Document VJXp7LO0372a3YVo0XE9mMdk8

AR AZ& - 07*7 TOXICITY TO AQUATIC PLANTS TEST SUBSTANCE___________________________________________ Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, FC-95, or as a component of FC-203. (1Octanesulfonic acid) (CAS # 2795-39-3). Remarks: The 3M production lot number was not noted. The test sample is FC-203. Current information indicates it is a mixture of 1.34% PFOS, 35% diethylene glycol butyl ether, 37.85% water, 20% ethylene glycol, 2.66 % Sultone foamer, 3% sodium octyl sulfate, 0.1% sodium lauryl sulfate, and 0.05% tolyltriazole. The following summary applies to a mixture with incompletely characterized concentrations of impurities. Data may not accurately reflect toxicity of the fluorochemical component of the test sample. METHOD:___________________________________________________ Method: OECD Guideline 201 Type: Acute Static GLP: No Year study performed: 1991 Species: Selenastrum capricornutum Source: In-house cultures. Originally obtained from the Culture Collection of Algae at the University of Texas at Austin. Element basis: Algal cell counts (cells/ml), growth rates. Exposure period: 96 hours Statistical Methods: The EC50 values were calculated using the computer program of Stephan, 1983. The NOEC was calculated using Dunnett's test. Analytical monitoring: pH and temperature. Test Conditions: Algal Nutrient Medium: Nutrient medium per U.S. EPA, 1978. This nutrient medium provided all mineral nutrients essential for algal growth and also served as the diluent for all algal operations. The pH of this synthetic algal medium was ~8.0. Stock and test solution preparation: A 1000 mg/L primary stock solution was prepared in nutrient medium. Aliquots were then added directly to the test vessels to create test solutions. Exposure vessels: 250 mi Erlenmeyers containing 100 ml test solution and capped with inverted glass beakers. Agitation: Shaken continuously at 100 rpm. 000639 Number of replicates: 3 Initial cell loading: 1.0 x 104 cells/mL Number of concentrations: five plus a blank control Lighting: 35 pEin/sec/m2from continuous cool-white fluorescent lighting. Water Chemistry: pH range (0-96 hours): 7.8 - 9.4 (control exposure) 7.8 - 8.0 (1000 mg/L exposure) Test temperature range (0-96 hours): 23.3 - 23.4 C (incubator) RESULTS____________________________________________________ Nominal concentrations: Blank control, 62.5, 125, 250, 500, and 1000 mg/L 96-Hour Algal Growth Response Values Nominal Cone. (mg/L) Control 62.5 125 250 500 1000 Mean Cell-Count (cells/mL) 399,000 645,000 347,000 69,000 51,000 12,000 Average Specific Growth Rate 0.038 0.044 0.037 0.020 0.017 0.002 Percent Change in Growth Rate from Control (96 hrs.) -- -16 3 47 55 95 Algal Growth Response ErC5ovalues: 96-hour ErC50 = 354 (323 - 389) mg/L 96-hour NOEC = 500 mg/L Element values are based on nominal concentrations. Reversibility of Growth Inhibition: Not give. Remarks: Testing was conducted on the mixture as described in the Test Substance Remarks field. The values reported apply to that mixture and not the fluorochemical proportion alone. CONCLUSIONS_______________________________________________ The test substance exhibits a 96-hour ErC50 growth rate value of 354 (323 389) mg/L. The 96-hour No Observed Effect Concentration (NOEC) is 500 mg/L for growth rate. