CTHF-14
LI.L LINE: CHO
CONTROL
BACKGROUND ) MEAN
VALUES VALUE
421 435
369
374
524
449
473
;ONCENTRATION REPLICATE MEAN
(UL/ML) VALUES VALUE
200 515 501
483
505
150 513 511
482
539
100 325 , 312
181
430
30 492 476
470
465
10 491 488
515
457
2 384 421
494
385
.2 484 466
473
440
RKAD AND I
159
IRE) r ___
DATE
1 £. /**'^.J_z2.
6/22/78
PAGE REF; 1214104
STANDARD
DEVIATION
60
STANDARD PERCENT
DEVIATION SURVIVAL
16 100
29 100
125 100
14 100
29 100
63 100
23 100
INOIM1TOOO BY DATE
-------
:*YTOTOXICITY DATA FOR CTHF-I5"
:ELL LINE: CHO
7/11/78
PAGE REF:
CONTROL
BACKGROUND)
VALUES
557
547
518
520
546
528
VALUE
536
STANDARD
DEVIATION
16
:ONCENTRATION
(LJL/ML)
,:>00
1 00
.,0
10
REPLICATE MEAN
VALUES VALUE
556 544
547
530
557 552
570
528
559 539
541
517
473 491
482
518
491 495
503
492
512 505
478
524
539 535
526
540
STANDARD
DEVIATION
13
22
21
24
7
24
8
PERCENT
SURVIVAL
100
100
100
92
92
100
100
160
-------
MONSANTO COMPANY
N2 1214114
SUBJECT
PREPARED BY (SIGNATURE)
-YTQTOXICITY DATA FOR CTHF-16
JELL LINE! ABORT
6/22/78
PAGE REFJ 1214105
CONTROL.
BACKGROUND) MEAN
VALUES VALUE
421 435
369
374
524
449
473
STANDARD
DEVIATION
60
ONCENTRATION REPLICATE MEAN STANDARD PERCENT
(UL/ML) VALUES VALUE DEVIATION SURVIVAL
200 354 309
254
320
150 472 475
483
471
100 462 405
373
379
50 395 355
386
283
10 420 441
452
450
2 460 378
272
403
•2 423 399
336
437
• READ AND UNDERIT
"7' '" ' -• -•-•-'. J ; :• 4 ^--i&t—
51 71
7 100
50 100
62 100
18 100
96 100
55 100
OODBV DATE
1
-------
APPENDIX CD
CHARACTERISTICS OF THE 14 WASTEWATER SAMPLES
AND RECONSTITUTED WATER AS DIRECTLY
REPORTED BY EG&G BIONOMICS
162
-------
Table OBI— Characteristics of CTHF-3 effluent measured on
16 June 1978, received from the Monsanto Research
Corporation, Dayton, Ohio on 15 June 1978. The re-
constituted water is also characterized.
Parameter Effluent3
Physical description: dark red-purple
liquid
pH: 10.1
DO (mg/H): 10.2
Temperature (°C): 18
Salinity (o/oo): 0
Specific conductance (ymhos/cm) ; 540
Reconstituted Water
pH: 7.6
Total hardness as CaCO3 (mg/X,) : 45
Total alkalinity as CaCO3 (mg/JL) : 31
Specific conductance (ymhos/cm): 145
a
Parameters measured before testing, after combining the four,
5-gallon containers.
163
-------
Table cbl— Characteristics of CTHF-4 effluent measured on 23
June 1978, received from the Monsanto Research
Corporation, Dayton, Ohio on 21 June 1978. The
reconstituted water is also characterized.
Parameter
Effluent
Physical description:
pH:
DO (mg/A) :
Temperature ( C):
Salinity (o/oo):
Specific conductance (ymhos/cm):
Reconstituted water
pH:
Total hardness as CaCO3 (mg/&):
Total alkalinity as CaCO3 (mg/£):
Specific conductance (ymhos/cm):
clear liquid
8.5
10.5
23
0
20
7.6
46
32
145
Parameters measured before testing, after combining the four,
5-gallon containers.
164
-------
Table
— Characteristics of CTHF-5 effluent measured on
23 June 1978, received from Clenson University,
Clemson, South Carolina on 21 June 1978. The re-
constituted water is also characterized.
Parameter
Effluent6
Physical description:
pH:
DO (mg/£) :
Temperature (°C):
Salinity (o/oo):
Specific conductance (ymhos/cm):
Reconstituted water
pH:
Total hardness as CaCO3 (mg/Jl) :
Total alkalinity as CaCO3 (mg/fc):
Specific conductance (ymhos/cm) :
clear liquid
7.9
11
23
0
11
7.6
46
32
145
a
Parameters measured before testing, after combining the four,
5-gallon containers.
165
-------
Table GDI— Characteristics of CTHF-6 effluent measured on
19 June 1978, received from the Monsanto Research
Corporation, Dayton, Ohio on 15 June 1978. The
reconstituted water is also characterized.
Parameter Effluent
Physical description: cloudy, orange-brown
liquid
pH: 9.9
DO fag/ft}: 10.8
Temperature ( C)s 13
Salinity
pH: 7.5
Total hardness as CaCO3 (mg/&): 44
Total alkalinity as CaC03 (rag/it) : 31
Specific conductance (ymhos/cm): 147
a
Salinity was measured with an American Optical refractometer.
166
-------
— Characteristics of CTHF-7 effluent measured on 19
June 1978, received from the Monsanto Research
Corporation, Dayton, Ohio on 15 June 1978. The re-
constituted water is also characterized.
a
Parameter Effluent
Physical description: clear liquid
pH: 10.7
DO (mg/A) : 10.3
Temperature (°C): 11
Salinity (o/oo): 0
Specific conductance (jjimhos/cm) : 1,003
Reconstituted Water
pH: 7'6
Total hardness as CaCO3 (mg/£): 45
Total alkalinity as CaCO3 (mg/fc): 31
Specific conductance (ymhos/cm) :
a
Parameters measured before testing, after combining the four,
5-gallon containers.
\alues of water used for test series conducted between 20-24 June
1978.
167
-------
Table GDI— Characteristics of CTHF-8 effluent measured on
23 June 1978, received from Clemson University,
Clemson, South Carolina on 21 June 1978. The
reconstituted water is also characterized.
Parameter
Effluent*
Physical description:
pH:
DO (mg/£) :
Temperature <°C):
Salinity (o/oo):
Specific conductance {pmhos/cm)
clear liquid
9.2
8.9
22
0
240
Reconstituted Water
pH:
Total hardness as CaCO3 (mg/£):
Total alkalinity as CaCO3 (mg/A):
Specific conductance {pmhos/cm):
7.6(7.7)J
46(43)b
32(33)b
145 (163)1
Parameters measured before testing, after combining all effluent
containers.
>
Parameters are for 100% effluent solution set up on 30 June 1978.
168
-------
Table GDI— Characteristics of CTHF-9 effluent measured on
23 June 1978, received from Clemson University,
Clemson, South Carolina on 21 June 1978. The
reconstituted water is also characterized.
Parameter Effluenta
Physical description: light yellow colored
liquid
pH: 9.3
DO (mg/£): 9.7
Temperature ( C): 22
Salinity (o/oo): 0.75
Specific conductance (pmhos/cm): 1,950
Reconstituted Water
pH: 7.6
Total hardness as CaCO3 (mg/£): 46
Total alkalinity as CaCO3 (mg/£): 32
Specific conductance (ymhos/cm): 145
a
Parameters measured before testing, after combining the five,
1-gallon containers.
169
-------
Table CDI— Characteristics of CTHF-10 effluent measured on
9 June 1978, received from Clemson University,
Clemson, South Carolina on 9 June 1978. The recon-
stituted water is also characterized.
Parameter
Effluent
3
Physical description:
pH:
DO
-------
TableCDli— Characteristics of CTHF-11 effluent measured on 9 June
1978, received from Clemson University, Clemson, South
Carolina on 9 June 1978. The reconstituted water is
also characterized.
Parameter
Effluent1
Physical description:
pH:
DO (mg/£):
Temperature ( C):
Salinity (o/oo) :
Specific conductance (ymhos/cm) :
Reconstituted water
pH:
Total hardness as CaCO3 (mg/Jl) ;
Total alkalinity as CaCO (mg/Jl) ;
Specific conductance (vimhos/cm) :
clear liquid
6.6
9..3
16
0
44
7.6
42
30
145
a
Parameters measured before testing, after combining the four,
5-gallon containers.
171
-------
l— Characteristics of CTHF-12 effluent measured on
9 June 1978, received from Clemson University,
Clemson, South Carolina on 9 June 1978.
The reconstituted water is also characterized.
Parameter Effluent
physical description: Clear liquid
6.5
DO
Temperature (°C) : ^g
Salinity (o/oo) : 0
Specific conductance (pmhos/cm) : 22
Reconstituted water
pH: 7.6
Total hardness as CaCO3 (mg/fc) : 30
Total alkalinity as CaCO3 (mg/A) : 42
Specific conductance (ymhos/cm) : X45
Parameters measured before testing, after combining the 4,
5-gallon containers.
172
-------
Table GDI— Characteristics of CTHF-13 effluent measured on 9 June
1978, received from the Monsanto Research Corporation,
Dayton, Ohio on 9 June 1978. The reconstituted water
is also characterized.
Parameter Effluent3
Physical description: a dark brown, slightly
cloudy liquid
pH: 7.7
DO (mg/£) : 8.4
Temperature ( C) : 16
Salinity (o/oo) : 0
Specific'conductance (pmhos/cm) : 1,520
Reconstituted Water
pH: 7-6
Total hardness as CaCO3 (nigA) : 42
Total alkalinity as CaCO3 (mg/£): 30
Specific conductance (ymhos/cm) : 145
Parameters measured before testing, after combining the two,
5-gallon containers.
173
-------
TableCDl— Characteristics of CTHF-14 effluent measured on
1.6 June 1978, received from the Monsanto Research
Corporation, Dayton, Ohio on 15 June 1978. The
reconstituted water is also characterized.
Parameter Effluent31
Physical description: pale yellow liquid
pH: 7.1
DO (rag/Jl): 4.6
Temperature ( C): 18
Salinity (o/oo): 0
Specific conductance (ymhos/cm): 680
i' -XT-'
Reconstituted Water
pH: 7.3
Total hardness as CaCO3 (mg/£): 43
Total alkalinity as CaCO3 (mg/&): 30
Specific conductance (ymhos/cm): 164
a
Parameters measured before testing, after combining the four,
5-gallon containers.
174
-------
Table dbi— Characteristics of CTHF-15 effluent measured on
16 June 1978, received from, Clemson University,
Clemson, South Carolina on 15 June 1978. The
reconstituted water is also characterized.
Parameter Effluent3
Physical description: a pale orange colored
liquid
pH: 8.3
DO (mgA): 9.7
Temperature (°C): 19
Salintiy (o/oo): 0
Specific conductance (ymhos/cm) : 95
Wi"
Reconstituted Water
pH: 7.6
Total hardness as CaCO3 (mg/&): 45
Total alkalinity as CaCO3 (mg/i): 31
Specific conductance (ymhos/cm): 145
a
Parameters measured before testing, after combining the four,
5-gallon containers.
