Document wrRKEdaM96Mw22BrdY2BRgxnQ

BACK TO MAIN Study Title Acuta Toxicity oi! N2 803-2 to tha Daphnid, Daphnla magna Data Requiresienti Guidalina Number U.s. EPA-TSCA, Guidalina 797.1300 Authors Timothy J. Hard Peter 1. Kowalski Robert L. Boeri Study Completed November 16, 1995 Report Amended March 25, 1996 Sponsor 3M Company Environmental Laboratory 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55144 Zesting Facility T.R. Wilbury Laboratories, Inc. 40 Doaks Lane Marblehead, Massachusetts 01945 T.R. wilbury' study Number 889-TH rr.j Page 1 of 22 BACK TO MAIN BACK TO MAIN Amended 3/25/96 II. QUALITY ASSURANCE STATEMENT Submitted byi T.R. Wilbury Laboratories, Inc. 40 Doaks Lana Marblehaad, Maasachuaatts Certification! T.R. Wilbury Laboratories, Inc. study number 889-TB, Acuta Toxicity of N2803-2 to tha Daphnid, Daphnia magna, was audited by the Quality Assurance unit for compliance with tha protocol, standard operating procedures, and applicable 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: Pinal Report: Amended Report: Audit Date 8/ 3/95 9/ 5/95 9/25/95 11/16/95 3/25/96 Reported to study Director 8/ 4/95 9/ 5/95 9/25/95 11/16/95 3/25/96 Reported to Management 8/ 4/95 11/16/95 11/16/95 11/16/95 3/25/96 VlilMlilllffMHBilllliPlIliROT BACK TO MAIN III. T A B U OP COHTKMTfl SECTIONI I. Good Laboratory Practica Compliance statement II. Quality Assurance Statement III. Tabla of contents IV. index of Tables and figuras v. summary VI. General information VII. Introduction VIII. Methods and Materials IX. Results X. References XI. Signature Page XII. Protocol Amendments and Deviations Appendix A. Hater Quality Data from Toxicity Test pAOE 2 3 4 5 g 7 8 8 13 17 18 19 20 T.R. wilbury study Number 8B9-TH Page 4 of 22 BACK TO MAIN BACK TO MAIN The acuta toxicity of N2803-2 to tho daphnid, Dtphnla aagna, is daacribad in this final raport. Tba taat waa conducted for 3M Company for 48 houra from September 5 to 7, 1995, at T.R. Hilbury Laboratories, Inc. in Marblehead, Maaaachuaetta. It waa conducted by Peter Kovalki, Jeanne Magazu, Jacqueline Neviua, Keven Steven, Peter Heviua, Robert Boeri, and Timothy Hard according to the protocol developed for T.R. Hilbury study Humber 889-T8. K2803-2 waa supplied by the sponsor. The teat waa performed under static conditions with five concentrations of test substance and a dilution water control at a temperature of 19.3-20.4*C. The dilution water was filtered deionized water collected at Marblehead, Massachusetts and adjusted to a hardness of 176 mg/L as caco,. Aeration was not required to maintain dissolved oxygen concentrations above an acceptable level. Nominal concentrations of N2803-2 were 0 (control), 13, 22, 36, 60, and 100 mg/L. Daphnid used in the test were less than 24 hours old at the start of the test. They were produced at T.R. Hilbury Laboratories and acclimated under test conditions for their entire life (cultures that produced test specimens were maintained at test conditions for more than 7 days). After 48 hours of exposure the control daphnids had an average wet weight (blotted) of 0.26 mg. All daphnids were in good condition at the beginning of the etudy. Exposure of daphnids to the test substance resulted in a 24 hour median lethal concentration (LC50) >100 mg/L N2803-2, a 48 hour LC50 of 93 mg/L H2803-2 (95 confidence interval 74 to >100 mg/L), a 24 hour median effective concentration (EC50; based on immobilization) of 72 mg/L (95% confidence interval 63 to 83 mg/L), and a 48 hour EC50 of 62 mg/L H2803-2 (95* confidence interval 36 to 100 mg/L). The 48 hour no observed effect concentration (NOEC) .is 13 mg/L. f BACK TO MAIN BACK TO MAIN VII. INTRODUCTION This atudy was sponsored by 3H