Nakahara Y, Kikura R, Takahashi K
National Institute of Health Sciences, Tokyo, Japan.
J Chromatogr B Biomed Appl. 1994 Jul 1;657(1):93-101. doi: 10.1016/0378-4347(94)80074-x.
A procedure for the detection and determination of 6-acetylmorphine (6AM) and morphine in rat and human hair is described. The efficiency of extraction of 6AM and morphine from hair was compared between five extraction methods using methanol, 0.1 M hydrochloric acid, methanol-5 M hydrochloric acid (20:1), helicase and methanol-trifluoroacetic acid (TFA) (9:1). Methanol-TFA was found to be the best solvent for extracting 6AM and morphine with minimum hydrolysis and maximum extraction efficiency of 6AM. The extraction rates of 6AM and morphine from heroin abuser's hair with methanol-TFA reached a plateau after 8-10 h. After the extraction from hair with deuterated internal standards added, the extract was purified by Bond Elut Certify in the usual manner. The 6AM and morphine were then derivatized with bis(trimethylsilyl)acetamide, which were characterized by GC-MS with electron impact ionization. Confirmation was accomplished by comparing their retention times and the relative abundances of major four selected ions with those of the standards. Quantification was based on the deuterated internal standards. Linearity was obtained in the concentration range 0.1-50 ng/mg. The overall recoveries after solvent extraction and solid-phase purification were 95% for 6AM and 97% for morphine. The limit of detection for confirmation was 0.2 ng/mg. The coefficients of variations for 6AM and morphine at concentrations of 4 ng/mg were generally less than 8%. 6AM was stable during extraction with methanol-TFA (9:1) at room temperature. This method was applied to the elucidation of heroin drug history by using hair sectional analysis.
描述了一种检测和测定大鼠及人毛发中6-乙酰吗啡（6AM）和吗啡的方法。比较了使用甲醇、0.1M盐酸、甲醇-5M盐酸（20:1）、解旋酶和甲醇-三氟乙酸（TFA）（9:1）的五种提取方法从毛发中提取6AM和吗啡的效率。发现甲醇-TFA是提取6AM和吗啡的最佳溶剂，水解最少且6AM的提取效率最高。用甲醇-TFA从海洛因滥用者毛发中提取6AM和吗啡的提取率在8-10小时后达到平稳状态。在添加氘代内标从毛发中提取后，提取物按常规方法用Bond Elut Certify进行纯化。然后6AM和吗啡用双（三甲基硅基）乙酰胺进行衍生化，通过电子轰击电离的气相色谱-质谱联用仪进行表征。通过将它们的保留时间和四个选定主要离子的相对丰度与标准品进行比较来完成确证。定量基于氘代内标。在0.1-50 ng/mg的浓度范围内获得线性关系。溶剂萃取和固相纯化后的总回收率6AM为95%，吗啡为97%。确证的检测限为0.2 ng/mg。6AM和吗啡在4 ng/mg浓度下的变异系数通常小于8%。6AM在室温下用甲醇-TFA（9:1）萃取期间稳定。该方法通过毛发分段分析用于阐明海洛因用药史。
