Study On Effective Substances In Xueshuan Xinmaining Tablet

In today’s world, as a kind of common and frequent disease, cardiovascular andcerebrovascular diseases were serious and affected people’s quality of life. Among them thecoronary heart disease (CHD) and stroke are main. With a growing and aging population, theincidence of the diseases and death rate will increase dramatically in China.Xueshuan Xinmaining Tablet (XXT) is composed of salvia miltiorrhiza Bge.,ligusticum chuanxiong Hort., ilex pubescens Hook.Et Arn., sophora japonica L.and somerare herbs such as Ginsenosides, artifical Calculus bovis, bufo bufo gargarizans Cantor,artifical Musk. etc. XXT can make the effect of aromatic resuscitation and promoting bloodcirculation for removing blood stasis. So it has been widely used in the treatment ofcardiovascular and cerebrovascular diseases such as CHD, angina, cerebal thrombosis whichbelong to the symptomatology of stagnation of vital energy and blood stasis.In this paper, XXT was studied by using HPLC-ELSD to study the fingerprints andusing UPLC-ESI-MS/MS to study the serum pharmacochemistry. This paper was not onlyexpected to study the transfer of blood-forming and active ingredients to improve the qualitycontrol standards of XXT, but also make its clinic application more effective and reliable,which also provide a new research method for quality control of XXT.First, the fingerprints of XXT was analyzed by Chromatographic fingerprint evaluationsystem (2004A version). Samples of10batches were of high similarity and the similiaritydegree were more than0.880. Seventeen common peaks were found in the HPLC-ELSDfingerprint of XXT, nine constituents were identified as rutin, ferulic acid, salvianolic acid B,ginsenoside Rg1, ginsenoside Re, cholic acid, cinobufagin, resibufogenin and tanshinone IIAby comparing with the peak retention time of reference substance. The fingerprint of XXTshowed good reproducibility, and the method is stable and reliable, which can be used forthe quality control of XXT.Second, the main constituents in XXT were analyzed. As XXT containes not onlyvolatile constituents but also nonvolatile constituents, so the coupling technique such asUPLC-ESI-MS/MS and GC-MS were used to analyze the main constituents in XXT. Byanalyzing the data obtained from UPLC-ESI-MS/MS, we identified47compounds includingtanshinones, salvianolic acids, ginsenosides, cholic acids cinobufotalin, rutin, ferulic acidand some other constituents in XXT. By searching the data obtained from GC-MS in NIST2008database,36volatile compouds were identified, and the major constituents wereborneol, isoborneol and muscone.Last, by using the established analysis method of UPLC-ESI-MS/MS, we optimized theadministration dosage of XXT and the pretreatment conditon of serum. Then we establishedthe analysis method of serum pharmacochemistry for XXT. From a comprehensive analysisof the UPLC-ESI-MS/MS chromatography of XXT, administration of serum and blankserum,23components were observed in rat’s serum. Among them,13of the original formcompounds from XXT and2of the metabolites were identified by comparing their retentiontime, MS and MS/MS spectra data and the reported literature. Most of these identifiedcompouds were reported to have effect on cardiovascular and cerebrovascular diseases. Weuse the methodology of serum pharmacochemistry, an analyzed method was established forthe study of the absorbed compounds in rat’s serum after oral administration of XXT, whichmade its real effective substances clear.Through HPLC fingerprints and serum pharmacochemistry, we have laid the foundationfor the improvement of quality control, provided the scientific basis for the clinicalapplication, made the accurate effective substances clear.