HPLC法测定柴胡皂苷前处理酸化条件的优化及相干分析方法的评价.doc

HPLC法测定柴胡皂苷前处理酸化条件的优化及 相关分析方法的评价 胡杰，刘晓节，秦雪梅*，张丽增，郭小青 （山西大学化学化工学院药学系，山西太原 030006；） [摘要] 目的:优选HPLC法测定柴胡药材中柴胡皂苷a、d前处理的酸化条件，并对相关分析方法进行评价。方法:采用单因素及正交实验优选柴胡皂苷a、d前处理的酸化条件最佳提取条件，并对相关分析方法进行比较评价。结果:最佳酸化条件为：8%盐酸，30℃反应18h；样品最佳提取条件为：取药材粉末0.5g，加8%氨-甲醇溶液25ml，超声提取3次，每次45 min。结果表明本方法灵敏度高，分离度好，色谱图杂质峰较小，目标峰面积较大。结论:[关键词] 柴胡皂苷；HPLC；酸化条件； 方法评价Optimization of acid conditions for HPLC determination of Saikosaponin and evaluation on related methods HU Jie, LIU Xiao-jie, QIN Xue-mei, ZHANG Li-zeng, GUO Xiao-qin （School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006, China;） Abstract：Objective: To optimize pre-treatment acid condition for determination of saikosaponin -a and -d in Radix Bupleuri by HPLC, and evaluate relevant methods. Methods: The single factor and orthogonal experimental designs were employed to optimize acid condition and extraction condition for saikosaponin -a and -d. Results: The optimal acid conditions are as follows:the sample acidified with 8% hydrochloric acid at 30 ℃ for 18 h, and then 8% KOH solution is added to adjust the pH neutral. And the best extraction condition is that the power 0.5g added with 8% NH3-CH3OH 25 ml ultrasound extract 3 times, each extraction lasting 45 min. The results show that this method is sensitive, with good resolution, less impurity peaks, and bigger object peak area. Conclusion: The optimized method for the determination of saikosaponin a and d could be used as the basis for quality control of Radix Bupleuri. Key words：saikosaponin；HPLC；Acidification conditions, Method Evaluation 柴胡为伞形科柴胡属植物北柴胡(Bupleurum chinense DC.)和狭叶柴胡(B.scozonerifolium Willd.)的干燥根 ，[1]。柴胡皂苷为其主要有效成分之一，其中以柴胡皂苷a、d（Saikosaponin-a and -d，简称SSa、SSd）的药理活性较强，具抗炎、保肝、降低血中胆固醇等作用。虽有较多文献报道HPLC法测定SSaSSd的含量[3][]，但因其分子结构中只含一个双键（见图1），处于紫外末端吸收，采用HPLC-UV法检测波长大多在210nm下，杂质峰干扰严重，检测灵敏度低，方法不稳定；而HPLC-ELSD法[]检测限高且仪器费用昂贵，不普及；另有报道[]SSa、SSd在酸性条件下易水解开环成共轭体系（见图1），一定温度下可定向转化为柴胡皂苷b1、b2（Saikosaponin-b1 and -b2，简称SSb1、SSb2），使检测波长红移至254nm，紫外吸收增强，检测灵敏度提高，但目前该方法的样品前处理酸