Sabença Rosa, Bicker Joana, Silva Rui, Carona Andreia, Silva Ana, Santana Isabel, Sales Francisco, Falcão Amílcar, Fortuna Ana
Laboratory of Pharmacology, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
Laboratory of Pharmacology, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal; CIBIT/ICNAS - Coimbra Institute for Biomedical Imaging and Translational Research, University of Coimbra, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Jan 1;1162:122491. doi: 10.1016/j.jchromb.2020.122491. Epub 2020 Dec 11.
Perampanel is a third-generation antiepileptic drug (AED), while lamotrigine is a second-generation AED. Both drugs are subject to extensive pharmacokinetic variability between different patients. Furthermore, it has been reported that perampanel and lamotrigine may be implied in pharmacokinetic drug-drug interactions with other AEDs such as carbamazepine or valproate, with consequent alterations of plasma concentrations. This emphasizes the relevance of therapeutic drug monitoring of perampanel and lamotrigine with appropriate bioanalytical methods. Herein, the development and validation of a bioanalytical techique for the simultaneous quantification of perampanel and lamotrigine in human plasma samples is described. The reported method is based on high-performance liquid chromatography coupled with diode-array detection (HPLC-DAD) and sample preparation consists of liquid-liquid extraction. Chromatographic separation of the analytes (lamotrigine and perampanel) and the internal standard (entacapone) was achieved in 12 min on a reversed-phase C column at 40 °C by applying a gradient elution program with a mobile phase composed of 0.1% ortho-phosphoric acid pH 2.79 (A) and acetonitrile (B) pumped at 1.0 mL/min. Perampanel was quantified at 320 nm while lamotrigine and the internal standard were monitored at 306 nm. Calibration curves were linear in the concentration range of 0.03-4.5 µg/mL (r = 0.9978) for perampanel and in the concentration range of 0.25-30 µg/mL (r = 0.9981) for lamotrigine. Overall precision did not exceed 14.3% and accuracy ranged from -6.08 to 12.66%. Some drugs potentially co-prescribed with perampanel and lamotrigine were tested and did not interfere with the retention times of the analytes and internal standard. The method was then successfully applied for the quantification of perampanel and lamotrigine in plasma samples obtained from 42 drug-resistant epileptic patients admitted to the Coimbra University Hospital Centre (CHUC.EPE, Coimbra, Portugal). In conclusion, it is a suitable method for the therapeutic drug monitoring of lamotrigine and perampanel in drug-resistant epileptic patients, as well as, for the assessment of drug-drug interactions. It can also be adopted by hospitals and laboratories, when HPLC with fluorescence and mass spectrometry detections are unavailable.
吡仑帕奈是一种第三代抗癫痫药物（AED），而拉莫三嗪是第二代AED。两种药物在不同患者之间都存在广泛的药代动力学变异性。此外，据报道，吡仑帕奈和拉莫三嗪可能与其他抗癫痫药物如卡马西平或丙戊酸发生药代动力学药物相互作用，从而导致血浆浓度改变。这突出了采用适当的生物分析方法对吡仑帕奈和拉莫三嗪进行治疗药物监测的重要性。本文描述了一种用于同时定量人血浆样品中吡仑帕奈和拉莫三嗪的生物分析技术的开发和验证。所报道的方法基于高效液相色谱-二极管阵列检测（HPLC-DAD），样品制备采用液-液萃取。在40℃下，通过应用由0.1%磷酸（pH 2.79）（A）和乙腈（B）组成的流动相的梯度洗脱程序，以1.0 mL/min的流速在反相C柱上12分钟内实现了分析物（拉莫三嗪和吡仑帕奈）和内标（恩他卡朋）的色谱分离。吡仑帕奈在320 nm处定量，而拉莫三嗪和内标在306 nm处监测。吡仑帕奈的校准曲线在0.03 - 4.5 μg/mL浓度范围内呈线性（r = 0.9978），拉莫三嗪的校准曲线在0.25 - 30 μg/mL浓度范围内呈线性（r = 0.9981）。总体精密度不超过14.3%，准确度范围为 - 6.08%至12.66%。对一些可能与吡仑帕奈和拉莫三嗪联合使用的药物进行了测试，这些药物不干扰分析物和内标的保留时间。该方法随后成功应用于对从葡萄牙科英布拉大学医院中心（CHUC.EPE，科英布拉）收治的42例耐药癫痫患者获得的血浆样品中吡仑帕奈和拉莫三嗪的定量。总之，它是一种适用于耐药癫痫患者中拉莫三嗪和吡仑帕奈治疗药物监测以及药物相互作用评估的方法。当没有配备荧光和质谱检测的HPLC时，医院和实验室也可采用该方法。
