Document G5VwR3MDYYypo6k0n3Xrn07Zq
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I. GOOD LABORATORI PRACTICE COMPLIANCE STATEMENT This study was conducted according to EPA Good Laboratory Practica Regulations (40 CFR 792), with the following exception. The stability of the teat substance was assumed but not verified at T.R. wilbury Laboratories. Deviations from the study protocol and protocol amendments are presented in Section XII of thia report.
MWfH
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II. QUALITY ASSURANCE STATEMENT
Submitted byi
T.R. wilbury Laboratori, m e 40 Doaks Lana Marblehead, Massachusetts
Certification:
T.R. Wilbury Laboratories, Inc. study number 888-TB, Acute Toxicity of N2803-2 to the rathead Minnow, Pimaphalaa promelas, was audited by the Quality Assurance Unit for compliance with the protocol, standard operating procedures, and applicatile Good Laboratory practices (U.s. EPA, 1993). Quality assurance audits were performed and the findings reported to T.R. Wilbury Laboratories management and the study director on the following dates:
Aspect Audited
Protocol : in-life: Draft Report: Final Report:
Audit Date
8/ 3/95 9/12/95 9/22/95 11/16/95
Reported to Study Director
8/ 4/95 9/12/95 9/22/95 11/16/95
Reported to
Management
8/ 4/95 11/16/95 11/16/95 11/16/95
Arthur P. Faradica Quality Assurance Officer '
T.R. wilbury study Number 888-t h
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IZZ. T A B U Or CONTEXTS
SECTION I
I. Good Laboratory Practice Compliance Statement II. Quality Assurance statement
III. Table of Contents
IV. Index of Tables and Figures
.
V. Summary
VI. General Information
VII. Introduction
vili. Methods and Materials
IX. Results
X. References
XI. Signature Page
XII. Protocol Amendments and Deviations
Appendix A. Hater Quality Data from Toxicity Test
PAGE
2 3 4 5 6 7 8 8 13 17 18 19
20
T.R. Wilbury study Number 888-th
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I V . INDEX or T A B U S AND rXOVJtSS
Tablo 1.
Chemical characterization of representative
sample of dilution water used for the toxicity teat with fathead uinnows, Plmephalea promelaa, and N2803-2
Table 2.
Test system summary for the toxicity test with N2803-2 and fathead minnows, Plmephalea promelaa
Table 3.
Survival and sublethal effect data from the toxicity test with fathead minnows, Plmephalea promelaa, and N2803-2
Table 4.
Median lethal concentrations (X.C50S) from the toxicity test with fathead minnows, Plmaphalea promelaa, and N2803-2
Table A.l.
Dissolved oxygen concentration, conductivity, pa, and temperature measured during the toxicity test with fathead minnows, Plmaphalea promelaa, and N2803-2
Figure 1. Survival of fathead minnows, Plmaphalea promelaa, exposed to N2803-2 for 96 hours
PACE 9
11 14 1.6 21
15
T.R. Wilbury Study Number 888-TH
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Sf The acuta toxicity of N2803-2 to tba fathoad minnow, Plmaphalea K proaalaa, ia dascribad in this final report. Tba tant was conducted for
$ 3H Company for 96 houra from saptembar 8 to 12, 1995, at T.R. Wilbury j Laboratories, Inc. in Marblahaad, Masaachuaatta. It waa conductad by ' Peter L. Kowalski, Jaanna Hagazu, Jacqueline Navius, Keven Stavana, Patar ' ........ Haviua, Robert Boari, and Timothy Hard according to tha protocol davalopad
for T.R. Wilbury Study Number 888-TH. N2803-2 waa auppliad by tha aponsor.
Tha taat waa performed under atatic conditiona with five concantrationa of teat substance and a dilution water control at a temperature of 21.6 to 22.0*C. The dilution water wan filtered deionized water collected at Marblehead, Masaachuaatta and adjusted to a hardness of 40 mg/L aa caco}. Aeration waa initiated at 72 hours to maintain dissolved oxygen concentrations above an acceptable level.
Nominal concentrations of N2803-2 were 0 (control), 160, 250, 400, 630, and 1,000 mg/L. Fathead minnows used in the test were procured from a commercial supplier and acclimated to test conditions at T.R. wilbury Laboratories. After 96 hours of exposure the control fish had an average wet weight (blotted dry) of 0.36 g and an average total length of 34.9 mm. All fish were in good condition at the beginning of the study.
