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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 HPLC method for the in vitro release of the active ingredient colonic pellets from Prunella vulgaris. Methods: Human gastrointestinal tract pH gradient was simulated. Rosmarinic acid was used as an index to determine the content of rosmarinic acid in three solutions of pH 1.0 HCl, pH 6.8 PBS and pH 7.6 PBS. Results: The detection wavelength of rosmarinic acid was 327 nm, 325 nm and 326 nm in pH 1.0 HCl, pH 6.8 PBS and pH 7.6 PBS respectively. The detection wavelength was 3.11 ~ 124.48 μg / ml for rosmarinic acid (R≥0.999 9). The intra-day precision RSD was less than 2%. Rosmarinic acid was more stable in pH 1.0 HCl and pH 6.8 PBS within 48 h, pH 7.6 PBS The solution should not be placed for a long time; the average recovery was 95% ~ 105% (RSD <2%). Conclusion: The HPLC method is specific, sensitive and accurate and can be used to determine the in vitro release of colonic pellets with effective components of Prunella vulgaris.