Document Eq6q9MnQGBNRMmYjO1ZJqGxg4

Effectof Potas Survivale,tc.( I .. TOXICITY TO AQUATIC INVERTABRATES (E.G.,DAPHNIDS) TEST SUBSTANCE Identity:Perfluorooctanesulfonatmea;y alsobe referredtoas PFOS or FC-95. (1-Octanesulfonica.c_id,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-p,otassitimsalt,CAS # 2795-39-3) Remarks field: The testsubstance isa whitepowder of uncharactedzed purity. The followingsummary isabbreviateddue tothefactthatthisstudy has been superceded by a more recenttest. METHOD Methods: ASTM, 1981, Proposed Standard PracticeforConduction Renewal Life-CycleToxicityTestswiththe Daphnia magna; OECD, 1981, Daphnia sp.,14-day ReproductionTest. Type: Acute staticand Chronicrenewal GLP: No Year completed: 1984 Species: Daphnia magna RESULTS 48-hour EC5o (immobilization):27 (25-28)mg/L 14-day EC5o (reproduction)*: 14.7(12-18)mg/L 21-day EC50 (reproduction)*: 12.4(11-14)mg/L 28-day EC5o (reproduction)*: 11.4(10-13)mg/L 28-day NOEC**: 7 mg/L 14,2 1,and 28-day MATC: 11.2 mg/L Cumulative(Young/Adult) This concentrationappliesto no observed effectson Cumulative results ofYoung/Adult and Broods/Adultand on the totalnumber ofYoung/Adult. DATA QUALITY ReliabilityK:limischranking= 2. This studysatisfieadllcriterifaor qualitytestingatthe time performed,butactualconcentrationswere not measured. Resultswere based on nominal concentrations.Additionally, sample puritywas notadequatelycharacterized. REFERENCES This studywas conducted by the 3M Company, Environmental Laboratory,1984. OTHER Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St.Paul,Minnesota,55133 Last changed: 5/3/00 Iutii.ii74 7 11-D TECHNICAL REPORT SUMMARY F-"L- -If)13 C-5--5- TO: TECHNICAL COMMUNICATIONS CENTER - 201-2C-12 tltii.oo-rtIafirieiporitsprinteodn bothsideosfpapers,endtwocopietsoTCC.)Guidelinaensreverseide. Division Environmental Laboratory (EE & PC) f'roject 5wi: r4uniber 0535 Praiect Nuse,ho, New Methods Develot)ment -Noport Till* 9970030000 u-b-,,, Effect of Potassium Perfluorooctanesulfonate on Survival, etc.i o- 49091 Period Covered or Date Bohon -Ih.,IL.-, 2/13/84 tmplofw r4umber(si X'i r SECURITY 10, KEYWORDS: Lab Code Open Report& SunvnarV 13 ClosedReport-OpenSumm (Company Confidential) (SiteciaAluthorization) CURRENT OBJECTIVE: V 3M CHEMICAL&, ;@V-@RFCGICS'TRY O@362 pa"@ ttictudinil 12, [:]Chock box ifnsvw chemicals at* fepof led.U P' calRegistryForm 6092 toreporiallnow su OtherKeywords Technology Transfer: To develop a daphnid chroni life cycle capability. ?INV,A,A@ .............. ............. REPORTAIMTRACT.ThiasbstraicntformatiistotnietribhyotlhoeTtedchniCcoamlmunicatiCoennstetrualer3tM'ertso CompanRy&D. ........ Abstract - Acute static and chronic renewal Daphnia magna te-stswere conducted to evaluate this organisms response to varying concentrations of potassium perfluoro- octanesulfonate. The acute 48-h EC was 27 mg/L, with 95% ........ C.L. of 25 to 28 mg/L. In 14, 21, )R-day chronic expos4re ...... studies the NOEC was 7 mg/L. ................. At the highest concentration tested (18 mg/L) reproductive indices; viz: cumulative live '''''-,-"""Younpger adult, average number of young per brood, and average number of broods per adult, were inhibited to a ...... significant degree p.@0.05. This concentration of potassium perfluorooctanesulfonate was several orders of magnitude ...........g.r.e.ater than levels anticipated under normal use .........c.onditions. The survival, growth, and reproduction of this species will not be impacted negatively by chronic 14, 21, or 28-day exposure to potassium perfluorooctanesulfonate. This study should be repeated since the two highest concentrations tested, 7 and 18 mg/L represent the NOEC and significant effect levels respectively. There is a need to develop information on concentrations lying between these two extremes. Key Words: cc: M. T. Elnabarawl,r E. A. Reiner Toxicity, acute, chronic, Daphnia magna lntntmatioLniaison