1、 API TR*402 95 U 0732270 0554648 O96 II II American Petroleum Institute Health and Environmental Sciences Department Tertiary-Amyl Methyl Ether (TAME) Toxicity to the Freshwater Alga, Selenas trurn capricornu turn FEBRUARY 1995 TOXICOLOGY REPORT NUMBER 402 CAIS ABSTRACT NO. 41 -5415 API TR*402 95 I
2、0732290 0554649 T22 I Enwmwuntaf Partnership One of the most significant long-term trends affecting the future vitality of the petroleum industry is the publics concerns about the environment. Recognizing this trend, API member companies have developed a positive, forward-looking strategy called STE
3、P: Strategies for Todays Environmental Partnership. This program aims to address public concerns by improving our industrys environmental, health and safety performance; documenting performance improvements; and communicating them to the public. The foundation of STEP is the API Environmental Missio
4、n and Guiding Environmental Principles. API ENVIRONMENTAL MISSION AND GUIDING ENVIRONMENTAL PRINCIPLES The members of the American Petroleum Institute are dedicated to continuous efforts to improve the compatibility of our operations with the environment while economically developing energy resource
5、s and supplying high quality products and services to consumers. The members recognize the importance of efficiently meeting societys needs and our responsibility to work with the public, the government, and others to develop and to use natural resources in an environmentally sound manner while prot
6、ecting the health and safety of our employees and the public. To meet these responsibilities, API members pledge to manage our businesses according to these principles: O To recognize and to respond to community concerns about our raw materials, products and operations. O To operate our plants and f
7、acilities, and to handle our raw materials and products in a manner that protects the environment, and the safety and health of our employees and the public. O To make safety, health and environmental considerations a priority in our planning, and our development of new products and processes. O To
8、advise promptly, appropriate officials, employees, customers and the public of information on significant industry-related safety, health and environmental hazards, and to recommend protective measures. 9 To counsel customers, transporters and others in the safe use, transportation and disposal of o
9、ur raw materials, products and waste materials. O To economically develop and produce natural resources and to conserve those resources by using energy efficiently. O To extend knowledge by conducting or supporting research on the safety, health and environmental effects of our raw materials, produc
10、ts, processes and waste materials. 9 To commit to reduce overall emission and waste generation. 9 To work with others to resolve problems created by handling and disposal of hazardous substances from our operations. O To participate with government and others in creating responsible laws, regulation
11、s and standards to safeguard the community, workplace and environment. + To promote these principles and practices by sharing experiences and offering assistance to others who produce, handle, use, transport or dispose of similar raw materials, petroleum products and wastes. API TRx402 95 0732290 05
12、54650 744 M FOREWORD API PUBLICATIONS NECESSARILY ADDRESS PROBLEMS OF A GENERAL NATURE. WITH RESPECT TO PARTICULAR CIRCUMSTANCES, LOCAL, STATE, AND FEDERAL LAWS AND REGULATIONS SHOULD BE REVIEWED. API IS NOT UNDERTAKING TO MEET THE DUTIES OF EMPLOYERS, MANUFACTURERS, OR SUPPLIERS TO WARN AND PROPERL
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15、554651 680 m ACKNOWLEDGMENTS THE FOLLOWING PEOPLE ARE RECOGNIZED FOR THEIR CONTRIBU- TIONS OF TIME AND EXPERTISE DURING THIS STUDY AND IN THE PREPARATION OF THIS REPORT: API STAFF CONT ACT Richard Rhoden, Ph.D., Health and Environmental Sciences Department MEMB ERS OF THE A IRAJE LS TOXICOLOGY HEALT
