Peng Aihua, Ye Haoyu, Li Xia, Chen Lijuan
State Key Laboratory of Biotherapy, West China Hospital and College of Pharmacy, Sichuan University, Chengdu, China.
J Sep Sci. 2009 Sep;32(17):2967-73. doi: 10.1002/jssc.200900151.
Capsaicin and dihydrocapsaicin are two main bioactive components of Capsicum frutescens and are widely used as food additives and drugs in China and India. Due to their similarity in structures, isolation of capsaicin and dihydrocapsaicin with traditional methods such as silica gel column chromatography, normal-phase thin-layer chromatography (TLC) becomes difficult. This study involves separating capsaicin and dihydrocapsaicin with sufficient purity and recovery using high-speed counter-current chromatography (HSCCC) with a solvent system composed of n-hexane-ethyl acetate-methanol-water-acetic acid (20:20:20:20:2, v/v/v/v/v). Separation parameters such as sample volume, and sample concentration were first optimized on analytical HSCCC, and then scaled up to preparative HSCCC. 0.65 g capsaicin and 0.28 g dihydrocapsaicin were obtained from 1.2 g crude extract and their purities were 98.5 and 97.8%, respectively. The recoveries of the two compounds were 86.3 and 85.4%, respectively. The purity of the isolated compounds was analyzed by high-performance liquid chromatography (HPLC) and their structures were identified by (1)H nuclear magnetic resonance (NMR) and (13)C NMR analysis.
辣椒素和二氢辣椒素是辣椒的两种主要生物活性成分，在中国和印度被广泛用作食品添加剂和药物。由于它们结构相似，采用硅胶柱色谱、正相薄层色谱（TLC）等传统方法分离辣椒素和二氢辣椒素变得困难。本研究采用高速逆流色谱（HSCCC），以正己烷 - 乙酸乙酯 - 甲醇 - 水 - 乙酸（20:20:20:20:2，v/v/v/v/v）组成的溶剂系统，分离出纯度和回收率足够高的辣椒素和二氢辣椒素。首先在分析型HSCCC上优化样品体积、样品浓度等分离参数，然后放大到制备型HSCCC。从1.2 g粗提物中获得了0.65 g辣椒素和0.28 g二氢辣椒素，其纯度分别为98.5%和97.8%。两种化合物的回收率分别为86.3%和85.4%。通过高效液相色谱（HPLC）分析分离得到的化合物的纯度，并通过氢核磁共振（NMR）和碳 - 13核磁共振（NMR）分析鉴定其结构。