Initial%: INTRODUCTION Ubel et al (1) reported that perfluorinated chemicals had been found in the serum of manufacturing plant workers* ?gi of these materials has been the subject of animal toxicity studies 0 Data on a similar compound, potassium perfluorooctanesulfonate, a off-white solid, indicates that this material is water soluble and preliminary information suggests that it will neither readily biodegrade nor bioconcentrate. Preliminary aquatic testing of potassium perflurorooctanesulfonate indicated that toxic signs could be elicited during acute exposure conditions. Of the aquatic species available for use as test organisms, the crustacean, Daphnia magna is being utilized widely in acute and chronic effects testing. Advantages associated with the use of daphnids are their size, comparatively short life cycle( eio;7gical importance, of laboratory cultoring, sensitivity, etc. 3 A, #1 Guidelines for (6) conducting acute 48-h tests have been published , and chr exposure protocols are being developed or have been adopted ?gic. A ease broad database consisting of both acute and chronic studies utilizing Di magna presently exists. Daphnids were used, therefore, to evaluate tWe7effect of acute (48-h) and 14, 21, or 28-day chronic exposures to varying concentrations of potassium perfluorooctanesulfonate. METHODS AND MATERIALS Water Diluent The dilution water used in all tests was an unchlorinated, carbon filtered well water, aerated to saturation before use. Mean values for some water quality parameters were: pH 7.6 (+0.1), total hardness as mg/L CACO of 256 (247-265), alkalinity ag mg7L CACO of 224 (219-229) ana conductivity as umhos/cm at 25 C of 349--(325-373). Test Species The laboratory stock culture of D. magna was maintained under intesmittant illumination, 16h light and 8h dark, at a temperature of 20+2 C. Stock and organisms used during chronic exposure tests were f@-d blended fish food of known nutritional quality and yeast at the rate of 30 mg solids/L of test solution. Acute Tests During these tests the dilution water temperature averaged 22+1,OC, while a photoperiod of 16-h light and 8-h darkness was maintained throughout the test. For the 48-h tests, 20 daphnids 12 + 12-h old were placed in duplicat @est beakers, 250 ml capacity, each containing ?6 200 ml of test solution . All acute tests were run without food additions. Five concentrations of potassium perfluorooctanesulfonater and an untreated control group were run in duplicate. The endpoint used in these tests was daphnid immobilization as characterized by a lack of coordinated movement. The calculated EC 52 value was used to define the range of concentrations to be used in he life-cycle renewal test. 2 Chronic Tests The chronic daphnid studies using first instars 12 + 12-h old were conducted in a temperature controlled growth chambe7r at 23 + IOC with photoperiod automatically fixed to a 16-h light, and 8-h dark cycle. This static renewal chronic test was conducted using ten 250 ml beakers containing 200 ml test solution at each of five concentrations of the test material plus an untreated control set of 20 beakers. Each test concentration was replicated three times with five Daphnia ma@na first instars per beaker for survival data and seven times with but one daphnid per beaker for reproductive measurements. Stock solutions were prepared in deionized water. Fresh solutions were made up thrice weekly (MWF) with food also being replenished at this time. Disposable glass pipettes having an I.D. of 5 mm were used to transfer the adult daphnids to fresh @iolution. The young daphnids were then counted and discarded. During the test the following data brood release, number of young per adul t, mortality of young or adult nomin* al concentrations of potassium were tabulated: day of initial adult per day, number of broods per daphnids. All data are based on.. perfluorooctanesulfonate. Statistics