摘要
目的建立超高效液相色谱串联三重四级杆质谱(UPLC-MS/MS)同时测定积雪草中7种有效成分(积雪草苷B、羟基积雪草苷、积雪草苷、羟基积雪草酸、积雪草酸、槲皮素和山柰酚)的含量。方法色谱柱:ACQUITYBEHC_(18)(2.1mm×100mm,1.7μm),流动相:乙腈-5mmol·L^(-1)醋酸铵,梯度洗脱,流速:0.4mL·min^(-1),柱温:35℃,正离子多反应监测模式。考察该方法的专属性、标准曲线和定量下限、精密度和回收率、稳定性和重复性。结果异构体积雪草苷B和羟基积雪草苷达到基线分离。定量分析的7种有效成分:积雪草苷B、羟基积雪草苷、积雪草苷、槲皮素、山柰酚、羟基积雪草酸和积雪草酸分别在考察的浓度范围内:20.00~400.00ng·mL^(-1),20.00~400.00ng·mL^(-1),20.00~400.00ng·mL^(-1),10.00~200.00ng·mL^(-1),10.00~200.00ng·mL^(-1),50.00~1000.00ng·mL^(-1),50.00~1000.00ng·mL^(-1),呈良好的线性关系(r>0.9902);回收率和精密度RSD分别在1.76%~5.69%和1.03%~4.67%内。稳定性和重复性良好,RSD范围分别为0.86%~3.21%和0.94%~3.45%。结论本研究所建立的同时测定雪草中7种有效成分(5个五环三萜和2个黄酮)的UPLC-MS/MS定量分析方法简便、快捷、准确,可为综合评价积雪草的质量提供参考。
Objective To develop a rapid and sensitive analytical method for the determination ofseven major compounds（ Asiaticoside B,madecassoside,Asiaticoside,madecassic acid,asiatic acid,quercetin,and kaempferol） in Centellaasiaticaby using ultra-performance liquidchromatography coupled with triple-quadrupole tandem mass spectrometry（ UPLC-MS/MS） in multiple-reactionmonitoring mode. Methods UPLC-MS/MS assay with positive ion mode was performed on a Waters ACQUITY BEHC18（ 2. 1 mm × 100 mm,1. 7 μm） with the mobile phase consisting of acetonitrile and 5 mmol·L^-1 ammonium acetate in gradient elution at a flow rate of 0. 40 m L·min^-1 and the column temperature was set at 35 ℃. The positive ion monitoring mode was adopted for MS. The selectivity,linear ranges and limit of quantitation,precision and recovery,repeatability and stability were investigated. Results Underthe optimized chromatographic conditions,good separation for seven target compounds including isomer asiaticoside B and madecassoside were achieved within 8. 5 min. Satisfactory linearities were achieved with wide linearranges,which were 20. 00-400. 00 ng · m L^-1 ,20. 00-400. 00 ng · m L^-1 ,20. 00-400. 00 ng · m L^-1 ,10. 00-200. 00 ng·m L^-1 ,10. 00-200. 00 ng·m L^-1 ,50. 00^-1 000. 00 ng·m L^-1 ,50. 00^-1 000. 00 ng·m L^-1 for the determined compounds respectively and fine determination coefficient（ r 0. 990 2）,the overall recoveries wereranged from 98. 3%^-1 00. 0% with the RSD ranging from 1. 76%-5. 69%. The RSD of precision was between1. 03%-4. 67%,for the repeatability and stability,which were between 0. 86%-3. 21%,0. 94%-3. 45%,respectively. Conclusion It was the first report about simultaneousanalysis of 7 major components including 5pentacyclic triterpenes and 2 flavonoids in Centellaasiatica by using UPLC-MS/MS method,which could be used for quality control of Centellaasiatica.
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2017年第19期1949-1953,共5页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金资助项目(81603066)
河北省自然科学基金资助项目(2015206116)
河北医科大学第二医院科学研究基金资助项目(NO2h2017022)
