Studies On The Chemical Constituents And Content Determination Of Mahuang Tincture Study On The Structural Modification Of Brevifolin And Their Pharmacological Activities

Part? Studies on the Chemical Constituents and Content Determination of Mahuang TinctureObjective: To isolate and identify the chemical composition and to establish a new HPLC method for the quantitative determination of Mahuang Tincture.Methods: Constituents were isolated and purified by sillca gel, Sephadex LH-20, ODS column chromatography, and TLC preparation, recrystallization technique. Their structures were identified on the basis of their chemical and physical properties as well as the spectra data of ESI-MS, 1H-NMR, 13C-NMR, etc, compared with data reported in literatures. HPLC method for the quantitative determination of geniposide in Mahuang Tincture was established.Results: 12 compounds were isolated and identified as follows: berberine(1), columbamine(2), coptisine(3), 8-oxo-epiberberine(4), 8-oxo-coptisine(5), geniposide(6), brucine(7), strychnine(8), vomicine(9), ?-amyrin(10), ?-sitosterol(11), daucosterol(12). HPLC method for the quantitative determination of geniposide in Mahuang Tincture was established. The linear ranges of geniposide was 0.149 ~ 0.894 ?g(r = 0.9997), the average recovery was 99.89%(RSD = 0.167%, n = 6).Conclusion: All compounds were isolated from this formula for the first time and alkaloids were the main compositions. HPLC method for detecting geniposide is fast, reliable, accurate, and can be used in the quality control of Mahuang tincture.Part? Study on the Structural Modification of Brevifolin and Their Pharmacological ActivitiesObjective: To modify the structure of brevifolin and evaluate antioxidant and anti-PVY effects for its derivatives in vitro and in vivo.Methods: Brevifolin was isolated as raw materials from the flowers of Ailanthus altissima(Mill.). Its derivatives were obtained through acetylation, epoxidation, reduction and addition reaction. Antioxidant activity was detected by DPPH method and anti-PVY activity was evaluated by DAS-ELISA and real-time quantitative PCR.Results: Acetylation, reduction and addition reaction were occurred and some corresponding products were obtained. In vitro antioxidant assay showed that brevifolin and its derivatives had a certain antioxidant activity. Methyl brevifolin carboxylate and brevifolin-11-oxime had obvious resistance to potato virus Y activity.Conclusion: Five brevifolin derivatives were successfully synthesized and in vitro antioxidant activity and anti-PVY effects in vivo for brevifolin derivatives were evaluated firstly.