The 48-h, acute and 14, 21, and 28-day chron*c EC values and-their associat 95% confidence analysis Ti) or the moving limits were dete@rmin gy either probit angle average method tg) Reproductive an d survival data were analyzed by analysis of variance. Throughout the' text "significant" will refer to statistical significance, having a p value of .05. Acute Test RESULTS The calculated 48-h EC value was 27 mg/L of potassium perfluorooctanesulfonaig with a 95% C.L. ranging from 25 to 28 mg/l. Based on this value the range of test concentrations selected for the daphnid chronic test were 0.26, 1.0, 2.6, 7.0, and 18 mg/l of potassium perfluorooctanesulfonate. Temperature and pH varied minimally while the dissolved oxygen level did not fall below 70% saturation. Chronic Test - 14, 21, ancl 28-Day Duration Indices of reproductive performance and survival of Daphnia magna exposed to the above listed concentrations of the test material are tabulated in Tables 1-4. Untreated control reproductive and survival indi Tib )as shown in Table.1 are in agreement with reported literature values . Literature values cited for the cumulative number of young per adult ranged from 0 0 30-122 at temperatures of 20-25 C while at temperatures of 18-20 C Daphnia magna survival varied from 80-92%. 3 The effect an reproductive indices elicited by chronic exposure to potassium perfluorooctanesulfonate are illustrated (Table 1). Over the 28-day exposure period and at chemical concentrations varying from 0.26 to 7.0 mg/L the mean values of the above cited parameters varied minimally when compared to control data. Thus, the absence of a concentration dependent response up to 7.0 mg/L is apparent. Interference with the normal reproductive process did occur at a toxicant concentration between 7.0 and 18.0 mg/L. At this latter concentration, the cumulative number of live young per adult were reduced by 80%, average number of live young per brood was inhibited by 55%, and the number of broods per adult was reduced by 62%. It has been reconukended(7)that three brood studies 'replace 21 or 28 day chronic exposure tests (Table 2). At potassium perfluorooctanesulfonate concentrations up to and including 7.0 mg/L the time to three broad release was either 14 or 15 days after initiation of the study. At IS mg/L it required approximately 17 days for release of the three broods, a statistically significant difference when compared to control values. Calculated values for 50% reproductive impairment at 14, 21# and 28 days of exposure to potassium perfluorooetanesulfonate are shown (Table 3). Adult mortality EC values could not be determined as they exceeded the highest teig concentration used in this study. -The 28-day EC values calculated for all reproductive parameters were similar, ragsing from 11.4 to 16.1 mg/l. Overlap of the 95% C.L. is indicative of the absence of statistically significant differences in these values. In this study statistically significant changes in alr reproductive indices did occur at a concentration of 18 mg/L potassium perfluorooctanesulfonate (Table 4). Exposure to concentrations of 0.26,-1.0, 2.6, and 7.0 mg/L of the test substance did not affect these indices or mortality to a significant degree. Thus, a concentration of 7.0 mg/L of potassium perfluorooctanesulfonate is considered to be the chronic NOEC or no observable effect concentration. Table 5 illustrates the derived acute/chronic ratios for those reproductive indices evaluated throughout the potassium perfluorooctanesulfonate exposure period. The 28-day acute/chronic ratios for all parameters studied were identical and were equivalent to the 14 and 21-day acute/chronic ratios derived from cumulative number of young per adult data. This may indicate a concentration dependent process which is not affected by increasing the exposure duration. Table 6 shows the chronic renewal toxicity of potassium perfluorooctanesulfonate expressed as the NOEC, MATC, and EC hich values were derived from reproductive indices. As in the preceg?nwg table, the derived values are chemical dependent and time independent. Thus, the NOEC and MATC are identical for all parameters measured at all exposure periods. The EC values differ slightly however there is no statistical diffgpence in these values based on variable exposure durations. 