高效液相色谱法测定巴旦木青皮提取物中齐墩果酸和熊果酸

Determination of Oleanolic Acid and Ursolic Acid in Almond Green Husk Extract by High Performance Liquid Chromatography

采用质谱仪对巴旦木青皮提取物中齐墩果酸（oleanolic acid,OA）和熊果酸（ursolic acid,UA）进行鉴定,进一步用高效液相色谱法测定OA和UA含量,优化提取条件。高效液相色谱条件：利用Platisil ODS C18色谱柱（250 mm×4.6 mm,5μm）,以甲醇-0.2%磷酸（85：15,V/V）溶液为流动相,流速0.8 mL/min;柱温20℃;检测波长210 nm。结果表明：OA在0.005-0.5 mg/mL范围内线性关系良好（R2=0.999 9）,回收率在89.67%-94.85%之间,相对标准偏差在1.28%-3.16%之间（n=3）;UA在0.005-0.5 mg/mL范围内线性关系良好（R2=0.999 9）,回收率在88.67%-94.77%之间,相对标准偏差在1.07%-1.92%（n=3）之间。在不同提取条件下,提取剂为体积分数95%乙醇、料液比1：40（g/mL）、提取时间40 min及提取3次为最佳条件。

Oleanolic acid（OA） and ursolic acid（UA） were identified by mass spectrometry（MS） in almond green husk extract. Further, a high performance liquid chromatographic（HPLC） method was established for the determination of the contents of OA and UA, and the extraction conditions were optimized using single factor and orthogonal array experiments. The HPLC conditions were set as follows： separation column, Platisil ODS C18（250 mm × 4.6 mm, 5 μm）; mobile phase, methanol-0.2% phosphorus acid（85：15, V/V）; flow rate, 0.8 mL/min; column temperature, 20 ℃; and detection wavelength, 210 nm. In the range of 0.005–0.5 mg/mL（R-2 = 0.999 9）, both OA and UA presented a good linear relationship. The average recoveries of OA varied from 89.67% to 94.85% with RSD of 1.28%–3.16%（n = 3）. The average recoveries of UA ranged from 88.67% to 94.77%, and the RSD was between 1.07% and 1.92%（n = 3）. The optimum conditions for extracting OA and UA from almond green husk extract were as follows： 95% ethanol as extractant; solid/solvent ratio, 1：40（g/mL）; extraction time, 40 min; and 3 extraction cycles.