Characterization Of Chemical Constituents In Gui-zhi-ma-huang-ge-ban-tang And Screenig The Constituents With Anti-inflammatory Activity

Gui-Zhi-Ma-Huang-Ge-Ban-Tang originates from Shang-Han-Lun, consisting of Cinnamomi ramulus, Paeoniae radix alba, Zingiberis rhizoma recens, Glycyrrhizae radix et rhizoma preparata, ephedrae herba, Jujubae fructus and armeniacae semen amarum. It’s a combination of Gui-Zhi-Tang and Ma-Huang-Tang, which is only one-third dosage of each them. This formula has the efficacy of slight sweating and relieving superficies acrid-warm, especially the unique therapeutic effects for allergic skin diseases.LC-Q-TOF-MS combined with LC-IT-MS method was applied to analyze the 124 constituents from Gui-Zhi-Ma-Huang-Ge-Ban-Tang, including classification of flavonoids, triterpenoids, monoterpene glycosides, alkaloids and organic acids, but also analysis their typical ions and fragmentation patterns. A total of 26 compounds were confirmed unambiguously by comparing with authentic standards.To minimize complex, the formula was dissected into several components through macroporous resin column chromatography and preparative liquid chromatography method. Evaluation of anti-inflammatory activity was performed by LPS-induced RAW 264.7 cells, and 21 of these components showed activity to some degree. Then, 11 compounds were selected to validate the anti-inflammatory activity following the qualitative analysis and active indexes calculation of active components, which were originated from armeniacae semen amarum, Jujubae fructus, ephedrae herba, Glycyrrhizae radix et rhizoma preparata and Paeoniae radix alba. As a result, isoliquiritin apioside, paeoniflorin and benzoylpaeoniflorin were proved to be the anti-inflammatory constituents of Gui-Zhi-Ma-Huang-Ge-Ban-Tang, and inhibition rate IC50 of benzoylpaeoniflorin was(155.7±7.7)μM.