Rabbit/NZ C14 – C16 Non irritating Rabbit/NZ White C14 – C18 Olefin/Paraffin Non irritating Rabbit/NZ C14 – C18 Olefin Non irritating Rabbit/NZ White C14 – C18 Olefin/Paraffin Non irritating Rabbit/NZ White C14 – C26 Olefin Non irritating Rabbit/NZ C14 – C26 Olefin/Paraffin Non irritating Rabbit/NZ White C18 – C24 Olefins Non irritating Rabbit/NZ White C18 – C26 Olefin/Paraffin Non irritating Rabbit/NZ White C18 – C30 Olefin/Alcohol Non irritating 9.1.7 Skin sensitisation 9.1.7.1 Landsteiner technique Guineapig/albino C12 – C16 Olefin Non sensiting 9.1.7.2 Buehler technique Guineapig/Hartley White C16/C18 Alpha Olefins, isomerised Non sensitising 9.1.7.3 Magnusson and Kligman Guineapig/Dunkin Hartley C20 - 24 Alkenes, branched and linear Non sensitising 9.1.7.4 Other Alkenes 9.1.7.4.1 Buehler technique Guineapig/strain not specified Neodene 16 Alpha Olefin (1-hexadecene) Non sensitising 9.1.7.4.2 Guineapig/strain not specified Neodene 18 Alpha Olefin (1-octadecene) Non sensitising 9.1.7.5 Other Alkenes Repeated Patch Insult Test Neodene 18 Olefin (1-octadecene) Non sensitising 9.2 Repeat Dose Toxicity 9.2.1 2-week, dermal Rat/Fischer 344 Gulftene 12-16 = 1 000 mg/kg 9.2.2 Alkenes 4-week, dermal Rabbit/NZ White C16 - C18 Slight hyperemia, exfoliation, oedema
9.2.3 Up to 51 days, oral Rat/ Sprague Dawley Crl:CD BR VAF/Plus 1-tetradecene (blend from three suppliers) NOAEL = 1 000 mg/kg/day for reproductive neurotoxic effects; NOAEL = 100 mg/kg/day for systemic effects in females. No for males 9.2.4 13-week, oral Rat/ Crl:CD BR C20 - C24 Alkenes, Branched and Linear NOAEL = 500 mg/kg/day 9.3 Genetic Toxicity 9.3.1 in vitro 9.3.1.1 Bacterial reverse mutation S.typhimurium, E.coli C20 - C24 Alkenes, branched and linear Negative with & w’out activation 9.3.1.2 Unscheduled DNA Synthesis Rat hepatocyte primary culture Gulftene 12 – 16 Negative 9.3.1.3 BALB/3T3 Transformation Test Mouse embryo cells BALB/3T3-A31-1-1 Gulftene 12 – 16 Negative 9.3.1.4 Assay Chinese Hamster cell Gulftene 12 – 16 Negative with & w’out activation 9.3.1.5 Chromosome aberration Human peripheral lymphocytes C20-C24 Alkenes, branched and linear Negative with & w’out activation 9.3.1.6 Other Alkenes Bacterial Reverse Mutation Assay S.typhimurium, E.coli 1-Octadecene Negative with & w’out activation Mitotic Recombination Rat liver RL1 cells. 1-Octadecene Negative with & w’out activation Chromosome AberrationH18 Rat liver RL1 cells. 1-Octadecene Negative with & w’out activation 9.3.2 in vivo
9.3.2.1 Micronucleus Assay, intraperitoneal injection Mouse/ Crl:CD-1 (ICR) BR Gulftene 12 – 16 Negative 9.3.2.2 Micronucleus Assay, dermal administration Mouse/Crl:CD-1 (ICR) BR C20 - C24 Alkenes, branched and linear Negative
Clinical observations: A few days dosing, animals had very coarse oily fur over the whole body, particularly severe in the hindquarter area; Animals developed a weakness in the hindquarters which caused them to drag around the cage. This observation persisted for the remainder of the observation period. Mortality: Nil Morphological findings: Not reported. LD50: > 5 000 mg/kg Result: C16 Olefin was of very low acute oral toxicity in rats. 9.1.1.3 Oral Toxicity of C16 Isomerised Olefin (Hill Top Biolabs Inc 1993) Test Substance: C16 Isomerised Olefin Species/strain: Rat/Sprague Dawley Number/sex of animals: 5/sex Observation period: 14 days Method of administration: Oral, gavage 5 050 mg/kg (dose volume of 5.92 mL/kg) Test method: OECD TG 420 – fixed dose method Clinical observations: One female lost weight between Days 0 and 7 and one female failed to gain weight between days 7 and 14. Prominent in life observations included activity decrease piloerection and polyuria, which were no longer evident by Day 7. Alopecia was observed in all animals on Days 7 through 14. Mortality: Nil Morphological findings: No abnormalities detected. LD50: > 5 050 mg/kg Result: C16 Isomerised Olefin was of very low acute oral toxicity in rats. 9.1.1.4 Acute Oral Toxicity of C18 Isomerised Olefin (Stillmeadow Inc 1993) Test Substance: C18 Isomerised Olefin
