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AR 236 - 072] ALGICIDAL ACTIVITY IESTSUBS~T ~~ ANC0E 000 Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, Octanesulfonic acid) FC-95, (CAS # or as a component 2795-39-3). of FC-203. (1- Remarks: The 3M production lot number was not noted. The test sample disieFtCh-y2l0e3n.e gCluycrorlenbtutiynlfeotrhmeart,io3n7.in8d5i%cawtaetseirt,is2a0%mixettuhryeleonfe1g.l3yc4o%l,P2F.O6S6,%35% Sultone foamer, 3% 0.05% tolyltriazole. sodium octyl sulfate, 0.1% sodium lauryl sulfate, and The following summary applies to a mixture with incompletely characterized concentrationsof impurities. Data may not accurately reflect toxicity of the fluorochemical component of the test sample. METHOO: 000000000 Method: Standard No. 22, Relating to Swimming Pool Water Treatment Chemicals and/or Processes (National Sanitation Foundation) Type (test type): Acute Static . GLP: No Year study performed: 1974 Species: Chlorella pyrenoidosa and Phormidium inundatum Source: In-house cultures Inc., Northbrook, IL. maintained at Industrial Bio-test Laboratories, Element basis: Cell populations turbidimetric measurement. calculated from hemocytometer counts, Exposure period: 14 days Statistical Methods: Not given. Analytical monitoring: Not given. Test Conditions: - Algal Nutrient Medium: Allen's nutrient medium. Stock and test solution preparation: Algal solutions prepared by blending algae with aliquots of 100 the mL test of Allen's medium using substance were handled a Waring as 100% Blender. One mL active and diluted with distilled water and added to Allen's medium. One mLof adjusted cell suspension was added to each test solution, for a final volume of 100 mL. ~~ Exposure Numberof vessels: Flasks replicates: 1 containing final volume of 100 mL. PInhiotiraml icdelilumloading: 7.5 x 10* cells/mL for Chlorella, 12.5 x 10* cells/mL for 000574 Number of concentrations: six plus a blank control for Chlorella, five plus a Lbilgahntkicnogn:troNlotfogriPvheonrmidium. Subculture: Inoculated bottles subcultured to fresh medium after oneday incubation, with final readings made after 14 days inoculation. Three dilutions of the test substance were subcultured (1:5, 1:10, and 1:100). Water Chemistry: Not given. R0 ESOWTS 0000 Nominal concentrations: Bk control, 1:5, 1:10, 1:100, 1:150, 1:1000, and 1:10000 dilution for Chiorella pyrenoidosa study. Bk control, 1:5, 1:10, 1:100, 1:1000, and 1:10000 dilution for Phorinidium inundatum study. Results of the Study withChlorella pyrenoidosa Sample Gross Appearance Turbidity [ [Co1 nt(d5 riluotioln)[[ngoene(Coelnor) --[[5--<2o(ab0 0sor1ba0 nce) ]| [EN 00 [nE o | -- -- n <oo|e r-- | [[7A0000000[geegrneen[20S20 00 | [T7B10susbuobcuullttourree|| 7700 subculture| nnoonnee | |----<--<000.1 01|| none |--<001T| Results of the Study with Phorinidium inundatum `Sample Gross Appearance Turbidity [ [C5 o(diolutilonT ) | (gCoelh eorn [2(e ab0 soorobarnce0 ) _ [ [Em00 0 [E1000 nnoonnee ---- |------<<o0oo0r1 | green 200 T0000 | [5subouttore | greentint none ||----<5020010 | [EF-OT0E0subeulture | Cnorne--|------<001-- Element values are based on dilutions of nominal concentrations. 000575 RSuebmsatraknsc:e TReesmtianrgkwsafsieldc.ondTuhcetveadlouenstrheepmoirxtteudraepapslydetosctrhiatbemdixitnutrheeaTnedstnot the fluorochemical proportion alone. Conclusions 0000 Algicidal rather than algistatic action of the test substance was obtained. Submitter: 3M Paul, Minnesota, Company, 55133 Environmental Laboratory, P.O. Box 33331, St. pa0 taQualry 000 Rmeeltiahboilditfyo:llKolwiemdi.scThhreanskaimnpgl=ep3u.ritSytwudayslnaoctksprdoepsecrrliyptcihoanraacntderriezceodrdasnodftthhee study in the lacks analytical solution. confirmation of the amount of fluorochemical proportion RE0 FERENCES0000 Test IL at was conducted by Industrial Bio-Test Laboratories, Inc., the request of the 3M Company, St. Paul, MN, 1974. of Northbrook, o0 rWE0 R 00000000000 Last changed: 6/27/00 000576 edly 5 Qle oneJ Zeon B1I91O0 F-RTOENTSATGELaRbOoAraDtories, Inc.E22) NoRiHBROOK, ILLINOIS 60062 _ ) REPORT TO 3M COMPANY - ALGICIDAL ACTIVITY OF TWO SAMPLES LABELED "A" AND "B" wn 203 JANUARY 23, 1974 : . IBT NO. 633-04473 > J ' 000577 Jniliistich BIO = TEST Lboralories, Ine. 5 REPORT TO 3M COMPANY TWO SAALMGPILCEISDALLAABCETLIEVDIT"YA"OAFND "B" :: JANUARY 23, 1974. - IBT NO. 633-0473 I Introduction - At the request of 3M Company, tests were conducted to determine ) the algicidal activity of two samples identified as "A" and "B". Two species of algae, Chlorells pyrencidosa and Phormidium inundatam, were used in the eveluation. . . 