夏枯草有效组分结肠定位微丸体外释放度HPLC测定方法

Determination on Release in vitro of Colon-specific Pellets of Effective Components of Prunella Vulgaris L. by HPLC

目的:建立夏枯草有效组分结肠定位微丸体外释放度的高效液相色谱法(HPLC)测定方法。方法:模拟人体胃肠道pH梯度变化,以迷迭香酸为指标,HPLC测定迷迭香酸在pH 1.0 HCl、pH 6.8 PBS、pH 7.6 PBS三种溶液中的含量。结果:迷迭香酸在pH 1.0 HCl、pH 6.8 PBS、pH 7.6 PBS三种溶液中的检测波长分别为327 nm、325 nm、326 nm;迷迭香酸三种溶液在3.11~124.48μg/ml浓度范围内线性关系良好(r≥0.999 9);日内精密度RSD均<2%;迷迭香酸在pH 1.0 HCl,pH 6.8 PBS中48 h内较稳定,pH 7.6PBS迷迭香酸对照品溶液不宜长时间放置;平均加样回收率均在95%~105%范围内(RSD<2%)。结论:HPLC方法专属、灵敏、准确,可用于夏枯草有效组分结肠定位微丸体外释放度测定。

Objective ： To establish a high performance liquid chromatography （ HPLC ） method for determination on release in vitro of colon-specific pellets of effective components of Prunella vulgaris L.. Methods ： Simulating the gradient variation of pH in human gastrointestinal tract,the contents of rosmarinic acid as the index in pH 1.0 HC1, pH 6.8 PBS and pH 7.6 PBS were determined by HPLC. Results ： The wavelength of rosmarinic acid is 327 nm, 325 nm, 326 nm in pH 1.0 HCl, pH 6.8 PBS and pH 7.6 PBS, respectively. The three kinds of solution show good linearity in the concentration range of 3.11-124.48 μg/ml （r≥0. 9999） ;The RSD of intraday precision are all less than 2% ;The contents of rosmarinic acid in pH 1.0 HCI, pH 6.8 PBS are more stable within 48 hours. The reference solution of rosmarinic acid in pH 7.6 PBS should not be placed for a long time. The average recoveries are all within the range of 95 %- 105 % （RSD 〈 2% ）. Conclusion： This method proves to be specific, sensitive and accurate and can be used in the determination on release in vitro of colon-specific pellets of effective components of Prunella vulgaris L. .