Alcohols, C6-24 and C6-24-unsatd., distn. residues

Endpoint summary

Additional information

Genetic toxicity in vitro:

A test of bacterial gene mutagenicity was conducted with the test item according to the OECD TG 471 as adopted 1983, with following Salmonella typhimurium strains: TA 98, TA 100, TA 1535, TA 1537 and TA 1538 (Henkel R 9300181, 1993). The test concentrations were 8, 40, 200, 1000 and 5000 µg/plate; Tween 80 in bidist. water was used as vehicle. Testing was based on the standard plate method and was conducted with and without S9 mix. Negative (buffer), solvent and positive controls were included. No mutagenic effect on bacteria was detected.

In a micronucleus test performed according to OECD TG 487 under GLP conditions the potential of the test substance (85.1% pure) to induce chromosomal damage in V79 cells was assessed (BASF 33M0276/12X184, 2012). Two independent experiments were performed with culture duplicates each. The concentration ranged from 11.5 to 5150 µg/mL (the evaluated experimental points were: 46.1, 92.2, 184.4 µg/mL without S9 mix and 1475.0, 2950.0, 5150.0 µg/mL with S9 mix) in the first experiment and from 20 to 500 µg/mL in the second experiment, which was only conducted in the absence of metabolic activation. Cyclophosphamide (+S9) and mitomycin C (-S9) were used as positive control substances (PC). The exposure duration was 4 h followed by and incubation period of 20 h. 1000 cells per culture were evaluated and the percentage of cells with micronuclei were reported. Cytotoxicity was observed from 368 µg/mL in experiment 1 and from 140 µg/mL in experiment 2 in the absence of metabolic activation and was expressed as proliferation index (PI). No precipitation but phase separation of the test item was observed at all applied concentrations. Since positive findings were observed in the absence of S9 mix, this experimental part was repeated with a top dose of 500 µg/mL (experiment 2). The number of micronucleated cells was statistically significantly higher than in the corresponding control at 92.2 µg/mL (2.65%) and 284.4 µg/mL (4.95%) in the first experiment (solvent control: 1.3%; PC: 3.3%). In the second experiment, the number of micronucleated cells was increased at 120.0 µg/mL (2.65%; solvent control: 1.1%, PC: 27.15%). Treatment in the presence of metabolic activation did not induce an increase in the number of micronucleated cells up to the limit concentration.

In conclusion, it can be stated that under the experimental conditions in this study, the test item induced micronuclei in V79 cells in vitro in the absence of metabolic activation. Therefore, the test item is considered mutagenic in this in vitro test system in the absence of metabolic activation, when tested up to the highest evaluable concentration.

In a gene mutation test performed according to OECD TG 476, the potential of the test substance (85.1% pure) to induce mutations at the HPRT locus was evaluated in V79 cells (BASF 50M0276/12X182, 2012). The test substance in DMSO as vehicle was applied at concentrations of 2.8, 5.6, 11.3, 22.5, 45.0, and 90.0 µg/mL (without S9 mix) and 5.6, 11.3, 22.5, 45.0, 90.0, and 180.0 µg/mL (with S9 mix) in the first experiment and at 1.3, 22.5, 45.0, 90.0, 135.0, and 180.0 µg/mL (without S9 mix) and 5.6, 11.3, 22.5, 45.0, 90.0, and 180.0 µg/mL (with S9 mix) in the second experiment. Ethylmethanesulphonate and 7,12-dimethylbenzanthracene were used as positive control substances. Exposure duration was 4 h (experiment 1 with and without S9 mix, experiment 2 with S9 mix) or 24 h (experiment 2 without S9 mix). Afterwards, cells were cultivated in growth medium for approx. 7 days and for another 7 - 10 days in selection medium supplemented with 6-thioguanine. Cytotoxicity (expressed as cloning efficiency; CE) was observed without metabolic activation at 46.9 - 3000 µg/mL following 4 h treatment and at 187.5 - 3000 µg/mL following 24 h treatment. Phase separation occurred from 93.8 µg/mL with and without S9 mix following 4 and 24 h treatment. No increase in the mutation frequency was observed at any test substance concentration compared to the vehicle control. Only at 22.5 µg/mL, the induction factor was 4.5 in the second experiment with metabolic activation, but as the increase was only slight and a single value without any dose-relation, the increase was not considered biologically significant. The positive controls gave the expected results.