16、HEFFEC TS WORKGROUP Russell D. White, Ph.D., Chuimn, Chevron Research i qmai YS %- rt:-zi use o+ the Springborr Laboratories Ini name IS permitlea excepl as eapressly autnorizeb in writ 19 Leners am repons apc y o-, 15 the SDCISC mite. a b p3cL?s 3 woieseo FCP- examined or surveyed ana are no1 neces
17、sarily indicative of the quaiil!es 01 8pparewy iaenlica or Slmi:ar maie*ias OCJCIS or D-OCSFS Tne 5t)i ,I, o QEnpt)- ;ab:.a!: CT IC: w#lh respect to se*mes rendered shall be iirnited IO the amom1 of the consloera1,on paio for sbcn sewces ano no1 inctde z-! coTiseaLell az-aqts API TRm402 95 = 0732290
18、 0559654 3T = Appendix il - The Certificate of Analysis for Lot No. 07905KZ was removed as this lot was not used during testing. Approval Signatures: r/z/91/ 4 James R. Hoberg fl Study Director - Supervisor, Quality Assurance Unit Page 1 of 66 TERT-AMYL METHYL ETHER (TAME) - TOXICITY TO THE FRESHWAT
19、ER ALGA, Selenastrum capricornutum GUIDEUNE REFERENCE NUMBER: 797-1 050 Submitted to: American Petroleum Institute 1220 L Street, Northwest Washington, D.C. 20005 SLI Report #93-11-5065 SU Study #12827.0692.6106.430 Study Director: James R. Hoberg Springborn Laboratories, Inc. Environmental Sciences
20、 Division 790 Main Street Wareham, Massachusetts 02571 12 August 1994 AMENDED FINAL REPORT API TRt402 95 0732290 0554b56 Lb2 = Report No. 93-1 1-5065 Page 2 of 66 GOOD LABORATORY PRACTICES COMPLIANCE STATEMENT The data and report prepared for “Ted-Amyl Methyl Ether (TAME) - Toxicity To The Freshwate
21、r Alga, Selenasfrum capricornutum“ were produced and compiled in accordance with all pertinent U.S. EPA Good Laboratory Practice Regulations (40 CFR, Part 792) with the following exceptions: routine water screening analyses for pesticides, PCBs and metals are conducted using standard U.S. EPA proced
22、ures by Lancaster Laboratories, Lancaster, PA. These data were not collected in accordance with Good Laboratory Practice procedures (.e. no distinct protocol, Study Director, etc.). Stability, characterization and verification of the test material identity and maintenance of these records on the tes
23、t material are the responsibility of the Study Sponsor. At the termination of the testing program, all remaining test material will be sent to the Study Sponsor. Archival of a sample of the test material is the responsibility of the Study Sponsor. SPRINGBORN IABORATORIES, INC. games R. Hoberg f Dte
24、Study Director Springborn Laboratories, Inc. API TR8402 95 0732290 O554657 OT Report No . 93-1 1-5065 Page 3 of 66 TABLE OF CONTENTS PAGE GOOD LABORATORY PRACTICES COMPUANCE STATEMENT . 2 UST OF TABLES . . . . 5 USTOFFIGURES . 6 SUMMARY 7 1.0 INTRODUCTION . 9 2.0 MATERIALS AND METHODS . 9 2.1 Protoc
25、ol . 9 2.2TestMaterial 9 2.3TestOrganism . 10 2.4 Test Dilution Water 11 2.5 Test Concentrations . 11 2.6 Preparation of Test Solutions 11 3.0 TEST PROCEDURES 12 3.1TestInitiation 12 3.2 Test Monitoring 12 3.2.1 Algal Growth 12 3.2.2 Recovery 13 3.2.3 Test Conditions . 13 3.2.4 Chemical Analysis 14
26、4.0 STATISTICAL ANALYSIS . 14 5.0 RESULTS AND DISCUSSION 15 5.1 Preliminary Testing 15 5.2 Definitive Testing . 17 PROTOCOL DEVIATIONS 20 QUAUTY ASSURANCE UNIT STATEMENT 21 REFERENCES 22 TABLES . 23 FIGURE 29 Springborn Laboratories. Inc . Report No . 93-1 1-5065 Paqe 4 of 66 SIGNATURES .AND APPROVA
27、L . 31 6.0 APPENDIX I . STUDY PROTOCOL 32 7.0 APPENDIX II . CERTIFICATE OF ANALYSIS . 43 8.0 APPENDIX 111 . DILUTION WATER ANALYSIS . 45 9.0 APPENDIX IV . ANALYTICAL METHODOLOGY 48 10.0 APPENDIX V . EXCERPTED COPIES OF RAW DATA . 62 Springborn Laboratories. Inc . API TR*402 95 m 0732290 0554659 971
28、m Report No. 93-1 1-5065 Page 5 of 66 LIST OF TABLES PAGE Table 1. Composition of algal growth medium (AAP medium) used during this study 24 Table 2. Conductivity, pH, temperature and light intensity measured during the 96-hour exposure of Selenastrum capricornufum to TAME. . . . . . . . . . . . . .
