基于HPLC-ESI-MS法续断发汗前后的成分分析

Component analysis of crude and sweated Dipsaci Radix based on HPLC-ESI-MS

目的 建立高效液相色谱-电喷雾-质谱（HPLC-ESI-MS）分析续断发汗前后成分变化的方法.方法 HPLC采用Agilent Eclipse XDB-C18 （250 mm×4.6 mm,5.0 μm）色谱柱；流动相为0.1％甲酸水溶液-乙腈,梯度洗脱：0～45 min,6％～～20％乙腈；45～90 min,20％～35％乙腈；90～110 min,35％～70％乙腈；110～120 min,70％乙腈；柱温为30℃,体积流量为0.8 mL/min；质谱采用电喷雾离子源,正负离子模式下全扫描检测,并通过[M-H]-、[M＋HCOO]-、[M＋H]＋、[M＋Na]＋等离子信息推断化合物.结果 建立了续断发汗前后的HPLC-ESI-MS图谱,均得到42个色谱峰,通过与对照品和文献对照,推断出了31个化合物；续断经产地加工发汗后,有些成分的量下降,如马钱苷酸、隐绿原酸、绿原酸、咖啡酸、马钱苷、异绿原酸B、续断苷A、续断苷B等；有些成分的量升高,如茶茱萸苷、川续断皂苷X、川续断皂苷Ⅵ、川续断皂苷Ⅶ、川续断皂苷乙、川续断皂苷XⅢ、川续断皂苷A等；其他成分的量几乎不变.结论 所建立的HPLC-ESI-MS图谱,便于全面控制续断发汗与未发汗药材的质量.

Objective A high performance liquid chromatography-electrospray tandem mass spectrometry （HPLC-ESI-MS） method was developed for the component analysis of the crude and sweated Dipsaci Radix. Methods Analyses were carried out on an Agilent Eclipse XDB-C18 （250 nun × 4.6 mm, 5.0 ~m） column by gradient elution using （A） 0.1% aqueous formic acid and （B） acetonitrile. The column was maintained at 30 ℃, and the flow rate was 0.8 mL/min. Chemical characteristics of the crude and sweated Dipsaci Radix were analyzed by MS techniques with an ESI source, the quasi molecular ions of compounds in both negative and positive modes were observed for molecule mass information as [M-H]-, [M ＋ HCOO]-, [M ＋ HI]＋, and [M ＋ Na]＋. Results An HPLC-ESI-MS spectrum of the crude and sweated Dipsaci Radix was established with 42 peaks respectively, and the potential structures of 31 characteristic components were identified by study on the MS of compounds and comparing with reference data and some of standards. After being sweated, the contents of some components in Dipsaci Radix decreased, such as loganic acid, 4-O-caffeoylquinic acid, chlorogenic acid, caffeic acid, loganin, isochlorogenic acid B, dipsanoside A, and dipsanoside B, but some ones increased, such as cantleyoside, dipsacoside X, dipsacoside VI, dipsacoside VII, dipsacoside B, dipsacoside XIII, and dipsacus saponin A, The amount of other ingredients are almost the same. Conclusion The HPLC- ESI-MS spectrum could comprehensively control the quality of the crude and sweated Dipsaci Radix.