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 000640 DATA QUALITY Reliability: Klimisch ranking = 2. This study meets the criteria for quality testing. However, sample purity was not properly characterized and the study lacks analytical confirmation of the amount of fluorochemical proportion in the solution. REFERENCES_______________________________________________ This test was conducted by EnviroSystems Division, Resource Analysts, Inc., or Hampton, NH at the request of the 3M Company, Lab Request number H2864-9, 1991. OTHER ______________________________ r____________________ Last changed: 6/27/00 000641 * BEST COPY AVAILABLE Study Title Static Acute Toxicity of PC 203 to the Alga, Seleoastrua capr-icornotit* Authors Robert I. Boeri Tiaothy J. Sard Study Coauleted June, 1991 r,> i* '.,:'Jt '* ^vV <, %' ... v' r'-\ ->V ' . i' '"i'-f/s.i.T/ .' r `V-V ^ a/ A^'" K ' `^ 1 v' i'M:**>V\, *' V1#* i'rSA 45 1 j / !'V - * .j/ iX K -,*j'v : Page l of 15 *'v ^ -'*,$vrii!svv* . -Perforuina Laboratory -v/ ~BnviroSysteas Division i- Resource Analysts, Incorporated . 1'cfcwr 2130 *"' /\t* Ro4 |jtarHaapshire/ ; <V./ . s fa ta * *^ Vf -vi Ky-i ' fi% V' .H.'irr'1, -, rv'V>.V*w??' *1`V v-psj nf /Jtt ._fc.*... ., , v$ ,<'-t` ^ f.Afcrf ReKx^AncXy^SubjkJaryoiM UJPOE fef` S^S fe'Vw ' i* f?vO"''Vr/; * j 000642 BEST COPY AVAILABLE I. SUHHARY The acute toxicity of FC 203 to the alga, Seleaastrua capricornuturn, is described in this final report. The test vas conducted for 3 H ' Conpany for 96 hours during Hay 6 to 10, 1991, at the EnviroSysteas Division of Resource Analysts, Inc. in Haspton, Hew Hampshire. It as conducted by Jeanne Hagazu, Peter Kowalski, Ellen Stanford, Robert Boeri, and Tiaothy Ward. The te&t vas performed under- static conditions with five concentrations of test substance and a dilution water control at a aean temperature of 23.6C. The dilution water was synthetic aedia prepared with sterile deionized water. Flasks were incubated on a rotary shaker. Light was adjusted to 35 uEin/sec/s2 with a photoperiod of 24 hours light and 0 hours dark. Hoainal concentrations of FC 203 were: 0 ng/L (control), 62.5 mg/L, 125 mg/L, 250 ag/L, 500 mg/L, and 1,000 ng/L. Hominal concentrations were used for all calculations. Algae used in the test was fros an in-house culture that was maintained under test conditions. Exposure of algae to the test substance resulted in a 72 hour EC50 of 338 mg/L FC 203 and a no observed effect concentration (HOEC) of 250 mg/L. The 96 hour EC50 and >0EC are 354 ng/L and 500 ag/L, respectively. 000643 BEST COPY AVAILABLE IT- TABLE OF CONTENTS SECTION: I. Suuary II. Table of Contents III. Index of Tables IV. Introduction V. Methods and Materials VI. Results m . References Appendix A. Vater Quality Bata fro* Toxicity Test PAGE 2 3 4 5 5 8 12 13 000644 BEST COPY AVAILABLE III. IHDEX OF TABLES Table 1. Chenical characterization of a representative sanple of deionized water used to prepare test nedia for toxicity test Table 2. Growth data froa toxicity test Table 3. Percent change fro control and average specific growth rate fro toxicity test Table 4. Median effective concentrations (EC50s) fro toxicity test Table A.l. Temperature easnred daring toxicity test V Table A.2. pH ensured during toxicity test PAGE 6 9 10 n 14 15 000645 BEST COPY AVAILABLE iv. iwtrodoctiow This study was sponsored by 3H Company, St. Paul, Hinnesota. The objective of the study was to detersine the acute toxicity of FC 203 to a freshwater alga. The report contains sections that describe the aethods and aaterials eaployed in the study, and the results of the investigation. The report also contains an appendix that presents the water quality data collected during the test. V. HETHODS 1XD HATERI1LS TEST SBBSTAHCE: FC 203 (EnviroSysteas Saaple Number 2699E) was delivered to EnviroSysteas on November 15, 1990. It was contained in a 250 aL glass bottle that was labelled with the following intonation: "H 2864-9, FC 203". FC 203 (a clear amber liquid} was supplied by 3H Company, 935 Bush Avenue, St. Paul, Hinnesota. Prior to use the test aaterial was stored in the dark at room teaperature. A reserve saaple (approxiaately 1.1 grans) will be archived at EnviroSysteas for a ainiaua of 10 years. DILUTION WATER: Water used for accliaation of test organises and for all toxicity testing was sterile enriched sedia (U.S. EPA, 1978) with a pH of approxiaately 8.0. Results of chemical analysis of a representative saaple of deionized water used to prepare the media are presented in Table 1. ; TEST ORGANISM: -- A ^ -w - Algae, used for the test was front a culture originally procured froa ; the .Culture .!Collection'-, of Algaeat the University of Texas at Austin and delivered to Envirosysteas on April 12,'?1990. The identity of the culture 4:was'verified ;uaing*V-an||appropriatey taxonoaie key.7.