Species/strain: Rat/Sprague Dawley Number/sex of animals: 5/sex Observation period: 14 days Method of administration: Oral, (gavage) 5 050 mg/kg (dose volume of 5.92 mL/kg) Test method: OECD TG 420 – fixed dose method Clinical observations: In life observations of note included diarrhoea, piloerection and polyuria, which were no longer evident by Day 11. Mortality: Nil Morphological findings: No abnormalities detected. LD50: > 5 050 mg/kg Result: C18 Isomerised Olefin was of very low acute oral toxicity in rats. 9.1.1.5 Acute Oral Toxicity of C20-24 Alkenes, branched and linear (Safepharm Laboratories Limited 1998) Test Substance: C20-24 Alkenes, branched and linear Species/strain: Rat/Sprague Dawley Crl:CD BR Number/sex of animals: 5/sex Observation period: 14 days Method of administration: Oral, gavage 5 000 mg/kg (dose volume of 6.29 mL/kg) Test method: OECD TG 420 – fixed dose method Clinical observations: Hunched posture and piloerection in all animals on day of dosing; Mortality: One female found dead one day after dosing Morphological findings: The female dead had haemorrhagic lungs, dark liver and dark kidneys; No abnormalities detected at terminal kill. LD50: > 5 000 mg/kg Result: C20-24 Alkenes, branched and linear was of very low acute oral toxicity in rats.
Method of administration: 10 000 mg/kg (dose volume 16 mL/kg) of test substance applied to an intact and abraded area of skin and held under semi occlusive dressing for 24 hours;. Observation period: 14 days Test method: US Federal Hazardous Substances Labelling Act. Mortality: Nil. Clinical observations: Skin became very taut, dry and scaly, with no regrowth of hair at the treatment site Morphological findings: No abnormalities detected at necropsy. LD50: > 10 000 mg/kg Result: C12-C16 Olefin was of low dermal toxicity in rats. 9.1.2.2 Acute Dermal Toxicity of C16 Alpha Olefin (Rinehart 1967) Test Substance: C16 Alpha Olefin (98.5% olefin,1.5% saturates) Species/strain: Rabbit/New Zealand Number/sex of animals: 4 males Method of administration: 10 000 mg/kg (dose volume 16 mL/kg) of test substance applied to an intact and abraded area of skin and held semi occlusive dressing for 24 hours. Observation period: 14 days Test method: US Federal Hazardous Substances Labelling Act Mortality: One death due to broken neck Clinical observations: Skin became very taut, dry and with no regrowth of hair at the treatment site. Morphological findings: No abnormalities detected at necropsy. LD50: > 10 000 mg/kg Result: C16 Alpha Olefin was of low dermal toxicity in rats. 9.1.2.3 Dermal Toxicity of C16 Isomerised Olefin (Stillmeadow Inc 1993) Test Substance: C16 Isomerised Olefin
Comment: Very slight to well defined erythema which persisted to the Day 7 observation. Very slight to defined oedema which cleared by the 48 hour observation. Result: C16/C18 Alpha Olefins, Isomerised was slight to moderately irritating to the skin of rabbits. 9.1.5.7 Acute Dermal Irritation of C20-24 Alkenes, branched and linear (Safepharm Laboratories Limited 1998) Test substance: C20-24 Alkenes, branched and linear Species/strain: Rabbit/New Zealand White Number/sex of animals: 6 Observation period: 1, 24, 48, 72 and 96 post exposure Method of administration: A single semi occluded application of 0.5 mL of the neat test substance to intact skin; after 4 hours residual test substance was removed by gentle swabbing with liquid paraffin; A contralateral area of untreated skin served as the control site. Control sites were similarly swabbed. Test method: OECD TG 404 Dermal effects: No evidence of skin irritation was observed, all individual scores were zero. Mean group score (24, 48 & 72 hour observation): Erythema/eschar formation: 0.0 Oedema: 0.0 Result: C20-24 Alkenes, branched and linear was non irritating to the skin of rabbits. 9.1.5.8 Acute Dermal Irritation of Other Alkenes (HEDSET 1-Octadecene) The following data, unless otherwise indicated, are from HEDSET data sheets for 1-tetradecene 1-hexadecene and 1-octadecene. The full study reports were not provided in the submission. The HEDSET documents indicate these tests were not conducted in accordance with GLP or OECD or EC testing guidelines.
