阅读说明：本技术 一种软枣猕猴桃根提取物、提取分离方法及其应用 (Actinidia arguta root extract, extraction and separation method and application thereof ) 是由 白乃生 王梦旭 郭雅欣 郭森 彭赛男 张姗姗 岳文平 高兵 于 2019-09-17 设计创作，主要内容包括：本发明属于植物提取分析技术领域,特别涉及一种软枣猕猴桃根提取物,提取分离方法及其应用；该提取物包含以下21种化学组分：7-O-[β-D-吡喃木糖-(1-6)-β-D-吡喃葡萄糖苷]-1,8-二羟基-3-甲氧基-酮,3-O-[β-D-吡喃鼠李糖-(1-6)-β-D-吡喃半乳糖]-5,7,3′,4′-四羟基黄酮,儿茶素,β-sitosterol,槲皮素,异槲皮苷,齐墩果酸,乌苏酸,2α,3β-二羟基齐墩果烷-12烯-28酸,2α,3α,24-三羟基乌苏烷-12烯-28-酸,胡萝卜苷等。本发明的提取物在制备改善癌症患者晚期生活水平的抗癌药物中具有美好的应用前景。(the invention belongs to the technical field of plant extraction and analysis, and particularly relates to a actinidia arguta root extract, an extraction and separation method and application thereof; the extract comprises the following 21 chemical components: 7-O- [ beta-D-xylopyranose- (1-6) -beta-D-glucopyranoside ] -1, 8-dihydroxy-3-methoxy-ketone, 3-O- [ beta-D-rhamnopyranose- (1-6) -beta-D-galactopyranose ] -5,7,3',4' -tetrahydroxyflavone, catechin, beta-sitosterol, quercetin, isoquercitrin, oleanolic acid, ursolic acid, 2 alpha, 3 beta-dihydroxyoleanane-12 ene-28 acid, 2 alpha, 3 alpha, 24-trihydroxy-ursane-12 ene-28-acid, daucosterol and the like. The extract has good application prospect in preparing anti-cancer drugs for improving the late life level of cancer patients.)

1. the actinidia arguta root extract is characterized by comprising the following components: 7-O- [ β -D-xylopyranosyl- (1-6) - β -D-glucopyranoside ] -1, 8-dihydroxy-3-methoxy-one; 3-O- [ β -D-rhamnopyranose- (1-6) - β -D-galactopyranose ] -5,7,3',4' -tetrahydroxyflavone; a catechin; beta-sitosterol; quercetin; isoquercitrin; oleanolic acid; 2 beta, 3 beta, 23 alpha-trihydroxyoleanane-12-ene-28-acid; arjunolic acid; euscaphic; 2 α,3 β,19 α, 23-aquahydroxyusu-12-en-28-acid-28-O- β -D-pyrangliosidase; ursolic acid; 2 α,3 β -dihydroxy oleanane-12 en-28 acid; 2 α,3 α, 24-trihydroxyursane-12-en-28-oic acid; 3 β -hydroxy-20-ene-ursane; corosolic acid; 2 α,3 β, 24-trihydroxyursane-12-en-28-oic acid; 2 α,3 β,19 α, 23-tetrahydroxy ursane-12-en-28-oic acid; 2 α,3 β, 23-trihydroxyursane-12, 20(30) -diene-28-acid; 2 α,3 α, 24-trihydroxyursane-12, 20(30) -diene-28-acid; daucosterol.

2. The method for extracting and separating actinidia arguta root extract as claimed in claim 1, comprising the steps of:

S1: pretreating Actinidia arguta Royle root, completely soaking in 90% ethanol solution for 48 hr, stirring every 2 hr, filtering to obtain extractive solution, and concentrating to obtain extract;

wherein, the material-liquid ratio of actinidia arguta root to ethanol solution is 3 g: 10 mL;

S2: completely dissolving the extract obtained in the step S1 in water, adding an ethyl acetate solvent with the volume being 3 times that of the extract, extracting for 4 times, combining all ethyl acetate extraction liquid, and concentrating under reduced pressure to obtain an ethyl acetate extraction extract;

s3: performing first-step coarse separation by using 100-200 mesh silica gel, naturally drying and crushing an ethyl acetate extract sample, sieving by using a 100-mesh sieve, performing dry-method sample loading, performing column chromatography by using a methanol-dichloromethane solution, analyzing by using a high performance liquid chromatography and a thin layer chromatography, and dividing the sample into 8 sections to obtain 8 fractions; and finally, carrying out chromatographic separation on 8 fractions to obtain 21 actinidia arguta root compound components.