Exposure of fathead minnows to the test substance resulted in a 96 hour median lethal concentration (LC50) of 960 mg/L N2803-2, with a 95% confidence interval of 830 to >1,000 mg/L. The 96 hour no observed effect concentration (NOEC) is 630 mg/L.
T.R. Wilbury Study Number 888-TH
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| VI. GENERAL INFORMATION
i
Material Testedi
N2803-2
| Sponsors
3M Company Environmental Laboratory 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55144
Test substance shipped From:
3m Company Environmental Laboratory 367 Grove Street Building 53-5S-02 St. Paul, Minnesota 55101
Reported Purity:
loot
study Initiation Date:
Experimental start Dates
Experimental Completion Dates
study Completion Date:
Archive Location for the Biological Raw Data and a Copy of the Final Reports
August 16, 1995 (protocol signed) September S, 1995
September 12, 1995 November 16, 1995
T.R. wilbury Laboratories, Inc. Marblehead, Massachusetts
T.R, Wilbury study Humber 888-TH
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VXI. INTRODUCTION
Thia study was sponsored by 3H Company, St Paul, Minnesota. Tha objactiva of tha study was to datarmina tha 24, 49, 72, and 96 hour madian lathal concantratlons (LC50s) of tha tast subatanca to tha fathaad minnow, Plmephalea promelaa, a freshwater fish, under static conditions. Tha tast
protocol was based on U.s. EPA methods (U.S. EPA, 1993). Tha report
contains sections that describe tha methods and materials employed in tha study, and results of tha investigation. The report also contains an appendix that presents water quality data collected during tha test.
VXXX. METHODS AMD MATERIAU
TEST SUBSTANCE I
The sample of N2803-2 (T.R. Wilbury Laboratories Sample Number 529) waa delivered to T.R. Wilbury Laboratories on August 29, 1995 in a 5 gallon metal can within a cardboard box at ambient temperature. The test substance was contained in an approximately 150 ml plastic jar. The attached label included the following information: "N2803-2, s. Beach, 23E-09, (612) 778-7452, 8/22/95".
N2803-2 (a clear liquid) was supplied by 3M Company, St Paul, Minnesota, prior to use the test substance was stored in the dark at room temperature. Test substance stock solutions were formulated in dilution water without the use of a solvent. The test substance was assumed to have a purity of 100% active ingredient and to be stable under storage and testing conditions. unused test substance will be returned to the sponsor.
DILUTION WATER:
Water used for acclimation of test organisms and for all toxicity
testing was deionized water collected at T.R. Wilbury Laboratories in
Marblehead, Massachusetts. Water was adjusted to a hardness of 40-48 mg/L
as CaCOj and stored in 500-gallon polyethylene tanks where it was aerated
and continuously passed through a particle filter, ultraviolet sterilizer,
and activated carbon. Results of chemical analysis of a representative
sample of water are presented in Table 1.
TEST ORGANISMI
\
Juvenile fathead minnows, Plmephalea prontelas, employed as test organisms were from a single source and were procured from Aquatic Biosystems, Fort Collins, Colorado on August 10, 1995. The species of the fish was verified using an appropriate taxonomic key.
T.R. Wilbury Study Number B88-TH
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IS'
.
Table 1.
Chemical characterization of a repraaentative ample of dilution water uaed for the toxicity teat with fathead minnowe, Plnaphalaa promalaa, and N2803-2.
