Studies On Constituents Of The Leaves Of Actinidia Kolomikta

Actinidia is a liane plant belonging to the family of Actinidiaceae. Approximately 107 species of Actinidia were mainly distributed in China, North Kora, Japan and Russia, and 105 species were found in Northeast region and Changbai mountain in China. Two species namely Actinidia Kolomikta (Rupr.et Maxim.) Planch and Actinidia arguta (Sieb. Et Zucc.) Planch were discovered widely growing in Changbai mountain of Jilin Province in China. The fresh fruit of Actinidia Kolomikta is riched in Vitamin C, and has been used to cure the lack of Vitamin C. Flavonoids, Leucoyandia, Lactone, Saponarin have been reported in the leaves of Actinidia Kolomikt.This paper further study on the chemical constituents and bioactivities of the leaves of Actinidia kolomikta on the base of pre-paper.Sixteen compounds were separated from the leaves of Actinidia Kolomikta and fourteen was identified by 1HNMR, 13CNMR, 1H-1HCOSY, HMQC, HMBC and MS as Kaempferol (L1), quercetin– 3– O–β– D - glucoside (L2), kaempferide– 7– O - (4" - O -acetyl)– rhamnoside(L3), kaempferol– 7– O - rhamnoside(L6), 4'– methoxyl– quercetin– 7– O - (4" - O -α-L- acetylrhamnosyl)– 3– O–β– D– glucopyranoside(L10), isorhamnetin– 3– O–β– D– glucoside(L11), kaempfero-l-3-O-β-D-galactopyranoside(L12), kaempferide– 7– O - (3"– O -α-L- acetylrhamnosyl)– 3– O– rutinoside(L13), kaempferol -7– O - (4"– O -α-L-acetylrhamnosyl) -3– O– rutinoside(L14), kaempferol– 3– O–α– L– rhamnopyranosyl - (1→3)–α– L - rhamn-opyranosyl - (1→6)–β– D– galactopyranoside (L16), Kaempferol- 7– O– rhamnosyl– 3– O– rutinoside (L17), 3-O-β-D-glucopyranosyl-2α, 3β, 6β, 19α, 21β, 23– hexahydroxylolean– 12– en– 28 - oic acid(L18),β-sitosterol(a) andβ-daucosterol(b).Except compound L1, all the compounds are obtained from Actinidia kolomikta for the first time. Compound L3, L10, L13, L14, L18 are new compounds.This paper studied the extract technology of the total flavonoids of the leaves of Actinidia Kolomikta through monofactor and orthogonal experiment.The suitable condition and optimal extraction technology of total flavonoids was determined as 4 times of 90% alcohol,refluxing 4 times and 2, 1.5 ,1 and 0.5 hours each.The optimum absorption conditions were the sample concentration of 200mg·ml-1, the PH of the sample at 6, the current velocity of 4BV·h-1 of eluant of 90% ethanol and the current velocity of 4BV·h-1. Combined the eluant and then removing the solvent under reduced pressure at 60℃, a dark residue was obtained. The residue was subjected to column chromatography on silica gel (EtOAc- MeOH-H2O, 10:1:0.3), and combined the same fractions on the basis of their TLC profiles. After the solvent was removed under reduced pressure at 50℃the total flavonoids in the leaves of Actinidia Kolomikt. was obtained.The quality standard of total flavonoids in the leaves of Actinidia Kolomikt was studied in this paper. We have identified the kaempferide -7– O - (4"– O - acetylrhamnosyl)– 3– O– rutinoside by TLC, determinated the water content and the residue at 500℃. The water content should below 5%, the residue at 500℃should below 0.2%. The content of total flavonoids was also determined by UV chromatophy, it showed that total flavonoidswas above 72%. We have also determinated the content of kaempferide -7– O - (4"– O - acetylrhamnosyl)– 3– O– rutinoside by HPLC. The content of kaempferide -7– O - (4"– O - acetylrhamnosyl)– 3– O– rutinoside and showed it was above 32%.This paper we also investigated the antioxidant effect of the isolated flavonoid compounds on the experimental system of free-radical-initiated peroxidation: the hemolysis of human erythrocyte induced by water-soluble initiator, 2,2'-azobis(2-amidinopronane hydrochloride) (AAPH). The isolated compounds can all inhibit the hemloysis induced by AAPH.The bioactivities of the total flavonoids (TFAK) has studied in this thesis, the results showed that high and midle dosage of TFAK can all decrease the raised content of TC,TG,LDL-c and TC/HDL-c, rise HDL-c.