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Nicotine and metabolites determination in human plasma by ultra performance liquid chromatography–tandem mass spectrometry: a simple approach for solving contamination problem and clinical application

Nicotine and metabolites determination in human plasma by ultra performance liquid... A quantitative method using ultra performance liquid chromatography–tandem mass spectrometry is described for simultaneous determination of nicotine and its metabolites (cotinine and trans‐3′‐ hydroxycotinine) in human plasma. Aliquots of 0.25 mL of plasma specimens were used for analysis, and 3 analytes were extracted by liquid–liquid extraction. The main problem was blank plasma contamination with environmental nicotine. Activated charcoal was used to avoid this analytical interference. For optimized chromatographic performance, a basic mobile phase consisting of 0.2% ammonia in water (mobile phase A, pH10.6) and acetonitrile (mobile phase B) was selected. The analytes were separated on a 50 mm × 2.1 mm BEH C18 column, 1.7 μm particle size, and quantified by MS/MS using multiple‐reaction monitoring (MRM) in positive mode. The chromatographic separation was achieved in 3 min followed by 1.2 min of column equilibration. The calibration curves were linear in the concentration range of 10–1000 ng/mL with correlation coefficients exceeding 0.99. Within‐day precisions and between‐day precisions (CV, %) were <15 %. The accuracy expressed as bias was within ±15% for all analytes. The recovery values ranged from 50% to 97%. The ions used for quantification of nicotine, cotinine and 3‐OH‐cotinine were 166.9 > 129.7; 176.9 > 79.7; 192.9 > 79.7 m/z, respectively. The original blank sample preparation solved the problem of contamination in a cost‐effective and efficient way. The validated method has been routinely used for analysis of nicotine and metabolites and determination of hydroxycotinine/cotinine metabolic ratio. This biomarker seems to be interesting at predicting response of nicotine patch replacement therapies. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Fundamental & Clinical Pharmacology Wiley

Nicotine and metabolites determination in human plasma by ultra performance liquid chromatography–tandem mass spectrometry: a simple approach for solving contamination problem and clinical application

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Publisher
Wiley
Copyright
Copyright © 2015 Société Française de Pharmacologie et de Thérapeutique
ISSN
0767-3981
eISSN
1472-8206
DOI
10.1111/fcp.12132
pmid
26118829
Publisher site
See Article on Publisher Site

Abstract

A quantitative method using ultra performance liquid chromatography–tandem mass spectrometry is described for simultaneous determination of nicotine and its metabolites (cotinine and trans‐3′‐ hydroxycotinine) in human plasma. Aliquots of 0.25 mL of plasma specimens were used for analysis, and 3 analytes were extracted by liquid–liquid extraction. The main problem was blank plasma contamination with environmental nicotine. Activated charcoal was used to avoid this analytical interference. For optimized chromatographic performance, a basic mobile phase consisting of 0.2% ammonia in water (mobile phase A, pH10.6) and acetonitrile (mobile phase B) was selected. The analytes were separated on a 50 mm × 2.1 mm BEH C18 column, 1.7 μm particle size, and quantified by MS/MS using multiple‐reaction monitoring (MRM) in positive mode. The chromatographic separation was achieved in 3 min followed by 1.2 min of column equilibration. The calibration curves were linear in the concentration range of 10–1000 ng/mL with correlation coefficients exceeding 0.99. Within‐day precisions and between‐day precisions (CV, %) were <15 %. The accuracy expressed as bias was within ±15% for all analytes. The recovery values ranged from 50% to 97%. The ions used for quantification of nicotine, cotinine and 3‐OH‐cotinine were 166.9 > 129.7; 176.9 > 79.7; 192.9 > 79.7 m/z, respectively. The original blank sample preparation solved the problem of contamination in a cost‐effective and efficient way. The validated method has been routinely used for analysis of nicotine and metabolites and determination of hydroxycotinine/cotinine metabolic ratio. This biomarker seems to be interesting at predicting response of nicotine patch replacement therapies.

Journal

Fundamental & Clinical PharmacologyWiley

Published: Oct 1, 2015

References