H. M. Albishri, Naflaa A. Aldawsari, D. Abd El-Hady
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The flow rate, wavelength and column temperature were fixed at 1.0 mL min−1, 214 nm and 40 °C, respectively. The proposed method was validated, giving a linearity within the concentration ranges 5–500 ng mL−1 for bromazepam and diazepam, 3–500 ng mL−1 for clonazepam and lorazepam and 1–500 ng mL−1 for nordiazepam with a determination coefficient (R2) more than 0.9992. The LOD values for the selected BZDs ranged from 0.54 to 2.32 and from 1.78 to 7.65 ng mL−1 for standard methanolic and plasma matrices, respectively. Precision, accuracy, selectivity, stability, and robustness were some of the terms considered in validating the current RPLC method. Based on these results, a simple and reliable RPLC method was successfully applied to quantify BZDs in human plasma matrix appearing with recoveries ranging from 96.5 to 107.5% and interday RSD less than 4%. 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引用次数: 1
摘要
苯二氮卓类药物(BZDs)是目前用于治疗神经心理疾病的重要药物之一。事实上,bzd被吸毒者滥用,无论其治疗用途如何。因此,寻找一种简便、可靠的方法对人血浆基质中BZDs的筛选和定量具有重要的法医学和临床毒理学意义。本工作采用简单可靠的RPLC方法,在人血浆基质中同时分离并检测溴西泮、氯硝西泮、劳拉西泮、去甲西泮和地西泮5种bzd。采用等温流动洗脱,由20 mmol L−1磷酸盐缓冲液(pH 7.0)和甲醇(50:50,v/v)在对称C18柱上组成。流速为1.0 mL min - 1,波长为214 nm,柱温为40℃。结果表明:溴西泮和地西泮在5 ~ 500 ng mL−1、氯硝西泮和劳拉西泮在3 ~ 500 ng mL−1、去硝西泮在1 ~ 500 ng mL−1的浓度范围内呈线性关系,测定系数(R2)均大于0.9992。对于标准甲醇和血浆基质,所选BZDs的LOD值分别为0.54 ~ 2.32和1.78 ~ 7.65 ng mL−1。精密度、准确度、选择性、稳定性和鲁棒性是验证当前RPLC方法所考虑的一些术语。建立了简便可靠的RPLC定量方法,回收率为96.5 ~ 107.5%,日间RSD < 4%。目前开发的方法可用于快速筛选市场上最常用的BZDs在其治疗浓度范围内。
A Simple and Reliable Liquid Chromatographic Method for Simultaneous Determination of Five Benzodiazepine Drugs in Human Plasma
Benzodiazepines (BZDs) are one of the most important drugs that have been used in the treatment of neuropsychological disorders. Indeed, BZDs are abused by drug addicts regardless of their therapeutic uses. Therefore, it was important in forensic and clinical toxicology to reach an easy and reliable method for the screening and quantification of BZDs in the human plasma matrix. In the current work, five BZDs, namely bromazepam, clonazepam, lorazepam, nordiazepam and diazepam were simultaneously separated and detected by a simple and reliable RPLC method in a human plasma matrix. Isocratic mobile elution consisting of 20 mmol L−1 phosphate buffer (pH 7.0) and methanol (50:50, v/v) on a Symmetry C18 column was employed. The flow rate, wavelength and column temperature were fixed at 1.0 mL min−1, 214 nm and 40 °C, respectively. The proposed method was validated, giving a linearity within the concentration ranges 5–500 ng mL−1 for bromazepam and diazepam, 3–500 ng mL−1 for clonazepam and lorazepam and 1–500 ng mL−1 for nordiazepam with a determination coefficient (R2) more than 0.9992. The LOD values for the selected BZDs ranged from 0.54 to 2.32 and from 1.78 to 7.65 ng mL−1 for standard methanolic and plasma matrices, respectively. Precision, accuracy, selectivity, stability, and robustness were some of the terms considered in validating the current RPLC method. Based on these results, a simple and reliable RPLC method was successfully applied to quantify BZDs in human plasma matrix appearing with recoveries ranging from 96.5 to 107.5% and interday RSD less than 4%. The current developed method was useful for rapidly screening the most commonly used BZDs in the market within their therapeutic concentration ranges.