反相高效液相色谱法测定木瓜中齐墩果酸和乌索酸的含量

Determining Oleanolic Acid and Ursolic Acid Contents in Fructus chaenomelis by RP-HPLC

[目的]建立木瓜中齐墩果酸和乌索酸含量的测定方法。[方法]色谱柱为Kromasil C18柱(250 mm×4.6 mm,5μm),流动相为甲醇∶0.2%磷酸(90∶10),流速0.8 ml/min,检测波长210 nm,柱温28℃。在此条件下测定木瓜中齐墩果酸和乌索酸含量。[结果]齐墩果酸进样量在0.66～5.94μg(r=0.999 6),乌索酸进样量在0.67～6.03μg(r=0.999 6)范围内具有良好的线性关系。在色谱检测中,乌索酸与齐墩果酸的分离度为1.86,齐墩果酸和乌索酸色谱峰对称因子分别为1.020和1.015,理论塔板数分别为13 687和14 290。在精密度试验中,齐墩果酸峰面积RSD为2.2%,乌索酸峰面积RSD为0.3%。在重现性试验中,齐墩果酸峰面积RSD为1.9%,乌索酸峰面积RSD为2.2%。齐墩果酸和乌索酸平均回收率分别为99.1%和97.5%,RSD分别为1.7%和2.3%。[结论]该方法简便,灵敏,结果准确,重现性好,可用于评价木瓜药材的质量。

[Objective] The purpose was to establish the determination method of oleanolic acid and ursolic acid contents in Fructus chaenomelis. [Method] The chromatography column was Kromasil C 18 column （250 min×4.6 mm, 5μm）. The mobile phase consisted of methanol and 0.2% phosphoric acid （90：10）,with flow velocity of 0.8 ml/min, detecting wavelength of 210 nm and column temperature of 28 ℃, under which the oleanolic acid and ursolic acid contents in F. chaenomelis were determined. [Result] The oleanolic acid with injection volume of 0.66-5.94μg （r= 0.999 6） and ursolic acid with injection volume of 0.67-6.03μg （r=0.999 6） had good linear relationship. In the chromatography detection, the separation of ursolic acid and oleanolic acid was 1.86 and their peak symmetry factors were 1.020 and 1.015 resp. and the theoretical plate numbers were 13 687 and 14 290 resp. In the precision test, the peak area RSDs of oleanolic acid and ursolic acid were 2.2 % and 0.3 % resp. In the repeatability test, the peak area RSDs of oleanolic acid and ursolic acid were 1.9 % and 2.2 % resp. Their average recoveries were 99.1% and 97.5 % resp, and RSDs were 1.7 % and 2.3 % resp. [Conclusion] The method was simple, convenient and sensitive with accurate resuh and good repeatability, and could be used to evaluate the quality of Chinese traditional crude drug F. chaenomelis.