Tabakspezifische Nitrosamine – Bestimmung von N‐Nitrosoanabasin, N‐Nitrosoanatabin, N‐Nitrosonornikotin und 4‐(Methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanol in Urin mittels LC‐MS/MS

Biomonitoring-Methode

Gerhard Scherer¹
Gerhard Gilch¹
Dominique Köhler¹
Wolfgang Völkel²
  Thomas Göen³
  Andrea Hartwig⁴
  MAK Commission<sup>5

1</sup> ABF Analytisch-Biologisches Forschungslabor GmbH, Goethestraße 20, 80336 München, Deutschland
² Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit (LGL Bayern), Pfarrstraße 3, 80538 München, Deutschland
³ Friedrich-Alexander-Universität Erlangen-Nürnberg, Institut und Poliklinik für Arbeits-, Sozial- und Umweltmedizin, Henkestraße 9–11, 91054 Erlangen, Deutschland
⁴ Institut für Angewandte Biowissenschaften, Abteilung Lebensmittelchemie und Toxikologie, Karlsruher Institut für Technologie (KIT), Adenauerring 20a, Geb. 50.41, 76131 Karlsruhe, Deutschland
⁵ Ständige Senatskommission zur Prüfung gesundheitsschädlicher Arbeitsstoffe, Deutsche Forschungsgemeinschaft, Kennedyallee 40, 53175 Bonn, Deutschland

Abstract

The working group “Analyses in Biological Materials” of the Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area developed and validated the presented biomonitoring method.

This analytical method permits the determination of tobacco‐specific nitrosamines (TSNA) in urine using liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). The parameters in question are N‐nitrosoanabasine (NAB), N‐nitrosoanatabine (NAT), N‐nitrosonornicotine (NNN) and 4‐(methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanol (NNAL). NNAL is a metabolite of 4‐(methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanone (NNK). Due to its sensitivity, this method is suitable for the detection of the aforementioned analytes in the urine of smokers. NNAL can also be quantified in the urine of passive smokers.

The analytes NAB, NAT, NNN and NNAL are present in urine in both free and glucuronidated forms. For the determination of the total TSNA level in urine, the glucuronides are cleaved by enzymatic hydrolysis and then the analytes are isolated and concentrated using solid phase extraction (SPE). Two sorbent materials are used for sample preparation via SPE, first a material based on molecularly imprinted polymers and then a mixed‐mode cation exchange polymer. Analysis is performed by LC‐MS/MS. Deuterated internal standards are used for calibration. Calibration standards are prepared in pooled urine obtained from non‐smokers and are processed in the same way as the samples to be analysed.

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