3. the method for extracting and separating actinidia arguta root extract as claimed in claim 2, wherein the volume ratio of the methanol-dichloromethane solution in S3 is 1: 100-1.

4. the method for extracting and separating actinidia arguta root extract as claimed in claim 2, wherein the chromatography and separation mode in S3 is as follows: repeatedly carrying out chromatographic separation by using one or more of silica gel, polyamide, CHP and Sephadex LH-20 packing;

The eluent used for the chromatographic separation in S3 was: one or more of petroleum ether-ethyl acetate solution, chloroform-methanol solution, dichloromethane-methanol solution, chloroform-acetone solution, methanol-water solution and dichloromethane-methanol solution.

5. The method for extracting and separating actinidia arguta root extract as claimed in claim 2, wherein the pretreatment in S1 comprises the following steps: drying Actinidia arguta, and pulverizing into granules;

In S1, the top of Actinidia arguta root powder is 10cm lower than the liquid level of ethanol solution.

6. The method for extracting and separating actinidia arguta root extract as claimed in claim 2, wherein the concentration under reduced pressure in S2 is performed at 40-45 deg.C, 0.09-0.10 Mpa, and 5.5-6.5 h.

7. the use of actinidia arguta root extract as claimed in claim 1, wherein said extract is used in the manufacture of an anti-cancer medicament for improving the later life level of a cancer patient.

8. the use of actinidia arguta root extract as claimed in claim 7, wherein 7-O- [ β -D-xylopyranose- (1-6) - β -D-glucopyranoside ] -1, 8-dihydroxy-3-methoxy-ketone, 3-O- [ β -D-rhamnopyranose- (1-6) - β -D-galactopyranose ] -5,7,3',4' -tetrahydroxyflavone, catechin, quercetin, isoquercitrin, oleanolic acid, ursolic acid in the preparation of a medicament for inhibiting α -glucosidase activity, acetylcholinesterase, tyrosinase activity.

Technical Field

The invention relates to the technical field of plant extraction and separation, and particularly relates to a actinidia arguta root extract, an extraction and separation method and application thereof.

background

Kiwi (Actinidia chinensis Planch) also known as "kiwifruit" is a large deciduous vine plant. Because of its rich nutritive value, it is known as "fruit king". The Actinidia plants are various in species, about 50 species, and are widely distributed in the market at present, the Actinidia arguta is a plant of Actinidia of actinidiaceae, and is a plant of the same genus and different from the Actinidia arguta, and the Actinidia arguta root is the root of Actinidia arguta (sieb. In ancient times, predecessors use actinidia arguta roots as medicines, and the actinidia arguta roots recorded in the ancient books have the effects of clearing heat, cooling blood and inducing diuresis. Modern researches show that actinidia arguta root has pharmacological effects of reducing blood sugar, reducing blood lipid and resisting tumor cells. Nowadays, scientific research mainly focuses on the research on the whole plant chemical components and the biological activity of actinidia chinensis planch of actinidia of actinidiaceae, and the research on actinidia arguta planch of the same genus is less, so that the extraction and separation method of actinidia arguta roots is necessary.

disclosure of Invention

In order to solve the problems, the invention provides a actinidia arguta root extract, an extraction and separation method and application thereof.

In order to achieve the purpose, the invention adopts the technical scheme that:

an actinidia arguta root extract comprising the following components:

7-O- [ β -D-xylopyranosyl- (1-6) - β -D-glucopyranoside ] -1, 8-dihydroxy-3-methoxy-one; 3-O- [ β -D-rhamnopyranose- (1-6) - β -D-galactopyranose ] -5,7,3',4' -tetrahydroxyflavone; a catechin; beta-sitosterol; quercetin; isoquercitrin; oleanolic acid; 2 beta, 3 beta, 23 alpha-trihydroxyoleanane-12-ene-28-acid; arjunolic acid; euscaphic; 2 α,3 β,19 α, 23-aquahydroxyusu-12-en-28-acid-28-O- β -D-pyrangliosidase; ursolic acid; 2 α,3 β -dihydroxy oleanane-12 en-28 acid; 2 α,3 α, 24-trihydroxyursane-12-en-28-oic acid; 3 β -hydroxy-20-ene-ursane; corosolic acid; 2 α,3 β, 24-trihydroxyursane-12-en-28-oic acid; 2 α,3 β,19 α, 23-tetrahydroxy ursane-12-en-28-oic acid; 2 α,3 β, 23-trihydroxyursane-12, 20(30) -diene-28-acid; 2 α,3 α, 24-trihydroxyursane-12, 20(30) -diene-28-acid; daucosterol.