175
-------
Table GDI— Characteristics of CTHF-16 effluent measured on
19 June 1978, received from the Monsanto Research
Corporation, Dayton, Ohio on 15 June 1978. The
reconstituted water is also characterized.
Parameter Effluent3
Physical description: a dark red liquid
pH: 8.5
DO (rag/*)* 9.8
Temperature f°c): 11
Salinity (o/oo): 0
Specific conductance (iimhos/cm): 2,255
'' -s»-
Reconstituted Water
pH: 7.6
Total hardness as CaCO3 (ing/2.) : 45
Total alkalinity as CaCO3 (mg/£): 31
Specific conductance (umhos/cm): 145
a
Parameters measured before testing, after combining the two,
5-gallon containers.
176
-------
APPENDIX CE
CHARACTERISTICS OF THE WASTEWATER SAMPLES AS A FUNCTION
OF TIME AND MORTALITY DOSE RESPONSE DATA AS
DIRECTLY REPORTED BY EG&G BIONOMICS FOR
FATHEAD MINNOR ACUTE TOXICITY TESTS
177
-------
Table cfe2— The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-3 effluent and fathead minnow (Pimephales promelas).
•
Nominal
concentration
(%) 0-hour 24-hour 48-hour 72-hour
PH
DO
(mg/fc)
specific
conductance
(umhos/cm)
•
total .hardness
as CaCOa (mg/Jl)
total alkalinity
as CaCO3 (mg/£)
53
5.3
0.24
control
53
5.3
0.24
control
53
5.3
0.24
control
53
5.3
0.24
control
53
5.3
0.24
control
. 9.9 9.7 9.2 8.7
8.7 7.5 7.0 7.0
7.4 7.3 7.1 7.0
7.2 7.1 7.0 7.0
9.4(nOO)a 9.0( 100) 0.2(2.3) 0.4(45)
9.0(>100) 6.1(69) 2.1(24) 1.9(22)
8.8(100) 7.8(89) 5.2(59) 5.3(60)
8.7(99) 7.5(85) 4.8(55) 5.2(59)
378
189
158
155
28
44
42
44
92
37
32
30
96-hpur
8.2
7.0
7.1
7.1
0.3(3.4)
2.0(23)
5.5(62)
5.4(61)
.
% of saturation at 22 C.
178
-------
Table <$4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-3 effluent for 24-, 48-, 72-
96-hour s.
Nominal
concentr at ion
(%)
53
24
11
5.3
2.4
1.1
0.53
0.24
control
2 4 -hour
100
90a
ob
ob
."• ,-*» .
0
0
0
0
0
% mortality
4 8 -hour 7 2 -hour
100 100
100 100
ob - ob
ob ob
0 0
0 0
0 0
0 0
0 0
and
9 6 -hour
100
100
0°
ob
ob
ob
0
0
0
a
One fish showed a complete loss of equilibrium.
b
Fish were lethargic.
c
Fish were at the surface.
179
-------
Table CE2-- The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-4 effluent and fathead minnow (Pimephales promelas).
1 —_-..- , t
PH
DO
(mg/Jl)
specific
conductance
(umhos/cm)
total hardness
as CaC03 (mg/Jl)
total alkalinity
as CaCO3 (mg/£)
•••••— •^•-••l— •— <•! ill •ll-il 1 • •!
Nominal
concentration
(%)
36
7.8
1.7
control
36
7.8
1.7
-."• ,«* .
control
36
7.8
1.7
control
36
7.8
1.7
control
36
7.8
1.7
control
— — — — — 1 1 "
0-hour 24-hour 48-hour 72-hour 96-hour
7.6 7.0 7.5 6.9 7.0
7.6 7.0 7.5 7.0 7.0
7.5 7.0 7.6 7.1 7.0
7.5 6.9 7.6 7.0 7.1
9.3(>100)a 6.9(78) 5.3(60) 4.5(51) 5.2(59)
9.1(>100) 6.9(78) 4.3(49) 3.7(42) 4.2(48)
9.0(>100) 6.8(77) 4.5(51) 3.6(41) 4.0(45)
8.9(>100) 6.7(76) 4.5(51) 3.6(41) 3.5(40)
105
141
147
149
28
42
44
44
21
29
29
30
% of saturation at 22 C.
180
-------
Table CE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-4 effluent for 24-, 48-, 72- and
96-hours.
Nominal
concentration
36
22
13
7.8
4.6
2.8
1.7
control
2 4 -hour
100
10a
Oa,b
0
0
0
0
0
%
mortality
4 8 -hour 7 2 -hour
100
30a
Ob
0
0
0
0
0
100
,b,c 4Qb'c
Ob
0
0
0
0
0
96-hour
100
40b'C
Ob
Ob
0
0
0
0
a
Some fish displayed a loss of equilibrium.
b
Pish were lethargic.
c
Fish displayed a dark coloration.
181
-------
TableCE2— The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-5 effluent and fathead minnow (Pimephales promelas).
•
Nominal
concentration
pH
DO
(mg/£)
specific
conductance
(ymhos/cm)
total hardness
as CaCO3 (mg/SL)
total alkalinity
as CaCO3 (mg/£)
(%)
100
36
7.7
control
100
36
7.7
control
100
36
7.7
control
100
36
7.7
control
100
36
7.7
control
0-hour 24-hour 48-hour
7.0 6.9 6.1
7.5 7.2 6.6
7.6 7.2 6.7
7.3 7.2 6.8
ll(>100)a 7.6(86) 5.6(64)
9.8(>100) 6.8(77) 3.8(43)
9.3(>100) 6.6(75) 3.9(44)
8.9(>100) 5.1(58) 3.7(42)
23
110
140
140
1
28
40
44
6
22
28
29
72-hour 96-hour
6.6 6.5
6.5 6.8
6.7 6.8
6.5 6.8
2.3(26) 1.6(18)
3.4(39) 4.1(47)
3.7(42) 3.6(41)
3.5(40) 3.5(40)
,
% of saturation at 22 C.
182
-------
Table CE3— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-5 effluent for 24-, 48-, 72- and
96-hours.
Nominal
concentrat ion
(%)
100
60
36
22
13
7.7
control
% mortality
2 4 -hour
oa
0
0
p
4*.'1"
0
0
0
4 8 -hour
30
0
0
0
0
0
0
7 2 -hour
30
0
0
0
0
0
0
96-hour
30
0
0
0
0
0
0
a
One fish displayed a complete loss of equilibrium.
183
-------
Table CE2~ The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-6 effluent and fathead minnow (Piroephales promelas)
'
Nominal
concentration
(%) 0-hour 24-hour
pH
DO
(mg/fc)
specific
conductance
(ymhos/cm)
total hardness
as CaC03 (mg/Jl)
total alkalinity
as CaCO3 (mg/i)
5.3
0.53
0.053
control
5.3
0.53
0.053
control
5.3
0.53
0.053
control
5.3
0.53
0.053
control
5.3
0.53
0.053
control
9.4 9.0
8.4 7.4
7.8 7.3
7.7 7.3
9.1{>100) 2.7(31)
9.1(>100) 6.2(70)
9.0(>100) 7.2(82)
9.1(>100) 7.3(83)
369
176
153
150
50
44
44
46
89
37
34
32
48-hour 72-hour 96-*hour
8.3 7.9 7.4
7.2 7.2 7.1
7.1 7.2 7.1
7.2 7.2 7.1
0.1(1.1) 0.2(2.3) 0.4(4.5)
4.7(53) 4.9(56) 4.8(55)
6.0(68) 6.0(68) 5.5(62)
6.1(69) 6.0(68) 5.6(64)
% of saturation at 22
184
-------
TableCE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-6 effluent for 24-, 48-, 72- and
96-hours.
Nominal
concentr at ion
(%)
5.3
2.4
1.1
0.53
0.24
0.11
0.053
control
2 4 -hour
100
30b'c
0
ob
- 0
0
0
0
%
mortality
4 8 -hour 7 2 -hour
100
100
0
ob
0
0
0
0
100
100
0
'd Ob
0
0
0
0
9 6 -hour
100
100
Od
0
0
0
0
0
a
All effluent test solutions were cloudy in proportion to the
concentration for the duration of the test.
b .
One fish displayed a dark coloration.
c
Some of the fish showed a complete loss of equilibrium.
d
Pish were lethargic.
185
-------
TableCE2— The pH, DO, specific conductance, total hardness and alklainity
measurements made during a 96-hour toxicity determination with
CTHF-7 effluent and fathead minnow (Pimephales promelas).
PH
DO
(mg/i)
specific
conductance
(ymhos/cm)
t
total hardness
as CaCO3 (mg/£)
total alkalinity
as CaCO3 (mg/A)
Nominal
concentration
100
46
7.7a
control
control
100
46
7.7a
control
a
control
100
46
a.
7.7
control
control
100
46
7.7a
control
control
100
46
7.7a
control
control
0-hour 24-hour 48-hour
10.6 10.1 9.7
10.2 9.8 9.2
9.1 8.3 7.4
7.6 7.2 7.1
7.0 7.0 7.0
11.7(>100)b 10.3(>100) 1.8(20)
10.3(>100) 9.2(>100) 0.3(34)
9.KXLOO) 5.0(57) 1.2(14)
9.1(>100) 6.4(73) 4.0(45)
8.6(98) 5.1(58) 4.3(49)
890
461
196
156
131
14
30
42
46
38
242
128
48
32
28
72-hour 96-hour
9.5 9.3
8.9 8.5
7.2 7.2
7.1 7.0
7.1 7.3
0.1(1.1) 0.1(1.1)
0.2(2.3) 0.3(3.4)
2.1(24) 3.2(36)
3.9(44) 4.0(45)
4.3(49) 4.3(49)
These test solutions were conducted between 20 and 24 June 1978.
% of saturation at 22°C.
186
-------
TableCE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-7 effluent for 24-, 48-, 72- and
96-hours.
Nominal
concentration
100
68
46
32
22
7.8C
control
control0
24-^hour
100
100
100
100
7bb
0
0
0
% mortality
4 8 -hour
100
100
100
100
100
10
0
10
7 2 -hour
100
100
100
100
100
10
0
10
96-hour
100
100
100
100
100
10
0
10d
a
All effluent test solutions were cloudy in proportion to the
concentration for the duration of the test.
b
Some fish showed a complete loss of equilibrium.
c . -••-•
There test solutions were conducted between 20 and 24 June 1978,
d
Fish were lethargic.
187
-------
TableCE2— The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-8 effluent and fathead minnow (Pimephales promelas).
1 Ill.-l. .. ••!•.. II • • • •••. — .. ••••it. .•• 1
PH
DO
100)C 8.0(91) 5.3(60) -a -&
9.3O100) 5.8(66) 2.7(31) 2.7(31) 2.7(31)
9.2O100) 6.6(75) 4.2(48) 4.1(47) 3.7(42)
8.8(100) 7.9(90) 5.7(65) -a -a
9.1O100) 7.0(80) 4.6(52) 4.6(52) 4.5(51)
237
160
150
157
150
1
38
42
41
44
109
39
33
31
29
.Measurements not made due to technician error.
cThis control set on 30 June with 100% effluent solutioni
% of saturation at 22 C.
