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The digest was extracted with dichloromethane and then 25 m<jats:sc>m<\/jats:sc> hydrochloric acid in methanol was added to the extract, to prevent loss of analytes. The solution was evaporated and redissolved in the mobile phase, methanol\/10 m<jats:sc>m<\/jats:sc> ammonium acetate (30\/70, v\/v). A 20 \u00b5L aliquot of redissolved solution was subjected to analysis. Nicotine and cotinine in human hair were quanti\ufb01ed by using deuterated analytes as internal standards. The quanti\ufb01cation limits were 8 \u00b5g\/L for nicotine and 0.9 \u00b5g\/L for cotinine. The proposed method was applied to measure the concentrations of nicotine and cotinine in hair of smokers and non\u2010smokers to evaluate their self\u2010reported smoking and exposure to environmental tobacco smoke. In both cases, the method provided good selectivity, accuracy and precision. 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