Toluenesulphonamide

Endpoint summary

Additional information

All key studies for this endpoint addressing bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity were all negative for either o/p-TSA itself or via cross-reading from both its individual constituents p-TSA and o-TSA. Furthermore, substance has no structural alerts for mutagenicity, and a large database of performed studies indicate that neither p-TSA nor o-TSA possess genotoxic properties.

 

Discussion:

o/p-TSA consists of a mixture of p-TSA and o-TSA. Both these substances show no structural alerts for mutagenicity, and a large database of performed studies indicate that p-TSA and o-TSA do not have genotoxic properties.

 

The table below lists all available data, including from literature:

Type of test	Substance	Test system	Doses	Results	Reference
Bacterial test
Ames test (reverse mutation)	o/p-TSA	S. typhimuriumstrains TA98, TA100, TA1535, TA1537, TA1538;Saccharomyces cerevisiaeD4	1.0 µg to 1000 µg/plate	No mutagenic effects observed (+/- S9).	Proprietary: Litton Bionetics, 1978 (Ketjenflex 9R)
Ames test (reverse mutation)	o/p-TSA	S. typhimuriumstrains TA98, TA100, TA1535, TA1537, TA1538;Saccharomyces cerevisiaeD4	0.5 µ to 250 µg/plate	No mutagenic effects observed (+/- S9).	Proprietary: Litton Bionetics, 1976
Ames test (reverse mutation)	p-TSA	S. typhimuriumstrains TA98, TA100, TA1535, TA1537, TA1538;Saccharomyces cerevisiaeD4	1, 10, 100, 500, 1000 µg/plate	No mutagenic effects observed (+/- S9).	Proprietary: Litton Bionetics, 1978 (o-TSA)
Ames test (reverse mutation)	p-TSA	S. typhimuriumstrains TA98, TA100, TA1535, TA1537;Escherichia coliWP2 uvrA	0, 312.5, 625, 1250, 2500, 5000 μg/plate	No mutagenic effects (+/- S9).	MHW, Japan; cited by OECD (1994)
Ames test (reverse mutation)	p-TSA	S. typhimuriumstrains TA98, TA100, TA102	TA98 &100: 0, 33, 100, 333, 1000 and 2000 or 3333 µg/plate ; TA102: 0, 100, 333, 1000, 3333 and 10000 µg/plate	No mutagenic effects observed (+/- S9).	NTP, 2007
Ames test (reverse mutation)	p-TSA	S. typhimuriumstrains TA1530, TA1535, TA1538, TA98, and TA100	4 x 10-7 mol up to 4 x 10-2 mol/plate.	No direct mutagenic effects were observed (only +S9).	Poncelet et al. (1980)
Ames test (reverse mutation)	p-TSA	S. typhimuriumstrains TA98, TA100, TA1535, TA1537	Up to 18000 μg/plate	No mutagenic activity (+/- S9). Evaluations included testing in ZLM agar (similar condition Eckhardt, 1981)	Herbold (1981)
Ames test (reverse mutation)	p-TSA	S. typhimurium –TA98, TA100, TA1535, TA1537 and TA1538.	Doses of up to 12 mg/plate were used.	(+/- S9) The only positive (marginally) increase in revertants were found intester strain TA98, with S9 and only using ZLM agar, and not in VB agar.	Eckhardt et al. (1980)
Gene mutation assay	o-TSA	S. typhimuriumstrains TA98, TA100, TA1535, TA1537, TA1538;Saccharomyces cerevisiaeD4	Up to 1000 ug/plate	Negative (+/- S9)	Proprietary, Litton Bionetics, 20838, 1978
Ames test (reverse mutation)	o-TSA	S. typhimurium (TA98, TA100, TA1535, TA1537) E. coli (WP2uvr A)	Up to 5000 ug/plate	Negative (+/- S9)	MHW, Japan: 1999
Ames test (reverse mutation)	o-TSA	S. typhimurium (TA1535, TA100, TA98, TA1537)	Up to 1000 ug/plate	Negative (+/- S9)	Stolts et al.: 1977
Ames test (reverse mutation)	o-TSA	S. typhimurium (TA1530, TA1535, TA1537, TA1538, TA100, TA98)	Up to 4 x 10-2 mol/plate	Negative (+/- S9)	Poncelet et al.: 1979
Ames test (reverse mutation)	o-TSA	S. typhimurium (TA1535, TA100, TA1538, TA98, TA1537)	Up to 18000 ug/plate	Weakly positive: TA98 (+ S9) Negative: TA98 (-S9) and other strains (+/- S9)	Eckhardt et al.: 1980