Parameter1
Unit of Measurement
Detection Limit
Measured Value
Metals Aluminum
Arsenic Boron Cadmium Chromium Cobalt Copper Iron Lead Mercury Nickel Silver Zinc
mg 'L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L
0.1 0.01 0.5 0.0002 0.01 0.03 0.005 0.03 0.005 0.0003 0.03 0.02 0.02
ND2
ND HD ND ND ND ND 0.03 ND ND ND ND ND
Nitrate
mg/L as N
0.05
ND
Chloride
mg/L
1
Fluoride Total organic
mg/L
0.1
ND
s,
Carbon
mg/L
1
ND
Total Phosphorus
mg/L
0.03
ND
Organochlorine Pesticides
Toxaphene
pg/L pg/L
0.5 2
ND ND
Organophospborus Pesticides
Dimethoate TEPP Monocrotophos
pg/L pg/L pg/L pg/L
0.5 2.0 2.0 2.0
ND ND ND ND
pg/L
0.5
ND
Notes
.1. Metals, nitrate, chloride, fluoride, total organic carbon, total phoaphorua, organochlorine pesticides, organophosphorua pesticides, and pcbs were measured in deionized water (used to
;j formulate dilution water) that was collected during August, - 1995 and analyzed by Pace, Inc. as part of routine water
quality testing. 2. HD not detected at or above the detection limit.
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Prior to testing tha fiah war* maintainod undar flow through conditiona in a 270 Xitar fiberglass tank or a 40 iitar glaaa aquarium. During acclimation fiah were not traatad for diaaaaa, tbay vara free of apparant sickness, injuries, and abnormalities at tba baginning of tha taat, and mortality during tha final 48 houra of acclimation was laaa than 3t.
During tha 14 day period before tha atart of tha taat the acclimation temperature range was 21.0 to 22.3*C and tha diaaolvad oxygen concentration vaa always at least 8.6 mg/L. During acclimation fish vara fed a dry commercial food (Trtra Hina staple rood. Lot # TM05) daily
except during the 48 houra immediately preceding the test, riah were not
fad during tha test. At the conclusion of tha test tha control fish had an average total length of 34.9 mm and an average vet weight of 0.36 g (loading during the definitive toxicity test was 0.24 g/L).
TOXICIT TESTMOl
A static screening test was conducted from August 18 to 22, 1995 with five concentrations of test substance and a dilution water control. Nominal concentrations of N2803-2 were 0.10, 1.0, 10, 100, and 1,000 mg,L. After 96 hours of exposure there was loot survival i.n the control and at 0.10, 1.0, 10, and 100 mg/L, and 0% survival at 1,000 mg/L.
The definitive toxicity test was performed from September 8 to 12, 1995, according to T.R. Hilbury Protocol 888-TH (Acute Toxicity of N2803-2 To The Fathead Minnow, PlmephaleB promalaa). The protocol, which was signed by the study director on August 16, 1995, is based on procedures of th U.S. Environmental Protection Agency (1993). A summary of the test system is presented in Table 2.
The test was conducted at a target temperature of 22 t 2*c with five concentrations of test substance and a dilution water control. Nominal concentrations of the test substance were: 0 mg/L (control), 160, 250, 400, 630, and 1,000 mg/L. Appropriate amounts of test substance were added directly to dilution water in test vessels to formulate test media without the use of a solvent.
Twenty fathead minnows were indiscriminately and equally distributed among two replicates of each treatment. . The test was performed in 20 liter glass aquaria that contained 15 liters of test solution (water depth was approximately 18 cm). Test vessels were randomly arranged in a water bath during the 96 hour test (a random numbers table was used to select the location of each vessel) and they were loosely covered. A 16 hour light and 8 hour dark photoperiod with a 15 minute transition period was automatically maintained with cool-white fluorescent lights that provided a light intensity of 25 footcandles. Aeration was initiated after the 72 hour dissolved oxygen concentration measurements to maintain dissolved oxygen concentrations above acceptable levels.
T.R. wilbury study Number 888-t h
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Tabi* 2
Teat system summary for tha toxioity taat with H2803-2 and fathaad minnows, Plmiphtla* promelaa.
Spaciaa >
Plmaphala* promelas
Age/Size of Teat organisms:
Juveniles/!}.36 grams, wet weight
Taat Duration:
96 hours
Taat Veaaala:
20 liter glass aquaria
Depth of Teat Media:
IS cm
Humber of Replicate Teat Veaaela: 2
Organisms per Replicate:
10
Temperature Range:
21.6-22.0*0
Minimum Dissolved oxygen:
4.6 mg/L (prior to aeration)
Photoperiod:
16 hours light and 8 hours dark with a 15 minute transition period.
Light Intensity:
25 footcandles
Water Quality Measurements:
Dissolved oxygen, pB, temperature, and conductivity daily in each test vessel. Temperature measured at least every hour in 1 teat vessel. Hardness, alkalinity, acidity, and particulate matter and TOC in the dilution water. Dilution water analysis is summarized in Table 1.
