Document zQYG45M5Rq7gKeNeL57MkwRxz

AR226-2967 Study Sponsor: ATOCHEM Issue Date: Unavailable (Data Only) Document Reference No.: Bacterial Growth Inhibition Author(s): J.C . Boutonnet A.M . Petit English Translation of Report Summary for DuPont Chemical Solutions Enterprise 27-Jan-03 . c TMMnySanteed0oes contain TSCa CBt ATOCHEM Levallois Applications Center Analysis - Environmental Services Eco-toxicology Laboratory TRANSLATION TEST REPORT DETERMINATION OF AN INHIBITING EFFECT ON BACTERIAL PSEUDOMONAS PUTIDA CELL GROWTH according to proposed standard ISO TC147/SC5/WG1/N111 Substance studied : Internarsample reference : Person responsible for te s t: J. C. BOUTONNET Person carrying out te s t: A. M. PETIT Date test started : Date report written : 22.10.91 Sample preparation Mother solution at 20 g/l in ultra-pure water Organism tested Pseudomonas putida MIGULA, STRAIN Berlin 33/2 (DSM 50025) Test conditions Equipm ent: Sterilized glassware Perkin-Elmer spectrophotometer Lamda 5 Thermostatically controlled agitator at 21 1 C Concentration range (each concentration is tested in trip lica te ): Voi. Substance solution (ml) 15 10 5 2.5 Voi. W ater (ml) 65 70 75 77.5 Voi. Medium (ml) 10 10 10 10 10 Voi. Inoculum (ml) 10 10 10 10 10 Substance concentration (g/l) 3 2 1 0.5 10 10 The flasks are incubated in a therm ostatically controlled agitator Company Sanitized. Does not contain TSCACBi The culture Is started on 22/10/91 at 16 h and stopped on 23/10/91 at 8 h 30 Incubation duration : 16 h 30 The turbidity is measured using a spectrophotometer by determination of the optical density at a wavelength of 436 nm (ISO standard 7027 -1984) after dilution : 10 before incubation Results Dilution at 1/10 to effect the spectrophotometer reading Substance concentration (mg/l) (Optical density of control = 0.0037) 3 2 1 05 OD in flask 12 0.151 0.150 0.247 0.202 0.241 0.277 0.412 0.427 3 0.154 0.312 0.267 0.414 Turbidity B 12 3 average = 4047ml average = 5877ml average = 7157ml average = 11477ml 10 flasks average = control O. D. = 0.019,1/10 5 average 0.228 Using the equation H = ((Bk - Bn)/(Bk - B0))x100 where H is percent cell growth inhibition H Bk is the turbidity in the control flasks Bn is the turbidity in the test flasks (after deduction of non-inoculated whites) B0 is the turbidity at t = 0 in the control flasks. We obtain: Substance concentration (mg/l) 3 2 1 0.5 H% 67.9 % 53% 42.6 % 7.9 % By drawing the logarithmic Gaussian curve (attached) we obtain : CE 10 = g/l CE 50 = g/l 0.47 g/l 1.75 g/l By calculation (linear regression) we obtain : CE 10 = CE 50 = 0.466 g/l 1.76 g/l Additional re su lts: The pH was measured at the start and end of the te s t: At 3 mg/l, at t = 0, pH = 7.03; at t = 16 h 30, pH = 7.10 At 0 mg/l, at t = 0, pH = 6.99; at t = -- h, pH = 7.06 .Company Sanitized. Does not contain TBCA Validity clauses : The test was carried out using the protocol described in docum ent: ISO TC147/SC5/WG1/N111 During the test, cell growth in the control series occurred at a factor o f: 12 (BO = 10 and Bk = 1299) The reference substance, 3,5-dichlorophenol gave under the test conditions : CE 50 = 8.55 mg/l Ce 10 = 0.427 mg/l T Company Sanitized. Does not contain TSCA CBI