Department of Forensic Medicine and Toxicology, School of Medicine, National and Kapodistrian University of Athens, Mikras Asias 75, Athens 11527, Greece.
J Pharm Biomed Anal. 2010 Aug 1;52(4):609-14. doi: 10.1016/j.jpba.2010.01.027. Epub 2010 Feb 1.
Benzodiazepines are used widely in daily clinical practice, due to their multiple pharmacological actions. The frequent problems associated with the wide use of benzodiazepines, as well as the multiple incidents of poisonings, led to the necessity for the development of a precise, sensitive and rapid method for the simultaneous determination of the 23 most commonly used benzodiazepines (diazepam, nordiazepam, oxazepam, bromazepam, alprazolam, lorazepam, medazepam, flurazepam, fludiazepam, tetrazepam, chlordiazepoxide, clobazam, midazolam, flunitrazepam, 7-amino-flunitrazepam, triazolam, prazepam, nimetazepam, nitrazepam, temazepam, lormetazepam, clonazepam, camazepam) in blood. A gas chromatographic method combined with mass spectrometric detection was developed, optimized and validated for the determination of the above substances. This method includes liquid-liquid extraction with chloroform at pH 9 and two stages of derivatization using tetramethylammonium hydroxide and propyliodide (propylation), as well as a mixture of triethylamine:propionic anhydride (propionylation). The recoveries were higher than 74% for all the benzodiazepines. The calibration curves were linear within the dynamic range of each benzodiazepine with a correlation coefficient higher than 0.9981. The limits of detection and quantification for each analyte were statistically calculated from the relative calibration curves. Accuracy and precision were also calculated and were found to be less than 8.5% and 11.1%, respectively. The developed method was successfully applied for the investigation of both forensic and clinical toxicological cases of accidental and suicidal poisoning.
苯二氮䓬类药物由于其多种药理学作用，在日常临床实践中广泛应用。由于苯二氮䓬类药物的广泛使用所带来的频繁问题，以及多次中毒事件，因此有必要开发一种精确、灵敏、快速的方法，同时测定 23 种最常用的苯二氮䓬类药物（地西泮、去甲西泮、奥沙西泮、溴西泮、阿普唑仑、劳拉西泮、美达唑仑、氟西泮、氟硝西泮、三唑仑、氯氮䓬、氯巴占、咪达唑仑、氟硝西泮、7-氨基氟硝西泮、三唑仑、普拉西泮、依替唑仑、硝西泮、替马西泮、劳拉西泮、氯硝西泮、卡马西泮）在血液中的含量。本研究建立了一种气相色谱-质谱联用方法，对上述物质进行了优化和验证。该方法包括 pH 值为 9 的氯仿液-液萃取和使用四甲基氢氧化铵和碘化丙基（丙基化）进行的两步衍生化，以及三乙胺：丙酸酐（丙酰化）混合物。所有苯二氮䓬类药物的回收率均高于 74%。各苯二氮䓬类药物的校准曲线在其动态范围内均呈线性，相关系数均高于 0.9981。每个分析物的检测限和定量限均从相对校准曲线统计计算得出。准确度和精密度也进行了计算，结果分别小于 8.5%和 11.1%。该方法成功应用于意外和自杀中毒的法医和临床毒理学案例的调查。
