Department of Clinical Laboratory, The First Hospital of Jilin University, Jilin University, Changchun 130021, China.
The Bethune Institute of Epigenetic Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130021, China.
Molecules. 2022 Feb 11;27(4):1224. doi: 10.3390/molecules27041224.
This study developed a detection method based on the strategy of HPLC/MS and verified its suitability by quantifying carbamazepine in human plasma. The high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS) system was performed using a Shimadzu UFLC XR liquid chromatography and a SCIEX QTRAP 5500 linear ion trap triple quadrupole mass spectrometer. The specific operation was as follows: the sample protein was firstly precipitated using methanol, then carbamazepine and carbamazepine-DN were separated on an ACQUITY UPLC HSS T3 column using the gradient elution with solvent A (0.1% formic acid) and solvent B (0.1% formic acid in acetonitrile) at a flow rate of 0.25 mL/min. Each sample was run for 7 min. This method was validated for various parameters including accuracy, precision, selectivity, linearity, LLOQ, etc. Only 5 μL of sample plasma could obtain the result of LLOD 0.5 µg/mL. The intra-day and inter-day precision was <8.23%, and accuracy was between -1.74% and 2.92%. This method was successfully used for monitoring the blood concentration of epilepsy patients after carbamazepine treatment.
本研究基于 HPLC/MS 策略开发了一种检测方法，并通过定量测定人血浆中的卡马西平验证了其适用性。高效液相色谱-串联质谱（HPLC/MS）系统采用岛津 UFLC XR 液相色谱和 SCIEX QTRAP 5500 线性离子阱三重四极杆质谱仪进行。具体操作如下：首先使用甲醇沉淀样品蛋白，然后使用 ACQUITY UPLC HSS T3 柱在流动相 A（0.1%甲酸）和流动相 B（0.1%甲酸乙腈）的梯度洗脱下，以 0.25 mL/min 的流速分离卡马西平和卡马西平-DN。每个样品运行 7 分钟。该方法对各种参数进行了验证，包括准确性、精密度、选择性、线性、LLOQ 等。仅需 5μL 样品血浆即可获得 LLOD 0.5µg/mL 的结果。日内和日间精密度<8.23%，准确度在-1.74%至 2.92%之间。该方法成功用于监测卡马西平治疗后癫痫患者的血药浓度。