Day 28: same procedure as induction phase except 5% w/v test substance in mineral oil was applied to a previously non treated site. 2nd challenge: Day 35: same procedure as for 1st challenge except previously non exposed naïve animals were used. Grading of dermal responses occurred 24 and 48 hours post exposure. Challenge outcome: 1st challenge - number of animals exhibiting positive responses: 1st Challenge Test Animals Naïve Control Animals Concentration 24 hoursT 48 hoursT 24 hoursT 48 hoursT 5% 9*/19 10**/19 10*/19 9**/19 7*/10 3**/10 8*/10 2**/10 T time after patch removal. * number of animals exhibiting Grade +/- reaction, slight, patchy erythema. ** number of animals exhibiting Grade 1 reaction, slight but confluent, or moderate patchy erythema. Comment: Test animals had a slightly greater incidence of Grade 1 reactions compared to the naïve control animals, suggesting that sensitisation may have been induced. To confirm these results a 2nd challenge was conducted 7 days later using the same test animals and challenge Ten non previously exposed naïve animals were also concurrently treated with the test substance. 2nd challenge - number of animals exhibiting positive responses: 2nd Challenge Test Animals Naïve Control Animals Concentration 24 hoursT 48 hoursT 24 hoursT 48 hoursT 5% 9*/19 10**/19 11*/19 7**/19 5*/10 5**/10 3*/10 7**/10 T time after patch removal. * number of animals exhibiting +/- reactions. ** number of animals exhibiting 1 reactions. Comment: Following rechallenge the incidence of 1 reactions was similar between test and control animals. The study authors concluded that sensitisation had not been induced.
Number/sex of volunteers: 31 females, 5 Induction procedure: Each volunteer received 0.2 mL (25% dilution in mineral oil) of test substance in a semi occluded exposure of 24 hours on 3 alternate days for weeks (9 induction exposures). A rest period of 10 – 17 days followed. Sodium lauryl sulphate (0.1% in water) served as the control. Challenge procedure: Each volunteer received 0.2 mL (25% dilution in mineral oil) of test substance in a semi occluded patch exposure to a naïve site on the upper arm for 24 hours. Challenge Outcome: No reaction to the to the test substance was reported. Result: Neodene 18 olefin was reported non sensitising to human skin 9.2 Repeat Dose Toxicity 9.2.1 Repeated Dose Dermal Toxicity: 2 Week Using Gulftene 12-16 (Gulf Life Sciences Center 1983) Test Substance: Gulftene 12-16 Species/strain: Rat/Fischer 344 Number/sex of animals: 5/sex/group duration: Doses of 0, 1.0, 2.0 g/kg applied once a day for a 6 hour exposure period, for a total of 9 doses over a 2-week period Method of administration: 2.60 mL/kg of test substance applied to a clipped site of the back. An Elizabethan collar prevented animals interfering with the dose site. The vehicle was corn oil. Test method: OECD TG 410 Mortality: Nil Clinical observations: Body weight gains were depressed in the 2.0 g/kg group. No test related toxic signs or symptoms were observed. Dermal Reactions: Individual Draize scores not provided. 1 000 mg/kg group:
9.3.1.6 In Vitro Genotoxicity of 1-octadecene (EU 1995) The following data are taken from the HEDSET data sheet for 1-octadecene. The full study reports were not provided in the submission. The HEDSET documents indicate tests were not conducted in accordance with GLP or OECD or EC testing guidelines. 1-Octadecene was considered not mutagenic or clastogenic in the following test systems: Test Comment Result Bacterial Reverse Mutation AssayH18 S. typhimurium TA98, TA 100, TA 1535, TA 1537, TA 1538. E. coli WP2, WP2uvrA. 0.2 to 2 000 g/plate; with and without metabolic activation. Negative Mitotic RecombinationH18 S.cerevisiae JD1. 0.01 to 5.0 mg/mL; with and without metabolic activation. Negative Chromosome AberrationH18 Rat liver RL1 cells. 0 to 500 g/mL as acetone solution. With and without metabolic activation. Negative 9.3.2 In Vivo 9.3.2.1 Mammalian Erythrocyte Micronucleus Test Using Gulftene 12-16 (Gulf Life Sciences Center 1983) Test substance: Gulftene 12-16 Species/strain: Mouse/Crl:CD-1 (ICR) BR Number and sex of animals: 5/sex/group of administration: Test substance: 1 000, 2 500 or 5 000 mg/kg; Vehicle control: corn oil; Test and vehicle control administered dermally to shaven of mice once per day for 2 days; Positive control, cyclophosphamide 75 mg/kg, administered via intraperitoneal injection; Sampling schedule: Test and vehicle control animals were sacrificed 24 or 48 hours after last dosing;