188
-------
Tabie O33— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-8 effluent for 24-, 48-, 72- and 96-
hours.
Nominal
concentration
(%)
100
88
46
24
13
6.8
3.6
1.9
control
control
2 4 -hour
0
Ob
0
0
0
•/t'l'
0
0
0
0
0
% mortality
4 8 -hour
Oa
0
0
0
0
0
0
0
0
7 2 -hour
0
10C
0
0
0
0
0
0
10
0
9 6 -hour
0
10
0
0
0
0
0
0
10
0
a
i
One fish displayed a complete loss of equilibrium.
b
Pish were lethargic.
c
Mortality was judged to be toxicant related.
This control set on 30 June with the 100% effluent solution.
189
-------
Table CE2-- The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-9 effluent and fathead minnow (Pimephales promelas)
PH
DO
(mg/£)
specific
conductance
(ymhos/cm)
total hardness
as CaCO3 (mg/Jl)
total alkalinity
as CaCOs (mg/fc)
Nominal
concentration
5.3
1.9
0.41
control
5.3
1.9
0.41
control
5.3
1.9
0.41
control
5.3
1.9
0.41
control
5.3
1.9
0.41
control
0-hour 24 -hour 48 -hour
8.9 7.9 7.6
8.4 7.4 7.6
7.6 7.4 6.9
7.6 7.5 6.9
9.0(>100)a 0.4(4.5) 0.3(3.4)
9.0O100) 4.8(55) 1.5(17)
8.9(>100) 6.3(72) 4.2(78)
8.9O100) 7.2(82) 5.7(65)
294
200
163
151
44
44
44
44
65
44
34
30
72-hour 96-hour
7.2 7.1
7.0 7.1
7.0 7.1
7.1 7.1
0.3(3.4) 1.7(19)
1.3(15) 1.0(11)
3.8(43) 3.5(40)
5.3(60) 5.3(60)
•
% of saturation at 22 C.
190
-------
TableCE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-9 effluent for 24-, 48-, 72- and 96-
hour s
Nominal
cone en tr at ion
(%)
5.3a
3.2a
1.9
1.1
0.68
0.41
control
•
% mortality
24 -hour 48-hour 72-hour 96-hour
100a 100a 100a 100a
20a'b'C 90a'c'd 90a'b 90a'b
0 Oe 10 20
oe o 10 10
-0 10 10 10
0 0 10 10
0000
a
Solutions were cloudy.
b
Fish displayed a dark coloration.
c
Some fish displayed a complete loss of equilibrium.
d
Fish were at the surface, gulping air.
e
Fish were lethargic.
191
-------
Table QE4-- The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-10 effluent and fathead minnow (Pimephales promelas).
PH
DO (mg/Jl)
specific
conductance
(ymhos/cm)
total hardness
as CaC03 (mg/£)
total alkalinity
as CaCO3 (mg/£)
Nominal
concentration
(%)
60
22
4.6
control
60
22
4.6
control
60
•*'• .*«.
22
4.6
control
60
22
4.6
control
0-hour 24-hour 48-hour
7.2 6.8 6.7
7.3 7.0 6.7
7.3 7.2 7.0
7.2 7.0 6.8
8.3(94)* 0.3(3.4) 0,2(2.3)
8.7(99) .4.1(47) 0.2(2.3)
8.8(100) 7.1(81) 3.9(44)
8.8(100) 7.5(85) 4.8(55)
186
160
142
142
30
36
42
42
42
37
33
30
72-hour 96-hour
6.7 6.7
6.6 6.8
6.9 6.9
6.9 6.9
0.1(1.1) 0.1(1.1)
0.2(2.3) 0.2(2.3)
3.7(42) 2.2(25)
4.4(50) 4.4(50)
% of saturation at 22 C.
192
-------
Table CE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-10 effluent for 24-, 48-, 72- and
96-hours.
Nominal a
concentration
60
36
22
13
7.8
4.6
control
24-hour
100
100
90b
10b
.V"
c
0
0
0
mortality
4 8 -hour 7 2 -hour
100
100
100
100
0
0
0
100
100
100
100
c oc
Ob,c
0
9 6 -hour
100
100
100
100
0
10°
0
a
All effluent solutions were cloudy in proportion to the concen-
tration for the duration of the test.
b
Some fish displayed a loss of equilibrium.
c
Fish were lethargic.
193
-------
Table CE2-- The pH, DO, specific conductance, total hardness and alkalinity measure-
ments made during a 96-hour toxicity determination with CTHF-11 effluent
and fathead minnow (Pimephales promelasj .
pH
DO
(mg/5.)
specific
conductance
(ymhos/cm)
total hardness
;as CaC03 (mg/Jl)
total alkalinity
as CaCOs (mg/£)
Nominal
concentration
100
36
7.8
control
100
36
7.8
control
100
36
7.8
control
100
36
7.8
control
100
36
7.8
control
0-hour 24-hour 48-hour
7.7 7.1 6.6
7.4 7.1 6.8
7.3 7.2 6.9
7.2 7.0 6.9
ll(>100)a 7.4(84) 4.2(48)
9.6(>100) 6.8(77) 3.9(44)
8.9(>100) 7.2(82) 4.3(49)
8.5(97) 6.6(75) '4.3(49)
36
95
°*130
142
1
28
40
42
7
21
29
30
72 -hour 96-hour
6.5 6.8
6.8 6.9
6.9 6.8
6.9 6.7
1.9(22) 3.0(34)
3.1(35) 3.6(41)
4.0(45) 3.8(43)
4.2(48) 3.3(38)
% of saturation at 22 C.
194
-------
Table CE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-11 effluent for 24-, 48-, 72- and 96-
hours.
Nominal
concentration
(%) 24-hour
100 20a
60 0
36 0
22 0
-vx
13 Ob
7.8 0
control 0
% mortality
48-hour
40a'b
0
0
b
0D
ob
0
0
72-hour
60b'c
20,
0
b
0°
ob
0
0
96-hour
60a
40b'd
0
10
10
10
0
a
Some fish displayed a loss of equilibrium.
b
Some fish displayed a dark coloration.
c
Fish were at the surface.
d
Fish were lethargic.
195
-------
Table CE2— The pH, DO, specific conductance, total hardness and alkalinity measure-
ments made during a 96-hour toxicity determination with CTHF-12 effluent
and fathead minnow (Pimephales promelas).
pH
DO
(mg/fc)
specific
conductance
(umhos/cm)
total alkalinity
as CaCOs (mg/£)
total hardness
as CaCOs (mg/£)
Nominal
concentration
100
46
15
control
100
46
15
control
100
46
15
control
100
46
15
control
100
46
15
control
0-hour 24-hour
7.5 6.9
7.4 7.0
7.3 7.1
7.2 7.0
ll(>100)a 8.5(97)
9.5(>100) 7.7(88)
8.9(>100) 7.2(82)
8.6(98) 7.4(84)
28
95
130
142
5
18
26
30
1
24
36
42
48-hour 72-hour 96-hour
6.8 6.6 6.4
6.8 6.8 6.9
7.0 6.9 6.9
7.0 6.9 6.7
6.8(77) 4.5(51) 2.8(32)
4.0(45) 4.0(45) 3.8(43)
4.7(53) 4.9(53) 4.7(53)
5.5(63) 4.2(48) 4.2(48)
% of saturation at 22 C.
196
-------
Table GE3— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-12 effluent for 24-, 48-, 72- and
96-hours.
Nominal
concentration
100
68
46
32
22
15
Control
2 4 -hour
10
10
0
ioa "
10
0
0
% mortality
4 8 -hour 7 2 -hour
10
20
oa
20
-------
Table CE2— The pH, DO, specific conductance, total hardness and alkalinity measure-
ments made during a 96-hour toxicity determination with CTHF-13 effluent
and fathead minnow (Pimephales promelas) .
PH
DO
(mg/£)
specific
conductance
(umhos/cm)
total hardness
as CaCOa (mg/£)
total alkalinity
as CaCOa (mg/5,)
Nominal
concentration
(%)
10
2.8
0.41
control
10
2.8
0.41
control
.*#<
10
2.8
0.41
control
10
2.8
0.41
control
10
2.8
0.41
control
0-hour 24-hour 48-hour 72-hour 96-houra
7.4 7.1 7.0 7.0
7.3 6.9 6.8 6.9
7.2 7.1 6.8 6.9
7.0 7.2 6.9 7.0
8.7(99)b 0.2(2.3} 0.2(2.3) 0.2(2.3)
8.9(>100) 1.0(11) 0.3(3.4) 0.7(8.0)
9.0(>100) 5.1(58) 2.4(27) 2.3(26)
8.6(98) 6.6(75) 4.2(48) 4.0(45)
293
185
j
148
142
56
46
42
42
63
41
34
30
Due to a scheduling oversight, pH and DO measurements were not made at 96-hours.
b
% of saturation at 22 C.
198
-------
TablecE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-13 effluent for 24-, 48-, 72- and
96-hours.
Nominal
concentration
(%) 24-hour
10 100
5.3 100
2.8 100
1.5 0~
0.78 0
0.41 0
control 0
% mortality
4 8 -hour
100
100
100
2Qb,c,d,e
0
0
0
72-hour
100
100
100
30b'd
0
0
0
96-hour
100
100
100
40b
0
0
0
a
All effluent test solutions were cloudy in proportion to the con-
centration for the duration of the test.
b
Fish displayed a dark coloration.
c
Fish were lethargic.
d
Some fish were at the surface.
e
Some fish were gulping at the surface.
199
-------
Table CE2— The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with CTHF-14
effluent and fathead minnow (Piroephales promelas).
Nominal
concentration '
^PV»««MBWMwaMHM^^H^^^flHM^^a^»i^MMi^HIHMIlallMWMBM
PH
DO
(mg/JO
specific
conductance
(ymhos/cm)
total hardness
as CaCO3 (mg/£)
total alkalinity
as CaCO3 (mg/2,)
MI"«IW^MIflHiM»h*AM«VHB»*«<4^M^H^
53
19
4.1
control
53
19
4.1
control
53
19
4.1
control
53
19
4.1
control
53
19
4.1
control
0-hour 24-hour
' "1 ii mi mi i •• f nmm •*•• m •• .^^••^•^•••i I i I — — ••
7.3 6.9
7.5 7.2
7.4 7.3
7.2 7.1
7.1(81)a 2.3(26)
8.5(87) 6.1(69)
9.0O100) 7.4(84)
8.8(100) 7.7](88)
471
265
183
142
36
40
42
44
40
35
32
30
48-hour 72-hour 96-hour
1 1 • i i i
6.7 6.7 6.7
6.8 6.8 6.8
6.9 6.8 6.9
6.9 6.9 7.0
0.2(2.3) 0.3(3.4) 0.5(5.7)
0.8(9.1) 1.2(14) 2.1(24)
3.1(35) 2.9(33) 3.4(39)
4.8(55) 4.7(56) 5.4(61)
% of saturation at 22 C.
200
-------
Table CE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed
9 6 -hours
Nominal
concentration
(%)
53
32
19
11
6.8
4.1
control
to CTHF-14
•
2 4 -hour
100
30
0
0
0
0
Od
effluent
%
4 8 -hour
100
50a/
Ob
0
0
0
0
for 24-, 48-, 72-
mortality
7 2-hour
100
b 100
10
0
0
0
0
and
96-hour
100
100
10
0
0
10C
0
a
Some fish displayed a loss of equilibrium.
b
Some fish were at the surface.
c
This mortality was judged not to be toxicant related.
d
Pish were lethargic.
