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AR226-3025 TRADE SECRET Study Title EVALUATIONS THE BIODEG^ADABILITY OF USING THE CLOSED BOTTLE TEST Author John T. Gannon Jr., Ph.D. Study Completed on 8/18/95 Report Revised on 9/6/95 Performing Laboratory E. I. du Pont de Nemours and Company Central Research & Development Applied and Environmental Microbiology Laboratory Glasgow Business Community 301, P.O. Box 6101 Newark, DE 19714-6101 AEM Project No. 204 Laboratory Project ID AEM Laboratory Report No. 205-95 Page 1 of 8 Compaq S@ni8Kc8. Does not eontain TSCA Cll8 DuPont AEMR 205-95 GENERAL INFORMATION Material Tested Svnonvms/Codes: Submitter's NotehookNo.- N/A Sponsor: |_| FLPR -3 Experimental Start & Completion: 7-11-95/ 8-8-95 Gsmpassg Sanitized. Does not ontain TSCA C^| -2- DuPont AEMR 205-95 EVALUATION^)? THE BIODEGRADABILITY OF I----------------PTPHSEINCGLOSED BOTTLE TEST SUMMARY The test cheniical|JUUJBflRJ was tested for biodegradability using the 28-day Closed Bottle Test. The results of the 28-day Closed Bottle Test indicated that a peak of 12.1 % Biodegradability was observed at Day 28. A minimum of 60 % biodegradability is required to pass this test. In addition, the "pass level" of 60 % must be reached within a 14 day tha^UmmUP^was windpwrfter^xceedine^.he 10 % level. Therefore, it may be concluded not "Readily Biodegradable." Please note that failure to pass a "Ready Biodegradability Test" does not imply that the test material will persist in the environment for an extended period of time. Study Conducted by: Wallace E. Bessicks (date) Microbiology Research Technician Study Conducted by: ^^. (QA.--^. Nicole L. DiMondi ^/W^ (date) Senior Microbiology Technician Study Conducted by: Q.^^k s^iry George M. Fisher (ffJ^ (date) Senior Microbiology Technician Report Prepared and ^ Approved for Issue By: y^\^-- I. AJa/^^t'~-~~- y6/yj J&n T Gannon Jr., Ph.D.(date) Research Microbiologist .OfflpanySanitiBed. Does not eentain TSCA cat 3- DuPont AEMR 205-95 Introduction/Purpose The Closed Bottle Test was performed to determine if flHBHHH^Ijwas "Readily Biodegradable". The Closed Bottle Test can be used for either water-soluble or water-insoluble and volatile or non volatile chemicals. In this test, the Time-0 DO (Dissolved Oxygen) is measured and is considered the starting point. The Closed Bottle Test is a closed system. Therefore, no additional dissolved oxygen should be added to the system for the duration of the test. If the bacteria metabolize the test chemical in this closed system, then they will use up the dissolved oxygen. Consequently, biodegradation is measured as the loss of dissolved oxygen within the closed bottle. If the DO (Dissolved Oxygen) loss at Day 28 >. 60 %, and this level of biodegradability has been achieved within 14 days of exceeding 10 % biodegradability, then the test chemical is considered "Readily Biodegradable" and no further testing is necessary. Materials/Method SOlfHH^BmBBHB----------HBHf\ The Closed Bottle Test determination was conducted in accordance with AEM The source of the inoculum for the Closed Bottle Test was the City of Wilmington POTW. See Table 1 for materials tested. Results/Discussion In the 28-day Closed Bottle Test|JJJHBBfi^^howed a peak of 12.1 % biodegradabijjty at Day 28 (Tables^2 & 3 and Figure 1). Therefore, it was determined t-hat|H|UU|Um|fvas not "Readily Biodegradable". The control chemical. Sodium Acetate, exceeded 60 % biodegradability IjBHiBHU^vas .(Figure 1), therefore^the test is valid. The Theoretical Oxygen Demand of calculated as 0.66 mg Oz per mg of active substance. Please note that failure to not imply that the test material extended period of time. pass 'a "Ready wilt persist in Biodegradability the environment Test" does for an . Does not eontain TSCA CBj -4- DuPont AEMR 205-95 JEVALUATIQMsQF THE BIODEGRADABILITY OP JSING THE CLOSED BOTTLE TEST TABLE 1 M-Namber 51000 46411 Description of Test and Control Substances Test Substance DescriptioiL 0 3 Sodium Acetate Gom^-my Sanitized. QQSS not eQatora TSCA CB| -5- DuPont AEMR 205-95 DuPont AEMR 205-95 % BIODEGRADABILITY 8- CHEM MOL FOR MOL WT :' J Linear Model Prediction : Does Hot Biodegrade Fast Non-Linear Model Prediction; Does Not Biodegrade Fast Ultimate Biodegradation Timeframe: Recalcitrant Primary Biodegradation Timeframe: Months TYPE -+j--N--UM--4._______-_------ __----.------.-. ------h.=----rFrag Frag MolWt| Const =+= RESULT folecular Weight Parameter Equation Constant LINEAR BIODEGRADATION PROBABILITY COEFF VALUE -+- -0.1839 -0.5204 -0.9196 -0.5204 -0.2048 0.7475 =+= -0.8973 |a A Probability Greater Than or Equal to 0.5 indicates --> Biodegrades Fast y A Probability Less Than 0.5 indicates --> Does NOT Biodegrade Fast Result Classification: (Primary & Ultimate) 5.00 -> hours 2.00 -> months 4.00 -> days 3.00 -> weeks 1.00 -> longer-