Determination of nicotine and cotinine in human plasma by liquid chromatography-tandem mass spectrometry with atmospheric-pressure chemical ionization interface.
Here we report a sensitive liquid chromatographic-tandem mass spectrometric (LC-MS-MS) method capable of quantifying nicotine down to 1 ng/ml and cotinine to 10 ng/ml from 1.0 ml of human plasma. The method was validated over linear ranges of 1.0-50.0 ng/ml for nicotine and 10.0-500.0 ng/ml for cotinine, using deuterated internal standards. Compounds were simply extracted from alkalinized human heparinized plasma with methylene chloride, reconstituted into a solution of acetonitrile, methanol and 10 mM ammonium acetate (53:32:15, v/v) after the organic phase was dried down, and analyzed on the LC-MS-MS, which is a PE Sciex API III system equipped with a Keystone BDS Hypersil CI8 column and atmospheric pressure chemical ionization (APCI) interface. The between-run precision and accuracy of the calibration standards were < or = 6.42% relative standard deviation (R.S.D.) and < or = 11.8% relative error (R.E.) for both nicotine and cotinine. The between-run and within-run precision and accuracy of quality controls, (2.5, 15.0, 37.5 ng/ml for nicotine and 25.0, 150.0, 375.0 ng/ml for cotinine), were < or = 6.34% R.S.D. and < or = 7.62% R.E. for both analytes. Sample stabilities in chromatography, in processing and in biological matrix were also investigated. This method has been applied to pharmacokinetic analysis of nicotine and cotinine in human plasma.
Xu
A S
AS
Harris Laboratories, Lincoln, NE 68502, USA.
Peng
L L
LL
Havel
J A
JA
Petersen
M E
ME
Fiene
J A
JA
Hulse
J D
JD
D002853:Chromatography, Liquid / Q000379:methods*; D003367:Cotinine / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D009538:Nicotine / Q000097:blood* / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D009538:Nicotine; D003367:Cotinine