| Purpose:The scutellariae syrup made from unicornolum unilaterally of Rhus chinensis has clinical efficacy in the treatment of coronary heart disease.However,its pharmacodynamic substance basis for the treatment of coronary heart disease is not yet clear.The purpose of this experiment is to screen the anti-coronary heart disease activity of different polar parts of Rhus chinensis.To lay the foundation for clarifying the pharmacodynamics and effective parts of Rhus chinensis against coronary heart disease.Method:(1)Prepare different polar parts of Rhus chinensis by macroporous resin method,including polysaccharide part(A fraction),macroporous adsorption resin water elution part(B fraction),10%ethanol elution part(C fraction),50%ethanol elution site(D fraction),90%ethanol elution site(E fraction)a total of 5 sites,The zebrafish heart failure model induced by verapamil hydrochloride was used to screen the anti-coronary heart disease activity of each chemical site of Rhus chinensis.(2)Enriching the effective parts of Rhus chinensis.against coronary heart disease,and combining various separation and purification methods such as conventional silica gel column chromatography,medium and low pressure chromatography,and high-pressure preparative liquid phase to separate and identify the chemical components of the active part of Rhus chinensis.(3)Aiming at the effective parts of Rhus chinensis against coronary heart disease,the peaks of the effective parts were scanned by liquid chromatography coupled with high-resolution time-of-flight mass spectrometry to perform primary and secondary mass spectrometry scans,and the characteristic peaks of the active site were identified by high-resolution mass spectrometry.Characterize its chemical profile.(4)The extraction process of the effective part of Rhus chinensis and the macroporous resin purification process were investigated by orthogonal test method,and the ultra-high performance liquid chromatographic tandem triple quadrupole mass spectrometry was used for the simultaneous determination of the effective fraction of Rhus chinensis.19 chemical components for quality control.Result:(1)The different parts of Rhus chinensis praecox were screened through the activity model of zebrafish anti-coronary heart disease.In the assessment of cardiac function improvement,10%ethanol eluted part(C part)and 50%ethanol eluted part(D part),The 90%ethanol elution site(E site)significantly improved zebrafish heart expansion,venous stasis,cardiac output,blood flow velocity,and heart rate.The polysaccharide(A site)and water elution(B site)groups were not significantly improved.(2)Twenty-one compounds were isolated and purified from the active parts of Rhus chinensis against coronary heart disease,they were gallic acid(1),protocatechuic aldehyde(2),sakakin(3),4-hydroxybenzoic acid(4),methyl gallate(5),catechin(6),ethyl gallate(7),ellagic acid(8),rutin(9),pentagalloylglucose(10),kaemferol-3-O-rutinose(11),dihydrofisetin(12),phloridzin(13),butin(14),phloretin(15),kaempferol(16),semialactone(17),rhuscholide A(18),teragalloylglucose(19),protocatechuic acid(20),fisetin(21).Among them,compounds 2,4,6,9,11,14 and 19 were isolated for the first time from Rhus chinensis.and their structures were determined by UV,mass spectrometry,and NMR.(3)A total of 46 components were identified by LC-TOF-MS on the active fractions of Rhus chinensis.,which contained 12 galloyl saccharides,8 phenolic acids,21 flavonoids,and 5 other classes.Inferred,the mass spectrometry properties of galloyl saccharides,phenolic acids,and flavonoids are summarized.(4)The optimum extraction process of the effective parts of Rhus chinensis was as follows:ratio of material to solution 1:20,proportion of ethanol solvent 70%,extraction times 2 times,extraction time 90 min;Purification of effective parts of Rhus chinensis:Use SP825L type resin column,ratio of diameter to height:1:5,sample volume:0.35 g·g-1 dried resin of crude drug,adsorption flow rate:1 BV·h-1,except miscible solvent acid Water consumption:4 BV,solvent removal flow rate:2 BV · h-1,enriched eluent concentration and dosage:50%ethanol,4 BV,enriched solvent flow rate:2 BV · h-1 The established ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was used to simultaneously determine the contents of 19 components in the effective fraction of Rhus chinensis,providing an effective detection method for the quality control of multi-components in Rhus chinensis.Conclusion:This experiment through the screening of the active parts of Rhus chinensis chinensis against coronary heart disease,carry out the separation and identification of the chemical composition of the active site,then,the use of liquid chromatography and mass spectrometry to identify the primary basis for the efficacy of the substance against coronary heart disease,elucidation of its efficacy components Most of them are polar phenols,and further researches on the extraction and purification of their effective parts and the determination of multi-components have laid the foundation for the development of effective parts of Rhus chinensis against coronary heart disease. |
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