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A gas chromatography–mass spectrometry method was developed and validated for the simultaneous automated solid‐phase extraction and quantification of cotinine and  trans ‐3‐hydroxycotinine in human urine. Good linearity was observed over the concentration ranges studied ( R  2  > 0.99). The limit of quantification was 10 ng/mL for both analytes. The limits of detection were 0.06 ng/mL for cotinine (COT) and 0.02 ng/mL for  trans ‐3‐hydroxycotinine (OH‐COT). Accuracy for COT ranged from 0.98 to 5.28% and the precision ranged from 1.24 to 8.78%. Accuracy for OH‐COT ranged from −2.66 to 3.72% and the precision ranged from 3.15 to 7.07%. Mean recoveries for cotinine and  trans ‐3‐hydroxycotinine ranged from 77.7 to 89.1%, and from 75.4 to 90.2%, respectively. This analytical method for the simultaneous measurement of cotinine and  trans ‐3‐hydroxycotinine in urine will be used to monitor tobacco smoking in pregnant women and will permit the usefulness of  trans ‐3‐hydroxycotinine as a specific biomarker of tobacco exposure to be determined. © 2014 The Authors.  Biomedical Chromatography  published by John Wiley & Sons Ltd.

biomedical chromatography

Simultaneous determination of cotinine and  trans ‐3‐hydroxycotinine in urine by automated solid‐phase extraction using gas chromatography–mass spectrometry