Simultaneous determination of the tobacco smoke uptake parameters nicotine, cotinine and thiocyanate in urine, saliva and hair, using gas chromatography-mass spectrometry for characterisation of smoking status of recently exposed subjects

Abstract

A method using gas chromatography (GC)-mass spectrometry (MS) for the simultaneous determination of the smoke uptake parameters thiocyanate, nicotine and cotinine in human tissues is reported. Nicotine, cotinine and thiocyanate, in combination with a phase-transfer catalyst, were extracted from urine, saliva and hair into dichloromethane (DCM). Thiocyanate was alkylated in the DCM-layer to form a pentafluorobenzyl derivative. The biochemical markers in DCM were directly injected into the GC system and separated on a DB-1MS column using a 9.4 min temperature program. The method was validated in urine and saliva between the limits of quantitation (1.0-15 microg ml(-1) thiocyanate, 0.010-3.0 microg ml(-1) nicotine and cotinine in urine, 0.010-1.0 microg ml(-1) nicotine and cotinine in saliva). The calibration curves were found to be linear (r > 0.996), the within- and between-day accuracy's were 83-120%, the repeatability coefficients of variation were 3-20% and the limits of detection were 0.060 ng ml(-1) thiocyanate and 0.60 ng ml(-1) nicotine and cotinine. The results of the analysis of the biomarkers in the urine of 44 volunteers were used to develop a predictive model for smoking status, using discriminant analysis. The classification model correctly classified 93.2% of cross-validated grouped cases. Saliva samples were used to confirm the results of the classification method.

Method validation for measurement of hair nicotine level in nonsmokers

“…Hair nicotine concentrations have been assessed using various chromatographic techniques, including high-performance liquid chromatography (HPLC) with an ultraviolet detector (Pichini et al, 1997a), an electrochemical detector (Mahoney and Al-Delaimy, 2001), and mass spectrometry (MS, MS/MS) (Chetiyanukornkul et al, 2004;Kronstrand et al, 2004;Pirkle et al, 2006); and gas chromatography (GC) with a nitrogen-phosphorus detector or mass spectrometry (Kintz, 1992;Zahlsen and Nilsen, 1994;Torano and van Kan, 2003). Immunoassay techniques, such as enzymelinked immunosorbent assay, fluorescence immunoassay and radioimmunoassay, have also been used (Eliopoulos et al, 1994(Eliopoulos et al, , 1996Klein and Koren, 1999;Florescu et al, 2007).…”

GC–MS determined cotinine in an epidemiological study on smoking status at delivery

“…To summarise, these methods include radioimmunoassay (6), high-performance liquid chromatography (7,8), as well as gas chromatography-mass spectrometry (GC-MS) (9)(10)(11)(12)(13)(14).…”

Higher hair nicotine level in children compared to mother living with smoking father in Malaysia

“…The accuracies and precisions within-and between-assays were less than 11.4 % [23]. The quantifying ions used were m/z 162 for nicotine and m/z 169 for internal standard [16,24].…”

“…Thiocyanate, carbon monoxide and PAH are not specific to SHS since they can be influenced by other dietary or environmental sources [15,16]. Nicotine and its metabolite, cotinine are widely used as specific biomarkers to investigate the exposure to tobacco smoke and they can be measured in hair, urine, blood or saliva [17,18].…”