Company St Paul, Minnesota. Tha objactlva of tha atudy was to datarmlna tha 24 and 48 hour median lathal concantrationa (LCSOa) and median affactlva concantrationa (EC50a) of tha taat aubatanca to tha daphnid, Daphala magna, a freshwater invartabrata, undar atatlo condltlona Tha taat protocol waa baaad on U.s. EPA methods (U.S. EPA, 1993). Tha raport containa aactlona that describe tha mathoda and materials employed in tha atudy, and raaulta of tha investigation. Tha report alao containa an appendix that praaanta water quality data collected during tha taat. VIII. METBODS AND MATERIALS TEST SUBSTANCE) The aample of N2803-2 (T.R. Hilbury Laboratoriea Sample Number 520) waa delivered to T.R. Hilbury Laboratoriea on Auguat 14, 1995 in a cardboard box at ambient temperature. It waa contained in an approximately 50 ml glaaa jar. The label attached to the bottle included the following information) "N2803-2". 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. 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 HATER) Hater used for acclimation of test organisms and for all toxicity testing was deionized water collected at T.R. Hilbury Laboratories in Marblehead, Massachusetts. Hater was adjusted to a hardness of 160 180 mg/L as CaCO, 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 ORGANISM) Juvenile daphnids, Daphala magna, employed as test organisms were less than 24 hours old at the start of the test. They were from a single aource and were identified using an appropriate taxonomic key. Test specimens were produced by adult daphnids that were maintained under test conditions at the T.R. Hilbury facility for more than 7 days. f j j j j } i i 1 j T.R. Hilbury Study Number 889-t h Page 8 of 22 BACK TO MAIN Tabla 1. chemical characterization of a raprazantativa sample of dilution water uaad for tha toxicity teat with daphnida, Daphnia magna, 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 ND1 HD ND HD HD ND HD 0.03 ND ND ND ND ND Nitrate Chloride Fluoride Total Organic Carbon Total Phosphorus mg/L as N mg/L mg/L mg/L mg/L 0.05 1 0.1 1 0.03 ND ND ND ND Organochlorlne Pesticides Toxaphene pg/L pg/L 0.5 2 ND ND Organophosphorus Pesticides Dimethoate TEPP Monocrotophos PCBS ' y'1<' pg/L pg/L pg/L pg/i* pg/L 0.5 2.0 2.0 2.0 0.5 ND ND ND ND ND Notasi 1. Metals, nitrate, chloride, fluoride, total organic carbon, total phosphorus, organochlorlne pesticides, organophosphorus pesticides, and PCBs were measured in deionized water (used to formulate dilution water) that was collected during August, ; 1995 and analyzed by Pace, Inc. as part of routine water quality tasting. 2. HO not detected at or above the detection limit. T.R. Wilbury Study Number 889-TH page 9 of 22 BACK TO MAIN The original cultura was obtained on Juna 1, 1994 from Aquatio Research organisms, Hampton, Haw Hampshire. Measurements made during tha 7 daya before tha atart of tha tast indicatad that tha cultura temperature rangad from 19.6 to 20.B*C and tha diaaolvad oxygan concantration was alwaya at laaat 6.0 mg/L. Tha cultura produced young prior to day 12 and a aubaampla of adulta producad at laaat 3 young/daphnid/day for 7 daya bafora tha atart of tha taat. During acclimation daphnida vara not traatad for diaeaaa, thay vara fraa of apparant aicknaaa, injuria, and abnormalitiaa at tha baginning of tha taat and thara waa no mortality during tha 48 boura pracading tha atart of tha taat. Tha cultura vaa auppliad with a yaaat/trout chow mix (Lott DC02) and tha fraahwatar alga, Salenaatrun caprlcornutua, daily during acclimation. Daphnida wara not fad during tha teat. Control daphnida, blottad and weighed at tha concluaion of the toxicity taat, averaged 0.26 mg and tha loading rate