201
-------
TableCE2— The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-15 effluent and fathead minnow (Pimephales promelas).
PH
DO
(rog/£)
specific
conductance
(umhos/cm)
total hardness
as CaCO3 (mg/Jl)
total alkalinity
as CaC03 (mg/Jl)
••^•^•^^•^^••^^^^••••••^•P^M^V^M^B^Hl^HA^M
Nominal
concentration
(%)
100
22
4.6
control
100
22
4.6
control
100
.*0L-
22
4.6
control
100
22
4.6
control
100
22
4.6
control
^^^••^•^^•^^^^^^••^•^^•^^••^^•••JMal •••.•PHI !•! ^•^•^^••^M^^
0-hour 24-hour
8.2 7.4
8.0 7.4
7.9 7.3
7.9 7.2
9.4(>100)a 7.6(86)
9.0O100) 7.5(85)
9.0O100) 7.6(86)
8.9O100) 7.3(83)
100
143
150
150
2
34
42
44
16
27
30
30
"""^ ^^^^^^^^^—^^^^^^••^•^••^^••^^.^ „ l MMIM ^
4 8 -hour 72-hour 96-hour
7.2 7.2 7.0
7.0 7.0 7.1
7.0 7.0 7.2
7.0 7.1 7.1
5.8(66) 4.4(50) 4.4(50)
4.8(55) 4.5(51) 4.9(56)
4.7(53) 4.9(56) 5.3(60)
4.7(53) 4.9(56) 5.5(63)
% of saturation at 22 C.
202
-------
Table CE3— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed
9 6 -hours
Nominal
concentration
100
60
36
22
13
7.8
4.6
control
to CTHF-15
2 4 -hour
0
0
0
o
0
0
0
0
effluent for 24-,
% mortality
4 8 -hour 72
0
0
0
0
0
0
0
0
48-, 72- and
-
-hour 9 6 -hour
10 20
Oa,b,c Qd
0 0
0 Oa
0 0
0 0
0 0
0 0
a
Some fish were lethargic.
b
Fish were at the surface.
c
Fish were gulping.
d
Fish displayed a dark coloration.
203
-------
TableCE2-- The pH, DO, specific conductance, total hardness and alkalinity
measurements made during a 96-hour toxicity determination with
CTHF-16 effluent and fathead minnow (Pimephales promelas).
PH
DO
(mg/Jl)
specific
conductance
(ymhos/cm)
total hardness
as CaCO3 (mg/£)
total alkalinity
as CaCOs (mg/i)
^^^^•^^^^^•^•^^M.^VHHVt^HM
Nominal
ncentratio
(%)
*ifmmtttt**im^mimtim*miii*mii~*imiiiiiii*iimim
36
3.6
0.36
control
36
3.6
0.36
control
36
3.6 ,,,
0.36
control
36
3.6
0.36
control
36
3.6
0.36
control
1 1^— ^^ 1 • 1 — — 1 II !••.- HI I'— 1 III. 1 .11 1 •..PI III
n
0-hour 24-hour
WW-mMMIMMIMMMIIIBBIIIIVtflMMM'MMal^^
8.0 7.2
7.6 7.1
7.5 7.0
7.5 7.0
9.7(>100)a 0.3(3.4)
9.3O100) 6.1(69)
9.4(>100) 6.3(72)
9.1O100) 6.2(70)
930
232
166
143
_b
50
44
46
58
35
33
32
48-hour 72-hour 96-hour
• • i "i
7.1 7.0 7.0
7.0 7.0 6.9
7.1 7.1 7.0
7.2 7.2 7.1
0.4(4.5) 0.2(2.3) 0.2(2.3)
2.2(25) 1.2(14) 1.6(18)
3.3(38) 3.1(35) 2.8(32)
4.1(47) 4.1(47) 3.9(44)
% of saturation at 22 C.
3
No measurement could be made due to similarity of effluent and titration end point
color.
204
-------
Table GE4— Concentrations tested and corresponding percentage
mortalities of fathead minnow (Pimephales promelas)
exposed to CTHF-16 effluent for 24-, 48-, 72- and
96-hours.
Nominal
concentration
36
17
7.8
3.6
1.7
0.78
0.36
control
% mortality
2 4 -hour
100
100
10
10
-0
oa
0
0
4 8 -hour
100
100
100
20
0
0
0
0
7 2-hour
100
100
100
20
ob
ob
ob
0
9 6 -hour
100
100
100
20
Ob
ob
0
0
a
Some fish displayed a dark coloration.
b
Fish were lethargic.
205
-------
APPENDIX CF
WATER QUALITY ANALYSIS OF THE 14 WASTEWATER SAMPLES AS A
FUNCTION OF TEST SOLUTION CONCENTRATIONS AND RAW MORTALITY
DOSE RESPONSE DATA AS DIRECTLY REPORTED BY EG&G BIONOMICS
FOR DAPHNIA ACUTE TOXICITY TESTS
206
-------
to
o
-J
TABLE CFI— Water quality analysis of CTHF-3 effluent test solutions during the static
acute exposure of the water flea (Daphnia magna).
Nominal
concentration
100
36
13
control
Dissolved3
oxygen
(mg/£) pHa
9.1-4.6 9.3-8.2
9.0-7.1 8.8-8.3
8.7-7.5 8.3-8.3
8.8-8.5 8.1-8.4
Total
hardness
(mg/£ CaCO3)
28
140
196
202
Specific
conductance
(ymhos/cm2)
648
668
711
669
V,
Alkalinity
(mg/S, CaCO3)
143
136
134
135
Measurements taken at 0- and 48-hours.
3
Measurements taken at 0-hour.
-------
TABLE CP2— Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-3 effluent. Each
mortality value represents the average of 3 repli-
cates .
Nominal concentration
(%>
100
60
36
22
13
control
Average percentage
2 4 -hour
93
73
0
0
0
0
mortality
4 8 -hour
100
93
87
20
13
0
208
-------
— Water quality analysis of CTHF-4 effluent test solutions during the static
acute exposure of the water flea (Daphnia mag'na).
N>
O
Nominal
concentration
(%)
100
36
7.8
control
Dissolved9
oxygen
(mgA)
12.0-8.0
9.6-8.1
9.2-8.2
8.9-8.1
Total
hardness
pH (rng/A CaC03)
t
7.6-7.5 6
8.1-8.2 138
8.2-8.2 196
8.1-8.2 212
Specific
conductance
(ymhos/cm2)
30
410
590
610
v,
Alkalinity0
(mg/Jl CaC03)
8
94
128
141
Measurements taken at 0- and 48-hour.
D
Measurements taken at 0-hour.
-------
TABLE CF2 — Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-4 effluent. Each
mortality value represents the average of 3 repli-
cates .
Nominal concentration
100
60
36
22
13
7.8
control
Average percentage
24-hour
93
0
0
0
0
0
0
mortality
48-hour
100
67
7
0
0
0
0
210
-------
TABLE CFI — Water quality analysis of CTHF-5 effluent test solutions during the static
acute exposure of the water flea (Daphnia tflagna).
to
M
H-1
Nominal
concentration
100
36
13
control
Dissolved
oxygen
(mg/Jl)
8.8-8.7
8.9-8.5
8.8-8.6
8.9-8.7
Total
hardness
pH (rag/ H CaC03)
9.3-7.8 4
8.7-7.8 136
8.4-7.6 184
8.2-8.1- 214 .
Specific
conductance
(ymhos/cm2)
249
460
590
600
Alkalinity
(mg/A CaCO3)
7
86
126
126
Measurements taken at 0- and 48-hour.
3
Measurements taken at 0-hour.
-------
TABLE. CF2 — Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia maqna) exposed to CTHF-5 effluent. Each
mortality value represents the average of 3 repli-
cates.
Nominal concentration
<%)
100
60
36
22
13
control
Average percentage
24 -hour
100
27
0
0
0
0
mortality
48-hour
100
93
13
7
0
0
212
-------
NJ
M
OJ
TABLE CFI — Water quality analysis of CTHF-6 effluent test solutions during the static
acute exposure of the water flea (Paphnia
3
Nominal Dissolved
concentration oxygen
(%) (mg/Jl)
13 8.9-4.3
4.6 8.9-5.7
1.5 8.6-6.9
control 8.8-8.5
Totalb
hardness
pH (mg/A CaCO3)
* '
9.0-8.3 206
8.7-8.2 226
8.5-8.1 204
8.1-8.4 202
Specific
conductance
(pmhos/cm2)
1165
821
762
669
K
Alkalinity0
(mg/A CaCO3)
260
178
149
135
Measurements taken at 0- and 48-hours.
:>
Measurements taken at 0-hour.
-------
TABLE CF2 — Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-6 effluent. Each
mortality value represents the average of 3 repli-
cates .
Nominal concentration
(%)
13
7.8
4.6
2.8
1.5
control
Average percentage
2 4 -hour
71a
27
0
0
0
0
mortality
4 8 -hour
100*
87
53
0
0
0
Data based on 14 daphnids. One daphnid could not be accounted
for.
214
-------
TABLE CFI — Water quality analysis of CTHF-7 effluent test solutions during the static
acute exposure of the water flea (Daphnia ittagna).
10
H1
Ui
Nominal
concentration
<%)
100
36
13
control
Dissolved3
oxygen
(mg/A) p!T
9.2-2.5 9.7-8.1
9.0-4.4 9.0-8.0
8.8-6.3 8.6-8.0
8.8-8.5 8.1-8.4
Total
hardness
(mg/fc CaC03)
14
136
192
202
Specific
conductance
(ymhos/cm2)
1000
801
780
669
Alkalinity
(mg/Jl CaC03)
239
172
147
135
Measurements taken at 0- and 48-hours.
>
Measurements taken at 0-hour.
-------
TABLE CF2— Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Paphnia magna) exposed to CTHF-7 effluent. Each
mortality value represents the average of 3 repli-
cates.
Nominal concentration
100
60
36
22
13
control
Average percentage
2 4 -hour
47
0
0
0
Q
0
mortality
4 8 -hour
100
100
73
'33
13
0
216
-------
10
TABLE CFI — Water quality analysis of CTHF-8 effluent test solutions during the static
acute exposure of the water flea (Daphnia irtagna).
Nominal
concentration
100
36
13
control
Dissolved
oxygen
(mg/JO pH
9
9
9
8
.7-6.
.2-7.
.0-7.
.9-8.
7
3
9
7
8
8
8
8
.2-8
.2-8
.2-7
.2-8
i.
.3
.1
.9
.1
Totalb
hardness
(mg/Jl CaC03)
2
142
186
214
Specific
conductance
(ymhos/cm2)
41
400
550
600
Alkalinity0
(mg/Jl CaCO3)
109
122
126
126
Measurements taken at 0- and 48-hours.
3
Measurements taken at 0-hour.
-------
TABLE CF2 — Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Paphnia magna) exposed to CTHF-8 effluent. Each
mortality value represents the average of 3 repli-*
cates.