000578 InduBsItOr-iTEaSlT Laboratories, Ine. 2 bh I. Summary i Two samples, identified as "A" and "B", were submitted on November 20, 1973, by 3M Company. The algicidal test was ` initiated on November 30, 1973, using two species of algae, Chlorella Pyrenoidosa Wisc. 2005, Phormidium inundatam Wisc. 1093. The test was conducted according to a method outlined and Prepared by the National Sanitation Foundation, Standard No. 22, Relating to Swimming Pool Water Treatment Chemicals and/or Processes. The turbidimetric determination canformed to the pro- cedure recommended by Fitzgerald*: A 1:100 dilution of Sample "A" or "B", effectively controlled the ) proliferation of Chlorella sp. and Phormidium sp. for a 14-day period. Cidal rather than static action of the candidate algacides was obtained. * WFiattzegreraanldd,SeGw.agPe. WBoirokass,say11f0o-r29Al6gi(c1i96d3a)l versus Algistatic Cheinicals, = *; Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC. Report prepased by; _ Deel) Clare GDarnoiuepl LCehaadn,erM.S. Report approved by: 1 A. D| aO videBaskin4 , Phe .D.ee Re January 23, 1974 Section Head, Microbiology . dp 000579 SuduBiIOb-iTaESlT LukorsoroTsays ~*~ 3 3 I. Procedure A. Preparation of Inoculum } Two species of algae, Chlorella pyrenoidosa Wisc. 2005, and Phormidium inundatum Wise. 1093, were used as the test organisms. Each organism was subcultured at weekly intervals in Allen's neutrient medium for three weeks, prior to inception of the test. The suspensions of Chlorella and Phormidium were adjusted to contatn 7.5 x 10 and 12.5 x 10 celta/ml respectively. They were prepared by shaking the algae growth in the flasks, to which was added 100 ml of Allen's medium and blenderized for one minute in aWaring * Blender. Cell populations were calculated from hemocytometer counts. y B. Preparation of Sample One ml aliquots of test samples handled as 100% active were diluted in distilled water and added to Allen's medium for a final concentration ranging from 1:5 to 1:10,000. These concentration, ; were achieved after the addition of 1 ml of the adjusted algae cell suspension for a final volume of 100 ml. The numbers of Chlorella and Phormidium cells used were 7.5 x 10% and 12.5 x 10* respectively. Six control flasks, two per species, were also prepared. All inoculated bottles were subcultured to fresh medium after one day incubation. Final readings for the presence of algal growth in all bottles were made fourteen days after inoculation. J 000550 Jodisticl BIO -TEST Laboraloris, Inc. 4 D IV. Results | The results of the growth examination and turbidimetric measure- `ment of all flasks inoculated with Chlorella pyrenocidosa and Phormidium inundatum used for the algicidal tests are presented in Table I and I respectively. ) : . " J 000581 Indushiok BIO-TEST Loboratonts, Ine. 5 ) TABLE I Results of the Gross and Turbidimetric [Examination of Chlorella pyrenoidosa y Algicidal Test - Sample "A" ete "Gross Appearance ------------ TT urbie dity t dilution) (color) (absorbance 1:5 1:10 : none none ' <o.01 <0.01 1:100 aso none . slight greentint _ <0.01 0.03 11::110,, 000000 green reen >2.00 >2.00 1:5 subculture 1:10 subculture none none: +. +1 <0.01 <o0.01 1:100 subculture none Control green <0.01 : >2.00 Sample "B" :) 1:5 none <0.01 1:10 none 1:100 none <o0.01 <o0.01 1:150 1:1,000 slight green tint green 0.03 >2.00 7 1:10,000 1:5 subculture green none >2.00 ; <0.01 1:10 subculture none 1:100 subculture none <0.01 <0.01 Control green ~ >2.00 ) o `Ssl1o,%192, EN , 29, fog, 000588. SidiBAiOt-TiESlTLokovatonsh, Ine 6 D . ' TABLE I Results of the: Gross and Turbidimetric Examination of Phorinidium inundatum Algicidal Test - Sample "A' (dilution) Gross Appearance color] `Turbidity (Absorbance] : 1:5 none <0.01 1:10 none <0.01. - 1.100 1:10,.000000 none 7 ggrreeeennttiinntt <0.01 - >2.00 52.00 1:5 subealtare none oar-- 1:10 subculture none <0.01 C1o:n1t0r0oslubculture gnroeneen <>02..0010 Sample "B" ) 1:5 none <0.01 1:10 none <0.01 1:100 1:1, 000 none green tint <>02..0010 11::150,su0b0c0ulture ngorneeen tint | <>o2..0010 " 111:11000ssuubcbuelatlutraere Control nnoonnee green << o0..0011 ! >2.00 rere---------------- 000583