A simple and rapid method for nicotine assay by hplc from cutaneous microdialysis samples

A simple reversed-phase high-performance liquid chromatographic method with UV detection has been developed for the quantification of nicotine in cutaneous microdialysis studies. Nicotine was analyzed using 5 mum Lichrospher(R) 60 RP-18 Select B column (125 x 4 mm i.d.). The mobile phase used was water-methanol-0.1 M buffer acetate (pH=4.5)-acetonitrile-acetic acid (74:3:20:2:1 v/v), adjusted to pH=4.2 with diethylamine at a flow-rate of 0.7 ml/min. Detection was carried out at 254 mn at room temperature. The limit of quantification was 20 ng/ml; linearity was obtained from 50 to 5000 ng/ml. The intra- and inter assay coefficient of variation was less than 5%. The present method was applied to analysis of nicotine in in vivo cutaneous microdialysis samples.