Zhu Yu, Tan Jia-Yi, Sun Yu-Qing
Department of Forensic Medicine, China Criminal Police College, Shenyang 110035, China.
Se Pu. 2002 Sep;20(5):394-7.
A highly sensitive method has been developed for the analysis of 7-aminonitrazepam (7-ANIZ), the major metabolite of nitrazepam, in urine by trimethylsilyl derivatization-gas chromatography/mass spectrometry. Urine samples were extracted with ethyl ether-ethyl acetate (99:1, volume ratio). The extracts were derivatized with N,O-bis (trimethylsilyl) trifluoroacetamide, and the total ion current chromatograms of derivatives were acquired. 7-ANIZ was identified by the relative abundance of major characteristic ions in the mass spectrum of its derivative and the retention time of the mass chromatogram peaks of these characteristic ions. Based on the mass chromatogram of the base peak ion, quantification was performed using 7-aminoclonazepam (7-ACLZ) as the internal standard. The extraction efficiency of 7-ANIZ was 82.8%. The linear range was 10 microg/L - 500 microg/L. The limit of detection was 1.2 microg/L and the limit of quantification was 3.5 microg/L. The recoveries were 94.7% - 103.5%, and the RSDs were 3.9% - 5.4%. 7-ANIZ in the urine sample excreted by the subject over 96 h period after oral administration of 10 mg nitrazepam was measured. It is demonstrated that the method can be applied to the forensic identification.
已开发出一种高灵敏度方法，通过三甲基硅烷基衍生化-气相色谱/质谱法分析尿液中硝西泮的主要代谢物7-氨基硝西泮（7-ANIZ）。尿液样本用乙醚-乙酸乙酯（体积比99:1）萃取。萃取物用N，O-双（三甲基硅烷基）三氟乙酰胺进行衍生化，并获取衍生物的总离子流色谱图。通过其衍生物质谱中主要特征离子的相对丰度以及这些特征离子质量色谱图峰的保留时间来鉴定7-ANIZ。基于基峰离子的质量色谱图，以7-氨基氯硝西泮（7-ACLZ）为内标进行定量。7-ANIZ的萃取效率为82.8%。线性范围为10μg/L - 500μg/L。检测限为1.2μg/L，定量限为3.5μg/L。回收率为94.7% - 103.5%，相对标准偏差为3.9% - 5.4%。测定了受试者口服10mg硝西泮后96小时内排出的尿液样本中的7-ANIZ。结果表明该方法可应用于法医鉴定。
