Content Determination of Oleanolic Acid and Ursolic Acid in Chaenomelis Fructus by HPLC with Chiral Mobile Phase
Objective To establish a high-performance liquid chromatography(HPLC)method with chiral mobile phase for simultaneous determination of oleanolic acid and ursolic acid in Chaenomelis Fructus.Methods The chromatographic column was Shimpack GIST C18 column(250 mm×4.6 mm,5 μm),and the mobile phase was acetonitrile-methanol-0.5% cyclodextrin ammonium acetate solution containing 12 mmol/L hydroxypropyl group-β-cyclodextrin(64∶12∶21,V/V/V),the flow rate was 1.0 mL/min,the detection wavelength was 210 nm,the column temperature was 18℃,and the injection volume was 20 μL.Results The separation degree of oleanolic acid and ursolic acid was higher than 1.5,the linear ranges of oleanolic acid and ursolic acid were 0.009-0.182 μg/mL and 0.016-0.325 μg/mL(r=0.999 8,0.999 9,n=6),respectively.The limits of detection were 0.86 and 0.58 μg/mL,respectively.The limits of quantification were 2.75 and 1.86 μg/mL,respectively.The RSDs of precision,stability,and repeatability test results were all lower than 2.0% (n=6).The recoveries of oleanolic acid and ursolic acid were 99.74% and 99.22%,with RSDs of 1.00% and 0.93% (n=6),respectively.The total content of oleanolic acid and ursolic acid in three batches of samples was in the range of 2.305%-2.357% .Conclusion The improved method is simple,accurate,and has good repeatability,it can effectively separate and determine the content of oleanolic acid and ursolic acid in Chaenomelis Fructus,which can be used for the quality control of Chaenomelis Fructus.
Chaenomelis FructusHPLCchiral mobile phaseoleanolic acidursolic acidcontent determination
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