Nominal concentration
(%)
100
60
36
22
13
control
Average percentage
2 4 -hour
0
0
0
0
0
0
mortality
48-hour
13
0
0
0
0
0
218
-------
TABLE CFI — Water quality analysis of CTHF-9 effluent test solutions during the static
acute exposure of the water flea (Paphnia ifiagna).
Nominal
concentration
(%)
19
6.8
1.5
control
Dissolved3
oxygen
(mg/H)
9.2-0.7
9.0-5.1
8.9-7.1
8.9-8.1
Totalb
hardness
pH ' (mg/A CaCO3)
8.9-8.9 190
8.6-8.2 198
8.2-8.2 212
8.1-8.2 212
Specific
conductance
(ymhos/cm2)
1000
750
610
610
Alkalinityb
(mg/S, CaCO3)
239
179
145
141
Measurements taken at 0- and 48-hours,
D
Measurements taken at 0-hour.
-------
TABLE CF2 — concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-9 effluent. Each
mortality value represents the average of 3 repli-?
cates.
Nominal concentration
19
11
6.8
4.1
2.4
1.5
control
Average percentage
24-hour
87
33
7
0
0
0
0
mortality
4 8 -hour
93
53
27
0
0
0
0
220
-------
TABLE CF1— Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-10 effluent. Each
mortality value represents the average of 3 repli-
cates .
Nominal concentration
(%)
100
60
36
22
13
control
Average percentage
2 4 -hour
33
27
. 0
0
0
0
mortality
4 8 -hour
100
93
87
0
0
0
221
-------
^ABLE :CFI — Concentrations tested and corresponding average
observed percentage mortalities for the water flea
{Daphnia magna) exposed to CTHF-11 effluent. Each
mortality value represents the average of 3 repli-
cates .
Nominal concentration
(%)
100
60
36
22
13
control
Average percentage
2 4 -hour
0
0
0
0
0
0
mortality
4 8 -hour
93
0
0
0
0
0
222
-------
•TSBLE. CFI— Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-12 effluent. Each
mortality value represents the average of 3 repli-
cates.
Nominal concentration
(%)
100
60
36
22
,13
control
Average percentage
2 4 -hour
100
0
0
0
0
0
mortality
4 8 -hour
100
0
0
0
0
0
223
-------
TABLE CFI— Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-13 effluent. Each
mortality value represents the average of 3 repli-
cates.
Nominal concentration
(%)
13
7.8
4.6
2.8
1.7
control
Average percentage
2 4 -hour
67
13
0
0
0
0
mortality
4 8 -hour
100
87
87
0
0
0
224
-------
TABLE CFI — Water quality analysis of CTHF-14 effluent test solutions during the static
acute exposure of the water flea (Paphnia irtagna).
10
K>
Ul
J
Nominal
concent rat ion
(%)
100
36
13
control
Dissolved3
oxygen
(mg/H)
9.0-4.7
7.5-7.1
8.3-7.5
8.8-8.5
Totalb
hardness
pHa (mg/Jl CaC03)
5-
7.0-7.5 38
7.8-8.1 148
7.9-8.2 194
8.1-8.4 202
Specific
conductance
(ymhos/cm2)
858
803
797
669
K
Alkalinity
(mg/£ CaC03)
40
94
122
135
Measurements taken at 0- and 48-hours.
3
Measurements taken a 0-hour.
-------
TABLE CF2 — Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-14 effluent. Each
mortality value represents the average of 3 repli-
cates.
Nominal concentration
(%)
100
60
36
22
13
control
Average percentage
2 4 -hour
60
0
0
0
0
0
mortality
48-hour
100
93a
0*
7*
O3
0
Some surviving daphnids were lethargic.
226
-------
TABLE-CFI— Water quality analysis of CTHF-15 effluent test solutions during the static
acute exposure of the water flea (Daphnia magna).
K>
--J
Nominal
concentration
(%)
100
36
13
control
Dissolved
oxygen
(mgA)
9.0-7.7
9.0-7.7
8.9-7.5
8.8-8.5
Total
hardness
pH (mg/£ CaCO3)
8.0-8.2 2
8.1-8.1 154
8.1-8.1 184
8.1-7.9 202
Specific
conductance
(ymhos/cm2)
138
524
672
669
V,
Alkalinity0
(mg/£ CaC03)
16
99
118
135
Measurements taken at 0- and 48-hours.
3
Measurements taken at 0-hour.
-------
TABLE CF2 — Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTEF-15 effluent. Each
mortality value represents the average of 3 repli-
cates.
Nominal concentration
(%)
100
60
36
22
13
control
Average percentage
2 4 -hour
47
Oa
Oa
Oa
Oa
0
mortality
4 8 -hour
87
7a
Oa
Oa
Oa
0
Some surviving daphnids became entrapped at the air-water
1 t1+•*»•!--Fa «•*« CC «*•••* nuwc.1.
interface.
228
-------
NJ
NJ
TABLE.CFI — Water quality analysis of CTHF-16 effluent test solutions during the static
acute exposure of the water flea (Daphnia magna).
Nominal
concentration
(%)
36
7.8
1.7
control
9.
Dissolved
oxygen
(mg/Jl) pH
9.0-4.7 8.1-7.7
8.7-8.0 8.1-8.0
8.6-8.5 8.1-8.1
8.6-8.6 7.8-8.0
X
Total
hardness
(mg/£ CaC03)
292
224
218
208
b
Specific
conductance
(y mhos /cm2)
1292
821
716
662
V.
Alkalinity0
(mg/£ CaC03)
122
132
134
132
Measurements taken at 0- and 48-hour.
3
Measurements taken at 0-hour.
-------
fcfc C.P2 — Concentrations tested and corresponding average
observed percentage mortalities for the water flea
(Daphnia magna) exposed to CTHF-16 effluent. Each
mortality value represents the average of 3 repli-
cates.
Nominal concentration
(%)
36
22
13
7.8
4.6
2.8
1.7
control
Average percentage
2 4 -hour
33
7
0
0
0
0
0
0
mortality
4 8 -hour
100
47
13
20
20
0
0
7
230
-------
APPENDIX CG
RAW DATA ON ACUTE ORAL TOXICITY STUDY
IN RATS PERFORMED BY LITTON BIONETICS, INC.
231
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-3
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-01
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-3
Test B-2
Date 6-12-78
Time 10:30
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2891.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 172 to 226 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain.[CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Litton 232
-------
4. RESULTS
The data have been summarized as follows.
Mean Body Weight
Dof (g) Deaths Total
Unl^2J DM Day. Mortality
5— '— I4— 0-14 Deaths/Treated
Males
0 216 265 343 - o/5
10 213 294 360 - n/5
Females
0 178 190 231 - 0/5
10 178 216 239 - 0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by: Reviewed by:
David R. Damske, B.A. Robert P. Beliles, Ph.D. Date
Toxicology Technician Director
Department of Toxicology Department of Toxicology
..-_ BIONETICS
Litton 233
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-4
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-02
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-4
Test B-2
Date 6-12-78
Time 1500
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2892.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 168 to 214 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Litton 234
-------
4. RESULTS
The data have been summarized as follows,
Dose
(ml/kg)
0
10
0
10
Mean Body Weight
(g)
Day
0
216
206
178
184
14
Deaths
Day
0-14
Total
Mortality
Males
265
288
343
348
Females
190
219
231
242
Deaths/Treated
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in.any of the treated or control animals. The lungs of two
treated females were observed to be slightly mottled. This finding
has previously been observed in this strain of animal in this
laboratory and was judged not to be related to compound adminis-
tration.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
n / r, /-)
Davia R. Damske, B.A.
Toxicology Technician
Department of Toxicology
robert P. BeTiles, Ph.D.
Director
Department of Toxicology
Date
Litton
BIONETICS
235
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-5
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LSI Project No. 20969-03
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-5
Test Sample B-2
Date 6-12-78
Time 1530
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LSI No. 2893.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 166 to 223 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRUCOBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad •libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Utt°n 236
-------
4. RESULTS
The data have been summarized as follows.
Dose
(ml/kg)
Mean Body Weight
(g)
Day
0 7 14
Deaths
Day
0-14
Total
Mortality
Males
0
10
0
10
216
214
178
179
265
305
343
364
Females
190
217
231
244
Deaths/Treated
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals. An enlarged, pitted,
fluid-filled right kidney was observed in one treated male rat.
This finding has previously been observed in this strain of
animal at this laboratory and was judged not to be related to
compound administration.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
Reviewed by:
^ay _a_gLLJJI_JIIlJ__SJ—•—••^•••^^••^•^^^^^•^-^•••^••^•^^••^
Robert P. Beliles, Ph.D.
Di rector
Department of Toxicology
Date
Utton
BIONET1CS
237
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-6
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-04
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-6
Test B-2
Date 6-12-78
Time 1400
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2894.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 164 to 225 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Litton 238
-------
4. RESULTS
The data have been summarized as follows.
Dose
(ml/kg)
Mean Body Weight
(g)
Day
0
10
0
10
0
216
211
178
175
7 14
Males
265 343
299 356
Females
190 231
216 240
Deaths
Day
0-14
Total
Mortality
Deaths/Treated
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
/ •-
*""
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
Reviewed by:
Robert P. Beliles, Ph.D. Date
Di rector
Department of Toxicology
Litton
BIONETICS
239
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-7
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-05
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-7
Test B-2
Date 6-13-78
Time 0900
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2895.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 179 to 220 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Litton 240
-------
4. RESULTS
The data have been summarized as follows.
Dose
Mean Body Weight
(g)
Day
V - ' f ' " J f
0
10
0
10
0
216
207
178
189
7 14
Males
265 343
292 343.
Femal es
190 231
222 247
Deaths
Day
0-14
Total
Mortality
Deaths/Treated
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in.any of the treated or control animals.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
Reviewed by:
Robert P. Beliles, Ph'.D.
Director
Department of Toxicology
Date
Litton
BIONETICS
241
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-8
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-06
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-8
Test B-2
Date 6-17-78
Time 2300
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2896
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 156 to 233 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow.were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
ffl
BIONETICS 242
Litton
-------
4. RESULTS
The data have been summarized as follows.
Dose
fa!/kg)
Mean Body Weight
(g)
Day
0
10
0
10
0
216
208
178
178
7 14
Males
265 343
285 335
Females
190 231 '
216 240
Deaths
Day
0-14
Total
Mortality
Deaths/Treatea
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
0 /)^k
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
Reviewed by:
)b'ert P. Bellies, Ph.D.
Director
Department of Toxicology
Date
Litton
BIONETICS
243
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-9
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-07
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-9
Test B-2
Date 6-17-78
Time 2300
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2897.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 166 to 246 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad. libitum rfith the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Litton 244
-------
4. RESULTS
The data have been summarized as follows.
Mean Body Weight
(ml/kg) $y °eaths Total
— ^ £§* _ Mortality
0-14 Deaths/Treated
0
10
Females
0 178 190 23i . Q/5
10 179 223 242 - Q/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated fo? male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals. One treated female was
noted to have an enlarged, fluid-filled kidney; the cortex was not
solid. Jhis finding has previously been observed in this strain of
animal in this laboratory and was judged not to be related to
compound administration.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was -judged to be
greater than 10 ml/kg.
