ANALYSIS OF OPIATES IN HUMAN HAIR BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY|
Pichini S., Altieri I., Pellegrini M., Pacifici R., Zuccaro P.
Clinical Biochemistry Department, National Institute of Health, Rome, Italy
An high-performance liquid chromatographic method with ultraviolet detection has been developed for the determination of the principal opiates (heroin, monoacetylmorphine, morphine, codeine) in human hair. An aliquot of 100 mg of hair was treated with 2 ml HCL 0.1N at 56° C overnight; it was then extracted by solid-phase extraction using Bond Elut Certify columns with the procedure provided for the opiates. Chromatography has been performed using a C18 reversed-phase column with a mobile phase consisting of water-acetonitrile (70:30 v/v) containing 0.001 M NaH2PO4 and 0.002 M sodium laurylsulphate at pH 2.1. The methodology was validated using reference material from National Institute of Standard and Technology consisting of human hair segments and powdered human hair soaked with opiates. Linearity of the method was obtained in the concentration range of 0.5-50 ng/mg hair for monoacetylmorphine , morphine and codeine, and of 10-200 ng/mg hair for heroin. Cocaine and benzoylecgonine did not interfere with the opiates in the chromatographic run, nor caffeine, nicotine and its metabolite cotinine, usually present in human hair. Using this methodology, hair from drug addicts were examined with a limit of quantitation of 0.5 mg morphine, codeine and monoacetylmorphine per mg hair and 10 ng heroin/mg hair.