Studies The Distribution Of Effective Components On Different Tissues Of Spatholobus Suberectus Dunn Based On The Chromatography-pharmacodynamics Relationship

Objective"Assessing quality by distinguishing the traditional Chinese medicinal materials(TCMM) features"is the essence of the experience of discrimination which determine the authenticity, pros and cons of Chinese traditional medicine based on traditional Chinese medicine appearance manifested features. The essence of "assessing quality by distinguishing the TCMM features" is the correlation between the appearance of TCM and inherent quality. The appearance of TCM is not only the shape characteristics, but also the external manifestation of organizational structures and chemical components. This study will conduct research of the chemical constituents of different tissues in Spatholobus suberectus Dunn in order to study the essence of "assessing quality by distinguishing the TCMM features".The color of xylem in Spatholobus suberectus Dunn is red brown or brown while the phloem is reddish brown to dark brown with resin secretions. They alternately arranged in a plurality of concentric or eccentric rings. The traditional Chinese medicine to the quality evaluation of Spatholobus suberectus Dunn that the number of eccentric ring-containing resin takes 3 to 8 rings, which means the ring number of phloem is 3 laps or more. However, is there a scientific basis to back up the idea? What are its effective components? What’re the differences and similarities of the chemical compositions between the xylem and phloem? These problems still need to be further explored.Thus, the study is to use the method of chromatographic-activity analysis for finding out the effective components in Spatholobus suberectus Dunn. Meanwhile the chemical compositions in the xylem and phloem is comparatively analyzed in order to definite the distribution of the effective components on the tissues and establish the correlation between shape characteristics and inherent quality. Currently, the chemical research in the xylem and phloem tissue of Spatholobus suberectus Dunn has not been reported.Method:1. Qualitative analysis of flavonoids on two tissues of Spatholobus suberectus DunnThe rapidly profiling, identifying and determining of chemical constituents of flavonoids of Spatholobus suberectus Dunn (SSDF) was achieved by using a Liquid Chromatography-Electrospray Ion Trap Mass Spectrometry. With analzying the mass spectrometry data of the various peaks and comparing the fragmentation regularity of flavonoids, comparing synthetically their retention time, MS data and reference data reported in the literature, each peak structures were identified, which will lay the material foundation for the oncoming experimental study.2. The hematopoiesis effect of SSDFAfter injection with APH (i.h.) and CTX (i.p.), blood-deficient model was established. Observe the SSDF’s function of enriching blood and the protective effect on the kidneys and spleens by measured the blood’s RBC, HGB, HCT and the pathological section after given drug treatment.3. Analysis of enriching blood effective components from SSDF based on chromatography-pharmacodynamics relationshipEstablishing the chromatogram-activity relationship by means of Grey Correlation Analysis and speculating the effective components according to the sorting of grey relational degree.Establishing the chromatogram-activity relationship by means of Pearson correlation analysis and speculating the effective components according to the correlation coefficient.4. HPLC fingerprint spectrum study of SSDF and the contents distribution of effective components and the total flavonoids in different tissues4.1 The contents distribution of effective components and the total flavonoids in the xylem and phloemMeasure the contents of effective components in different batches of SSDF in order to analyze the contents distribution between the xylem and phloem.4.2 The contents distribution of effective components and the total flavonoids in different rings of Spatholobus suberectus DunnScreen different number of rings of Spatholobus suberectus Dunn and measure the contents of effective components in order to analyze the distributional differences with the rings changed.Result:1. Qualitative analysis of flavonoids on two tissues of Spatholobus suberectus DunnA total of 23 compounds were isolated and 10 of which were identified or tentatively characterized by LC-ESI-MS based on standard components comparison, UV absorption curve and MS spectra data in both negative and positive ion modes, including one of which was firstly reported from Spatholobus suberectus Dunn.2. The hematopoiesis effect of SSDFThe mice injected with APH (i.h.) and CTX (i.p.) can be blood-deficient, after given the extract of SSDF, it can be significantly increase the amount of RBC, HGB, HCT in the blood and perfect the damages of kidneys and spleens.3. Analysis of enriching blood effective components from SSDF based on chromatography-pharmacodynamics relationshipIt was demonstrated from the Grey Correlation Analysis that pharmacodynamic substances of enriching blood were mainly for 10、1、8、6、2、4,while the result of Pearson correlation analysis indicated that the peak of 5、10、14 are highly correlated with the promotion of RBC, the peak of 2、4、5、12、14 with HGB, the peak of 5、10、 14 with HCT.4. HPLC fingerprint spectrum study of SSDF and distributional differences of effective components’ content4.1 The contents distribution of effective components and the total flavonoids in the xylem and phloemCompared of chromatographies, there were some similarities and differences between the xylem and phloem. There were 17 common peaks. The contents of five kinds of effective components and the total flavonoids in the xylem were higher than the phloem, especially the contents of formononetin, ononin, daidzin. From the result, we could infer that these three components were mainly distributed in the xylem.4.2 The contents distribution of effective components and the total flavonoids in different rings of Spatholobus suberectus DunnCompared with the Spatholobus suberectus Dunn in 2 rings, the contents of catechin, epicatechin, daidzein, formononetin, ononin and the total flavonoids had the characteristics of tendency that contents were positively related with rings.Conclusion:The rapidly profiling, identifying and determining of chemical constituents of flavonoids of Spatholobus suberectus Dunn (SSDF) were achieved by using LC-ESI-MS; The SSDF has significant effect of enriching blood which combined its’ chromatographic fingerprint with the enriching blood activity and the effective components were screened; The contents distribution of effective components and the total flavonoids in the xylem and phloem were different and positively related with the rings of Spatholobus suberectus Dunn.