大川芎方制剂类天麻素类、苯酞类以及酚酸类成分测定方法研究

Study on determination method of gastrodin, phthalide and phenolic acid in Dachuanxiong prescription

目的建立3个内标、3个波长法测定大川芎方不同制剂的天麻素类、苯酞类以及酚酸类3类成分的含量。方法采用Agilent ZORBAX Eclipse XDB-C18色谱柱,流动相为乙腈-0.05%磷酸水溶液,梯度洗脱,检测波长为220、278、322 nm,柱温为35oC,体积流量为1 mL/min。以天麻素、藁本内酯、阿魏酸为内标,建立天麻素类(巴利森苷、巴利森苷B)、苯酞类(洋川芎内酯I、H、A和欧当归内酯A)以及酚酸类(绿原酸)成分的相对校正因子(RCF),在不同进样体积、不同色谱柱、不同添加剂条件下验证其耐用性,利用RCF值计算2种大川芎方制剂中10个成分的含量,并与外标法测定结果进行比较,验证方法的可行性。结果所建立的方法相对校正因子耐用性良好(RSD≤3.5%),与外标法的含量测定结果基本一致。结论该3内标、3波长法可用于大川芎方制剂的药效成分测定。

Objective Three internal standards and three wavelength methods were established to determine the content of gastrodin,phenolics and phenolic acids in different preparations of Dachuanxiong prescription. Methods The separation was carried out on Agilent ZORBAX Eclipse XDB-C18 column. The mobile phase consisted of acetonitrile-0.05% phosphoric acids aqueous solution with gradient elution. The detection wavelength was at 220, 278, 322 nm with flow velocity at 1.0 mL/min and column temperature at 35 oC. Firstly, gastrodin, Z-ligustilide, ferulic acid were used as references to get the average relative correction factors(RCF value) of gastrodin analogues(parishin B, parishin), phthalides(senkyunolide I, senkyunolide H, senkyunolide A and levistolide A) and phenolic acids(chlorogenic acid) respectively. Secondly, durability test of RCF was carried out in different factors including inject volume,columns, and additives. Finally, the contents of 10 compounds in two kinds of Dachuanxiong prescription preparations was calculated based on the RCF value, and compared with the determination results of external standard method to verify the feasibility of the method.Results The result showed that durability of established method was good(RSD ranging below 3.5%), which is basically consistent with the content determination results of external standard method. Conclusion The determination method of three internal standards and three wavelengths can be used to determine the pharmacodynamic composition of Dachuanxiong prescription.