Bis[[R-(R*,S*)]-β-hydroxy-α-methylphenethyl)methylammonium] sulphate

Endpoint summary

Additional information

Ames test

The substance was tested for its mutagenic potential based on the ability to induce point mutation in several strains of Salmonella typhimurium (TA 1535, TA 100, TA98, TA 97) in a reverse mutation assay. Strains were exposed to 0, 100, 333, 1000, 3333, and 10,000 µg/plate. An increase in the number of his+ revertants was not observed in the preincubation test either without S9 mix or after the addition of a metabolizing system. It was therefore concluded that the test substance is not mutagenic under these experimental conditions.

In addition, two bacterial reverse mutation studies performed with two different structural analogues of ephedrine sulphate are available. The mutagenicity of (-)-Norephedrin S krist has been tested with4 Salmonella typhimurium strains (TA 1535, TA 1537, TA 98, and TA 100) and one E. Coli strain (WP2 uvrA) in astandard plate test and a preincubation test (BASF 2000). The mutagenicity of (+)-Pseudoephedrin-HCl has been tested with 4 Salmonella typhimurium strains (TA 1535, TA 1537, TA 98, and TA 100) in a standard plate test and a preincubation test (BASF 1991). In none of these studies an increase in the number of revertants was observed either without S9 mix or after the addition of a metabolizing system.

In the study performed with ephedrine sulphate, Salmonella typhimurium strains TA 1535, TA 100, TA 98, and TA97 were used. In accordance with OECD Guideline 471, mutagenicity in bacteria should also be tested in Salmonella typhimurium strain TA102 or E.coli WP2. In the studies performed with the two analogues of ephedrine sulfate no mutagencity was observed in the different Salmonella strains and E.coli WP2. It was therefore concluded that these test substances are not mutagenic in bacteria and as a result ephedrine sulphate is considered to be not mutagenic to bacteria.

CHO sister-chromatid exhange

The substance was tested for its mutagenic potential based on the ability to induce sister-chromatid exchanges in Chinese hamster ovary cells. Cells were exposed to 6500, 7000, 7500, and 8000 µg/mL with S9 -mix and to 1000, 1250, 1490, 1740 µg/mL without metabolic activation. Cytotoxicity was observed at the highest concentration tested for both experiments with and without metabolic activation. No increase in the number of sister-chromatid exchanges was observed with metabolic activation. An increase in the number of sister-chromatid exchanges was observed without metabolic activation. However this increase was less than two-fold and only at one dose tested and therefore this effect is considered to be a weak evidence of activity.

CHO chromosomal aberrations

The substance was tested for its mutagenic potential based on the ability to induce chromosomal aberrations in Chinese hamster ovary cells. Two chromosomal aberrations tests have been performed. In experiment 1, cells to which the metabolizing system was not added were exposed to 0, 1490, 1740, 1990, 2490, 2760, and 3000 µg/mL, and cells to which the metabolizing was added were exposed to 0, 5600, 6000, 6400, 7000, 7600, and 8000 µg/mL. In experiment 2, cells to which the metabolizing system was not added were exposed to 0, 503, 1081, and 2325 µg/mL, and cells to which the metabolizing was added were exposed to 0, 1081, 2325, and 5000 µg/mL. Only in experiment 1 in the presence of S9-mix an increase in the percentage of cells with aberrations was observed (6400 and 6400 µg/mL). However the concentrations at which the effects were observed were much higher than the recommended ones in OECD Guideline 473. Therefore it is concluded that there is no evidence of induction of chromosomal aberrations by the test substance and its metabolites in Chinese hamster ovary cells.

Short description of key information:
Three different genetic toxicity in vitro studies are available, performed with two different cell lines. According to these studies ephedrine sulphate does not induce the number of revertants and the number of chromosomal aberrations. However a weak positive response is observed in the sister-chromatid exchange assay in the absence of metabolic activation. In addition two bacterial reverse mutation studies are available, performed with two different structural analogous of ephedrine sulphate. These two structural analogues: (-)-Norephedrin S krist, and (+)-Pseudoephedrin-HCl do not induce the number of revertants either without S9 mix or after the addition of a metabolizing system.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

The bacterial reverse mutation assay with ephedrine sulphate and two structural analogues were negative. In addition a chromosome aberration assay with ephedrine sulfate showed that ephedrine sulfate is not clastogenic. However a weak positive effect has been observed in a sister chromatid exchange assay. Therefore classification for genetic toxicity is accordance with Directive 67/548/EEC (DSD) and EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008 is not possible and additional data is necessary.