-- Theculture .*was transferred r.,tovfsterileWiriched aedia ^identical to media used for this [i^^^The3';5definitivei|ltoxirity;Stes^^^^erfoned during April 22 to 26, ' ^ 1 9 9 1 ^ ^ `Xt^is^bai^^e)iii^ jirocrtniri^ -(1984). `The test'.was ^conducted'^atj.'f.ajtargntr*t e n ^ irith five concentrations of fj!'\fV '.test2substahceffluidia-dilutioitwatif|TOntrbl^A 1,000 ag/L stock solution > '} was 'prepared`iin7dilution water^jThe2stocksolution was added directly to ^ i,J.,dilution^waterijcontained Jin ^theSi.test^.vessels without the use of a solvent. ^ 2 NNoommiinnaall:^i'concen-tr-a-t-io-n-s-|ioftheigtest- aaterial were: 0 ag/L '* & (control) /;62.5 W ^ | l ? S ^ / l ' ^ 2 5 g ^ ^ . 5 0 0 j g / L ; and.1,000 ag/L. "V k1'>& . 1 - - ~.* i 000646 BEST COPY AVAILABLE Table 1 Cheaical characterization of a representative sanple of deionized water used to prepare test aedia for toxicity test Paraaeter Organochlorine pesticides Unit of Heasureaent Reporting Liait ug/L 2 Measured Value HD Organophosphorus ug/L 0.5 HD pesticides Polychlorinated ug/L 0.5 HD biphenyls Hote: HD * not detected at or above the reporting limit. 000647 BEST COPY AVAILABLE Algae was randomly and equally distributed among three replicates of each treatment at.the rate of 10,000 cells/ml. The test was performed in 250 ml glass Krlenmeyer flasks that contained 100 ml of test solution. The flasks were capped with inverted glass beakers. Test vessels were randomly arranged on a rotary shaker in an incubator during the 96 hour test (a random numbers table was used to select the location of each vessel). A 24 hour light and 0 hour dark photoperiod was maintained with cool-white fluorescent lights that provided a light intensity of 35 uEs-*mr*. The number of algal cells/ml in each test vessel was determined visually every 24 hours by means of direct microscopic counts with a hemacytometer. The pH (Beckman model pHl 12 meter) was determined in each test flask at the beginning and end of the test, and temperature (ASTM mercury thermometer) was measured and recorded daily in the incubator. STATISTICAL KETHODS: Results of the toxicity test were interpreted by standard statistical techniques (Stephan, 1983). The binomial or moving average method was used by the author to calculate EC50 values using nominal concentrations of test substance. The MOEC was calculated using Dunnett's test, which includes a parametric one-way analysis of variance (ANOVA)- . The average specific growth rate was calculated as the natural log of the number of cells per ml at 96 hours minus the natural log of the number of cells per ml at 0 hours divided by the exposure period. Thep e r c e n t change from control is calculated by subtracting the sample :average specific growth rate from the control average specific 'growth ^rate, dividing that value by the average* specific growth rate in the control and multiplying that value by 100. The percent change,; from -control was used to compute EC50 and H0EC values. ' ' A . - -i* v* c . *.. . w r i **<* > r '' , JA V i ' A- "i < i ' 'vv BEST COPY AVAILABLE VI. RESULTS Biological and water quality data generated by the acute toxicity test are presented in Table 2 and Appendix A,- respectively, and the percent of control and average specific growth rate information is presented in Table 3. The 24, 48, 72, and 96 hour EC50s for algae exposed to FC 203 