during the definitive toxicity test waa approximately 10.4 mg/L. TOXICITY TESTING s A static screening teat was conducted from August 23 to 25, 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 48 hours of exposure there was 100% survival in the control and at 0.10, 1.0, 10, and 0% survival at 100 and 1,000 mg/L. The definitive toxicity test waa performed from September 5 to 7, 1995, according to T.R. Wilbury Protocol 889-TH (Acute Toxicity of N2803-2 To The Daphnid, Daphnla magna). Tho protocol, which waa signed by the study director on August 16, 1995, is based on procedures of the U.s. Environmental protection Agency (1993). A summitry of the test system is presented in Table 2. The test was conducted at a target temperature of 20 1*C with five concentrations of test substance and a dilution water control. Nominal concentrations of the test substance were: 0 mg/L (control), 13, 22, 36, 60, and 100 mg/L. A 200 mg/L stock solution was formulated by combining 0.200 g test substance and dilution water in a Class A volumetric flask and adjusting the final stock solution volume to 1,000 ml. Appropriate amounts of this stock solution were added to dilution water in teat vessels to forsnilate test media without the use of a solvent. Twenty daphnida were indiscriminately and equally distributed among two replicates of each treatment. The test was performed in 300 ml glass beakers that contained 250 ml of test solution (water depth was approximately 9 cm). Test vessels rare randomly arranged during the 48 hour test (a random numbers table was used to select the location of each vessel) and they were loosely covered. T.R. wilbury Study Number 889-TH Page 10 of 22 BACK TO MAIN Table 2. Taat system summary for the toxiolty taat with N2803-2 and daphnids, Daphaia magna. species: Daphnla magna Age/Size of Taat Organlamai <24 hours old Taat Duration! 48 hoars Taat Vassals i 300 ml glass beakers Taat Madia Volumai 250 ml Depth of Test Media: 9 cm Number of Replicate Test Vessels! Organisms per Replicate: 2 10 Temperature Range! 19.3-20.4'C Minimum Dissolved oxygen: Photoperiod: 8.6 mg/L 16 hours light and 8 hours dark. Light Intensity: 110 footcandles Hater Quality Measurements: Dissolved oxygen, pH, temperature, and conductivity daily in each test vessel. Hardness, total organic carbon, and alkalinity once in the dilution water. Residual chlorine daily. Dilution water analysis is summarized in Tabic 1. End Points: 24 and 48 hour LC50 and EC50 (based on immobilization), 48 hour no observed effect level. Acceptability Criteria: slot control mortality and acceptable temperature range. , T.R. wilbury study Number 8B9-TH Page 11 of 22 BACK TO MAIN BACK TO XX. RESULTS Insoluble material was not noted during the teat. Biological and water quality data generated by the acute toxicity test are presented in Table 3 and Appendix A, respectively. The measured concentrations of dissolved oxygen ranged from 8 . 6 to 9.0 ng/L, the temperature ranged from 19.3 to 20.4*C, conductivity ranged from 670 to 720 pmhos/cm, and pa ranged from 8.3 to 8 .6 . one hundred percent survival occurred in the control exposure and these daphnids did not exhibit any sublethal effects. The 24 and 48 hour LC50s and EC50s for daphnids exposed to N2803-2 are presented in Table 4. The 48 hour LC50 for daphnids exposed to N28032 is 93 mg/x> (95% confidence interval 74 to >100 mg/L) and the 48 hour EC50 is 62 mg/X. (95% confidence interval 36 to 100 mg/L). The slope of the 48 hour dose response curve (based on mortality and immobilization) is 6.1. The 48 hour no observed effect concentration (NOEC) is 13 mg/L. T.X. Wilbury study Number 889-TH Page 13 of 22 BACK TO MAIN Table 3. survival and aublathal effect data iron tha toxicity taat with dapbnida, Dtphnla aagna, and M2803-2. Nominal Concentration of N2803-2 (mg/L) 0 (control) 13 22 36 60 100 Rep. Number Alive 0 24 48 hr hr hr 1 10 10 10 2 10 10 10 1 10 10 10 2 10 10 10 1 10 10 9 2 10 10 10 1 10 10 10 2 10 10 9 1 10 10 2 10 9 9 9 1 10 8 4 2 10 9 3 Number Affected 0 24 48 hr hr hr 000 000 000 000 003 004 009 009 0 2 9b 0 3 9* 0 6 4b 0 8 3* Note: Affected daphnids were immobilized at 24 hours and less active than control daphnids at 48 hours. 