Analytical and Bioanalytical Chemistry

, Volume 407, Issue 12, pp 3417–3423 | Cite as

Rapid and sensitive LC–MS–MS determination of 2-mercaptobenzothiazole, a rubber additive, in human urine

  • Wolfgang GriesEmail author
  • Katja Küpper
  • Gabriele Leng
Research Paper


2-Mercaptobenzothiazole (MBT) is one of the most important vulcanization accelerators in the industrial production of rubber, especially car tires. Given its wide use in household articles and industrial rubber products it has a high potential to migrate into the environment. Humans can be exposed by dermal, oral, or inhalative routes. Incorporated MBT is excreted in urine, mainly as conjugates to glucuronide, sulfate, and mercapturic acid. On the basis of these facts MBT has been selected as a substance of high interest in the large scale 10-year German project on human biomonitoring (HBM); a cooperation between the German Federal Ministry for the Environment (BMUB) and the German Chemical Industry Association (VCI) with the objective of developing new analytical methods for relevant chemicals. The presented method was developed to determine MBT in human urine to reliably investigate the internal human MBT dose. Total MBT is measured after enzymatic hydrolysis followed by application of high-pressure liquid chromatography tandem mass spectrometry (HPLC–MS–MS) in positive-electrospray-ionization mode (ESI+) using isotope-dilution quantification. High sample throughput could be obtained by use of the column-switching technique. Optimization yielded an analytical method with a low and reproducible limit of detection (LOD) of 0.4 μg L−1 and a limit of quantification (LOQ) of 1 μg L−1, and low relative standard deviations in the range 1.6–5.8 %. A small biomonitoring study covering unexposed humans and occupationally exposed workers was performed to establish the feasibility and reliability of the method. MBT was found in only one urine sample from the unexposed humans, at a value of 10.8 μg MBT per liter, whereas it was found in all samples from the tested workers at values of up to 6210 μg MBT per liter.


2-Mercaptobenzothiazole MBT Urine Conjugates Excretion LC–MS–MS Biomonitoring 



The development of this method of analyzing MBT found in the human body (including the synthesis of the internal standard) was funded by the German chemicals industry. The method was developed during an ongoing 10-year project on human biomonitoring. The project is a cooperation agreed in 2010 between the Federal Ministry for the Environment, Nature Conservation, Building and Nuclear Safety (BMUB) and the Verband der chemischen Industrie e.V. (German Chemical Industry Association – VCI); it is administered by the Federal Environment Agency (UBA). Experts from government authorities, industry, and science accompany the project in selecting substances and developing methods.


