yzer :f::::::\\ \\.~::::::: FINAL REPORT "ACUTE ORAL TOXICITY IN RATS - FIXED DOSE" F R.1 Unreported

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1 grupo # yzer, ec. :f::::::\\ \\.~:::::::..... FNAL REPORT "ACUTE ORAL TOXCTY N RATS - FXED DOSE" Sponsor of Study: Address: Ecolyzer Protocol: Receipt of Subst. Test: Start of Trial: End of Trial: Emission Report: Test Substance: Declared Chemical Composition: Quantity (ml ou g): LotVal./Fab. Declared: Chemical Name of Subst. Test: Purity: Uniformity: Study nitiation Study Termination: GREENSUN ND. COM. DE PROD. LlMP. LTOA R. DA FONTE, NOVA CDADE RO DAS OSTRAS - RJ R 10/05/ /05/ /06/ /06/2011 DESENGORDURANTE NDUSTRAL GSF 200 Unreported 200, /04/ /04/2011 UNREPORTED UNREPORTED Liquid 16/05/ /06/2011 Operational unit: Laboratórios Ecolyzer Ltda. Rua Romão Puiggari, Vila das Mercês Phone/Fax: (Oxx11) ecolyzer@ecolyzer.com.br CEP: São Paulo - SP Study Director: André Luiz Figueiredo Júnior Biologist Rua Romão Puiggari, Vila das Mercês São Paulo - SP CEP: Page 1 of7 LABORATÓROS ECOLYZER LTDA. Rua Romão Puiggari, 898 Vila das Mercês- São Paulo- SP Brasil- CEP Tel. (55 11) ecolyzer@ecolyzer.com.br.

2 grupo "... ec ;f:::::~\\ yzer,,,. '. FNAL REPORT "ACUTE ORAL TOXCTY N RATS - FXED DOSE" RESUME The test was conducted Acute Oral Toxicity - Fixed Dose for rats to study the possible toxic effects of the test. The test substance used was pure and administered orally to rats from 8 to 12 weeks of age, at a dose of 2000 mg / kg body weight on the pretest. The animais were observed at the time that they had died behavioral changes, clinical signs and gross pathological findings. The definitive test was conducted with a dose of 2000 mg / kg P. V., in which the animais showed clinical signs of toxicity, which classifies the test substance in the toxicological class V. NTRODUCTON Average Oral Lethal Dose (DL500ral) is defined as the amount of a substance expressed in mg of product per kg of body weight required to kill 50% of a population of animais after a single oral administration within a short period of time. Acute oral toxicity constitutes the sum of adverse effects occurring in a short time after application of an oral single dose of the test substance. Dose is the amount of substance administered. The dose is expressed as weight of substance per unit weight of test animal (ex mg / kg). OBJECTVE The trial of acute oral toxicity to rats - a fixed dose is as purpose to evaluate the toxicological characteristics of a substance, providing information about the health risks of an acute oral exposure. MATERALS E METHODS Materiais and Equipment Digital Scale Glass of common laboratory Rigid Probe Disposable Syringe Page 2 of7 LABORATÓROS ECOLYZER LTDA. Rua Romão Puiggari, 898 Vila das Mercês- São Paulo- SP Brasil- CEP Tel. (55 11) ecolyzeneecolyzer.corn.br-

3 ~:""""'" yze r..... ec grupo,... f::::::~~\ \ ",,,... FNAL REPORT "ACUTE ORAL TOXCTY N RATS - FXED DOSE" Teste substance and dose levels. The test substance in liquid form was originally used pure. Pre test: concentration of 1000 mg / ml (2000 mg / kg). Definitive test: concentration of 1000 mg / ml (2000 mg / kg) Test System. Have been used albino rats (Rattus norvegicus) nulliparous and not pregnant Wistar, adult with aged 8 to 12 weeks at the beginning of the experiment, having 172 to of body weight. Condições de teste Test Conditions The animais were acclimated to laboratory conditions for at least five days before the start of the experiment. Ventilation were maintained with 10 to 15 air changes per room per hour, temperature between 19 and 23 o C, relative humidity between 30 and 70% and photoperiod of 12 hours light and 12 hours in the dark. The diet was constituted of commercial feed, supplemented with filtered water, both provided ad libitum. The animais were randomized into polypropylene boxes, covered by a metal grid and covered with wood shavings, with a maximum of five animais per cage. Methodology The animais were fasted nutritional approximately 12 hours (at night). After this stage were weighed individually, and then after the calculations to determine the doses used, the test substance was administered orally to animais in a single dose. The animais were observed daily for 14 days after application of the substance tested clinically, and we noted the onset, degree and duration of symptoms. These findings were based mainly on examinations of skin, hair, eyes, mucous membranes, system circulatory, respiratory and nervous, somatomotor activity and behavior. The animais were weighed at the beginning and end of the experiment. Survivors were euthanized and necropsied at 14 days of testing. The autopsy of ali animais in the test. was performed, with special attention to the macroscopic examination of organs heart, lung, liver, stomach, intestines, kidneys, spleen and adrenal glands. Pretest Firstly it held a pre-analysis, using an animal for each dose. The test is started in one of the following doses: 5, 50, 300 and 2000mg/kg of PV (body weight). From the result of the observed parameters during the test, such as mortality and clinical signs of toxicity, new dose is applied or will already be set to initial dose of the ultimate test Definitive Test For this second step are used five animais per dose. The test will start in one of the following doses; 5, 50, 300 and 2000mg/kg; from the results of the pretest. The parameters are observed in animal mortality and clinical signs of toxicity. From these observations the test item is classified according to the table of the GHS. (Annex 1) Page 3 of 7 LABORATÓROS ECOLYZER LTOA. Rua Romão Puiggari, 898 Vila das Mercês- São Paulo- SP Brasil- CEP Tel. (55 11) ecolyzeneecolyzer.corn.br-