the invention also provides an extraction and separation method of the actinidia arguta root extract, which comprises the following steps:

S1: pretreating Actinidia arguta Royle root, completely soaking in 90% ethanol solution for 48 hr, stirring every 2 hr during soaking, filtering to obtain extractive solution, and concentrating to obtain extract;

wherein, the material-liquid ratio of actinidia arguta root powder to ethanol solution is 3 g: 10 mL;

S2: completely dissolving the extract obtained in the step S1 in water, adding an ethyl acetate solvent with the volume being 3 times that of the extract, extracting for 4 times, and combining all ethyl acetate extract liquor; concentrating under reduced pressure to obtain ethyl acetate extract;

S3: performing first-step coarse separation by using 100-200 mesh silica gel, naturally drying and crushing an ethyl acetate extract sample, sieving by using a 100-mesh sieve, performing dry-method sample loading, performing column chromatography by using a methanol-dichloromethane solution, analyzing by using a high performance liquid chromatography and a thin layer chromatography, and dividing the sample into 8 sections to obtain 8 fractions; and finally, carrying out chromatographic separation on 8 fractions to obtain 21 actinidia arguta root compound components.

Preferably, the volume ratio of the methanol-dichloromethane solution in the S3 is 1: 100-1.

preferably, the separation mode in S3 is as follows: repeatedly separating with one or more of silica gel, polyamide, CHP, Sephadex LH-20 filler

the eluent used for the chromatographic separation in S3 was: one or more of petroleum ether-ethyl acetate solution, chloroform-methanol solution, dichloromethane-methanol solution, chloroform-acetone solution, methanol-water solution and dichloromethane-methanol solution.

Preferably, the specific process of the pretreatment in S1 is as follows: drying Actinidia arguta, and pulverizing into granules;

In S1, the top of Actinidia arguta root powder is 10cm lower than the liquid level of ethanol solution.

preferably, the conditions of the reduced pressure concentration in S2 are 40-45 ℃, 0.09-0.10 Mpa of vacuum degree and 5.5-6.5 h of time.

the invention also protects the application of the actinidia arguta root extract, in particular to the application of the actinidia arguta root extract in preparing an anti-cancer drug for improving the late life level of cancer patients.

preferably, the extract of the actinidia arguta root contains 7-O- [ beta-D-xylopyranose- (1-6) -beta-D-glucopyranoside ] -1, 8-dihydroxy-3-methoxyl-ketone, 3-O- [ beta-D-rhamnopyranose- (1-6) -beta-D-galactopyranose ] -5,7,3',4' -tetrahydroxyflavone, catechin, quercetin, isoquercitrin, oleanolic acid and ursolic acid, and can be applied to the preparation of medicines for inhibiting the activity of alpha-glucosidase, acetylcholinesterase and tyrosinase.

the invention has the following beneficial effects:

The actinidia arguta root extract is separated by using solvents such as ethyl acetate and the like for extraction and combining column chromatography technologies such as silica gel, CHP-20P, macroporous resin, Sephadex LH-20 and the like, wherein part of the extract has good tyrosinase, acetylcholinesterase and alpha-glucosidase inhibitory activity, and can be applied to research on anticancer pharmacological action for improving the late life level of cancer patients by combining with medicines.

Drawings

FIG. 1 is a partial extraction and separation diagram of Actinidia arguta root extract of the present invention;

FIG. 2 shows the chromatographic separation process of Fr2, Fr4 and Fr7 in 8 fractions of Actinidia arguta root extract;

FIG. 3 shows the chromatographic separation process of Fr1 and Fr3 from 8 fractions of Actinidia arguta root extract;

FIG. 4 shows the chromatographic separation process of Fr5 and Fr6 from 8 fractions of Actinidia arguta root extract;

FIG. 5 is a high performance liquid chromatogram of the invention when 5 compounds in Actinidia arguta root extract are analyzed;

FIG. 6 is a high performance liquid chromatogram of the Actinidia arguta root extract of the present invention when 5 compounds are analyzed.

Wherein, 5 compounds in fig. 5 and fig. 6 are respectively: 7-O- [ β -D-xylopyranosyl- (1-6) - β -D-glucopyranoside ] -1, 8-dihydroxy-3-methoxy-ketone (1); 3-O- [ β -D-rhamnopyranose- (1-6) - β -D-galactopyranose ] -5,7,3',4' -tetrahydroxyflavone (2); a catechin (3); quercetin (5); isoquercitrin (6).

Detailed Description

In order that the objects and advantages of the invention will be more clearly understood, the invention is described in detail below by way of specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.