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Clinical Chemistry 45: 2164-2172, 1999;
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(Clinical Chemistry. 1999;45:2164-2172.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Analysis of Nicotine, 3-Hydroxycotinine, Cotinine, and Caffeine in Urine of Passive Smokers by HPLC-Tandem Mass Spectrometry

Tapani Tuomia,1, Tom Johnsson1 and Kari Reijula1

1 Finnish Institute of Occupational Health (FIOH), Indoor Air & Environment Program of the FIOH, Uusimaa Regional Institute, Arinatie 3A, 00370 Helsinki, Finland.
a Author for correspondence. Fax 358-9-5061087; e-mail tapani.tuomi{at}occuphealth.fi

Background: A method is described for the simultaneous analysis of nicotine and two of its major metabolites, cotinine and 3-hydroxycotinine, as well as for caffeine from urine samples. The method was developed to assess exposure of restaurant and hotel workers to environmental tobacco smoke.

Methods: The method includes sample pretreatment and reversed-phase HPLC separation with tandem mass spectrometric identification and quantification using electrospray ionization on a quadrupole ion trap mass analyzer. Sample pretreatment followed standard protocols, including addition of base before liquid-liquid partitioning against dichloromethane on a solid matrix, evaporation of the organic solvent using gaseous nitrogen, and transferring to HPLC vials using HPLC buffer. HPLC separation was run on-line with the electrospray ionization-tandem mass spectrometric detection.

Results: The detection limits of the procedure were in the 1 µg/L range, except for nicotine (10 µg/L of urine). Still lower detection limits can be achieved with larger sample volumes. Recoveries of the sample treatment varied from 99% (cotinine) to 78% (3-hydroxycotinine).

Conclusions: The method described is straightforward and not labor-intensive and, therefore, permits a high throughput of samples with excellent prospects for automation. The applicability of the method was demonstrated in a small-scale study on restaurant employees.




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