Simultaneous determination of cotinine and trans-3'-hydroxycotinine in human serum by high-performance liquid chromatography.
A high-performance liquid chromatographic method with ultraviolet photometric detection has been developed for the quantitation of cotinine and trans-3'-hydroxycotinine in human serum. A solid-phase extraction procedure was performed for the analytes and the internal standard, N-ethylnorcotinine, before chromatography. The use of a 30-cm reversed-phase column and a mobile phase of water-methanol-0.1 M sodium acetate-acetonitrile (67:24.5:6.5:2, v/v), pH 4.3, prevented the co-elution of caffeine with cotinine. The limit of quantitation observed with this method was 5 ng/ml for both cotinine and trans-3'-hydroxycotinine. The present method proved useful for the determination of serum levels of these metabolites, correlating with nicotine daily intake.
Pichini
S
S
Clinical Biochemistry Department, Istituto Superiore di Sanità, Rome, Italy.
Altieri
I
I
Pacifici
R
R
Rosa
M
M
Ottaviani
G
G
Zuccaro
P
P
D002851:Chromatography, High Pressure Liquid; D003367:Cotinine / Q000031:analogs & derivatives* / Q000097:blood*; D006801:Humans; D009538:Nicotine / Q000008:administration & dosage; D010947:Plants, Toxic; D012907:Smoking / Q000097:blood; D013056:Spectrophotometry, Ultraviolet; D014026:Nicotiana
C001381:hydroxycotinine; D009538:Nicotine; D003367:Cotinine