Zezulak M, Snyder J J, Needleman S B
Navy Drug Screening Laboratory, Great Lakes, IL.
J Forensic Sci. 1993 Nov;38(6):1275-85.
Analysis of the opiates, morphine and codeine, often proceeds by way of acid hydrolysis for release of the parent morphine from its glucuronide formed during metabolism. Following use, heroin is rapidly deacetylated to 6-monoacetylmorphine (6-MAM), which can be detected in the urine for a short time following injection of heroin. Only a small amount of 6-MAM may be further metabolized to morphine glucuronide. Thus, in general, the urine specimen has not been hydrolyzed prior to analysis for heroin, using a separate procedure from morphine and codeine. Simultaneous analysis of morphine, codeine, 6-MAM and heroin would be complicated by loss of identity between morphine and heroin when heroin converts to morphine following acid hydrolysis for removal of the glucuronide moiety from morphine glucuronide. Another significant problem in simultaneous analysis is the relative disparity in concentration between morphine/codeine and 6-MAM/heroin (which might be present in the urine specimen). In the proposed method of analysis, free morphine resulting from B-glucuronidase rather than acid hydrolysis of morphine glucuronide is derivatized with propionic anhydride to form dipropionylmorphine. Heroin that does not react with B-glucuronidase remains unhydrolyzed as the diacetylmorphine derivative. Some of the more exacting steps for the acid procedure are eliminated altogether making overall costs for the enzyme procedure comparable to those of the acid hydrolysis method. The enzyme reaction mixture is purified through a solid phase column system. The optimal conditions for concentration of enzyme, temperature of hydrolysis and pH are individually characterized for B-glucuronidase hydrolysis and the ions which identify the propionyl derivatives are characterized for the simultaneous analysis of morphine, codeine, 6-MAM and heroin.
对阿片类药物吗啡和可待因的分析，通常通过酸水解的方式进行，以从代谢过程中形成的葡糖醛酸吗啡中释放出母体吗啡。使用海洛因后，它会迅速脱乙酰化生成6-单乙酰吗啡（6-MAM），在注射海洛因后的短时间内可在尿液中检测到。只有少量的6-MAM可能会进一步代谢为吗啡葡糖醛酸。因此，一般来说，在对海洛因进行分析之前，尿液标本不会像对吗啡和可待因那样进行水解处理。当海洛因在酸水解以从吗啡葡糖醛酸中去除葡糖醛酸部分后转化为吗啡时，吗啡和海洛因之间的身份丧失会使吗啡、可待因、6-MAM和海洛因的同时分析变得复杂。同时分析中的另一个重大问题是吗啡/可待因与6-MAM/海洛因（可能存在于尿液标本中）之间浓度的相对差异。在提议的分析方法中，由β-葡糖醛酸酶而非吗啡葡糖醛酸的酸水解产生的游离吗啡用丙酸酐衍生化形成二丙酰吗啡。不与β-葡糖醛酸酶反应的海洛因仍以二乙酰吗啡衍生物的形式未被水解。酸法的一些更严格的步骤被完全省去，使得酶法的总体成本与酸水解法相当。酶反应混合物通过固相柱系统进行纯化。针对β-葡糖醛酸酶水解分别确定了酶浓度、水解温度和pH的最佳条件，针对吗啡、可待因、6-MAM和海洛因的同时分析确定了鉴定丙酰衍生物的离子。
