Department of Laboratory Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130021, China.
Vascular Surgery, General Surgery Center, The First Hospital of Jilin University, Jilin University, Changchun 130021, China.
Molecules. 2022 Sep 2;27(17):5670. doi: 10.3390/molecules27175670.
A sensitive and rapid bioanalytical method based on the LC-triple-stage fragmentation (LC-MS) strategy on a hybrid triple quadrupole-linear ion trap mass spectrometer in combination with protein precipitation extraction for sample pretreatment has been developed and validated for the simultaneous determination of the antiepileptic drug oxcarbazepine (OXC) and its main active metabolite (MHD) in human serum. The separation was performed on a Waters XBridge BEH C18 column (2.5 µm, 2.1 × 50 mm) in isocratic elution with 0.1% formic acid in water and methanol (50:50, ) as the mobile phase. The run time for each sample was 2.0 min. The calibration curves ranging from 25 to 1600 ng/mL for OXC and from 0.5 to 32 μg/mL for MHD showed correlation coefficients (r) better than 0.99. All of the validation data, such as precision, accuracy and other parameters, fit the requirements of the current bioanalytical method validation guidelines. The LC-MS method for quantitation of OXC and MHD was compared with the LC-MRM based method. Passing-Bablok regression coefficients and Bland-Altman plots showed that the developed LC-MS method is a reliable method for quantitative analysis of OXC and MHD. The proposed LC-MS method was successfully applied to determine the serum concentrations of OXC and MHD to support a clinical study.
建立并验证了一种基于 LC-三重碎裂（LC-MS）策略的灵敏、快速的生物分析方法，该方法在混合型三重四极杆-线性离子阱质谱联用仪上使用，结合蛋白沉淀萃取进行样品预处理，用于同时测定人血清中的抗癫痫药物奥卡西平（OXC）及其主要活性代谢物（MHD）。采用 Waters XBridge BEH C18 柱（2.5 µm，2.1 × 50 mm），以水和甲醇（50:50，v/v）中的 0.1%甲酸为流动相，进行等度洗脱，实现分离。每个样品的运行时间为 2.0 min。OXC 的校准曲线范围为 25-1600 ng/mL，MHD 的校准曲线范围为 0.5-32 μg/mL，相关系数（r）均大于 0.99。所有验证数据，如精密度、准确度和其他参数，均符合当前生物分析方法验证指南的要求。所建立的 LC-MS 方法用于定量分析 OXC 和 MHD，并与基于 LC-MRM 的方法进行了比较。通过 Passing-Bablok 回归系数和 Bland-Altman 图表明，所开发的 LC-MS 方法是一种用于定量分析 OXC 和 MHD 的可靠方法。该方法成功地应用于测定奥卡西平和 MHD 的血清浓度，以支持一项临床研究。
