Screening Of Efficient Constituents Of Compound Biejiarangan Tablets Based On Different Levels And A Discussion Of Some Ralated Issues

Efficient constituents recognition is the foundation of the mordernization of traditional Chinese medicine (TCM). Screening the efficient constituents (EC) of TCM objectively can provide important scientific evidence and technical support for the illustration of mechanism, improvement of quality control, guarantee of clinical efficiency and security, and can lay a solid foundation of the development of multiple-component Chinese medicine. Due to the complexity and the limitations, the EC of TCM formulas have not been successfully recognized.In order to get a breakthrough of EC-recognization, taking Compound Biejiaruangan Tablets (CBTs) as a model medicine, a new channel was established: learning serum pharmaco chemistry of TCM (SP-TCM), the constituents derived from in-vitm, animal and clinical samples were detected in turn On this basis, a tentative pattern for screening EC of TCM named "in vitro-animal-clinical" which is progressive and clinical-combination was established. This aimed at screening EC of TCM more effectively.First, to characterize the compounds of CBTs in vitro, an analytical method was established:adopting HPLC, ginsennoside Rgl, paeoniae, forsythin and formic acid were detected. Utilizing UPLC-Q-TOF/MS,38possible compounds were characterized, including saponins (ginsennosides, astragalosides), monoterpenes (paeoniae series), flavonoids (fermononetin), phenylethanoid glycosides (Forsythosides), volatile oil (ligustilide), organic acids (gallic acid) etc, which paved the way to the EC-screening of CBTs, the establishment of the method of quality control, illustration ofmechandsm.Second, for further screening of EC, based on the previous result that13constituents including Ginsennosides, Notoginsennosides, Paeoniae, has been detected in the rats’blood, and tracking metabonomics, using UPLC-Q-TOF/MS, the difference of plasma between normal rats and rats which suffered from hepatic fibrosis were compared. Meanwhile, based on GC/TOF-MS, contrastive study of their endogenous substance was carried out. The PCA analysis result suggest that the metabolits of the two groups could be distinguished, which indicates that the ADME process of drug-induced constituents differs from each other, causing the different acquisition of EC.Third, to avoid the species variation derived from in-vitro to in-vivo, from animal to human body, and to screen EC more scientifically, the blood collected from the persons who have taken normal dose of CBTs in advance has been analyzed using both UPLC-TOF/MS and MS/MS. The results showed that only Paeoniae have been captured. Reviewing the dose of administration, the sample preparation and detection methods etc, some problems of current SP research pattern have appeared before our eyes, and remained to be further and deep study.Making a general observation of these results above, we could know:1, the captured compounds successively decreased from to human body, indicating that the constituents in-vitro and from animal are not the EC of CBTs.2, the variation of plasma between normal and model rats indicates that EC screening based on model rats should be more focused.3, the analysis of clinical samples exists problems such as administration dose and sample preparation etc. The phenomenon above shows current SP applied in the EC-recognization has some problems including clinical correlation, feasibility and universality etc. These issues have been discussed one by one in this paper.Overall, through the triune avenue of "in vitro-animal-clinical" for EC screening, the possible EC of CBTs was initially interpreted, which can serve as a research foundation for the final formation of the novel pattern of EC recognition of TCM. Meanwhile, the discussion of the reflected problems and the proposed suggestions can provide some references for the research of EC of TCM.