End Points:
24, 48, 72, and 96 hour LC50; 96 hour HOEC.
Acceptability Criteria:
slO% control mortality and acceptable temperature range.
T.R. Wilbury study Humber 888-TH
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Ths number of surviving organisms and tbs occurrsnco of sublsthal offsets (immobilization, loss of equilibrium, loss of reflex, excitability, discoloration, or change in behavior) were determined visually and rscordad initially and after 24, 48, 72, and 96 hours (mortality and sublethal effects vera monitored twice each day). Dead test organisms were removed when first observed. Dissolved oxygen (YSI Model 57 Mter; instrument numbers 1), pH (Beckman model pBI 12 meter; instrument number 124), conductivity (Cole Parmer sieter, instrument number 94), and temperature (Beckman model pHl 12 meter; instrument number 124, probe pHT-1) were measured and recorded daily in each test chamber that contained live animals. The temperature in one test vessel was recorded at least hourly during the test.
STATISTICAL MSTHODSl
Results of the toxicity test were interpreted by standard statistical techniques. Computer methods (Stephan, 1983) were used to calculate the median lethal concentrations (LC50s). All calculations were performed by the author using nominal concentrations of test substance and the number of live fathead minnows. The no observed effect concentration is the highest concentration of test substance that did not cause any toxicant-related mortality or sublethal effects.
T.R. Wilbury Study Number 888-TH
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Table 3.
Survival and aublathal affect data from tha toxicity taat with fathead minnow*, Pinaphala* promtltt, and N2803-2.
Nominal Concentration
of N2803-2 (mg/L)
0 (control)
160
250
400
630
1,000
Rep.
Number alive ------------------
0 24 48 72 96 hr hr hr hr hr
Number affected ----------------- .
0 24 48 72 96 hr hr hr hr hr
1 10 10 10 10 10 2 10 10 10 10 10
1 10 10 10 10 10 2 10 10 10 10 10
1 10 10 10 10 10 2 10 10 10 10 10
1 10 10 10 10 10 2 10 10 10 10 10
1 10 10 10 10 10 2 10 10 10 10 8
1 10 6 4 3 3 2 10 8 7 6 6
00000 00000
00000 00000
00000 00000
00000 00000
00000 00000
00000 00000
T.R. wilbury Study Humber 88S-TH
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Figure 1. survival of fathead minnows, Pimephalea pzomelaa, exposed to N2803-2 for 96 hours.
T.R. wilbury Study Number 888-TH
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Table 4.
Median lathal concentrationa (LC50a) from the toxicity taat with fathead minnova, Plmuphalaa pcomolaa, and N2803-2.
I*C50
95 percent confidanca limit
Calculation method
Binomial
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ASTM. 1986. standard Practice for Conducting Acuta Toxicity Taata with Fishes, Kacroinvertebrates, and Amphibian*. E-729-80. XU Annual Book of Standard*.
Stephan, C.E. 1983. Computer Program for tha Calculation of LC50 Valuaa. U.s. EPA. Duluth, m n . Personal communication.
U.S. EPA. 1993. Toxic Substances Control Act Test Guidelines; Final Rules. Section 797.1400.
U.S. EPA. 1993. 40 CFR Part 792. Toxic Substances Control Act (TSCA); Good Laboratory Practice standards. Code of Federal Regulations, Title 40, July 1, 1993.
T.R. Milbury study Number 888-th
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_____
XIX. PROTOCOL AMnnJWjrt AMD deviations
PROTOCOL AMENDMENTS!
The protocol was amended to pacify the nominal concentrations of test substance for the definitive teat and the eource of the test organisms.
PROTOCOL DEVIATIONS t The dissolved oxygen concentrations were not always maintained above
60 saturation (aeration was initiated as soon as dissolved oxygen concentrations below 60% saturation wore observed). This deviation did not affect the outcome of the test and no other deviations were made from the protocol.
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Appendix A. WATER QUALITY DATA FROM TOXICITZ TEST
T.R. wilbury study Nunber 888-th
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Tab! A.l. Dissolved oxygan concentration, conductivity, pH, and
temperature meaeured during the toxicity teat with fathead # minnowa, Piaephalea promeiaa, and N2803-2. *
Nominal
Concentration
of N2803-2
(mg/L)
Rep.
