Quantitative Analysis of 6 Components in Hebra Sarcandrae by Ultra Performance Liquid Chromatography

KeyWord:Hebra Sarcandrae  ultra performance liquid chromatography(UPLC)  external standard method  quantitative analysis of multicomponents by single marker  quantitative analysis  caffeoyl derivatives  coumarin

Abstract:

      An ultra performance liquid chromatographic(UPLC) method was developed to quantitatively analyze six components,iª±e.,neochlorogenic,chlorogenic acid,4 dicaffeoylquinic acid,caffeic acid,rosmarinic acid and isofraxidin in Hebra Sarcandrae.The UPLC analysis was carried out on a Waters Acquity BEH C18 column(2.1 mm¡Á50 mm£¬1.7 ¦Ìm) by gradient elution with acetonitrile and water(both containing 0.1% formic acid) as mobile phase at a flow rate of 0.5 mL?min-1.The column temperature was maintained at 35 ¡æ and the detection wavelength was set at 330 nm.Isofraxidin was used as an internal reference substance to get the average relative correction factors(RCFs) of the other 5 components(neochlorogenic,chlorogenic acid,4-dicaffeoylquinic acid,caffeic acid,rosmarinic acid),and chromatographic column,mobile phase,flow rate,column temperature were investigated to validate the durability of the achieved RCFs.The contents of 6 components were determined by both quantitative analysis of multicomponents by single marker(QAMS) and external standard method(ESM),and the quantitative results were compared between the two methods by using paired t-test.The results showed that the linear ranges for neochlorogenic,chlorogenic acid,4-dicaffeoylquinic acid,caffeic acid and,rosmarinic acid and isofraxidin were 0.522 0-26.09£¬1.482-74.13£¬0.591 0-29.49£¬0.632 5-31.29£¬2.612-130.5£¬0.970 4-48.73 ¦Ìg?mL-1(r¡Ý0.999 7),respectively.The RSD values(n=6) of precision,reproducibility,and sample stability were not more than 1.9%.The average recoveries of 6 components were between 98.6% and 101.5% with RSDs not more than 1.9%.The achieved RCFs are good(RSD£¼5.0%).There is no significant difference in the contents obtained by the two methods with P values greater than 0.05,so both of the methods could be applied in the simultaneous determination of five caffeoyl derivatives and one coumarin in Hebra Sarcandrae.