| Thlaspi arvense Linn, also called "da-ji","e-lan" vegetable, was the cruciferous plant in a family of dicotyledons, and it is widely edible in the southeast border gansu province. Thlaspi arvense Linn has active effects on liver clearing, vision improving and diuresis for folk records, and can be used for treating nephritis and endometriosis.In order to optimize the flavonoids extraction process of Thlaspi arvense, investigate its antioxidant and anti-inflammatory activity, screen active fractions, purify chemical compounds and analysis it’s content, Thlaspi arvense provided by Hua-hong pharmaceutical was selected as the research object, then eight fractions was obtained by water extraction and alcohol sedimentation, different solvents extraction, D101macroporous resin separation successively. Isovitexin was prepared by High performance liquid chromatographic(HPLC). Finally, antioxidant and anti-inflammatory activity-tracking were carried out. The mainly research aspects content as the following:1. Optimization of The Flavonoids Extraction Process of Thlaspi Arvense Linn Herb Single factor experiments (extraction temperature, extraction time, material liquid ratio, ethanol concentration) and the orthogonal experiment were successively carried out to optimize the flavonoids extraction process of Thlaspi arvense herb. Results showed that the order of the factors was temperature of heating mantle> extraction time> soild-liquid ratio> ethanol concentration, the raw material(20g) was extracted at180℃, in50%ethanol with the ratio of material to liquid at1:13for2.5h, under these conditions the average yield of total flavonoids came up to89.05mg.2. Antioxidant Activity ResearchCompared with the ascorbic acid, antioxidant activities of these8fractions were evaluated in vitro including scavenging DPPH free radical and hydroxyl free radical, reducing Fe3+, and inhibiting peroxide oxidation of peanut oil. It showed that all fractions were found to have marked different antioxidant activities but weaker than ascorbic acid, Thlaspi arvense Linn herb existed antioxidant compositions indeed. Furthermore, the data generated by this study showed that30%ethanolic elution of n-butanol extract by D101macroporous resin (fraction VII) and the ethyl acetate extract (fraction II) had better antioxidant activity obviously than other fractions but slightly weaker than ascorbic acid, the remaining water layer(fraction IV) had the weakest antioxidation. It also explained that different solvent extraction, macroporous resin separation can enrich more antioxidant contents.3. Anti-inflammatory Activity Research Compared with the Hua-hong tablet and dexamethasone, anti-Inflammatory activities of these fractions were evaluated using murine models of xylene induced ear edema and carrageenin induced paw edema. The result showed that the ethyl acetate extract (fraction II), the n-butyl alcohol extract (fraction III) and70%ethanolic elution of n-butanol extract by D101macroporous resin (fraction IX) had no significant anti-inflammatory effects for no significant difference (P>0.05) compared with blank, the others had obviously anti-inflammatory effects for significant difference (P<0.05or P<0.01) compared with blank, in the two models. Furthermore, in the model of xylene induced ear edema,30%ethanolic elution of n-butanol extract by D101macroporous resin (fraction VII, inhibition ratio:81.55%) had marked anti-inflammatory ability for significant difference (P<0.05) compared with Hua-hong tablet (inhibition rate:49.51%).4. Separation and Content DeterminationIsovitexin was prepared from fraction VI and fraction VII by High performance liquid chromatographic (HPLC) and the structure was identified by the spectrum analysis. With HPLC, the standard curves of isovitexin was obtained by gradient sample injection, relationship between the content of isovitexin in X (μg) and peak area Y is:Y=7.7×105X-1.4×105, R2=0.9999, and it showed a good linear relationship in the range from0.32μg to3.2μg. According to the regression equation, the isovitexin contents in fraction I, III, VI and VII were0.280%,0.259%,44.059%,6.983%respectively. |
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