Diaaolved oxygen (mg/L)
--------------------------
0 24 48 .,,73 h r . 96 hr hr hr b a hr
Conductivity (pmhoa/cm) ----------------------
0 24 48 72 96 hr hr hr hr hr
0 (control) 160 250 400 630
1,000
i 2
1 2
1 2
1 2
1 2
1 2
8.4 7.1 6.6 6.3 7.1 8.6 8.4 7.2 6.7 6.3 7.2 8.6
8.4 7.0 6.6 6.2 7.0 8.4 8.4 6.9 6.6 6.3 7.2 8.7
8.4 6.9 6.5 4.9 6.8 8.8 8.4 6.9 6.4 5.8 7.0 8.5
8.4 6.7 6.2 5.2 6.6 8.3 8.4 6.8 6.2 5.3 6.9 8.6
8.4 6.6 5.8 4.6 6.8 8.6 8.4 6.6 5.8 4.7 7.1 7.8
8.4 6.8 5.6 4.8 6.7 8.0 8.4 6.7 5.5 4.6 6.9 8.4
150 140 150 150 150 150 150 150 150 150
150 140 150 150 150 150 140 150 150 150
150 150 150 150 150 150 150 150 150 150
150 150 150 150 160 150 150 150 150 ISO
160 150 150 150 160 160 160 160 150 170
170 170 170 170 180 170 170 170 170 180
Note:
All test vessels were aerated following the 72 hour dissolved oxygen measurements. Measurements made at 72 hours before aeration are noted above in the column headed "b" and the measurements made after aeration are noted in the column headed "a".
T.R. wilbury study Number 888-TH
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Tabla A.l. continuad.
Nominal
PB
Concantratlon
of N2803-2
0 24 48 72 96
(mg/L)
Rap. hr hr hr hr hr
Temperatura (C)
0 24 48 72 96 hr hr hr hr hr
0 (control) 1 2
160 1 2
250 1 2
400 1 2
630 1 2
1,000
1 2
7.5 7.4 7.6 7.4 7.6 7.5 7.4 7.6 7.4 7.7
7.5 7.4 7.6 7.4 7.7 7.5 7.5 7.6 7.5 7.7
7.5 7.5 7.6 7.5 7.8 7.5 7.5 7.6 7.5 7.7
7.5 7.5 7.6 7.5 7.7 7.5 7.4 7.6 7.5 7.8
7.5 7.5 7.6 7.5 7.7 7.5 7.5 7.6 7.5 7.8
7.5 7.5 7.6 7.5 7.7 7.5 7.5 7.6 7.5 7.7
21.9 21.9 21.8 21.6 21.6 21.9 21.9 21.9 21.6 21.9
21.9 21.9 21.8 21.6 21.9 21.9 21.9 21.8 21.7 21.8
21.9 21.9 21.8 21.7 21.8 21.9 21.9 21.8 21.7 21.7
21.9 21.9 21.8 21.6 21.9 21.9 21.8 21.8 21.7 21.9
22.0 21.9 21.8 21.7 21.9 21.9 21.9 21.8 21.6 21.9
21.8 21.8 21.8 21.6 21.6 21.9 21.9 21.8 21.6 21.8
T.R. wilbury Study Nimber 888-TH
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04/ 01/2001
ENVIRONMENTAL LABORATORY ARCHIVED FINAL REPORT
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LAB REQUEST NO. N2803 REQUESTOR NAME: SAB/GOEBEL/PICKETT DEPARTMENT: 3048 PROJECT NO: BI703004UQ DATE RECEIVED: 08/01/1995 DESC: FC-143 AND RELATED PRODUCTS ECOTOX WORK
CONTRACT LAB(S): TRWIL EXP COMP DATE: 10/04/1995 DATE COMPLETED: 03/27/1996 PROJECT LEAD: PHONE NO: 3M FAX NO: 651-778-6176
Samples received from S.J. Martin.
Samples 1-5 sent to TRWIL on 8/11/95. An additional 4L of sample 5 sent on 8/22/95.