Ames test (reverse mutation)	o-TSA	S. typhimurium (TA98, TA 100, TA 1535, TA1537)	Up to 18000 ug/plate	Negative (+S9)	Herbold: 1981
Ames test (reverse mutation)	o-TSA	S. typhimurium (TA1535, TA1538, TA98, TA100)	Up to 2500 ug/plate	Negative (+S9)	Ashby et al.: 1978
Non-bacterial in vitro test
forward mutations (TK locus)	p-TSA	L5 178Y mouse lymphoma cells	Up to 5000 µg/mL	(+/- S9) An increased frequency of mutations was only observed at the highest, cytotoxic, dose level with metabolic activation, involving an increase in small colonies.	Proprietary. Covance, 2000
Chromosomal aberration	p-TSA	CHL cells	Without S9: 0, 0.33, 0.65, 1.30 mg/mL; with S9: 0, 0.43, 0.85, 1.70 mg/mL.	No chromosomal aberrations (+/- S9).	MHW, Japan; cited by OECD (1994)
Chromosomal aberration	p-TSA	CHO-K1 cells	Cells were treated with 14, 200, and 400 μg/mL p-TSA for 24 h.	(- S9) Negative.	Masubuchi (1978a abstr)
Development of ouabain resistance	p-TSA	RSa human cell line	900 and 1800 μg/mL [5.26 and 10.51 mM], for 24 h	(- S9) No increase in ouabain resistance was observed inp-TSAtreated cells, relative to controls.	Suzuki and Suzuki (1988)
forward mutations (TK locus)	o-TSA	mouse lymphoma L5178Y cells	Up to 1712 μg/mL	Negative (+/- S9)	Proprietary, TNO Triskelion, V20203/05, 2013
Chromosomal aberration test	o-TSA	CHL cells	Up to 3000 ug/mL	Negative (+/- S9)	MHW, Japan:1999
Ouabain-resistant mutation assay	o-TSA	Human RSa cells	Up to 1800 ug/mL	Negative (-S9)	Suzuki & Suzuki: 1988
Chromosomal aberration test	o-TSA	CHO-K1 cells	Up to 400 ug/mL	Negative (-S9)	Masubuchi et al.: 1978
In vivotest
Micronucleus assay	p-TSA	Mouse Crl:CD-1, adult, M and F	Oral 187.5, 375, 750, 1500 mg/kg bw	Negative	Proprietary. Covance, 2002
Micronucleus assay	p-TSA	Mouse Crl:CD-1, adult, only males	i.v. 80, 120, 160 mg/kg bw,	Negative	Proprietary. Covance, 2000
Micronucleus assay	p-TSA	Mouse, NMRI, adult, number n.p., M and F	p-TSA administered both orally 2 x 855 mg/kg and i.p. 2 x 428 and 2 x 855 mg/kg	Negative	Eckhardt et al. (1980)
Mammalian spot test	o-TSA	Mouse	1000 mg/kg bw (oral)	Inconclusive	Fahrig: 1982
Micronucleus assay	o-TSA	Mouse	1026 mg/kg bw (by gavage)	Negative	Eckhardt et al.: 1980
Micronucleus assay	o-TSA	Mouse	Up to 1026 mg/kg bw (i.p.)	Negative	Eckhardt et al.: 1980
SLRL (Sex-linked recessive lethal) test	p-TSA	D. melanogaster(Berlin K [wild-type] and Basc strains)	Conc. 2.5 mM in 5% sucrose solutions used as feed for 3 days.	A small but statistically significant increase in the frequency of recessive lethal mutations was observed only in the first brood, corresponding to mature sperm. (about double the control frequency)	Eckhardt et al. (1980)
SLRL (Sex- linked recessive lethal) test	p-TSA	D. melanogaster(Oregon-K [wild type] and Basc strains)	Adult flies injected abdominal with about 0.2 μL of a 5 mM solution (~0.170 μg)	There were no or marginal positive effects observed in first brood under the test conditions. (frequency was about 2.5 times higher than of the control)	Kramers (1977)

 

References:

o/p-TSA

·   Proprietary: Litton Bionetics (Jagannath, D. R. & Brusick, D.), 20838, 1978.(Ketjenflex 9R)

·   Proprietary: Litton Bionetics (Brusick, D.), 2547, 1976.

o-TSA

·   Proprietary: Litton Bionetics (Jagannath, D. R. & Brusick, D.), 20838, 1978.(o-TSA)

·   Proprietary: TNO Triskelion, V20203/05, 2013

·   Ashby, J. et al. (1978) Fd. Cosmet. Toxicol., 16, 95-103.