29、 25 Table 3. Concentrations of TAME measured in the exposure solutions during the 96-hour toxicity test with Selenastrum capricornufum , . , . . . . . . . . . . . 26 Table 4. Cell densities (x lo4 cellslml) of Selenastrum capricornufum after 24, 48,72 and 96 hours of exposure to TAME. , . . . . . .
30、. . . . . . . . . . . . . . . . . . 27 Table 5. EClO, ECSO and EC90 values for TAME calculated from results (cell density) of the 96-hOur toxicity test with Selenastrum capricornutum. . . . . 28 Table 1A. Analytical results for the recovery of TAME from AAP media . . . . . . . . . , . 55 Table 2A. A
31、nalytical results for the recovery of TAME from filtered seawater . . . . . , . 56 Table 3A. Analytical results for the recovery of TAME from freshwater (reconstituted to increase hardness) . . . . . . . . . . . . . . . . . . , . , . . . . . . . . . 57 Table 4A. Repeatability of TAME analysis from A
32、STM Type II water at 0.026 mg/L 58 Springborn Laboratories, Inc. Report No. 93-1 1-5065 Page 6 of 66 Figure 1. LIST OF FIGURES PAGE Graphical illustration of the relationship between cell densities and time during the 96-hour toxicity test exposing Selenastrum capricornutum to TAME. . 30 Figure 1A.
33、A representative chromatogram of TAME purge and trap GC/FID analysis . 59 Figure 2A. A representative linear regression analysis from standard TAME analysis . 60 Figure 3A. A representative polynomial regression analysis from standard TAME analysis. 61 Springborn Laboratories, Inc. API TR1402 95 073
34、2270 0554663 52T Report No. 93-1 1-5065 Page 7 of 66 SUMMARY Teh-Amyl Methyl Ether FAME) - Toxicity to the Freshwater Alga, Selenastrum capricornutum SPONSOR: PROTOCOL TITLE: REPORT NUMBER: STUDY NUMBER: TEST MATERIAL: DATE RECEIVED: DESCRIPTIO N : EXPERIMENTAL START DATE: EXPERIMENTAL TERM IN AT1 O
35、N DATE: TEST ORGANISM: DILUTION WATER: TEST CONDITIONS: NOMINAL TEST CONCENTRATIONS: American Petroleum Institute 1220 L Street, Northwest Washington, DC 20005 “Protocol for Conducting a 96-Hour Acute Toxicity Test with the Alga, Selenasfrum capricornufum, Following TSCA 797- 1 050,“ Springborn Labo
36、ratories Protocol #:O91 1 92/TSCA 797.1050 and Protocol Amendment #1 dated 18 November 1993. This protocol conforms to U.S. EPA TSCA Guideline S797-1050 (U.S. EPA, 1985, 1987) 93-1 1-5065 12827.0692.61 06.430 Tert-Amyl Methyl Ether (TAME) Lot #O281482 17 August 1993 A clear liquid, 98.8% active ingr
37、edient (Certificate of Analysis, Appendix Il). 18 November 1993 22 November 1993 Selenastrum capricornufum, inoculum - three days since previous transfer, source - Springborn culture Algal Assay Procedure (AAP) medium 96-hour duration, 25 OC, continuous illumination at 3200 to 4800 lux (300 to 450 f
38、ootcandles), shaking rate of 100 rpm 0.040, 0.080, 0.16, 0.31, 0.63, 1.3, 2.5 and 5.0 mg A.I./L Springborn Laboratories, Inc. API TR*402 95 0732290 0554662 466 Report No. 93-1 1-5065 Pawe 8 of 66 MEAN MEASURED CONCENTRATIONS: 0.017, 0.037, 0.067, 0.23, 0.48, 0.52, 1.4 and 3.7 mg A.I./L EFFECT CRITER
39、ION: Inhibition of cell growth (culture density relative to the control) RESULTS: The 96-hour EC50 value was calculated to be 0.11 mg A.I./L (95% confidence limits of 0.018 and 0.64 mg A.I./L). The 96- hour No-Observed-Effect Concentration (NOEC) was determined to be 0.01 7 mg A.I./L. Results from t