  1. 1.
    Hanssen HW, Henderson ND (1991) A review of the environmental impact and toxic effects of 2-MBT. Prepared for Environmental Protection Division, B.C. Environment Victoria, BC, 31 pGoogle Scholar
  2. 2.
    Bundesanstalt für Arbeitsschutz und Arbeitsmedizin (2001) Benzothiazol-2-thiol (MBT): Accessed 10 Jun 2013
  3. 3.
    Bundesamt für Risikobewertung BfR (2008) Hilfsstoff zur Herstellung von Gummiluftmatratzen hat allergenes Potential. Stellungnahme Nr. 033/2008 des BfR, 6 pGoogle Scholar
  4. 4.
    Environmental Protection Agency (1994) Sodium and zinc salts of 2-mercaptobenzothiazole. EPA-738-F-94-024 Assessed 20 Jun 2014
  5. 5.
    Manninen A, Aurolia S, Vertiainen M, Liesivuori J, Turunen T, Pasanen M (1996) Determination of urinary 2-mercaptobenzothiazole (2-MBT), the main metabolite of 2-(thiocyanomethylthio)benzothiazole (TCMTB) in humans and rats. Arch Toxicol 70:579–584CrossRefGoogle Scholar
  6. 6.
    Nawrocki ST, Drake KD, Watson CF, Foster GD, Maier KJ (2005) Comparative toxicity evaluation of 2-(thiocyanomethylthio)benzothiazole and selected degradation products using Ceriodaphnia dubia. Arch Environ Contam Toxicol 48(3):344–350CrossRefGoogle Scholar
  7. 7.
    Fukuoka M, Satoh M, Tanaka A (1995) Metabolism of 2-thiobenzothiazoles in the rat Urinary, fecal and biliary metabolites of 2-benzothiazyl sulfenamides. Arch Toxicol 70:1–9CrossRefGoogle Scholar
  8. 8.
    el Dareer SM, Kalin JR, Tillery KF, Hill DL, Barnett JW Jr (1989) Disposition of 2-mercaptobenzothiazole and 2-mercaptobenzothiazole disulfide in rats dosed intravenously, orally, and topical and in guinea pigs dosed topically. J Toxicol Environ Health 27(1):65–84CrossRefGoogle Scholar
  9. 9.
    OECD (2004) Emission scenario document on additives in rubber industry Assessed 24 Feb 2014
  10. 10.
    Fiehn O, Reemtsma T, Jekel M (1994) Extraction and analysis of various benzothiazoles from industrial wastewater. Anal Chim Acta 295:297–305CrossRefGoogle Scholar
  11. 11.
    Haroune N, Combourieu B, Besse P, Sancelme M, Kloepfer A, Reemtsma T, de Weaver H, Delort AM (2004) Metabolism of 2-Mercaptobenzothiazole by Rhodococuus rhodochrous. Appl Environ Microbiol 70(10):6315–6319CrossRefGoogle Scholar
  12. 12.
    European Commission (2005) Opinion on 2-Mercapaptobenzothiazole (sensitization only), SCCP/0883/05. Accessed 20 Jun 2014
  13. 13.
    Sorahan T, Pope D (1993) Mortality study of workers employed at a plant manufacturing chemicals for the rubber industry: 1955-86. Br J Ind Med 50:998–1002Google Scholar
  14. 14.
    Sorahan T, Hamilton L, Jackson JR (2000) A further cohort study of workers employed at a factory manufacturing chemicals for the rubber industry, with special reference to the chemicals 2-mercaptobenzothiazole (2-MBT), aniline, phenyl-ß-naphthylamine and o-toluidine. Occup Environ Med 57:106–115CrossRefGoogle Scholar
  15. 15.
    Sorahan T (2009) Cancer risks in chemical production workers exposed to 2-mercaptobenzothiazole. Occup Environ Med 66(4):269–273CrossRefGoogle Scholar
  16. 16.
    Sorahan T (2008) Bladder cancer risk in workers manufacturing chemicals for the rubber industry. Occup Med 58:496–501CrossRefGoogle Scholar
  17. 17.
    Dost A, Straughan TJK, Sorahan T (2007) A cohort mortality and cancer incidence survey of recent entrants (1982-91) to the UK rubber industry: findings for 1983-2004. Occup Med 57:186–190CrossRefGoogle Scholar
  18. 18.
    Agostini M, de Vocht F, van Tongeren M, Cherrie JW, Galea KS, Kromhout H (2010) Exposure to rubber process dust and fume since 1970s in the United Kingdom; influence of origin of measurement data. J Environ Monit 12:1170–1178CrossRefGoogle Scholar
  19. 19.
    de Vocht F, Vermeulen R, Burstyn I, Sobala W, Dost A, Teager D, Bergendorf U, Straif K, Swuste P, Kromhout H, on behalf of the EU-EXASRUB consortium (2008) Exposure to inhalable dust and its cyclohexane soluble fraction since the 1980s in the rubber manufacturing industry in the European Union. Occup Environ Med 65:384–391CrossRefGoogle Scholar
  20. 20.
    Ginsberg G, Toal B, Kurland T (2011) Benzothiazole toxicity assessment in support of synthetic turf field human health risk assessment. J Toxicol Environ Health A 74(17):1175–1183CrossRefGoogle Scholar
  21. 21.
    ECHA (2014) Justification for the selection of a candidate CoRAP substance. Assessed 12 Sep 2014

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Health Protection, Institute of BiomonitoringCurrenta GmbH & Co. OHGLeverkusenGermany

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