Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, 380009, India.
Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, 380009, India.
J Pharm Biomed Anal. 2020 Sep 10;189:113452. doi: 10.1016/j.jpba.2020.113452. Epub 2020 Jul 2.
A simple, fast and environmental friendly supercritical fluid chromatography-photodiode array (SFC-PDA) method was developed for the simultaneous determination of metformin and three sodium-glucose co-transporter 2 (SGLT-2) inhibitors (canagliflozin, dapagliflozin and empagliflozin). The impact of different stationary phases, co-solvents, column temperature and outlet pressure was extensively evaluated for selective separation and quantitation of the drugs. Amongst several achiral and chiral stationary phases tested, the best results were obtained on amylose based Chiralpak IG column using CO and 0.1 % diethylamine in methanol and isopropanol 50:50 (v/v) in 60:40 ratios as the mobile phase. The separation efficiency was mainly dependent on the nature of stationary phase and the mobile phase composition. The limit of detection (LOD) and quantitation (LOQ) of the method were 0.155 and 0.469, 0.062 and 0.187, 0.015 and 0.045, 0.028 and 0.084 μg/mL for metformin, canagliflozin, dapagliflozin and empagliflozin, respectively. The chromatographic resolution between metformin-canagliflozin, metformin-dapagliflozin and metformin-empagliflozin pairs was 6.87, 3.44 and 6.47, respectively. The intra-batch and inter-batch precision of the method was ≤1.64 % while the accuracy was in the range of 96.2-103.3 %. The method was used to analyze metformin and SGLT-2 inhibitors in their binary fixed-dose formulations with acceptable accuracy (recovery) and precision. To support the experimental results, molecular docking was performed on Schrodinger software to provide a deeper insight into the interactions between the analytes and the chiral stationary phase and to understand their elution orders under optimized conditions.
建立了一种简单、快速、环保的超临界流体色谱-光电二极管阵列（SFC-PDA）法，用于同时测定二甲双胍和三种钠-葡萄糖共转运蛋白 2（SGLT-2）抑制剂（卡格列净、达格列净和恩格列净）。考察了不同固定相、共溶剂、柱温和出口压力对药物选择性分离和定量的影响。在所测试的几种手性和非手性固定相，在基于直链淀粉的 Chiralpak IG 柱上，使用 CO 和 0.1%二乙胺在甲醇和异丙醇 50:50（v/v）的 60:40 比例作为流动相，得到了最佳结果。分离效率主要取决于固定相和流动相的组成。该方法的检出限（LOD）和定量限（LOQ）分别为 0.155 和 0.469、0.062 和 0.187、0.015 和 0.045、0.028 和 0.084μg/mL 用于二甲双胍、卡格列净、达格列净和恩格列净。二甲双胍-卡格列净、二甲双胍-达格列净和二甲双胍-恩格列净对的色谱分辨率分别为 6.87、3.44 和 6.47。该方法的批内和批间精密度均≤1.64%，准确度在 96.2-103.3%范围内。该方法用于分析其固定剂量复方制剂中的二甲双胍和 SGLT-2 抑制剂，具有可接受的准确度（回收率）和精密度。为了支持实验结果，在 Schrodinger 软件上进行了分子对接，以更深入地了解分析物与手性固定相之间的相互作用，并了解在优化条件下它们的洗脱顺序。