In conclusion it can be stated that under the experimental conditions chosen in this study the test item did not induce gene mutations at the HPRT locus in V79 cells. Therefore, the test item is considered to be non-mutagenic in this HPRT assay.

For the homologue substance "Alcohols, C6-C24, distn residues" (CAS 102242-49-9), three in vitro genetic toxicity test are available which are considered suitable for read across. The bacterial mutagenic potential was investigated in the Ames test according to OECD 471 (Cognis (now BASF) TBD880157, 1988) using the five strains TA 1535, 1537, 1538, 98 and 100 with and without metabolic activation. Five test substance concentrations from 8 to 5000 µg/plate and adequate positive controls were applied in agar. No enhanced mutation rate in the S9 treated or untreated cells was observed in the Ames test. Therefore, the test substance was considered not to be mutagenic in bacteria. The substance was further assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster (Cognis (now BASF) C-0901283-4, 2010). The study was performed in two independent experiments with and without metabolic activation. Treatment duration was 4 or 24 hours. Precipitation was observed at concentrations equal or higher than 35.0 µg/mL. Cytotoxic effects occurred only in concentrations showing very strong test item induced precipitation. In both experiments, all mutant frequencies remained well within the historical range of solvent controls. The induction factor did not exceed the threshold of three times the corresponding solvent control. All controls were valid. Thus, the test substance did not induce gene mutations at the HPRT locus in V79 cells (with and without metabolic activation). Therefore, it is considered to be non-mutagenic in this HPRT assay. The test substance was also assessed for its potential to induce micronuclei in human lymphocytes in vitro in the absence and presence of metabolic activation by S9 mix according to the OECD TG 487 (Cognis (now BASF) C-0901283 -3, 2010). Two independent experiments were performed, both with and without S9, with exposure periods of 4 or 20 hours. The chromosomes were prepared 40 hours after start of treatment with the test item. In each experimental group two parallel cultures were analysed. 1000 binucleate cells per culture were scored for cytogenetic damage on coded slides. The highest treatment concentration in this study was chosen with regard to the solubility properties of the test item. While no cytotoxicity was observed, precipitation occurred at 13.6 µg/mL or higher. In both experiments, in the absence and presence of S9 mix, no biologically relevant increase in the number of cells carrying micronuclei was observed. All controls were valid. In conclusion, under the experimental conditions reported, the test substance did not induce micronuclei in human lymphocytes in vitro, when tested up to precipitating concentrations.

Genetic toxicity in vivo

No data on genetic toxicity in vivo is available. As there is a positive result in one of the in vitro genetic toxicity tests (micronucleus test without metabolic activation; BASF 33M0276/12X184, 2012), conduction of a test to evaluate the mutagenic potential in vivo is mandatory according to REACH Annex IX. As the test substance is only handled under strictly controlled conditions, exposure is considered negligible. Based on the strictly controlled conditions as well as on the 3R principle and animal welfare, further in vivo testing is not considered necessary.

Short description of key information:
Three in vitro tests are available to evaluate the mutagenic potential of the test substance. The first study was an Ames test conducted according to the OECD TG 471 (Henkel R 9300181, 1993) with a negative result. A micronucleus test performed according to OECD TG 487 (BASF 33M0276/12X184, 2012) revealed a positive result without metabolic activation. The gene mutation potential was assessed in a HPRT assay performed according to OECD TG 476 (BASF 50M0276/12X182, 2012), which was negative with and without metabolic activation.
Further data are available for a close analogue substance, "Alcohols, C6-C24, distn residues", (CAS 102242-49-9) that can be considered for read-across. In fact, "Alcohols, C6-C24, distn residues" (CAS 102242-49-9) was examined in an Ames test according to the OECD TG 471 (Cognis (now BASF) TBD880157, 1988), in the in vitro micronucleus test according to the OECD TG 487 (Cognis (now BASF) C-0901283-3, 2010) and in the HPRT test according to the OECD TG 476 (Cognis (now BASF) C-0901283-4, 2010). All three tests were negative.
No data on genetic toxicity in vivo is available.

