| Schisandrae Chinensis Fructus(wuweizi)is the dried ripe fruit of the magnoliaceae plant Schisandra chinensis(Turcz.)Baillon.Wuweizi was a folk medicine in China,Korea and Russia,which was used as a sedative and tonic to treat seminal emission,excessive sweating,diarrhea,insomnia,fatigue and neurasthenia.Lignans with dibenzocyclooctadiene skeleton and organic acids are major kinds of bioactive compounds in wuweizi.It has been demonstrated that lignans have abundant bioactivities,including antihepatotoxic,antioxidant,antitumor,protecting the central nervous system and anticancer.Organic acids had beneficial pharmacological effects of arresting coughs and removing phlegm.Most reports of quantitative analysis were focused on each particular class of compounds or certain active ingredients for the quality control of wuweizi.To the best of our knowledge,there have been little previous reports on the simultaneous determination of lignans and organic acids in wuweizi.Not only ecological environment,but also harvesting,processing,storage and other factors,which results in the uneven quality of commercial Chinese medicine on the market.Therefore,how to ensure the quality and uniformity of wuweizi has become an essential problem in the production process,which needs to be solved.This project was supported by the National Traditional Chinese Medicine Standardization Project:the construction of Shengmai injection standardization(ZYBZH-C-JS-32),to carry out the research about quality formation and evaluation of wuweizi and the improvement of the quality standard of,wuweizi,the raw material of Shengmai injection.In order to explore the quality formation of wuweizi,ultra-fast performance liquid chromatography coupled with triple quadrupole-linear ion trap mass spectrometry(UFLC-QTRAP-MS/MS)was used to simultaneous detection of 15 components,including 11 lignans and 4 organic acids in samples from different product specifications,ecological environments,harvest time,processing and storage methods.To formulate the standard of wuweizi,harvesting,processing and storage,the raw material of Shengmai injection,wuweizi,which were collected from the base,were identified,fingerprinted and quantitatively analysed by TLC and UPLC.The research is mainly divided into the following parts:1.Literature research The systematic reviews of relevant literature from the domestic and abroad,and the study of literature reviews.2.Quality analysis of wuweizi from the product specifications.An useful method based on UFLC-QTRAP-MS/MS was optimized and exploited for the simultaneous detection of 15 constituents.Not only 11 lignans(schisandrol B,schizandrin C,angeloylgomisin H,gomisin D,schizantherin B,schisantherin,schisanhenol,gomisin J,deoxyschizandrin,schisandrin and y-schisandrin)but also 4 organic acids(protocatechuic acid,(S)-malic acid,quinic acid,and D(-)-tartaric acid)were detected in wuweizi.Combine with gray relational analysis(GRA)and technique for order preference by similarity to ideal solution(TOPSIS),wuweizi from different product specifications were analysed and evaluated.The protocol was reliable and accurate.It was fit for the simultaneous detection of multiple substances.And it may provide a new basis for the comprehensive assessment and overall control of wuweizi.3.Study on the effects of different ecological environments to wuweizi.A reliable method based on UFLC-QTRAP-MS/MS was used to simultaneously determine the samples.After detennining the concentrations of 15 different metabolites in each of 12 different samples(6 wild and 6 cultivated).Following the detection of the compounds in these samples,principal components analysis(PCA),partial least squares discrimination analysis(PLS-DA),independent sample t-test and GRA have been applied to determine differences between cultivated and wild samples and evaluated the samples comprehensively.The results showed that the differential compounds(i.e.quinic acid,L-(-)-malic acid,protocatechuic acid,schisandrol B)were significantly related to the classification of wild and cultivated wuweizi.The quality of wild wuweizi was better than the quality of cultivated wuweizi.The protocol laid the way for the basis of the effects of different ecological environments of wuweizi.4.Study on the dynamic accumulation and variation of indicators of wuweizi.UFLC-QTRAP-MS/MS was established to simultaneously determine the contents of 15 constituents in wuweizi at various harvest times,and to investigate the variation of dynamic accumulation.GRA was carried out for the comprehensive evaluation.The results demonstrated that the wuweizi collected from mid august to mid september were superior.Our protocol provided the foundation and support for optimizing suitable harvesting time of wuweizi.5.Study on the effects of different processing methods on wuweizi.UFLC-QTRAP-MS/MS was established to simultaneously determine the contents of 15 constituents in wuweizi from different processing methods.Hierarchical cluster analysis(HCA)and GRA were used to comprehensively analyze and evaluate the different samples.GRA results indicated that the quality of oven drying after steaming samples not as good as the oven drying samples and the quality of S7 was the best.Our protocol may lay the way for the optimization of wuweizi from different processing methods,standardization and normalization.6.Analysis of compounds in wuweizi under different storage conditions.An useful method based on UFLC-QTRAP-MS/MS was applied to the simultaneous detection of 15 constituents including lignans and organic acids in wuweizi under different storage conditions(various temperatures and package materials).Besides,GRA and TOPSIS were used to comprehensively evaluate the samples under different storage conditions.The results of these two kinds of methods showed that the samples of polyethylene,ceramic cylinder and gunny bag were of good comprehensive quality.The optimum storage condition for wuweizi was storing with a polyethylene bag in a cool storage warehouse.This protocol may provide the foundation and support for optimizing fit storage conditions of wuweizi.7.Standardized research of wuweizi,which is one of the raw material of Shengmai injection.According to the efficacy characteristics of shengmai injection and the active constituents of wuweizi,and the research of wuweizi quality formation,the quality standard of the raw material of shengmai injection,wuweizi,was improved on the basis of the quality standard of wuweizi in the Chinese Pharmacopoeia(2015).After optimization,the thin-layer chromatography(TLC)which could identify standard wuweizi,deoxyschizandrin,schisandrin and schisandrol B were established.The UPLC fingerprint of the high quality raw material of wuweizi were optimized and established,and 8 lignans were identified from 13 common peaks.To establish an optimized method for the determination of schisandrin and schisandrol B,and to establish the content limit.Based on the actual conditions of harvesting,processing and storage in the producing area,schisandrin and schisandrol B were used as the indexes to investigate the harvesting time,optimized processing methods and storage conditions,and to establish the enterprise standards for the harvesting,processing and storage of wuweizi,the raw material of shengmai injection. |
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