Determination of Oleanolic Acid and Ursolic Acid in Almond Green Husk Extract by High Performance Liquid Chromatography
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 (R2=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.
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