Bhatt Nejal M, Chavada Vijay D, Sanyal Mallika, Shrivastav Pranav S
Chemistry Department, School of Sciences, Gujarat University, Ahmedabad, Gujarat, India.
Chemistry Department, St. Xavier's Science College, Ahmedabad, Gujarat, India.
Chirality. 2017 Feb;29(2):80-88. doi: 10.1002/chir.22673. Epub 2016 Dec 28.
A simple and efficient high-performance thin-layer chromatographic method was developed for chiral separation of rac-bupropion (BUP) and its active metabolite rac-hydroxybupropion (HBUP). Design of experiment (DoE)-based optimization was adopted instead of a conventional trial-and-error approach. The Box-Behnken design surface response model was used and the operating variables were optimized based on 17 trials design. The optimized method involved impregnation of chiral reagent, L(+)-tartaric acid, in the stationary phase with simultaneous addition in the mobile phase, which consisted of acetonitrile : methanol : dichloromethane : 0.50% L-tartaric acid (6.75:1.0:1.0:0.25, v/v/v/v). Under the optimized conditions, the resolution factor between the enantiomers of BUP and HBUP was 6.30 and 9.26, respectively. The limit of detection and limit of quantitation for (R)-BUP, (S)-BUP, (R,R)-HBUP, and (S,S)-HBUP were 9.23 and 30.78 ng spot , 10.32 and 34.40 ng spot , 12.19 and 40.65 ng spot , and 14.26 and 47.53 ng spot , respectively. The interaction of L-tartaric acid with analytes and their retention behavior was thermodynamically investigated using van't Hoff's plots. The developed method was validated as per the International Conference on Harmonization guidelines. Finally, the method was successfully applied to resolve and quantify the enantiomeric content from marketed tablets as well as spiked plasma samples.
开发了一种简单高效的高效薄层色谱法，用于手性分离外消旋安非他酮（BUP）及其活性代谢物外消旋羟基安非他酮（HBUP）。采用基于实验设计（DoE）的优化方法，而非传统的试错法。使用Box-Behnken设计表面响应模型，并基于17次试验设计对操作变量进行了优化。优化后的方法包括在固定相中浸渍手性试剂L(+)-酒石酸，并同时添加到流动相中，流动相由乙腈:甲醇:二氯甲烷:0.50% L-酒石酸（6.75:1.0:1.0:0.25，v/v/v/v）组成。在优化条件下，BUP和HBUP对映体之间的分离度分别为6.30和9.26。(R)-BUP、(S)-BUP、(R,R)-HBUP和(S,S)-HBUP的检测限和定量限分别为9.23和30.78 ng/斑点、10.32和34.40 ng/斑点、12.19和40.65 ng/斑点以及14.26和47.53 ng/斑点。使用范特霍夫图对L-酒石酸与分析物的相互作用及其保留行为进行了热力学研究。所开发的方法按照国际协调会议指南进行了验证。最后，该方法成功应用于拆分和定量市售片剂以及加标血浆样品中的对映体含量。
