A rapid LC-MS-MS method for the determination of nicotine and cotinine in serum and saliva samples from smokers: validation and comparison with a radioimmunoassay method.
The development and validation of a rapid liquid chromatography (LC)-tandem mass spectrometry (MS-MS) method for determination of nicotine and cotinine in smokers' serum is described. The method is based on solid-phase extraction in a 96-well plate format and requires only 100 microL of serum. Using normal-phase chromatography, both analytes elute in less than 1 min, which permits high sample throughput applications. The calibrated range is 2-100 ng/mL nicotine and 20-1,000 ng/mL cotinine. For known samples, recovery is 95-116% for nicotine and 93-94% for cotinine. The method is extended to rat serum and human saliva (cotinine only) using partial validation techniques. When compared with an existing radioimmunoassay method in our laboratory, the LC-MS-MS method gives improved accuracy, precision, and sample throughput.
Byrd
G D
GD
Product Evaluation Department, R.J. Reynolds Tobacco Company, Winston-Salem, NC 27102, USA. byrdg@rjrt.com
Davis
R A
RA
Ogden
M W
MW
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D003367:Cotinine / Q000032:analysis* / Q000097:blood; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D009538:Nicotine / Q000032:analysis* / Q000097:blood; D011786:Quality Control; D011863:Radioimmunoassay; D051381:Rats; D015203:Reproducibility of Results; D012463:Saliva / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012907:Smoking / Q000097:blood / Q000378:metabolism*
D009538:Nicotine; D003367:Cotinine