Shanghai Key Laboratory of Forensic Medicine, Department of Forensic Toxicology, Institute of Forensic Science, Ministry of Justice, Guangfu Xi Road 1347, Shanghai 200063, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Dec 12;911:140-6. doi: 10.1016/j.jchromb.2012.10.018. Epub 2012 Oct 31.
Zolpidem (ZPD) is an imidazopyridine derivative used as a new type of hypnotic and is commonly used in drug-facilitated crimes. A rapid, sensitive, and specific ultra-performance liquid chromatography-tandem mass spectrometry (UPLC/MS/MS) assay method for the simultaneous determination of zolpidem and its main metabolites zolpidem 6-carboxylic acid (ZCA) and zolpidem phenyl-4-carboxylic acid (ZPCA) in biological fluids was developed and validated. Aliquots of 0.1mL blood or urine specimens were used for the analysis, and zolpidem and its metabolites were extracted in a single step using acetonitrile (containing 0.1% formic acid) precipitation. The supernatant was then dried, and 100μL methanol was added. The separation was performed on an Acquity™ UPLC HSS T3 (100mm×2.1mm, 1.8μm) analytical column by API 4000Q ultra-performance liquid chromatography-tandem mass spectrometry. Positive ionisation tandem MS detection in the multiple reaction monitoring (MRM) mode was used. The mobile phases consisted of either acetonitrile or water containing 20mmol/L ammonium acetate and 0.1% formic acid, and the flow rate was 0.5mL/min. The chromatographic separation time was 4min, and calibration curves for human blood were generated over the range of 0.1-300ng/mL for ZPD, 0.1-500ng/mL for ZPCA and 0.1-200ng/mL for ZCA. For urine, the linear range was 0.1-600ng/mL for ZPD and ZPCA, and 0.1-300ng/mL for ZCA. The limit of detection was 0.05ng/mL and the limit of quantitation was 0.1ng/mL for ZPD, ZCA and ZPCA. The linear correlation coefficients were greater than 0.9995. Both the inter-day and intra-day precisions were less than 15%, the recoveries were in the range of 70.0-98.3%, the matrix effects were approximately 79.5-99.0%, and the process efficiency was between 60.7% and 94.4%. This method allowed for the determination of zolpidem and its metabolites in human blood and urine and may be applied to forensic toxicological analyses.
唑吡坦（ZPD）是一种咪唑并吡啶衍生物，用作新型催眠药，常用于药物辅助犯罪。本研究建立并验证了一种用于同时检测生物体液中唑吡坦及其主要代谢物唑吡坦 6-羧酸（ZCA）和唑吡坦苯-4-羧酸（ZPCA）的超高效液相色谱-串联质谱（UPLC/MS/MS）分析方法。分析时取 0.1mL 血或尿标本，采用乙腈（含 0.1%甲酸）沉淀一步法提取唑吡坦及其代谢物。上清液干燥后，加入 100μL 甲醇。采用 Acquity™ UPLC HSS T3（100mm×2.1mm，1.8μm）分析柱，在 API 4000Q 超高效液相色谱-串联质谱仪上以正离子化串联质谱检测，多反应监测（MRM）模式检测。流动相为乙腈或含 20mmol/L 乙酸铵和 0.1%甲酸的水，流速为 0.5mL/min。色谱分离时间为 4min，人血的校准曲线范围为 ZPD 0.1-300ng/mL，ZPCA 0.1-500ng/mL，ZCA 0.1-200ng/mL。尿液的线性范围为 ZPD 和 ZPCA 0.1-600ng/mL，ZCA 0.1-300ng/mL。检测限为 0.05ng/mL，定量限为 0.1ng/mL，ZPD、ZCA 和 ZPCA 的线性相关系数均大于 0.9995。日内和日间精密度均小于 15%，回收率为 70.0-98.3%，基质效应约为 79.5-99.0%，过程效率为 60.7%-94.4%。该方法可用于检测人血和尿中的唑吡坦及其代谢物，可应用于法医毒理学分析。
