Studies On Chemical Constituents And Content Determination Of Total Saponins From Kalopanax Septemlobus

Kalopanax septemlobus (Thunb.) Koidz.(Araliaceae) recorded in the Pharmacopoeia of the People's Republic of China(1977), is widely distributed in east of Asia. The stem barks of Kalopanax septemlobus (Thunb.) Koidz. have been used to treat neuralgia, rheumatic arthritis, lumbago, furuncle, carbuncle, wound, diarrhea and scabies for many years. In order to develop medical resource and evaluate the quality control of this crude drug, the chemical constituents and content determination of total saponins from Kalopanax septemlobus (Thunb.) Koidz. were studied in this thesis.Firstly, the advances on the chemical constituents and pharmacological effects of Kalopanax septemlobus (Thunb.) Koidz. were reviewed. 45 triterpenoids were found, including mono-saccharides and bisaccharides. The main aglycones of these compounds were hederagenin and oleanane. Pharmacological investigations performed so far on Kalopanax septemlobus (Thunb.) Koidz. have shown that extracts and saponins from this herb possesses several biological activities, such as antitumor, antibacterial, anti-inflammatory, anti-rheumatoid arthritis and hypoglucemic effect. Kalopanaxsaponin A is the main pharmacological constituent.Six known compounds have been isolated from the barks of Kalopanax septemlobus (Thunb.) Koidz. The dried barks of Kalopanax septemlobus (Thunb.) Koidz. (8kg) obtained in Jingyu, Jilin Province of China were reflexed with 75% ethanol. The extract was combined and concentrated to dryness. The residue was dissolved in water and extracted with CHCl3, EtOAc and n-BuOH saturated with water to afford three fractions. The CHCl3 fraction (40g) was chromatographed on a silica gel column and eluted with mixtures of petroleum ether and EtOAc (3:1) to afforded hederagenin (31mg). The EtOAc fraction was also chromatographed on a silica gel column and eluted with gradient mixtures of CHCl3-MeOH (9:1) to afford kalopanaxsaponin A (66mg) and 3-O-α-L-arabinopyranosyl hederagenin(41mg), with CHCl3 -MeOH-H2O (70:30:1.5) to afford kalopanaxsaponin I (38mg), with CHCl3 -MeOH-H2O (7:3:1) to afford syringin (27mg). The n-BuOH fraction was subjected to silica gel column chromatography using CHCl3 -MeOH-EtOAc-H2O (2:2:4:1) as elutents to afford kalopanaxsaponin B (51mg).Additionally, the author obtained total saponins through macroporous adsorbing resin. An HPLC method was established for determination of kalopanaxsaponin A in total saponins. The determination was achieved on a stainless steel Kromasil C18 column (4.6 mm×250 mm, 5μm) at 25℃, and carried out with acetonitrile and 0.05% phosphoric acid (55:45) at a flow rate of 1.0mL/min. Detection wave was set at 203nm. Three batches of total saponins obtained from Kalopanax septemlobus (Thunb.) Koidz. were determined and the average content of kalopanaxsaponin A was 9.66mg/g. In conclusion, this thesis provided systematic research on the chemical constituents of Kalopanax septemlobus (Thunb.) Koidz., obtained total saponins through macroporous adsorbing resin, and established an HPLC method to determine pharmacological constituent kalopanaxsaponin A in total saponins. It offered experiment data for further development of Kalopanax septemlobus (Thunb.) Koidz.