UPLC-MS/MS法测定中药材中齐墩果酸与熊果酸的含量

Determination of Oleanolic Acid and Ursolic Acid in Chinese Herbal Medicine by UPLC- MS/MS

建立了一种同时检测中药材中齐墩果酸(OA)和熊果酸(UA)含量的超高效液相色谱串联质谱方法。采用超高效液相色谱-三级四极杆质谱(UPLC-TQMS)法对样品进行测定,Waters Acquity UPLC BEH C_(18)色谱柱(50 mm×2.1 mm,1.7μm)进行分离,以乙腈-5 mmol/L乙酸铵水溶液(氨水调至pH 9.24)为流动相,梯度洗脱;负离子模式下检测。结果表明,齐墩果酸和熊果酸在0.5~50.0 ng/mL浓度范围内线性关系良好,相关系数(r^2)分别为0.999 8和0.999 7;检出限(S/N=3)分别为0.006 6,0.012 8 ng/mL,定量下限(S/N=10)分别为0.002 0,0.003 8 ng/mL;对OA和UA进行加标回收实验,平均回收率分别为101.1%和100.8%,相对标准偏差(RSD,n=9)分别为1.8%和0.04%。对10种不同中药材中齐墩果酸和熊果酸含量进行检测,结果表明该方法快速简便、准确度高、重现性好,可用于含有齐墩果酸和熊果酸的中药材含量测定。

An ultra performance liquid chromatography coupled with tandem mass spectrometric（ UPLC- MS / MS） method was established for the simultaneous determination of oleanolic acid（ OA） and ursolic acid（ UA） in Chinese herbal medicine. The sample was detected by ultra performance liquid chromatography coupled with tandem mass spectrometry（ UPLC- MS / MS）. The separation of two compounds was achieved on a Waters Acquity UPLC BEH C_（18）column（ 50 mm × 2. 1 mm,1. 7 μm）with a mobile phase of 5 mmol / L ammonium acetate in water（ ammonium hydroxide adjust pH 9. 24）and acetonitrile in gradient elution mode. The detection of the compounds was performed in negative ion mode. The result indicated that the concentration of OA and UA had good linear relationships with peak area in the concentration range of 0. 5- 50. 0 ng / mL,and the correlation coefficients（ r^2）were 0. 999 8 and 0. 999 7,respectively. LODs（ S / N = 3） of the two compounds were 0. 006 6 ng /mL and 0. 012 8 ng / mL,respectively. LOQs（ S / N = 10） were 0. 002 0 ng / mL and 0. 003 8 ng / mL,respectively. The recoveries of OA and UA were 101. 1% and 100. 8%,with RSDs（ n = 9） of 1. 8%and 0. 04%, respectively. The method showed the advantages of simpleness, high accuracy and good reproducibility,and could be used for the detection of OA and UA content in the Chinese herb.