4 DISCUSSION Reproductive measurements have been used as indicators of toxicant effects following exposure to sublethal co?stntrations of PCB, p,p'DDT, or AtrazinO. Furthermore, Parkhurst et al investigated the reproducibility of the Daphnia magna chronic test and concluded that, of the toxicity criteria studied, the number of young produced per* brood or per adult were sensitive indicators of a chronic toxicant effect. Our resyl@i are in agreement wit@)those reported by Maki and Johnson and Parkhurst et al that changes in indices of reproduction are sensitive parameters of a toxicant effect. Changes occuring at concentration levels of IS mg/L of potassium perfluoro- octanesulfonate iddluded reductions in: 1) the average number of broods per parent, 7 to 3; 2) number of live young per parent, 190 to 37; 3) average number of young per brood 27 to 13. An increase in the. number (to 54) of dead young per parent was also noted. When combining the number of live and dead young a 52% reduction in total number of young released was observed. The most noteworthy result of daphnid_ exposure to high levels of potassium perfluorooctanesulfonate has been the attenuation of all reproductive indices. On the other hand, at this concentration maturation was not delayed nor was longevity of the parent affected. Water quality guidelines have specified the use of two protective concentrations, namely a 24-h average equivalent to the no observable effect concentration (NOEC) and either a 24-h maximum or 48-h EC level. Neither of these protective concentrations should be exc-e52ded over the indicated time period. Utilizing this concept, protective concentrations of 7.0 and 27 mg/L would be established-for potassium perfluorooctanesulfonate. Over the 28-day exposure period no observable effects occurred using either mortality or reproductive indices as the response criteria. A NOEC of 7 mg/L and the toxicant. level-which elicited an effect, 18 mg/L, give us a gross approximation of the'GM-'MATC, equal to 11 mg/L. Conversely, during a 28-day chronic exposure to 18 mg/L of potassium perfluorooctanesulfonate, reproductive impairment was noted. This phenomenon was observed as early as the second week and remained at the same EC value for the duration study. so of the Based on these findings and considering the concentrations of test chemical necessary to elicit a chronic effect, it is concluded that potassium perfluorooctanesulfonate will not impact negatively on the survival, growth and reproduction of Daphnia magna under present environmental release conditions. REFERENCES 1. Ubel, F. A., Sorenson, S. D., and Roach,-D. E. 1980. Health Statu's of plant workers exposed to fluorochemicals-a preliminary report. J. Am. Ind. Hyg. Assoc. 41:584-589. 2. Griffith, F. D. and J. E. Long, 1980. Animal Toxicity Studies with Ammonium Perfluorooctanesulfonate. Am. Ind. Hyg. Assoc. 41:576-583. 5 3. Parkhurst, B. R., J. L. Forte and G. P. Wright, 1981. Reproducibility of a Life-Cycle Toxicity Test with Daphnia magna. Bull. Environ. Contam. Toxicol 26:1-8. 4. Presing, M. 1981. On the Effects of Dikonirt (sodium salt of 2,4Dichlorophenoxi-acetic acid) on the Mortality and Reproduction of Daphnia magna Hydrobiologia 83:511-516. S. Adema, D. M. M., Kuiper, J., Hantsveit, A. 0., and Canton, J. H. 1982. Consecutive System of Tests for Assessment of the Effects of Chemical Agents in the Aquatic Environment. National Technical Information Service, Springfield, VA. 6. American Society for Testing Materials, 1981, Proposed Standard practice for conducting renewal life-cycle toxicity tests with the Daphnia magna. American Society for Testing and Materials, Philadelphia, PA. 7, OECD. 1981. Daphnia sp., 14-day reproduction test (including an acute immobilisation test). Organization of Economic Cooperation -and Development. Paris, France. 