毛细管电泳法分离丹参水溶性有效成分——丹参素、原儿茶醛和原儿茶酸

Separation of Water-Soluble Active Principles of Danshen (Salvia miltiorrhiza) Danshensu, Protocatechuic Aldehyde and Protocatechuic Acid by HPCE

毛细管电泳法分离丹参水溶性有效成分丹参素 (DSS)、原儿茶醛 (PAH)和原儿茶酸 (PA )。分别考察电渗流改向剂十六烷基三甲基溴化铵 (CTAB)的浓度、电泳缓冲液 p H值及电解质浓度、有机添加剂种类及浓度、运行电压、样品溶剂组成 ,以及检测波长等因素对有机阴离子类分析对象的峰迁移时间、分辨率、柱效的影响 ,优化选择的分离条件为 :以 p H为 7.0 ,含 0 .5 m mol/ L CTAB,浓度为 2 0 0 mm ol/ L的磷酸盐溶液与乙腈按 2 0∶ 5混合配制电泳缓冲液 ,在 75 μm(id)× 34 .5 (eff. 30 ) cm的毛细管柱 (柱温控制于 2 0℃ )上 ,10 k V(6 s)电迁移进样 ,运行电压为 8k V。在 8min内能将以 2 0 %乙腈 -水为溶剂的样品 ,即内标 (对羟基苯甲酸 ,p- HBA) ,DSS、PAH和 PA完全基线分离 ,柱效达 40万 /米理论塔板数。

Water soluble active principles of Salvia miltiorrhize Bge. —Danshensu (DSS), protocatechuic aldehyde (PAH) and protocatechuic acid (PA) were separated by HPCE. Many factors that can affect the resolution, sensitivity and column efficiency of organic anion, including the concentration of cetyltrimethyl ammonium bromide (CTAB), pH value, concentration of phosphate buffer, percentage of organic modifier in the buffer, running voltage, composition of solvent used for the sample, and detector wavelength were investigated to optimize the experimental conditions. Good separation of internal standard (p hydroxy benzoic acid), DSS, PAH and PA can be achieved in less than 8 min under the following conditions: capillary column, 75 μm (id)×34.5 (eff. 30) cm, uncoated; column temperature controled at 20 ℃; detector, UV 200 nm; injection, 10 kV/6 s; running voltage, 8 kV; running buffer, 200 mmol/L phosphate buffer (pH 7.0, containing 0.5 mmol/L CTAB) and acetonitrile 5 mL in 20 mL (20∶5, v/v). Sample solvent was acetonitrile water (2∶8 v/v). The column efficiency reached 400 000 per metre in thoretical plate number.