Mulé S J, Casella G A
New York State DSAS Testing and Research Laboratory, Brooklyn 11217.
J Anal Toxicol. 1989 May-Jun;13(3):179-84. doi: 10.1093/jat/13.3.179.
We report a reliable, rapid, sensitive, and quantitative method for the N- and O-trimethylsilyl (TMS) derivatives of the diazolo- and triazolobenzodiazepines after enzymatic incubation (2 h) and liquid-liquid extraction of 0.5-mL urine volumes. Each analyte was identified by gas chromatography/mass spectrometry through the retention time for the total ion current (TIC) and selected ion monitoring (SIM) of four ion currents. Quantitation of the diazolobenzodiazepines was obtained by the ratio of the base peak ion of the TMS analyte to that of the internal standard bromazepam in the concentration range 50-500 ng/mL (1-10 ng injected into the GC/MS). The limit of detection (LOD) at p less than 0.01 was 50 ng/mL for all the diazolobenzodiazepines. The assay was not quite as sensitive for triazolobenzodiazepines (5-20 ng injected in the GC/MS). The extraction efficiency of the assay ranged from 75 to 92% for all the analytes. The coefficient of variation (CV) for the diazolobenzodiazepines ranged from 5.4 to 9.4% for within-day runs and from 11.1 to 13.9% for between-day runs. For the triazolobenzodiazepines the values were 3.8 to 18.9% for a single day and 3.4 to 19.9% between days. The selected ion current ratio for each analyte was determined for a single day and over a one-week period. There was no statistical difference in the ratios during this time. The confirmation of diazolobenzodiazepines in urine by this method was relatively easy after screening by the immunoassay technique. Identification of triazolobenzodiazepines appeared to be more difficult by both the screening technique and the GC/EI/MS analysis.
我们报告了一种可靠、快速、灵敏且定量的方法，用于测定经酶孵育（2小时）和对0.5毫升尿液进行液液萃取后的重氮并和三氮并苯二氮䓬的N-和O-三甲基硅烷基（TMS）衍生物。通过气相色谱/质谱法，依据总离子流（TIC）的保留时间以及四个离子流的选择离子监测（SIM）来鉴定每种分析物。重氮并苯二氮䓬的定量是通过TMS分析物的基峰离子与内标物溴西泮的基峰离子的比值来实现的，浓度范围为50 - 500纳克/毫升（注入气相色谱/质谱仪的量为1 - 10纳克）。所有重氮并苯二氮䓬在p小于0.01时的检测限（LOD）为50纳克/毫升。该测定法对三氮并苯二氮䓬的灵敏度稍低（注入气相色谱/质谱仪的量为5 - 20纳克）。所有分析物的该测定法萃取效率在75%至92%之间。重氮并苯二氮䓬日内运行的变异系数（CV）在5.4%至9.4%之间，日间运行的变异系数在11.1%至13.9%之间。对于三氮并苯二氮䓬，单日的值在3.8%至18.9%之间，日间的值在3.4%至19.9%之间。针对每种分析物的选择离子流比值在单日和一周时间内进行了测定。在此期间，这些比值没有统计学差异。通过免疫分析技术筛选后，用该方法确认尿液中的重氮并苯二氮䓬相对容易。通过筛选技术和气相色谱/电子轰击质谱（GC/EI/MS）分析鉴定三氮并苯二氮䓬似乎更困难。
