Kamal Husein, Gandhi Varun, Akil Lina, Al-Tannak Naser F, Rattray Nicholas J W, Khadra Ibrahim
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kuwait University, P.O. Box 24923, Safat 13110, Kuwait.
Molecules. 2025 Aug 22;30(17):3451. doi: 10.3390/molecules30173451.
A simple and reliable method was developed using LC-MS/MS to quantify alprazolam, bromazepam, clonazepam, diazepam, and flunitrazepam in clinical samples. This method was validated for the simultaneous determination of alprazolam, bromazepam, clonazepam, diazepam, and flunitrazepam. It was applied to human urine samples collected from people suspected of drug abuse in the Kuwaiti region. Formic acid in water and acetonitrile was used in mobile phase with a gradient mode of elution using C18 reverse-phase column. The instrument was operated in a positive mode with an electrospray ionization source using multiple reaction monitoring. For sample extraction, the liquid-liquid extraction technique was used. The method was validated for limit of detection, limit of quantitation, selectivity, linearity, accuracy, and precision. The concentration for limit of quantitation was 6.0 ng/mL, the linearity ranged from 2.0 to 300 ng/mL for each of the analytes, and the r values were ≥0.99. The accuracy was found to be within a range of 80-120% and precision had a %RSD of ≤15% for each of the analytes. The method was applied to 48 urine samples collected from those suspected of drug abuse by the Toxicology Department of the General Department of Criminal Evidence, Kuwait, and alprazolam, bromazepam, clonazepam, diazepam and flunitrazepam were identified commonly in the samples. The overall drug positivity rate obtained considering 48 samples was 93.75%. Based on these results and successful determination of alprazolam, bromazepam, clonazepam, diazepam and flunitrazepam in human urine samples from those suspected of drug abuse, this method is deemed to be suitable for its routine analysis.
开发了一种简单可靠的方法，使用液相色谱-串联质谱法（LC-MS/MS）对临床样本中的阿普唑仑、溴西泮、氯硝西泮、地西泮和氟硝西泮进行定量。该方法经验证可同时测定阿普唑仑、溴西泮、氯硝西泮、地西泮和氟硝西泮。它被应用于从科威特地区涉嫌药物滥用的人群中收集的人体尿液样本。流动相使用水和乙腈中的甲酸，采用梯度洗脱模式，使用C18反相柱。仪器采用电喷雾电离源，在正模式下运行，使用多反应监测。对于样品提取，采用液-液萃取技术。该方法在检测限、定量限、选择性、线性、准确度和精密度方面进行了验证。定量限浓度为6.0 ng/mL，各分析物的线性范围为2.0至300 ng/mL，r值≥0.99。发现各分析物的准确度在80-120%范围内，精密度的相对标准偏差（%RSD）≤15%。该方法应用于科威特刑事证据总局毒理学部门从涉嫌药物滥用者中收集的48份尿液样本，样本中常见阿普唑仑、溴西泮、氯硝西泮、地西泮和氟硝西泮。考虑48个样本得出的总体药物阳性率为93.75%。基于这些结果以及在涉嫌药物滥用者的人体尿液样本中成功测定阿普唑仑、溴西泮、氯硝西泮、地西泮和氟硝西泮，该方法被认为适用于常规分析。