·   Eckhardt, K. et al. (1980) Toxicology Letters, 7, 51-60.

·   Fahrig, R. (1982) Mutat. Res., 103, 43-47.

·   Herbold, B.A. (1981) Mutat. Res., 90, 365-372.

·   Masubuchi, M. et al. (1978) Mutat. Res., 54, 242-243.

·   MHW, Japan (1999) Ministry of Health and Welfare, Toxicity Testing Reports of Environmental Chemicals, 7, 125-160.

·   Poncelet, F. et al. (1979) Fd. Cosmet. Toxcol., 17, 229-231.

·   Stolts, D. R. et al. (1977) J. Environ. Pathol. Toxicol., 1, 139-146

·   Suzuki, H. & Suzuki, N. (1988) Mutat. Res., 209, 13-16.

p-TSA:

·   Covance, 2000, Proprietary: in vitro-Mouse Lymphoma, report# 21198-0-431-ICH

·   Covance, 2000, Proprietary: in vivo-micronucleus, report# 21198-0-455OECD

·   Covance, 2002, Proprietary: in vivo-micronucleus, report# 3464-0-455OECD

·   Eckhardt K., M.T. King, E. Gocke, and D. Wild. 1980. Mutagenicity study of Remsen-Fahlberg saccharin and contaminants. Toxicol Lett 7:51-60

·   Herbold B.A. 1981 . Studies to evaluate artificial sweeteners, especially Remsen-Fahlberg saccharin, and their possible impurities, for potential mutagenicity by the Salmonella mammalian liver microsome test. Mutat Res 90:365-372.

·   Kramers P.G. 1977. Mutagenicity of saccharin in Drosophila: The possible role of contaminants. Mutat Res 56:163-167

·   Litton Bionetics, 1978, Proprietary: in vitro-Ames, report# 20838

·   Masubuchi, M., S. Nawai, M. Hirokado, and K. Hiraga. 1978a abstr. Lack of the cytogenetic effects of saccharin impurities on CHO-K1 cells. Mutat. Res. 54[2], 242-243.

·   MHW, Japan; cited by OECD (1994) – Ames: Hatano Research Institute, Food and Drug Safety Center, Japan, Reverse Mutation Test of 4 -Methylbenzenesulfonamide on Bacteria (http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF70-55-3e.pdf)

·   MHW, Japan; cited by OECD (1994) – CHL: Hatano Research Institute, Food and Drug Safety Center, Japan, In vitro Chromosomal Aberration Test of 4-Methylbenzenesulfonamide on Cultured Chinese Hamster Cells (http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF70-55-3f.pdf)

·   NTP, 2007: The Salmonella Mutagenicity Test on p-Toluenesulfonamide, report A84238 (http://tools.niehs.nih.gov/ntp_tox)

·   Poncelet F., M. Mercier, and J. Lederer. 1980. Saccharin: Para forms of some impurities are not mutagenic inSalmonella typhimurium. Food Cosmet

·   Suzuki H., and N. Suzuki. 1988. Mutagenicity of saccharin in a human cell strain. Mutat Res 209:13-16.

Justification for selection of genetic toxicity endpoint
No one specific study is selected. All key studies for this endpoint addressing bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity were all negative.

Short description of key information:
All key studies for this endpoint addressing bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity were all negative for either o/p-TSA itself or via cross-reading from both its individual consituents p-TSA and o-TSA. Furthermore, substance has no structural alerts for mutagenicity, and a large database of performed studies indicate that neither p-TSA nor o-TSA possess genotoxic properties.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Both p-TSA and o-TSA have no structural alerts for mutagenicity, and available studies covering bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity all indicate that p-TSA and o-TSA do not have genotoxic properties. Consequently their mixture o/p-TSA does not need to be classified either.