Submitted by: Reviewed by:
David R. Damske, B.A. Rtfb'ert P. Beliles, Ph.D. Date
Toxicology Technician Director
Department of Toxicology Department of Toxicology
re BIONETICS
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-10
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-08
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-10
Test B-2
Date 6-2-78
Time 1200
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2898.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 155 to 252 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Litton 246
-------
4. RESULTS
The data have been summarized as follows.
Dose
Mean Body Weight
(g)
Day
y*" ' f '^3 /
0
10
0
10
0
216
224
178
169
7 14
Ma 1 es
265 343
301 358
Females
190 231
199 215
Deaths
Day
0-14
Total
Mortality
Deaths/Treated
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
Reviewed by:
£55ert P. Beliles, Ph.D. Date
Director
Department of Toxicology
Litton
BIONETICS
247
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-11
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-09
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-11
Test B-2
Date 6-2-78
Time 1300
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2899.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 178 to 229 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cyrle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -H).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
_- BIONETICS 248
Litton 248
-------
4. RESULTS
The data have been summarized as follows.
Dose
(ml/kg)
Mean Body Weight
(g)
Day
0
T
"IT
Deaths
Day
0-14
Males
Total
Mortality
Deaths/Treated
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals.
0
10
0
10
216
214
178
186
265 343
301 359
Females
190 231
220 244
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
Reviewed by:
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
v^rj »r~ _ - .—^ -^m__9_m-,__ ly* in ^f***^^^ ^\
Robert P. Beliles, Ph.I
Director
Department of Toxicology
Date
Litton
BIONET1CS
249
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-12
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-10
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-12
Test Sample B-2
Date 6-2-78
Time 1300
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2900.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 157 to 239 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Litton
-------
4. RESULTS
The data have been summarized as follows.
Dose
(tin/leg)
Mean Body Weight
(g)
Day
0
14
Deaths
Day
0-14
Males
0
10
0
10
216
222
178
184
265 343
305 361
Females
190 2.31
226 245
Total
Mortality
Deaths/Treated
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
Reviewed by:
R
Director
Department of Toxicology
ate
Litton
BIONETICS
251
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-13
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-11
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-13
Test B-2
Date 6-2-78
Time 1300
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2901.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 161 to 218 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
LJtton 252
-------
4. RESULTS
The data have been summarized as follows.
Dose
(ml/kg)
0
10
0
10
Mean Body Weight
(g)
Day
0
216
209
178
184
14
Deaths
Day
0-14 •
Males
265
280
343
336
Females
190
217
231
238
Total
Mortality
Deaths/Treated
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals. The uterus of one
treated female was noted to be distended. This finding has been
previously observed in this strain of an-imal in this laboratory
and was judged not to be related to compound administration.
CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
_
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
Reviewed by:
_____
Dert P. Beliles, Ph.D.
Director
Department of Toxicology
Date
ffl
Litton
BIONETICS
253
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-14
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-12
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-14
Test B-2
Date 6-9-78
Time 1500
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2902.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 175 to 222 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS
Litton 254
-------
4. RESULTS
The data have been summarized as follows.
Mean Body Weight
Dose (g) Deaths Total
(ml/kg) Day Day Mortality
^— Z—14 0-14 Deaths/Treated
Males
0 216 265 343 - 0/5
10 217 308 364 - 0/5
Females
0 178 190 231 - 0/5
10 185 230 248 - , 0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values greater than 10 ml/kg were estimated for both male
and female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg)
of the test compound to fasted young adult rats, no mortalities
were observed. Therefore, the median lethal dose was judged to
be greater than 10 ml/kg.
Submitted by: Reviewed by:
-: /O
David R. Damske, B.A. Robert P. Bellies, Ph.D. Date
Toxicology Technician Director
Department of Toxicology Department of Toxicology
BIONET1CS 255
Utton
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-15
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-13
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-15
Test B-3
Date 6-12-78
Time 0730
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2903.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 167 to 228 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BRJ were obtained from the
Charles River Breeding Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
Litton BIONETICS 256
-------
4. RESULTS
The data have been summarized as follows.
Dose
(ml/kg)
Mean Body Weight
(g)
Day
0
7
Deaths
Da
-------
SPONSOR: Monsanto Research Corporation
MATERIAL: CTHF-16
SUBJECT: FINAL REPORT
Acute Oral Toxicity Study in Rats
LBI Project No. 20969-14
1. OBJECTIVE
The objective of this study was to evaluate the acute toxicity
of the test compound when administered by oral gavage to male
and female rats.
2. MATERIAL
A glass bottle containing one liter of a liquid labeled:
CTHF-16
Test B-2
Date 6-12-78
Time 0730
was received from Clemson University by Litton Bionetics, Inc.
(LBI) on June 20, 1978 and designated as LBI No. 2904.
3. EXPERIMENTAL DESIGN
Young adult rats (weighing 180 to 219 g and eight to nine weeks
of age at the time of treatment, July 26, 1978) of the Charles
River CD strain [CRL:COBS CD (SD) BR] were obtained from the
Charles River Breedi-ng Laboratories, Inc., Portage, Michigan,
and acclimated to laboratory conditions for six days. The
animals were individually housed in wire-bottom cages in
temperature-controlled quarters under artificial illumination
controlled to provide a 12-hour light cycle. Water and Purina
Laboratory Chow were provided ad libitum with the exception of
the night before treatment when food was removed from the cages.
The test material was given undiluted. A single dose (10 ml/kg)
of the test material was administered by oral gavage to five
rats of each sex. A group of 10 untreated rats (five of each
sex) served as a control for all materials tested in this project
(LBI Project Nos. 20969-01 through -14).
The rats were observed frequently on the day of treatment and
daily thereafter. The animals were weighed on the day of treat-
ment and on Days 7 and 14 following treatment. Necropsies were
performed on the surviving animals killed 14 days after treatment.
BIONETICS 258
Litton
-------
4. RESULTS
The data have been summarized as follows.
Deaths
Dose
(ml/kg)
Mean Body Weight
(g)
Day
0
10
0
10
0
216
209
178
187
7 14
Males
265 343
293 352
Femal es
190 231
213 250
0-14
Total
Mortality
Deaths/TreateQ
0/5
0/5
0/5
0/5
Based on the absence of deaths in the 14 days following treatment,
LD50 values of greater than 10 ml/kg were estimated for male and
female rats.
No signs of toxicity or abnormal necropsy findings were observed
in any of the treated or control animals.
5. CONCLUSION
Following the oral administration of a single dose (10 ml/kg) of
the test compound to fasted young adult rats, no mortalities were
observed. Therefore, the median lethal dose was judged to be
greater than 10 ml/kg.
Submitted by:
David R. Damske, B.A.
Toxicology Technician
Department of Toxicology
Reviewed by:
Robert P. Beliles, Ph.D.
Director
Department of Toxicology
)ate
Litton
BIONETICS
259
-------
CONVERSION FACTORS AND METRIC PREFIXES (15)
CONVERSION FACTORS
To convert from To Multiply by
Degree Celsius (°C) Degree Fahrenheit (°F) t0p =1.8 t0(, + 32
Grams/meter3 g/m3) Milligrams/liter 1.0
Kilogram (kg) Pound-mass (avoirdupois) 2.205
Meter (m) Inch 3.937 x 101
Meter3 (m3) Gallon (U.S. liquid) 2.642 x 102
Meter3 (m3) Liter 1.0 x 103
METRIC PREFIXES
Multiplication
Prefix Symbol factor Example
Kilo k 103 5 kg = 5 x 103 grams
Centa c 10-2 5 cm = 5 x 10~2 meters
Milli m 10"3 5 mg = 5 x 10~3 gram
Micro p 10-6 5 yg = 5 x 10"6 gram
(15) Standard for Metric Practics. ANSI/ASTM Designation:
E 380-76e, IEEE Std 268-1976, American Society for Testing
and Materials, Philadelphia, Pennsylvania, February 1976.
37 pp.
260
-------
APPENDIX D
HYPERFILTRATION OF NONELECTROLYTES:
DEPENDENCE OP REJECTION ON SOLUBILITY PARAMETERS
H. G. Spencer, Department of Chemistry, Clemson University, Clemson,
SC 29631 (USA)
J. L. Gaddis, Department of Mechanical Engineering, Clemson University,
Clemson, SC 29631 (USA)
JUMMARY
The dependence of hyperfiltration rejection of nonelectrolyte solutes
in single-solute water solutions on solubility parameters is demonstrated
using hyperfilfcration results reported in the literature. The hyperfiltra-
tion systems are characterized by a solubility parameter derived empirically
from the rejection-solubility parameter dependence. A criterion for high
r<;joction follows.
INTRODUCTION
Hyperfiltration possesses high potential for separating toxic solutes
in industrial unit operation effluents (1) . Some of the nonelectrolytes
of concern are quite volatile and many are only slightly soluble in water.
Thus, the direct experimental measurement of the salt rejection .R^ of the
approximately 100 nonelectrolyte priority pollutants would be difficult and
a reliable metiiod for predicting ^ in a hyperf iltration system from a few
reference measurements and molecular properties of thn solutes i would h<->
valuable.
The most detailed model developed for this purpose has been provided
by Sourirajan and coworkers (2). Using one or mort? molecular properties
(acidity, basicity, Hammett and Taft numbers, steric parameters and Small's
number) and the permeability and rejection of a reference solution one can
relate these properties to Rj_. Other model:; use flux equations to relate
the measurable properties of a hyperfiltration system (3, 4). All
approaches include^both a transport property of the hyperfiltration system
and a coefficient for the distribution of the solute between the bulk solu-
tion and tho barrier.
W« hav- previously pointed out the value of solute molecular weights
in or-di-fi'.y U- of nonelectrolytes (1). Moat high rejection hyperfiltra-
'-on nK-mbr:mr.-''iff.v-tively reject nonoloctrolytos with molecular wciohts
greatpr than about 30. Cellulose acetate is an exception to this generali-
zation. Although scatter can be large in plots of rejection vs_. molecular
-------
weights, when the molecular weight is the most reliable or perhaps the only
molecular property available it can be used to estimate R^ in systems char-
acterized by a few measurements.
This report demonstrates a dependence of R^ for nonelectrolytes on
the solute solubility parameter introduced by Hildebrand and Scott (5)
and characterizes the hyperfiltration system by solubility parameters. It
also provides an empirical method for the rough estimation of R^ for a
solute of known solubility parameter in a hyperfiltration system from values
of R. obtained for a few reference solutes without explicitly considering a
transport property for the solute, providing its molecular volume is not
vastly larger than those of the reference solutes.
Chian and Fang (6) proposed the difference between the solubility para-
meters of the solute and membrane plays the major role in determining ^ of
nonelectrolyte solutes. Klein et^ al. (3) qualitatively related solute
permeabilities in the absence of hydraulic flux with two-dimensional
solubility parameters and used the experimental permeabilities to predict
specific separations of organic solutes under hyperfiltration conditions.
A quantitative correlation of R. with solubility parameters was not attempted
in either report.
DEFINITIONS, CONCEPTS, AND CALCULATIONS
- - k - -
The solubility parameter is defined by <5 = (AE/V) where AE/V is the
energy of evaporation per unit volume, called the cohesive energy density.