40、he supplemental exposure conducted at test termination demonstrated that TAME, at a concentration of 3.7 mg A.i./L has an algistatic (reversible) rather than algicidal (non-reversible) effect on the growth of Selenastrum capricornutun? once it is diluted to a non-inhibitory concentration (0.040 mg A
41、.I./L). Springborn Laboratories, Inc. _ _ API TRx402 95 0732290 0554663 3T2 Report No. 93-1 1-5065 Page 9 of 66 1 .O INTRODUCTION The objective of this study was to determine the effect of Tert-Amyl Methyl Ether (TAME) on the growth of the freshwater alga Selenastrum capricornutum. The exposure syst
42、em was specifically modified to minimize the loss of test material due to volatilization. The resuits are based on mean measured concentrations and are reported as the 96-hour EClO, EC50 and EC90 values (.e., the concentrations of test material that reduce culture density by 10, 50 and 90%, respecti
43、vely, as compared with the control). The study was initiated on 16 October 1992, the day the Study Director signed the protocol, and was completed on the day the Study Director signed the final report. The experimental phase of the 96-hour acute toxicity test was conducted from 18 to 22 November 199
44、3 at the Environmental Sciences Division of Springborn Laboratories, Inc. (Stl), Wareham, Massachusetts. A final report for this study was issued to American Petroleum Institute dated 24 February 1994. This amended final report, 12 August 1994, incorporates changes as made in Final Report Amendment
45、#1. 2.0 MATERIALS AND METHODS 2.1 Protocol The toxicity test was performed according to the Springborn protocol entitled “Protocol for Conducting a 96-Hour Acute Toxicity Test with the Alga, Selenastrum capricornutum, Following TSCA 797-1 050,“ Springborn Laboratories Protocol #:O91 192fTSCA 797.1 0
46、50, and Protocol Amendment #1 dated 18 November 1993 (Appendix I). This protocol conforms to US. EPA TSCA Test Guideline 797-1050 (U.S. EPA, 1985, 1987) with modifications approved by the US. EPA to minimize volatilization of the test material from the exposure vessels. 2.2 Test Material A sample of
47、 Tert-Amyl Methyl Ether (TAME), CAS #994-05-8, Lot 0281 4BZ, a clear liquid was received from Experimental Pathology Labs, Inc., Herndon, Virginia, on 17 August 1992. This sample was identified by Aldrich Chemical Company to contain 98.8% active ingredient (Certificate of Analysis, Appendix Il). Upo
48、n receipt at Springborn, the sample was stored in a dark, ventilated cabinet at room temperature (approximately 20 OC). This sample was used to Springborn Laboratories, Inc. API TRt402 95 m 0732290 0554664 239 m Page 10 of 66 Report No. 93-1 1-5065 prepare test solutions for the preliminary and defi
49、nitive exposures. Additionally, it was used for analytical standards and quality control samples. Test concentrations are expressed as milligrams of TAME as active ingredient per liter of solution (mg A.I./L), 2.3 Test Organism The alga used in this toxicity test was the freshwater green alga Selenastrum capricornutum, strain 1648, class Chlorophyceae. The alga was originally obtained from the Carolina Biological Supply Company, Burlington, North Carolina, and was maintained in stock culture at Spring
