HPLC法同时测定牛大力中豆甾醇和β-谷甾醇含量

Simultaneous Determination of Stigmasterol andβ-Sitosterol in Millettia speciosa Champ.by HPLC

【目的】建立一种同时测定牛大力中豆甾醇和β-谷甾醇含量的高效液相色谱法。【方法】采取超声提取法,提取溶剂为100%甲醇,提取料液比为1∶20 g/mL,提取时间为60 min;检测色谱柱为CAPCELL PAK C18(4.6 mm×250 mm,5μm),流动相为甲醇-水(97∶3),流速为1.0 mL/min,柱温为35℃,检测波长为210 nm,进样量为20μL。【结果】豆甾醇、β-谷甾醇分别在0.0202~1.0120 mg/mL(r=0.9997)、0.0202~1.0075 mg/mL(r=0.9998)范围内与峰面积呈现良好的线性关系,平均回收率分别为98.3%、96.8%,RSD分别为1.20%、1.64%,方法精密度、重复性、稳定性均良好。【结论】该方法简便快速、准确稳定、灵敏度高、专属性强,适用于牛大力中豆甾醇和β-谷甾醇含量的测定;该方法可为牛大力进行质量控制提供参考方法,可为牛大力进行深入开发提供科学借鉴。

【Objective】The purpose of this study was to establish a high performance liquid chromatography method(HPLC)for the determination of stigmasterol andβ-sitosterol in Millettia speciosa Champ.【Methods】Ultrasonic method was adopted to extract the herbs.The extraction effect was good when the extraction solvent was 100%methanol,the material-to-liquid ratio was 1∶20 g/mL,and the ultrasonic time was 60 min.The optimized HPLC conditions included that the chromatographic column was CAPCELL PAK C18(4.6 mm×250 mm,5μm)column,the mobile phase was methanol-water(97∶3),the flow rate was 1.0 mL/min,the column temperature was maintained at 35℃,the detection wavelength was set at 210 nm and the injection volume was 20μL.【Results】Under these conditions,the calibration curve of stigmasterol was in good linearity over the range of 0.0202—1.0120 mg/mL(r﹥0.9997)and 0.0202—1.0075 mg/mL(r﹥0.9998),respectively.The average recoveries of stigmasterol were 98.3%and 96.8%,respectively.RSDs were 1.20%and 1.64%,respectively.The precision,repeatability and stability of the method are good.【Conclusion】The method is simple,accurate,high sensitivity,and can be adopted to determine the content of stigmasterol andβ-sitosterol in Millettia speciosa Champ.This method could provide a reference for quality control and utilization of Millettia speciosa Champ.