Number and Sex of Animals Dose mg/kg bw Mortality I 5 per sex 600 0 II (Control) 5 per sex 0 0 LD50 > 600 mg/kg bw Signs of Toxicity No clinical signs were noted up to 2 hours after the initial administration of the test substance. After 4 hours, hypocinesia was observed in all the animals treated with the test substance. The general behaviour and body weight of the animals were not influenced by the treatment after 24 hours and to day 15. Effects in Organs Macroscopic examination of the animals sacrificed at the end of the revealed no abnormalities. The notified biopolymer is of low toxicity via the oral route up to maximum practicable dose tested. Rhone-Poulenc (1988a) 7.1.2. Acute toxicity – oral Notified biopolymer OECD TG 401 Acute Oral Toxicity. Species/Strain Mice/Swiss Crl CD-1 Vehicle Water Remarks – Method Statement of GLP. No significant deviations from standard protocol. The dose 600 mg/kg, corresponded to the maximum dose level it was technically possible to administer. The dose was given over a period of 5 hours in 3 equal fractions, each in a volume of 20 ml/kg. Group Number and Sex of Animals Dose mg/kg bw Mortality 1 5 per sex 600 0 Control 5 males and 5 females - 0 LD50 > 600 mg/kg bw Signs of Toxicity General behaviour and body weight of the animals were not influenced by the treatment. Effects in Organs Macroscopic examination of animals sacrificed at the end of the revealed no abnormalities. Remarks – Results The notified biopolymer is of low toxicity via the oral route. Rhone-Poulenc (1988b) 7.2. Acute toxicity – dermal Not determined. The biopolymer has a high molecular weight and is unlikely to pass biological membranes. Daily application of a 1% solution of xanthan gum for 15 days to rat skin produced no sign of irritation (JECFA (2006)). 7.3. Acute toxicity – inhalation Xanthan gum - analogue
The lowest No Observed Adverse Effect Level (NOAEL) for the analogue chemical was established as 250 mg/kg bw/day. JECFA (2006) 7.8. Genotoxicity – bacteria Notified biopolymer In house protocol – plate incorporation method. Species/Strain S. typhimurium: TA1535, TA1537, TA1538, TA98, TA100. Metabolic Activation System Liver fraction (S9 mix) from rats pretreated with Aroclor 1254 Concentration Range in Main Test a) With metabolic activation: 0 – 1000 µg/plate. b) Without metabolic activation: 0 - 1000 µg/plate. Vehicle Water Remarks – Method Maximum dose selected based on solubility limit of the test substance observed in the preliminary test. Deviations from OECD TG 471 Bacterial Reverse Mutation test: Strains E. coli WP2 uvrA, or E. coli WP2 uvrA (pKM101), or S. typhimurium TA102 not used. These strains can detect certain cross linking mutagens not detected by the other strains. 2-Aminoanthracene used as the sole indicator of the efficacy of the S9-mix. No other significant deviations. Statement of GLP. Metabolic Activation Test Substance Concentration (µg/plate) Resulting in: Cytotoxicity in Preliminary Test Cytotoxicity in Main Test Precipitation Genotoxic Effect Present > 1000 Test 1 > 1000 not stated negative (all concentrations) Test 2 > 1000 not stated negative (all concentrations) Absent > 1000 Test 1 > 1000 not stated negative (all concentrations) Test 2 > 1000 not stated negative (all concentrations) Remarks – Results The test substance did not cause a marked increase in the number of revertants per plate of any of the tester strains either in the presence or absence of activation. Positive controls confirmed the sensitivity of the test system. The notified biopolymer was non-mutagenic to bacteria the conditions of the test. Rhone-Poulenc (1990) 7.9. Genotoxicity – in vitro Not determined. An in vivo mouse micronucleus test has been conducted and therefore an in vitro chromosome aberration test is not required.