Chinese Formula Mahuangtang Chemical Composition Analysis And Spectrum-Effect Relation Research

Objective:This experiment using spectrum-effect combination method, by chromatographic mass spectrometry, the compounds of Mahuangtang will be identified, and the fat-soluble and water-soluble parts of the fingerprint will be established. At the same time, through combining with pharmacological experiment result of diaphoresis and asthma. Followed by studying the relationship of the "spectrum-effect" using partial least squares regression (PLSR) method. Therefore the effective substance of Mahuangtang will be revealed.Methods:1、GC-EI/MS was applied to establish 41 groups of Mahuangtang different does fat-soluble fingerprints. Comparing the differences of chromatograms, the common peak mass spectrogram were compared with NIST standard mass spectrogram, then its chemical information was identified, and analysis the component herbs ownership with the literature.2、UPLC-Q-TOF/MS was applied to establish 41 groups of Mahuangtang different does water-soluble fingerprints.Comparing the differences of chromatograms, the common peak’s quasi molecular ion peaks m/z values were be extracted automatically by MZmine-2.9.1 software, and compared with m/z values under [M+H]+ model in self-built chemical components database, then its chemical information and component herbs ownership was identified.3、At 40min、60min、80min、100min、120min points, the influence of the sweat gland secretion of rats toes of Mahuangtang different does groups and single herbs was investigated by coloring the drops, with sweat percentage increase as the evaluation index; guinea pig isolated trachea spiral strip was use to investigate the compound’s relaxation effect on the trachea smooth muscle, with spasmolysis percentage as the evaluation index.4、In the research of the "spectrum-effect" relation, the independent variables were the common peak area of 41 fat-soluble and water-soluble fingerprints, the dependent variables 1 were sweat drops number of rats toes at different time points, the dependent variables 2 was the difference value of guinea pig isolated trachea smooth muscle. The PLSR method was used to establish "spectrum-effect" relationship mathematical mode, according to the distribution of scattered points in scored diagrams classifying chromatographic peak, according to the distance between independent variables and origin, and the size of the weight coefficient in load ing diagram, screening the characteristic peaks which were significantly correlated with diaphoresis and asthma, then its chemical information was cleared.Results:1、Forty chemical compositions were identified, including a-terpineol, cinnamic aldehyde, clove aldehyde, butanol, etc. From these, volatile oil content was the main chemical composition, which come from Ephedrae Herba and Cinnamomi Ramulus mainly.2、Thirty nine chemical compositions were identified, including ephedrine, cinnamic acid, latrile and glycyrrhizin. The main chemical composition was alkaloids from Ephedrae Herba, organic acids from Cinnamomi Ramulus and Armeniacae Semen Amarum, and flavonoids from Glycyrrhizae Radix et Rhizoma.3、The results from pharmacodynamic experiments showed Ephedrae Herba is the major influencial element, different proportions of Mahuangtang were diaphoresis and asthma action in rats. In addition, Cinnamomi Ramulus and Armeniacae Semen Amarum proved to be significantly promoted diaphoresis and asthma pharmacodynamic results of Mahuangtang, but Glycyrrhizae Radix et Rhizoma proved to be significantly inhibited diaphoresis results of Mahuangtang.4、The research results of "spectrum-effect" relation between Mahuangtang and diaphoresis and asthma pharmacodynamic results showed, the explanation capacity for diaphoresis of principal components Compl-Comp4 have been reached 85.1%, explanation capacity for diaphoresis of principal components Compl～Comp5 have been reached 85.8%, Compl has the strongest predict ability to model; Chromatographic peaks that were correlated larger with diaphoresis and asthma pharmacodynamic results in loading diagram come from fat-soluble fingerprint; Screening 121 chromatographic peaks as characteristic peaks of Mahuangtang diaphoresis,21 chromatographic peaks had been identified, Screening 180 chromatographic peaks as characteristic peaks of Mahuangtang asthma,20 chromatographic peaks had been identified; including 43 chromatographic peaks as characteristic peaks of Mahuangtang diaphoresis and asthma.Conclusion:The research of "spectrum-effect" relation between Mahuangtang and diaphoresis and asthma pharmacodynamic results had been carried out with chromatography-mass spectrometry linked technique.79 chemical compositions were identified, mainly including volatile oil content of fat-soluble part, alkaloids/organic acids/flavonoids of water-soluble part; Screening 121 chromatographic peaks as characteristic peaks of Mahuangtang diaphoresis, and 180 chromatographic peaks as characteristic peaks of Mahuangtang asthma,43 chromatographic peaks as characteristic peaks of Mahuangtang diaphoresis and asthma,27 chromatographic peaks had been identified; Chromatographic peaks that were correlated larger with diaphoresis and asthma pharmacodynamic results in loading diagram come from fat-soluble fingerprint, prompting volatile oil content is the the main effective substance of Mahuangtang diaphoresis and asthma.