8. Finney, D. J. 1971. Probit Analysis. University Press, Cambridge. 9. Stephan C. E. 1977. Methods for calculating an LC In F. L. Mayer and J.L. Hamelink, eds. Aquatic toxicology an59 hazard evaluation. American Society for Testing and materials, Philadelphia, pp. 65-84. 10. U.S. EPA 1982. Environmental Effects Test Guidelines, EPA 560/6.82-002 ES-1, PP. 1-42. 11. Nebeker, A. V., C; K. McAuliffe, R. MBhar, and G. Stevens, 1983. Toxicity of silver to steelhead and rainbow trout, fathead minnows and Daphnia magna. Environ, Toxicol. Chem. 2:95-104. 12. Maki, A. W., and H. E. Johnson, 1975. Effects of PCB (Aroclor R54) and p,p'DDT on production and survival of Daphnia magna Strauss. Bull Environ Contam. Toxicol 13:412-416. 13. Schober, V. and W. Lampert: 1977. Effects of Sublethal Concentrations of the herbicide AtrazinO on growth and reproduction of Daphnia I>ulex. Bull. Environ Contam. Toxicol: 269-277. TABLE 1 Chronic Exposure of Daphnia magna to Potassium Perfluorooctanes 9 lfonate: Young Reproduction Treatment mqZL Control Cumulative Live Young/Adult Average Number Avg. Number n Number Young/Brood Broods/Adult 11 190 27 7 +36 +4 +1 0 .26 11 169 26 7 +33 +4 +1 1.0 7 183 25 7 +40 +4 +1 2.6 7 186 26 7 +29 +4 +1 7.0 6 199 27 7 +19 +3 +1 18.0 7 37 b 12 b 3 b +28 +8 +1 a 28-day exposure data* b p.@O 0 5. Mean + S,D, T,KBLE 2 ChroniC Exposure of Daph nia magna to Potassium Perfluorooctanesultonate: Comparison of lSt and 3rd Brood Release Days Treatment mg/L a Control n 0.26 17 1.0 10 2.6 1D 7.0 9 18.0 10 a b Beakers in tesi. p.io.05. Initial Brood Third Brood Release Day Release Day 9 is +1 +1 10 is +2 +2 9 15 +1 +2 9 15 +0.3 +2 9 14 +0.3 +1 10 17 b +2 +2 Table 3 Acute and Chronic Toxicity of Potassium Perfluarooctanesulfonate to Daphnia magna: Acute and Chronic EC so Exposure Period YoaRg Reproduction--EC mg/L Cumulative Live - (95% C.L.)' Avg. No, Immobilization Young/Adult Per Brood Broods/Adult 48-hr 26.6 (25-28) 14 Day 14.7 a a (12-18) 21 day 12.4 (11-14) 28 day 11.4 (10-13) 16.1 (13-20) 14.7 (13-18) a Not calculated. TABLE 4 Comparison of the Chronic Effects of 7 and 18 mg/L Potassium Perfluorooctanesulfonate to Da hnia magna 14-Day 21-Day 28-Day A. 7.0 mg/L: Number Live Young 59 /Adult Number Dead/Adult 3 Total.Number Young 369 Adult 14ortality 0 129 3 793 4.5 199 4 1220 4.5 B. 18.0 mg/L: .Number Live Young /Adulta Number Dead/Adult a Total Number Younga Adult Mortality 24 12 249 4.5 33 51 591 9.1 a Comparison of 7.0 and 18.0 mg/L piO.05. 37 54 641 - 18.2 TABLE 5 Acute/ChroniC EC Ratios Following Exposure of a Daphnia magna to PoiRssium Perfluorooctanesulfonate Day of Exposure Cumulative Young/Adult Cumulative Broods/Adult Number of Young/Adult 14 1.5 21 1.5 28 1.5 1.5 1.5 a Acute/Chronic EC so Ratio 48h Concentration Eliciting an Acute Effect in 50% of Organisms (EFx-p-osure Duration) ConFentr-ation Eliciting a Chronic Effect in 501 of Organisms TABLE 6 Chronic Renewal Toxicity of Potassium PerfluorooctanesulEonate to Daphnia magna .Day@ Cumulative Young/Adult 14 21 28 Cumulative Broods/Adult 28 Number of Young/Adult 28 b NOEC 7 7 7 7 7 c,d MATC 11.2 11.2 11.2 11.2 11.2 e EC 50 -14.7 12.4 11.4 (12-18) (11-14) (10-13) 14.7 (13-18) 16.1 (13-20) a Data in mg/L. b No observable effect concentration. c Maximum acceptable toxicant concentration. d MATC - Geometric mean between highest test conccentration having no significant (p>0.05) effect and the lowest test concentration havii@g-a significant (piO.05) effect. e Dose at which 50% of organisms are affected by-the toxicant. Expressed as Mean (95% C.L.).