Black D A, Clark G D, Haver V M, Garbin J A, Saxon A J
Department of Veterans Affairs, Medical Center, Seattle, WA 98108.
J Anal Toxicol. 1994 Jul-Aug;18(4):185-8. doi: 10.1093/jat/18.4.185.
A solid-phase extraction and GC-MS confirmation method was developed for certain urinary diazolo- and triazolobenzodiazepines, including the metabolites of lorazepam, clonazepam, alprazolam, and triazolam. The latter two do not form benzophenones, and the others are not readily confirmed by conventional thin-layer chromatography or GC-MS techniques. Samples were hydrolyzed with glucuronidase at 37 degrees C, adjusted to pH 4.5, extracted with Bond Elut Certify columns, dried, and derivatized using BSTFA with 1% TMCS. Sample preparation time averaged 4 hours. A GC-MS selected-ion-monitoring acquisition method targeting retention time, molecular ion abundances, and qualifier ion ratios was used to determine positive results. The recovery of 7-NH2-clonazepam was 95%, and recoveries of alpha-hydroxyalprazolam, alpha-hydroxytriazolam, and lorazepam were greater than 66%. Linearity was demonstrated from 0.1 to 1.0 microgram/mL for each drug. Within-run CVs were less than 11%, and between-run CVs were less than 16%. Using this technique, we have been able to confirm suspected cases of abuse that had not been confirmed by previous techniques.
开发了一种固相萃取和气相色谱-质谱联用确证方法，用于检测某些尿样中的二氮杂卓类和三氮唑苯二氮卓类药物，包括劳拉西泮、氯硝西泮、阿普唑仑和三唑仑的代谢物。后两者不会形成二苯甲酮，而其他几种药物难以通过传统的薄层色谱法或气相色谱-质谱联用技术进行确证。样品在37℃下用葡萄糖醛酸苷酶水解，调节至pH 4.5，用Bond Elut Certify柱萃取，干燥，并用含1%三甲基氯硅烷的N,O-双(三甲基硅基)三氟乙酰胺进行衍生化。样品制备时间平均为4小时。采用气相色谱-质谱联用选择离子监测采集方法，以保留时间、分子离子丰度和定性离子比率为目标来确定阳性结果。7-氨基氯硝西泮的回收率为95%，α-羟基阿普唑仑、α-羟基三唑仑和劳拉西泮的回收率大于66%。每种药物在0.1至1.0微克/毫升范围内均显示出线性关系。批内变异系数小于11%，批间变异系数小于16%。使用该技术，我们能够确证先前技术未能确证的疑似滥用病例。
