Study On The Analysis Methods Of Major Functional Components In Crataegus Pinatifida By HPLC And HPCE

Three reversed-phase high-performance liquid chromatographic (HPLC) methods and two capillary zone electrophoretic (CZE) methods for the analysis of functional components in Crataegus Pinatifida were developed and comparison of some properties between capillary electrophoresis and high-performance liquid chromatography was made. Changes in functional components during development of Cratagus Pinatifida were monitored by the developed HPLC methods.The separations of components in Crataegus by HPLC were obtained with a reversed-phase column (4.6 X 200mm) filled with 5 u rn HYPERSIL BDS as the solid phase. The flow rate of mobile phase was 0.6 ml/min. Detection at 350 nm with CH3CN: H2O (20:80,V/V) as the mobile phase was found to be the most suitable approach for the analysis of vitexin-2"-O-rhamnose, rutin, hyperside, and vitexin, the four flavones were baseline separated in llminites. For cholorogenic acid, epicatechin and B2, the mobile phase was CH3OH: H2O (23:77,V/V) at pH 3.10 (adjusted by 50% phosphoric acid) with detective wavelength 280 nm. The mobile phase for ursolic acid and oleanolic acid was methanol and water (90:10) at pH 3.10 (adjusted by 50% phosphoric acid) and 210 nm as the detective wavelength. The purified extract process of proanthocyanidin B2 in Crataegus was primarily established and the extract was characterized by H NMR and I3C NMR.In HPCE system, the four flavones were separated on baseline within 15 minites by use of 50mM borax buffer contained 15% acetonitrile and adjusted to pH 8.15 with50% phosphoric acid. The condition for the seven major components in Crataegus which was baseline separated with 14 minutes was 40 mmol/L borax buffer solution and 10% acetontrile as the additive at pH (adjusted by 50% phosphoric acid) 8.21.These methods either by HPLC or CZE revealed good accuracy and precision by method validations. By comparison, it showed better sensitivity and higher efficiency in CZE system.The contents of functional components in different part of Crataegus were different by the analysis of developed HPLC methods and hyperside was not detected. With the development of Crataegus, the functional components changed in different stages.