The units of 5 are (J/ra3)^. Solubility parameter theory predicts that the
best solvent for a given solute, e.g. a polymer, is one whose solubility
parameter is equal to or close to that of the solute (5).
The rejection R^ of a solute is defined as 1 - Cp/Cjj, where Cp and C^
are concentrations in the permeate and bulk feed solutions respectively.
An intrinsic rejection, 1 - Cp/Cw based on the concentrations of permeate
and that occurring at the feed-membrane interface Cw is commonly defined.
Normally the intrinsic rejection is projected as the infinite-velocity
asymptote of the rejection Rj., and this intrinsic rejection is the property
logically addressed in this-'study- Because most investigations are conducted
30 as to preclude large differences in the two rejections, and virtually
no data exist projecting the intrinsic rejection, the observed rejection is
used throughout. Errors produced by this simplification may be substantial
and are largest near rejection of 0.5.
In hyperfiltration the distribution of solute between the bulk solution
and the barrier is assumed to be important in determining R^. Further,
assuming the concentration of solute available for transport across the
hyperfilter depends on AJJJ, » 8^ - 6m, where 5ra characterizes the hyper-
filter, Ri should be a function of Aim. of course, an attempt to relate B^
to ^im alone is imcomplete because a transport property characterizing the
hyperfiltration system is not explicitly included.
The group contribution method of Konstram and Fairheller (5) was used
to calculate Small's number (8) S^ and the $j_ were obtained by 5 • = S^/v^,
where v^ is the molar volume. This method is used although tables containing
6^ for many solutes are available (5, 9). It is desirable to use a consis-
tent method for as many compounds as possible. Even with the use of this
262
-------
•
general approach some values of Si are not available, especially those for
polyfunctional molecules. Values of v^ were calculated by dividing mole-
cular weight of the solute J^ by its density pj[ in the liquid state at the
temperature of the experiment, with Mi and pt obtained from commonly used
- -
tables (10).
DEPENDENCE OF REJECTION ON SOLUBILITY PARAMETERS
Figures 1-5 show the dependence of R^ on 5i for the six hyperfiltra-
tion systems described in Table 1 (11 - 15?. The 5i occurring at R. » 0 is
assumed to be the solubility parameter characterizing the membrane^system
and is designated 5m. The <5ffl were obtained by a visual linear extrapolation
of the plots. A slope of -10 x 103 (m3/J)^ was satisfactory for all graphs
in the region A^ < 0. Insufficient data are provided to determine ^ by
extrapolation in the poly(ether/amide) thin film composite (PA-300) systems,
Figure 4. Using the slope observed in the remainder of the graphs, 5m should
be in the interval 34 x 103 < 6m < 36 x 10 (J/m3P- It should be noted that
the scatter is very large in the cellulose acetate systems.
In the Permasep B-9 and cellulose acetate systems, Figures 2 and 5,
several values of Rj_ at A^ > 0 are available. It is clear in Figure 2 that
Ri increases monotonically with increasing A^ in the region AJJ, > 0. The
dependence is not well defined in the cellulose acetate case where in this
region several of the solutes listed contain two functional groups and the
calculations of <5i is less reliable.
The molecular weight, or v. , may be used to estimate fa in many systems,
however this dependence appears to be absent in the cellulose acetate system.
Figure 6 is provided to illustrate this observation and it should be compared
with Figure 5, where ^ are plotted vs. 5^.
DISCUSSION
The dependence of £i on Si has been illustrated in several hyperfiltra-
tion systems. It is evident that high rejection occurs when JAimj is large.
Figure 7 provides a graph of £j_ vs. Aj^ for all the membranes other than the
cellulose acetate membranes and Figure 5 provides a graph of £j_ vs. 6^ for
the cellulose acetate systems, where the scatter is greater.
Thia empirical treatment of rejection of nonelectrolytes in hyperfiltra-
tion systems also provides a method for estimating £j_ for any solutes of
known 6- from a few reference experiments. The value of 5m is determined
from a graph of Rj_ vs. Oi obtained for the reference solutes. Solutes
having ^ < -10\ 103 (J/m3P should have £i > 0.90. In the region
-10 x lO3^ < Aim < '0 (J/m3)^ £t a - 10 x IB- Aim, this estimate being better
for membranes other than cellulose acetate than for cellulose acetate.
The value of 5 appears to characterize the hyperfiltration system with
respect to its rejection of nonelectrolytes and provides an attractive
criterion for selecting membranes for hyperfiltration applications. Although
the nonelectrolyte priority pollutants possess a broad range of solubility
parameters, the maximum appears to be about 28 x 103 (J/mJ) and many are^
much smaller. Hyperfiltration systems with 6^ greater than about 36 x 10-
or 38 x 103 (J/m5P should provide high rejections of these solutes.
263
-------
Table Dl -CHARACTERISTICS OF HYPERFILfRATION SYSTEMS
Pressure
Membrane (MPa)
Aromatic poly amide 1.72
Aromatic poly amide.
Per mas ep B-9, hollow
fiber 3.10
NJ
Cellulose acetate 1.72
Cellulose acetate 1.72
US- 100 5.52
Poly (ether/amide)
PA- 300 6.89
Temperature. l53 fiu. ,
<«e) <*/»*)*
25 29.0
20 30.0
25 25.0
23-25 25.0
25 34.5
25 CW34) *
Classes of Solutes
alcohols , aldehyes ,
ethers, ketones
alcohols , acids
alcohols , aldehydes
alcohols , aldehydes ,
esters , ethers , hydro-
carbons
acids , alcohols , alde-
hydes, esters, ketones,
amines
acid, alcohols, alde-
hydes, esters, ketone,
ch lorohydrocarbons
Reference
11
12
11
15
13
14
* Not enough data for extrapolation.
-------
Refinement of this approach using solubility parameters as a measure
of the membrane-solute interaction will likely require incorporation in a
flux model.
ACKNOWLEDGEMENT
The authors wish to acknowledge the generous financial support of this
work by the U.S. Environmental Protection Agency, Industrial Environmental
Research Lab, Research Triangle Park, North Carolina, EPA Grant Number
R805777-1.
265
-------
REFERENCES
1. H. G. Spencer, J. L. Gaddis, and C. A. Brandon, Membranes for
Toxic Control, presented at the Membrane Separation Technology
Seminar, Clemson University, Clemson, SC 1977.
2. Summary of the method: S. Sourirajan and T. Matsuura, in
"Reverse Osmosis and Synthetic Membranes," S. Souriragan, ed.,
National Research Council of Canada Publications, Ottawa,
Canada, 1977, Chapter 2.
3. Examples: K. S. Spiegler and 0. Kedera, Desalination, ^ (1966)
311? H. K. Lonsdale, U. Merten, and R. L. Riley, J. Appl.
Polymer Sci., 9 (1965) 1341; and L. Dresner and J. S. Johnson,
Jr., in "Principles of Desalination," 2nd ed., K. S. Spiegler
and A. D. K. Laird, eds., Academic Press, New York (in press).
4. E. Klein, J. Eichelberger, C. Eyer, and J. Steith, Water Res.,
9 (1975) 807.
•v
5. J. H. Hildebrand and R. L. Scott, "The Solubility of Non-
electrolytes," Rheinhold, New York, 1950.
6. E. S. K. Chian and H. H. P. Fang, AIChE Symposium Ser., 70
(1973) 497. ^
7. H. H. Konstam and W. R. Fairheller, Jr., AIChE J, 16 (1970) 837.
' ~" v\i
8. P. A. Small, J. Rppl. Chem., 3 (1953) 71.
^
9. J. L. Gordon, in Encyclopedia of Polymer Science and Technology,
3 (1965) 833; H. Burrell, J. Paint Technol., 27 (1955) 726;
C. M. Hansen, Ind. Eng. Chem., Prod. Res. Dev^ 8 (I960) 2.
10. "Handbook of Chemistry and Physics," R. E. Weast, ed., CRC Press,
Inc., Cleveland, Ohio.
11. J. M. Dickson, T. Matsuura, P. Blais, and S. Sourirajan, J. Appl.
Polymer Sci., 19 (1975) 801. ~~
—•—— -Wi
12. V. a. Caracciolo, N. W. Rosenblatt, and V. J. Tomsic, in "Reverse
Osmosis and Synthetic Membranes," S. Sourirajan, ed., National
Research Council of Canada, Ottawa, Canada, 1977, Chapter 16.
13. L. T. Rozelle, J. E. Cadotte, K. E. Cobian, and C. V. Kopp, Jr.,
ibid., Chapter 12.
266
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14. R. L. Riley, R. L, Fox, C. R. Lyons, C. E. Milstead, M. W. Seroy,
and M. Togami, Spiral-wound Poly (ether/amide) Thin-film Composite
Membrane Systems, presented at the Membrane Separation Technology
Seminar, Clemson University, Clemson, SC 1976.
15. T. Matsuura and S. Sourirajan, J. Appl. Polymer Sci., 15, 2905
(1971); ibid., 16, 1663, 2531 (1972); ibid., 17, 1043,<3683 (1973).
267
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"WA
Figure Dl Solute rejection vs. solubility
parameter: Polyamide membrane,
1.72 MPa, 25°C (11).
Figure D2 solute rejection vs. solubility
parameter: Polyaaide, Permasep
B-9, 3.10 WPa, 20°C (12).
0 It 20 22 24 26 2t SO 32 34
vnuiinirf *•»««« rr« toM,u/mV '
z
e
—o
O Q O
o
o
(PHi18 2022 Z4 K 28 30 M 94 36
SOLUBILITY MftAMETCft (0* if i (J/ms)l/*
Figure D3 Solute rejection vs. solubility
parameter: NS-100, 5.52 MPa,
25°C (11).
Figure
Solute rejection vs. solubility
parameter: poly(ether/amide),
PA-300, 6.89 MPa, '25°C (14).
-------
00W,
14 16 18 30 22 426
SOLUBILITY PARAMETER 10*
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APPENDIX E
1.0 IDENTIFICATION OP COLLECTION SAMPLES
Hyperfiltration will be performed on two process fluids, a
scour waste and a dye drop, selected and obtained as described
in the Program Plan. Sixteen collection samples, identified in
Table 1, will be obtained, separated and bottled as test samples,
and shipped to the various laboratories designated for testing
and analysis. In addition, one-gallon contingency collection
samples will be obtained and stored at Clemson University until
the project is completed.
270
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Table El. COLLECTION SAMPLES FOR BIOASSAY TESTS AND CHEMICAL ANALYSES
Sample
CTHF-1
CTHF-2
CTHF-3
CTHF-4
V
CTHF-5
CTHF-6
CTHF-7
CTHF-8
CTHF-9
CTHF-10
CTHF-11
CTHF-12
CTHF-1 3
CTHF-14
CTHF-1 5
CTHF-16
Description
Plant water
Apparatus water
Scour-1, feed for PEA and CA hyperfiltration
Scour-1, permeate from PEA hyperfiltration
Scour-1, permeate from CA hyperfiltration
Scour-1, concentrate from PEA and CA
hyperfiltration
Scour-2, feed for DM hyperfiltration
Scour-2, permeate from DM hyperfiltration
Scour-2, concentrate from DM hyperfiltration
Dye-1, feed for PEA and CA hyperfiltration
Dye-1, permeate from PEA hyperfiltration
Dye-1, permeate from CA hyperfiltration
Dye-1, concentrate from PEA and CA
hyper fi It ration
Dye-2, feed for DM hyperfiltration
Dye-2, permeate from DM hyperfiltration
Dye-2, concentrate from DM hyperfiltration
Volume
(gallons)
5
5
25
25
25
ioa
25
25
ioa
25
25
25
ioa
25
25
ioa
a concentrate samples will be 2 - 5 gallons, containing equivalent
solids to the feed sample.
