Zeid Abdallah M, Nasr Jenny Jeehan M, Belal Fathalla, Walash Mohamed I, Baba Yoshinobu, Kaji Noritada
Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan; Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt.
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt.
Spectrochim Acta A Mol Biomol Spectrosc. 2021 Feb 5;246:119021. doi: 10.1016/j.saa.2020.119021. Epub 2020 Oct 3.
In this study, a facile, sensitive, and precise lab-on-a-chip electrophoretic method coupled with light-emitting diode induced fluorescence (LED-IF) detection was developed to assay three antiepileptic drugs, namely, vigabatrin, pregabalin, and gabapentin, in pharmaceutical formulations. The analytes were derivatised offline for the first time with fluorescine-5-isothiocyanate (FITC) to yield highly fluorescent derivatives with λ of 490/520nm. The FITC-labelled analytes were injected, separated, and quantitated by a microfluidic electrophoresis device using fluorescence detection. The labelled analytes were monitored using a blue LED-IF system. The separation conditions were significantly optimised adding specific concentrations of heptakis-(2,6-di-O-methyl)-β-cyclodextrin (HDM-β-CD) and methylcellulose to the buffer solution (40mM borate buffer). HDM-β-CD acted as a selective host for the studied antiepileptic drugs, rendering a high separation efficiency. Methylcellulose was used as an efficient dynamic coating polymer to prevent the labelled drugs from being adsorbed on the inner surfaces of the poly (methylmethacrylate) microchannels. A laboratory-prepared ternary mixture of the three antiepileptic drugs was separated within 100s with acceptable resolution between all the three analytes (R>3) and a high number of theoretical plates (N) for each analyte (N≈10 plates/m). The sensitivity of the method was enhanced approximately 80-fold by stacking to yield a detection limit below 0.6ngmL in the concentration range of 2.0-200.0ngmL. The method was successfully validated for analysing the studied drugs in their pharmaceutical formulations.
在本研究中，开发了一种简便、灵敏且精确的芯片实验室电泳方法，并结合发光二极管诱导荧光（LED-IF）检测，用于分析药物制剂中的三种抗癫痫药物，即氨己烯酸、普瑞巴林和加巴喷丁。首次将分析物离线用异硫氰酸荧光素（FITC）衍生化，以产生λ为490/520nm的高荧光衍生物。将FITC标记的分析物注入微流控电泳装置中，通过荧光检测进行分离和定量。使用蓝色LED-IF系统监测标记的分析物。通过向缓冲溶液（40mM硼酸盐缓冲液）中添加特定浓度的七（2,6-二-O-甲基）-β-环糊精（HDM-β-CD）和甲基纤维素，显著优化了分离条件。HDM-β-CD作为所研究抗癫痫药物的选择性主体，实现了高分离效率。甲基纤维素用作有效的动态涂层聚合物，以防止标记药物吸附在聚甲基丙烯酸甲酯微通道的内表面上。三种抗癫痫药物的实验室制备三元混合物在100秒内分离，所有三种分析物之间具有可接受的分辨率（R>3），且每种分析物具有大量理论塔板数（N）（N≈10塔板/米）。通过堆积将该方法的灵敏度提高了约80倍，在2.0-200.0ng/mL的浓度范围内检测限低于0.6ng/mL。该方法已成功验证可用于分析药物制剂中的所研究药物。
