建立测定积雪草苷溶解度的高效液相色谱法

Determination＇on the solubility of Asiaticoside by RP-HPLC

目的建立高效液相色谱法测定积雪草苷，为积雪草苷溶解度的研究提供基础。方法采用反相高效液相法，色谱柱：BDSHYPERSILC18（150mmx4．6mm，5μm），流动相为乙腈：水（30：70，v/v），流速0．8m1．min-1，柱温35℃，检测波长220nm，进样量20μl。结果积雪草苷的线性方程为：表明在26．65～426．4μg·ml-1范围内线性关系良好。低、中、高三种浓度的积雪草苷的日内精密度RSD〈1．4％（n=5）；连续五天的日间精密度RSD〈3．6％。积雪草苷的最低检测浓度约为0．5μg·ml-1。积雪草苷在水和缓冲液中的溶解度相近，生理盐水中溶解度小。结论本研究建立的RP—HPLC方法简便、快速、准确，适用于积雪草苷的溶解度研究。

Objective To establish reversed-phase high＇performance liquid chromatography （RP-HPLC） for the equilibrium solubility so as to provide a basic study for the exploitation of Asiaticoside preparation. Methods HPLC method was established to determine the concentration of Asiaticoside in water as well as phosphate buffer solutions. BDS HYPERSIL C18 column （150 mm×4.6 mm, 5 μm） was used. The mobile phase was consisted of aeetonitril and water （30： 70, v/v）. The flow rate was 0.8 ml·min-1, and the UV detection wavelength was set at 220 nm. The column temperature was 35℃ and the sample size was 20 μl. Results The linear calibration equation of Asiaticoside was： A=2287.7C-3934.5 （r=-l） , it is shown that the linear relationship was good when the concentration of Asiaticoside from 26.65 μg·ml-1 to 426.4 μg· ml-1. The intra-day and inter-day RSD were within 1.4% and 3.6%, respectively. The minimum detectable concentration was 0.5 μg· ml-1. The equilibrium solubility of Asiaticoside was similar in water and in buffer （pH 8.0）, it solubility in saline was the smallest of all the solvent. Conclusion The RP-HPLC is simple, rapid and suitable method to study solubility of Asiaticoside.