are presented in Table 4. The 72 hour EC50 is 338 mg/b, and the 72 hour KOEC is 250 ng/L FC 203. The 96 hour EC50 is 354 mg/L and the 96 hour NOEC is 500 ng/L. 000649 BEST COPY AVAILABLE Table 2. Growth data fro* toxicity test lfoninal Concentration (ff/L) rep. Vuaber of Cells per Hilliliter x 10 (hour) 0 24 48 72 96 0.0 (control) 62.5 125 250 500 1 2 3 ean 10 10 10 10 26 94 32 34 30 116 29 81 1 10 30 92 2 10 52 94 3 10 48 92 ean 10 43 93 1 10 26 52 2 10 26 76 3 10 34 46 ean 10 29 58 1 10 28 48 2 10 26 62 3 10 20 42 ean 10 25 51 1 .: ' 10 32 2 :>.;;10 '.fT 44 3 v-ilO./;^:.: 28 ean -V..10. . 35 *v V ! 1 10 . .24 2 ;/10 , * 20 '.'.3 ,'>4wjfe*S|-.X0 3- 26 * .) .23 '-- v. -1** < ^ . 32 28 64 41 36 36 32 35 188 394 172 418 282 386 214 399 346 604 256 596 296 734 299 645 100 350 82 214 94 476 92 347 62 76 42 82 64 50 56 69 30 58 28 58 26 36 28 51 !>;,`'Sw*''V 28 V 4 .12 24 10 30 14 27 12 u<-*,**1.. Z' ^" >s^T\'^v ; ; sLtg/ - Uv h -v* V - * *'^<-;*C'/~5>-4 * ' *' ,v N * Tr - _c u V'V,t/* r \ _* Rwoi^yWv^^^SuMdfciyofMllJ^ `fUb'M tC"fc *&g.tVj&?r` $i 000650 BEST COPY AVAILABLE Table 3. Percent change fro* control and average specific growth rate fro* toxicity test Noainal Concentration of Test Substance (g/L) Average specific growth rate -----------------------24hr 48hr 72hr 96hr Percent change fro* control ---------------- . 24hr 48br 72hr 96hr 0.0 (control) 62.5 125 250 500 1,000 0.045 0.044 0.043 0.038 0.062 0.047 0.047 0.044. 0.044 0.037 0.031 0.037 0.038 0.034 0.024 0.020 0.052 0.030 0.014 0.017 0.036 0.026 0.014 0.002 0000 -38 -7 -9 -16 2 16 28 3 16 23 44 47 -16 32 67 55 20 41 67 95 000651 BEST COPY AVAILADLE Median effective concentrations {EC50s> fro toxicity test Exposure period EC50 95 percent confidence limits Calculation ethod 24 hours 48 hours 72 hours 96 hours >1,000 9/1. >1,000 wgfL 338 g/I> 354 mg/h -- -- _ 293 - 395 mg/h 323 - 389 mg/h -- -- ovina average oving average 000652 BEST COPY AVAILABLE! v ii. nnm os OECD. 1984. Guidelines for Testing of Chemicals. Section 2: Effects on Biotic Systems. 4, 1984. Method 201, Alga, Growth Inhibition Test. Adopted June Stephan, C.E. 1983. Cospnter program for calculation of LC50 values. Personal conaunication. .S. EPA. 1978. The Selenastrua capriccrrnutua Printz Algal Assay Bottle Test. EPA-600/9-78-018. July, 1978. lW m flV t' I ->Lv-*v .-> 1 ./-. ' % :*'<--/< * 1 ^ ^ t "W =V'f^J:&;Y-s5*^tr^iv^-4Y^- *? s"h jk.*,.> T-Me f J'VV*- ijMSSiS* 4'vr- ^ ' Kr`: ?, S;`X--''vKt--'C<''T"'&s'fAZtK}H*'- v--- , t**ssstth .- '> ,, ., "^ V -'* - .* * SO ..-Yvi . .'-A..- r - ' r ^ C v r >r r i* \m& irW**-AV, *' ^ ~ a' Page 12 of 15 "*.* ?,.* I" *. f ...*>.'' .uV.-.F^H v^ : i *, ?" . ;' .JJY,J` ' ..J <. v , t *Anci|(*infcSu&^ i ^ ? i - ^*5H<=',/ 'Vvl." . ^-h ?. 000653 BEST COPY AVAILABLE Appendix A. VATER Q0ALIT7 DATA BLOW TOXTCTTT TEST 000654 BEST COPY AVAILABLE I Table A.l. Temperature measured during toxicity test Day of Exposure 0 1 2 3 4 Temperature of Incubator,C 23.3 _ 23.3 23.3 23.4 23.4 ia}* Via<! "-V' -w ij . -5 / " A' " v4**^V2s*-v?,!.i *.,j',v$v*-** ; . w x- ....-..... . *->''v>V*",*^** r, . ..^..'.v;..v:-v-' ;.v-'r ' ' V- V ,\ '' - t/t-'i4'-> - v,'. ' jin's v> * . < ^ ... ' Page 14 of 15 '* 's*-k : ;u V- '1i?*, *, (.Tli'-x-*'^*.c-*.*-' s < y-Ai ; I> '>iHV V n >" J< "l 1 *t* 4if' *i -4,J > u - * ,-i - 't3 ' .; "*? J " / ' , ? S". ;vx . *,*-\ '*i ; -v; * ,,'*'' -J'a '^^r" -- .'?..*..i . ..'-1..'.j^-'.' y -.Y. ' . ~*w'..*1%- i > *. *wy>.- <v-v>f ' /s% . - . i y BewucmAnatra Inc, Subddoy of MUJPCUE ->,> `v --a 000655 E BEST COPY AVAILABLE Table A.2. pH Measured during toxicity test Noninal concentration (g/L) 0 (control) 62.5 125 250 500 1,000 Replicate 1 2 3 1 2 3 1 2 3 1 2 3 1 3 1 2 3 pH initial final 7.8 8.0 7.8 8.0 7.8 9.4 7.7 9.5 7.7 10.0 7.7 9.5 7.7 8.7 7.7 8.7 7.7 8.7 7.7 8.3 7.7 8.3 7.7 8.3 7.6 8.3 7.7 8.2 7.7 8.1 7.8 8.0 7.8 8.0 7.8 8.0 000656