4 ec ~:"""\' yzer f:::::::~\ grupo,... ' ". "...~~::::::::. FNAL REPORT "ACUTE ORAL TOXCTY N RATS - FXED DOSE" RESULT Classification of the test substance. The test substance was classified according to the table in Annex 1. This classification was performed according to the doses applied to animais and observed in these parameters. Number of Deaths Dose 1 - Pre test (2000 mg kg): O Dose 1 - Final (2000 mg kg): O Median lethal dose ( (DL~ - Female: > 2000 mg/kg (PV.). Confidence nterval to 95%. - Not calculated. Clinical signs. Dose 1 - Pre test: The animal not showed clinical signs of toxicity. Dose 1 - Definitive: The animal not showed clinical signs of toxicity. Pathological (macroscopic). Animais that eventually died: - Any animais died after administration of the test substance. Survivors: - Respiratory system: NN until the dose of 2000 mg/kg of PV. - Cardio vascular system: NN until the dose of 2000 mg/kg of PV. - Central Nervous System: not investigated _ - Digestive system: NN until the dose of 2000 mg/kg of PV. - Genitourinary system: NN until the dose of 2000 mg/kg of PV. - Other (liver and splee): NN until the dose of 2000 mg/kg of PV. Note: NN = Nothing noteworthy Page 4 of7 LABORATÓROS ECOLYZER LTOA. Rua Romão Puiggari, 898 Vila das Mercês- São Paulo- SP Brasil- CEP Tel. (55 11) ecolyzer@ecolyzer.com.br.

5 grupo ec yzer :f:::::~\\.. \\~:::::::.. '. FNAL REPORT "ACUTE ORAL TOXCTY N RATS - FXED DOSE" CONCLUSON The test substance showed median lethal dose (DLso) to white rats, greater than 2000 mg kg body weight when administered orally pure, which classifies the test substance in the toxicological class V. The animais showed clinical signs of toxicity after exposure to the substance test. FLE ANO STORAGE This test report was issued in two copies, one being sent to the Sponsor and one saved. The Raw Data, Study Plan, Final Report, comments relating to this study sample and the test substance are archived and available at Ecolyzer Laboratories Ltda. The records will be maintained during the period of five years and substance test within six months, both frem the date of entry of the test substance in the laboratory. OECLARATON, the undersigned, declare that this study represents a true and accurate record of results and meets the requirements of standard Nit-Dicla Rev Princípio das Boas Práticas de Laboratório BPL. NMETRO, Paqe 5 of 7 LABORATÓROS ECOLYZER LTDA. Rua Romão Puiggari, 898 Vila das Mercês- São Paulo- SP Brasil- CEP Tel. (55 11) ecolyzer@ecolyzer.com.br

6 grupo 't,,\.,,..., ec f::::::~~\yzer '"'.,\",~::::::::. FNAL REPORT "ACUTE ORAL TOXCTY N RATS - FXED DOSE" F R.1 DECLARATON, the undersigned, declare that this Final Report has been evaluated by the Unit for Quality Assurance and truthfully reflects the Raw Data and Study Plan. declare that audits and inspections were performed, as described below Process Audit Date of completion Critical phases of the study 06/10/09-23/10/09 23/10/09 23/10/09 Reporting date the Reporting date to nspections Audit i Date of. completion Study Director Manage Operational 11/11/09 11/11/09 06/06/ /06/ /06/2011 Operational Unit (Laboratory) 26/10/09-:30/10/0'9 Raw Data, Study Plans, Final i Report,t ~ ~:.1(j)u ~lanmion U it Manager Quality Assuran Ecolyzer e DECLARATON, the undersigned, declare that this study was conducted under our supervision, according to the procedures described herein. The results presented refer only to test the substance tested. Sampling is the responsibility of Sponsor. This document must not be reproduced. Page 6 of 7 LABORATOROS ECOLYZER LTDA. Rua Romão Puiggari, 898 Vila das Mercês- São Paulo- SP Brasil- CEP Tel. (55 11) ecolyzer@ecolyzer.com.br.

7 ec!::::::~~\yzer grupo ":,,\;. \".,\. "'. FNAL REPORT "ACUTE ORAL TOXCTY N RATS - FXED DOSE" F R.1 METHOD USED OECO, Guídelíne For Testíng Of Chemícals, Acute Oral Toxicity - Fixed Dose Procedure, Sectíon 4: Health Effects, 420,17/12/2001. Pág Annex 1: Classífícatíon table of the second acute toxícologícal GHS (Globally Harmonised Classification System- OECD). (> O - 5 mg/kg ) (> 5-50 mg/kg ) 111 (> mg/kg ) V (> mg/kg ) V (> 2000 mg/kg ) Page 7 of7 LABORATÓROS ECOLYZER LTDA. Rua Romão Puiggari, 898 Vila das Mercês- São Paulo- SP Brasil- CEP Tel. (55 11) ecolyzer@ecolyzer.com.br

FINAL REPORT "ACUTE ORAL TOXICITY IN RATS - FIXED DOSE" F R.1

FINAL REPORT ACUTE ORAL TOXICITY IN RATS - FIXED DOSE F R.1 grupo 4- ',.-,..., ec f::::::~~~yzer \.,\.. FNAL REPORT "ACUTE ORAL TOXCTY N RATS - FXED DOSE" Sponsor Address: of Study: Ecolyzer Protocol: Receipt of Subst. Test: Start of Trial: End of Trial: Emission

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