Endpoint Conclusion: Adverse effect observed (positive)

Exposure based adaptation of information requirements:

According to REGULATION (EC) No 1907/2006, Annex IX and Annex X, in vivo mutagenicity testing may be omitted, if relevant human exposure can be excluded in accordance with Annex XI section 3. Furthermore and in accordance with section 3.2 (b) of Annex XI (as amended by Regulation 134/2009), testing for in vivo mutagenicity can be omitted when the substance is not incorporated in an article and the manufacturer can demonstrate and document for all relevant scenarios that throughout the life cycle strictly controlled as well as rigorously contained conditions as set out in Article 18(4)(a) to (f) (Regulation 1907/2006) apply.

 

Life-cycle stage(s) covered:

1.Production of distillation residues (PROCs 1, 8b)

2.Use of distillation residues in production of energy products (PROC 1, 3, 8b)

3.Use of distillation residues as renewable energy source (PROC 1, 8b, 16)

 

Classification:

H341: Suspected of causing genetic defects.

The substance is not harmful after single ingestion and single dermal application. Additionally, the substance is not considered to be corrosive to skin as well as the eyes. The substance is not a skin sensitizer. No effects occurred in the bacterial reverse mutation assay and the HPRT assay. Based on effects in the micronucleus test the test substance is considered to be mutagenic in the absence of a metabolic activation system.

 

Process description:

Saturated and/or unsaturated fatty acids or triglycerides are transferred to methyl esters by esterification/transesterification processes with Methanol. Methylesters are transferred to saturated and unsaturated fatty Alcohols by partial catalytic reduction with hydrogen. The Fatty alcohols are purified by vacuum distillation, delivering fatty alcohols in the distillate and the distillation residues, which are topic of this registration in the distillation sump.

After the destillation is completed, the distillation residue is transferred via a closed dedicated pipeline to a buffer tank, where the residue is stored at 40-50°C. Sampling is not required for this process step. The reactor can immediately be used for the next destillation.

The process is operated discontinuously at elevated temperature. The reactor is located outdoors. Due to the closed system, exposure to workers will not take place under normal operation conditions.

Thereafter, the distillation residue is transported from the storage tank to the filling station under strictly controlled conditions. Here, the residue is filled in a transport vehicle under strictly controlled conditions. Exposure to workers will not take place under normal operation conditions. Then, the residue is transported to the powerplant via street where it is blended with other residues or directly pumped into the powerplant under strictly controlled conditions.

The substance is used as fuel in powerplants in closed systems similar to mineral oil products.The use is dedicated to large powerplants industrial style with the confirmation of strictly controlled conditions similar to the use of on site resprective transported intermediates. Any widespread use by professional workers or consumers (e. g. in neighbourhood small block heat and powerplants) is a use advised against.

 

 

 

Rigorous containment measures:

The substance is manufactured and used under strictly controlled conditions over the entire lifecycle. Transport, storage tanks, reactors and processing equipment operate in fully closed systems.

Procedural and control technologies are used to minimise residual emissions/exposure as well as qualitative risk considerations:

Operational and technical conditions and measures affecting and controlling workers exposure, as well as personal protective equipment, such as goggles, chemically resistant gloves, and respiratory protection where potential exposure may occur as reported in the CSR are followed (see chapters 9 & 10).

On the basis of the described process conditions, testing of the distillation residue in an in vivo mutagenicity test was not performed since the criteria of exposure based adaptation of information requirements are met.

Justification for classification or non-classification

Based on the available data classification of "alcohols, C6-C24 and C6-C24 unsat., distn residues" (CAS 102242-48-8) for mutageniciy is warranted as "R 68" according to the EU Directive 67/548/EEC and as "category 2" according to the CLP regulation (EC) No. 1272/2008.