Acetylcodeine as a marker of illicit heroin in human hair: Method validation and results of a pilot study

Acetylcodeine (AC), which is an impurity of illicit heroin synthesis, was suggested as a marker of heroin abuse. A procedure for simultaneous quantitation of 6-monoacetylmorphine (6-MAM), which is the major metabolite of heroin, morphine, codeine, and AC in hair was developed. Fifty-milligram hair samples were incubated in 0.01M HCl overnight at 60°C. The resulting hydrolyzed solutions were extracted by an automated solid-phase extraction procedure and drugs were analyzed by gas chromatography-mass spectrometry in selected ion monitoring mode (SIM). This required prior derivatization with propionic anhydride. Different validation parameters, such as linearity, intra-assay accuracy, extraction recoveries, and limit of quantitation, were described. Seventy-three hair samples from heroin abusers and 43 hair samples from subjects who had completed a heroin-maintenance program were analyzed. AC was detected in 92% of the first sample group and in only 12% of the second sample group. In the two groups, about 98% of AC-positive samples were found. These results prove that AC can be considered as a suitable marker of illicit heroin use, along with 6-MAM detection.