Sample 1 = Tetrabutyl ammonium hydroxide, 40% in water. Sample 2 = L-13492 (45% solids, 27% water, 27% IPA) Sample 3 = FC-26, 100% solids. Dissolve in 50:50 water:IPA solution. Sample 4 = FC-143, 100% solids. Sample 5 = L-13856, 15% solids. Sample 6 = L -13856, diluted by a factor of 15, tested as an effluent.
Sample #1 (Tetrabutylammonium hydroxide) - a new bottle was ordered from Aldrich because TR Wilbury needed more sample. This was lot 05922BN, red label, cat. # 17,878,0. sab 9/11/95
NOTE: All data for sample 3 (FC-26) is reported in terms of FC-26, and not
as a 50/50 mixture with isopropanol.
sab 11/21/95
NOTE: For sample #5, only a range-finding test was run for toxicity to algae. The requestor canceled the request for testing so no definitive test was run. The results for the range finder are as follows:
Test Cone.
Percent Difference in Avg. Specific Growth Rate
mg/1
96-hrs Expsure
0.1 2 1. 0 18
10 4
100 32
1000
100
EC50 (95% C.I.)
160 (100 - 1000) mg/L.
sab 2/16/96
For N2803-6, the testing was canceled before a definitive study with fish could be completed. Results reflect testing with a single concentration of 13% v/v, which showed no mortality after 96-hours, sab 3/14/96
SAMPLE DATE
CODE
DESCRIPTION RESULT
1 08/11/95 TB NH40H
Lot 10317KQ
TRWIL 48-HR-EC50 *DAPHNIA ACUTE TEST
27 MG/L
MDL or (95% C.I .) (23 - 32)
* = CONTRACT LAB DATA
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04/01/2001
LAB REQUEST NO. N2803
SAMPLE DATE
CODE
ENVIRONMENTAL LABORATORY ARCHIVED FINAL REPORT
CONTINUED
DESCRIPTION RESULT
2 MDL or (95% C.I.)
1 08/11/95 TB NH40H
Lot 10317KQ
TRWIL 96-HR-LC50 *FISH ACUTE TEST - 96 HOUR 720 MG/L
TRWIL IC50-ALGAE *ALGAE ASSAY - IC50
3.7 MG/L
(600 - 1000) (1.8 - 7.7)
2
L-13492
Lot 2
TRWIL 48-HR-EC50 *DAPHNIA ACUTE TEST TRWIL 96-HR-LC50 *FISH ACUTE TEST - 96 HOUR TRWIL IC50-ALGAE *ALGAE ASSAY - IC50
62 MG/L 960 MG/L 8.4 MG/L
(36-100) (830->1000) (5.9 - 14)
3
FC-26
Lot 135
TRWIL 48-HR-EC50 *DAPHNIA ACUTE TEST TRWIL 96-HR-LC50 *FISH ACUTE TEST - 96 HOUR TRWIL IC50-ALGAE *ALGAE ASSAY - IC50
360 MG/L 140 MG/L 90 MG/L
(300 - 500) (110 - 180) (63 - 130)
4
FC-143
Lot 427
TRWIL 48-HR-EC50 *DAPHNIA ACUTE TEST TRWIL 96-HR-LC50 *FISH ACUTE TEST - 96 HOUR TRWIL IC50-ALGAE *ALGAE ASSAY - IC50
720 MG/L 740 MG/L >1000 MG/L
5 08/16/95 L-13856
As-is
(600 - 1000) (660 - 830) (-)
TRWIL 48-HR-EC50 *DAPHNIA ACUTE TEST TRWIL 96-HR-LC50 *FISH ACUTE TEST - 96 HOUR TRWIL IC50-ALGAE *ALGAE ASSAY - IC50
190 MG/L
(130-600)
>1000 MG/L
(-)
SEE COMMENT ABOVE
6
L -13856
1 :15 diluted
TRWIL 48-HR-EC50 *DAPHNIA ACUTE TEST TRWIL 96-HR-LC50 *FISH ACUTE TEST - 96 HOUR TRWIL IC50-ALGAE *ALGAE ASSAY - IC50
1.2 % V/V
(1.0 - 1.7)
>13 % V/V
(-)
TESTING CANCELED
* = CONTRACT LAB DATA