271
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2.0 BIOASSAY TESTS AMD CHEMICAL ANALYSES
The planned bioassay tests and chemical analyses for the
129 consent decree priority pollutants are listed in Table 2.
This table includes the test-sample container requirements and
designates the collection samples for which each test is planned.
272
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Table E2. BIOASSAY TESTS AND CHEMICAL ANALYSES, TEST-SAMPLE CONTAINERS,
AND TESTS DESIGNATED FOR COLLECTION SAMPLES
U)
Test
#
B.I
B.2
B.3
C.I
C-2
C.3
C.4
C.5
C.6
Description
Microbial mutogenicity
(Ames)' and cytotoxicity
(hamster ovary cells)
Acute toxicity (rat)
Freshwater static bio-
assay (Daphnia and
Fathead minnows)
Volatile solutes
Nonvolatile solutes
Metals
Cyanide
Phenols
Pesticides
Sample
Volume
500 m£,
500 m£
20 galsa
2 x 40 m£
2x1 gal
500 mJl
500 m£
500 m£
(use part
Required for
Collection Samples
Container ( CTHF- )
amber glass,
Teflon- lined caps
glass. Teflon- lined caps
5 gallon, plastic
cubitainers
glass vials.
Teflon- lined septa
amber glass, Teflon-
lined caps
plastic bottles
plastic bottles
amber glass
of test sample C.2)
3-16
3-16
3-16
1-16
1-16
1-16
1-16
1-16
a concentrate samples will be 2 - 5 gallons, containing equivalent solids to the feed sanqple.
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3.0 PREPARATION OF TEST-SAMPLE CONTAINERS
3.1 Containers
The containers listed in Table 3 will be used for the test
samples. Unless otherwise specified the caps will be lined with
Teflon tape, 2 mils thick.
3.2 Cleaning Procedures
v a. Narrow-mouth glass and amber glass test-sample bottles
and caps
Hash with strong acid (50% H2SOtf and 50% HNOg) and rinse several
times with tap water and deionized water. Heat the bottles for thirty
minutes at 4OO°C in a glass annealing oven then cool to room tempera-
ture and cap.
b. Cubitainers and caps
Rinse cubitainers and caps several times with deionized water,
drain at room temperature, and cap.
c. Plastic bottles and caps
Wash with acid (5 m£ of redistilled HN03 per liter of deionized
water) and rinse several times with deionized water. Cap after
draining at room temperature.
d. Glass vials and Teflon-lined septa
The glass vials are prepared as in 3.2 (a) . Rinse the Tef on-
lined septa several times with deionized water and dry at room
temperature. Cap vials after cooling.
274
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Table t3. DESCRIPTION OP TEST-SAMPLE CONTAINERS
Bottles
480 mZ, amber glass
480 m&, glass
\
480 m£, polyethylene
5 gal, 1 gal
cubitainers
1 gal, amber glass
40 m£, vials and
septa
Supplier and Catalog Number
A. H. Thomas
A. H. Thomas
A. H. Thomas, high density
polyethylene, Nalge 2002
series
Cole-Parmer Instrument Co.
Fisher Scientific Co.
ring jugs
Pierce, Inc.
1702-N43
1702-F70
1702-K63
6100-40,
6100-30
2-884-5BB
13075
12722
275
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e. Teflon sheets for lining caps
Wash with acid (5 a£ of redistilled HN03 per liter of de-
ionized water) and rinse with deionized water.
276
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4.0 PREPARATION OF HYPERFILTRATION APPARATUS AND FEED TANK
The tanks will be scrubbed, washed with a detergent, rinsed
several times with plant water and drained. The process fluid will
be transferred through a polypropylene filter to the feed tank using
an existing industrial tube using a stainless steel pump. The process
fluid will cool to room temperature in the feed tank until operation
commences with the hyperfiltration apparatus. The feed tank will
be kept covered to minimize escape of volatiles.
The hyperfiltration unit will be cleaned of residual material
using a sequence of washes. A detergent operation followed by a
base wash is expected to remove most greases, waxes, and organic
materials. The unit will be rinsed with plant water. The unit will
be operated with plant water to indicate whether materials are evolved
within the plumbing or feed system. A sample (CTHF-2) will be analyzed
for comparison with the plant water sample.
277
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5.0 SAMPLING PROCEDURE
5.1 Collection Samples
A sample of the plant water will be obtained from the tap
providing water for the scour operation (CTHF-1).
The feed samples will be collected during the transfer of
the test fluids from the feed tank to the hyperfiltration apparatus
(CTHF-3, CTHF-7, CTHF-10, CTHF-14).
The total permeate from each hyperfilter will be collected in
a stainless steel container (prepared as described in 3.2 (c). The
test samples will be taken from this collection container by draining
or siphoning. The permeate samples are a composite of the permeate
from each hyperfilter (CTHF-4, CTHF-5, CTHF-8, CTHF-11, CTHF-12,
CTHF-15).
The concentrate samples will be obtained by draining the
hyperfiltration apparatus at the completion of each of the four
experiments (CTHF-6, CTHF-9, CTHF-13, CTHF-16).
5.2 Test Samples
Each collection sample will be divided into the specified test
samples.
a. Bioassay test (B.I, B.2, B.3), nonvolatile solutes and
pesticide (C.2 and C.6) and metals (C.3)
Remove sample container cap, fill completely by draining or
siphoning from the collection container, cap immediately and cool to
4°C as rapidly as possible. No preservative is added.
278
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b. Volatile solutes (C.I)
Collect two 40 tal samples. Slowly fill each vial to overflowing.
Carefully set the vial on a level surface. Place the septum (Teflon
side down) on the convex sample tniniscus. Seal the sample with the
screw cap. To insure the sample has been properly sealed, invert the
sample and lightly tap the lid on a solid surface. The absence of
entrapped air bubbles indicates a proper seal. If air bubbles are
present, open the bottle, add additional sample, and reseal. Cool
to 4°C as rapidly as possible. No preservative will be added.
c. Cyanide (C.4)
r
Collect sample as described in 5.2 (a). A preservative is
required at the time of collection. Add 1.0 mJl of 10 N[ NaOH, to
obtain pH = 12. Oxidizing agents such as chlorine decompose most
cyanides. Test a drop of the sample at the time of collecting using
Kl-starch paper; a blue color indicates a need for chlorine treatment.
Add ascorbic acid, a few crystals at a time, until a drop of sample
produces no color on the indicator paper. Then add 0.3 g of
ascorbic acid.
279
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d. Phenols (C.5)
Collect sample as described in 5.2 (a). A preservative is
required at the time of collection. Acidify the sample to pH * 4
by addition of phosphoric acid. Determine pH with pH paper. Note
volume of acid added and its concentration on the sample tag.
5.3 Labels
Labels are to be waterproof and information written with
India ink. Each label will indicate:
Collection Sample Number
Test Sample Designation
Sampler ______________
Date
Time
Preservatives
280
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6.0 SHIPPING PROCEDURES
All samples will be refrigerated upon collection. Cardboard
cartons, suitably insulated, will be charged with water ice or
dry ice and the samples placed therein. Care will be taken to
insure the non-freezing of samples if dry ice is used. A refrigera-
tion life of forty hours will be used as a design criterion.
Notice will be given in advance of testing to the recipients
of test fluids and approximate scheduling. A definite notice
including bill of lading number and anticipated flight schedule
will be relayed upon shipment.
Shipments will be by air freight. Parcels will be marked
"Contents under Refrigeration," "Perishable," "Handle with Care -
Fragile," and "Contains Dry Ice" or "Contains Ice" as appropriate.
281
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7.0 DEFINITIONS
PEA - poly(ether amide) hyperfilter
CA - cellulose acetate hyperfilter
DM - ZrO-PAA dynamic membrane hyperfilter
Collection Sample - designated samples in Table 1, e.g., CTHF-3
Test Sample - Sample of collection sample sent for a specific
test. Designations are listed in Table 2. Example: Test
sample CTHF-3, C.I is the sample of collection sample CTHF-3
bottled in two 45 m£ vials for volatile solids analyses.
282
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
REP'
EPA-600/2-79-118
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
t. 1 M UC MINU OUB I I I I.C
Evaluation of Hyperfiltration for Separation of Toxic
Substances in Textile Process Water
5. REPORT DATE
June 1979
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
J.L. Gaddis and E.G. Spencer
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Clemson University
Department of Mechanical Engineering
Clemson, South Carolina 29631
10. PROGRAM ELEMENT NO.
1LA760
11. CONTRACT/GRANT NO.
Grant R805777
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
Final; 1/78 - 4/79
14. SPONSORING AGENCY CODE
EPA/600/13
15. SUPPLEMENTARY NOTES JERL-RTP project officer is Max Samfield, Mail Drop 62, 919/
541-2547.
report gives results of an evaluation of hyperfiltration for separation
of toxic substances in textile process water. Three membranes (cellulose acetate,
polyether /amide , and dynamic zirconium oxide /polyacry lie acid) were used to separ-
ate process water from scour and dye operations into permeate and concentrated
streams. Feed, permeate, and concentrate samples from each run were analyzed.
Chemical analyses for organic and metal toxic pollutants and bioassays for rat acute
toxicity, fathead minnow and daphnia acute toxicity, microbial mutagenicity , and
hamster ovary clone cytotoxicity response were conducted. The minnow and daphnia
tests showed active results , with good correlation. The other bioassays produced no
response. Toxicant rejections of 55 to 100% were observed: the relative rejection by
the three membranes was almost exclusively counter to the relative rejection of
salt. Mass balances of biological toxicant were excellent, suggesting high confidence
in the result. Chemical analysis for organic compounds sensed 19 of the organic
toxic pollutants in low levels, <300 mg/cu m. The results were difficult to interpret
for mass balance and membrane rejection of particular solutes. Except for a few
compounds , the data appear to suggest membrane separation. Metal toxic pollutant
concentrations were low: only three were concentrated enough for valid estimations.
. ABSTRACT
17.
KEY WORDS AND DOCUMENT ANALYSIS
a.
DESCRIPTORS
b. IDENTIFIERS/OPEN ENDED TERMS
c. COS AT I Field/Group
Pollution
Fluid Filters; Membranes
Toxicity Scouring
Textile Industry Dyeing
Industrial Water Analyzing
Water
Pollution Control
Stationary Sources
Hyperfiltration
Process Water
13B
13K;11G
06T
11E
07B
13H
14B
18. DISTRIBUTION STATEMENT
Release to Public
19. SECURITY CLASS (ThisReport)
Unclassified
21. NO. OF PAGES
291
20. SECURITY CLASS (Thispagef
Unclassified
22. PRICE
EPA Form 2220-1 (9-73)
283
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