3 immobilized daphnids b 4 immobilized daphnids T.R. Wilbury study Number 889-TH Page 14 of 22 Amended 3/25/96 BACK TO MAIN Figure 1. survival and subletbal effects of daphnids, Daphnla jnagna, exposed to N2803-2 for 48 hours. T.R. tfilbury study Number 889-TH Page 15 of 22 BACK TO MAIN BACK TO MAIN Z. RXrZHZMCZS ASTH. 1986. Standard Practice for Conducting Acute Toxicity Teats with rishea, Macroinvertebrates, and Amphibians, e -729-80. Jfl Annual Book of Standards. Stephan, C.E. 1983. Computer Program for the Calculation of LCSO Values. U.S. EPA. Duluth, MN. Personal Communication. U.S. EPA. 1993. Toxic Substances Control Act Test Guidelines; Pinal Rules, section 797.1300. I BACK TO MAIN Amended 3/2S/96 XI. SIOMAXURZ PACK "Otff r L . Kowalski Study Director Aquatic Toxicologist 3 )<;*<. 0-p.m frfofou M b Jeanne P. Magaiu Aquatic Toxicologist 'fne,uu.t*e Jaceqgueeline M. Nevius Biologist 7? jcC fo4y:? Peter C. Nevius Biologist 2 05<l ^ C r v f/1. 3 g > ^ Keven M. Stevens Biologist 5 /^r/?G B. wilbury study Number 089-th Page 18 of 22 BACK TO MAIN ZXZ. PROTOCOL AMENDMENTS AND DEVIATIONS PROTOCOL AMENDMENTS t The protocol was amended to specify the nominal concentrations for the definitive test. No other changes were made to the protocol. PROTOCOL DEVIATIONS! No deviations were made from the protocol. T.R. wilbury study Number 889-TH Page 19 of 22 BACK TO MAIN BACK TO MAIN Table A.l. Dissolved oxygen concentration, conductivity, pH, and temperature neasured during the toxicity ta*t with daphnids, Daphnla magrut, and N2803-2. Nominal Dissolved oxygen (mg/L) Conductivity (pmhos/cm) c o n c e n t r a t i o n ----------------------- --- -------------- .------ - of N2803-2 0 24 48 0 24 48 (mg/L) Rap. hr hr hr hr hr hr 0 (control) 1 8.7 9.0 8.6 2 8.7 9.0 8.6 13 1 8.7 9.0 8 .8 2 8.7 9.0 8.8 22 1 8.7 9.0 8.8 2 8.7 9.0 8.8 36 1 8.7 9.0 8.8 2 8.7 9.0 8.8 60 1 8.7 9.0 8.8 2 8.7 9.0 8.8 100 1 8.7 9.0 8.7 2 8.7 9.0 8.7 670 670 690 670 670 690 670 670 710 670 670 710 670 670 710 670 670 710 670 670 710 670 670 710 670 670 710 670 670 720 670 670 720 670 670 720 T.R, Wilbury study Number 889-t h Page 21 of 22 BACK TO MAIN Tabla A.l. Continuad. Nominai Concantration of N2803-2 (mg/L) 0 (control) 13 22 30 60 100 Rap. pii ----------- 0 24 hr hr -- --. 48 hr Tamparatura (*C) .-- -- ... 0 24 48 hr hr hr 1 8.5 8.5 8.3 2 8.5 8.5 8.4 1 8.5 8.5 8.4 2 8.6 8.5 8.4 1 8.6 8.5 8.4 2 8.6 8.5 8.4 1 8.6 8.5 8.4 2 8.6 8.5 8.4 1 8.6 8.5 8.4 2 8.6 8.5 8.4 1 8.6 8.5 8.4 2 8.6 8.5 8.4 20.2 20.2 20.2 20.2 20.3 20.3 20.3 20.3 20.4 20.4 20.4 20.4 19.4 19.5 19.6 19.5 19.4 19.5 19.5 19.6 19.6 19.4 19.3 19.5 19.3 19.4 19.4 19.3 19.4 19.4 19.4 19.5 19.4 19.3 19.3 19.3 T.R. Hilbury Study Number 889-TH Paga 22 of 22 03/29/2001 ENVIRONMENTAL LABORATORY ARCHIVED FINAL REPORT BACK TO MAIN 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 11000 4 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) k = CONTRACT LAB DATA BACK TO MAIN 03/29/2001 LAB REQUEST NO. N2803 SAMPLE DATE CODE ENVIRONMENTAL LABORATORY ARCHIVED FINAL REPORT CONTINUED DESCRIPTION RESULT 2 M D L o r (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 * = CONTRACT LAB DATA 1.2 % V/V (1.0 - 1.7) >13 % V/V (-) TESTING CANCELED