CN101591325B

CN101591325B - Method for extracting catechin

Info

Publication number: CN101591325B
Application number: CN200810049869XA
Authority: CN
Inventors: 康文艺; 宋艳丽; 姬志强; 王金梅
Original assignee: Henan University
Current assignee: Henan University
Priority date: 2008-05-26
Filing date: 2008-05-26
Publication date: 2012-02-29
Anticipated expiration: 2028-05-26
Also published as: CN101591325A

Abstract

The invention discloses a method for extracting catechin, comprising the following steps: (1), a raw material of rubiaceae mitragynaKorth plant cap core wood is subjected to cold soaking by using a solvent at room temperature after being dried and pulverized, an extract is obtained after the reagent is recovered by decompression, the extract is suspended in water, and sherwood oil, ethyl acetate and normal butanol are sequentially used for carrying out extraction; (2), an extract of the ethyl acetate is packed by silica gel of 60-100 meshes according to the weight ratio of 1 to 30 after sample stirring, the sherwood oil and acetone are eluted according to the volume ratio of 9:1-10:1, thin-layer detection is carried out, the similarities are combined, the solvent is recovered by decompression, and a crude product of catechin is obtained; and (3), the acetone is added into the crude product, the mixture is heated, the acetone is gradually added into the mixture until the crude product is dissolved, aether with the volume of one tenth to one ninth of the volume of the acetone is added into the mixture, and crystals are separated out by standing and are filtered to obtain a pure product of the catechin. The invention has simple extracting method, abundant sources of the raw materials and low cost, a catechin compound obtained by extracting has extensive bioactivity, a catechin monomer is obtained by extracting, the solvent used in the extracting process can be recycled, the extracting quantity is large and industrial production can be realized. By adopting the method, the extracting efficiency is high, and the yield reaches 50-90 percent.

Description

A kind of method of extracting catechin

Technical field

The present invention relates to a kind of process for extracting of catechin, relate in particular to a kind of from medicinal plant Rubiaceae cap stamen fogfruit cap stamen wood the method for extraction separation compound catechin.

Background technology

Catechin compounds is of a great variety; Be representative wherein with flavanoid compoundss such as catechin, l-Epicatechol, l-Epigallocatechols; In plant, distributing extensively, is many with pulse family (like catechu, acacia farnesiana), Rubiaceae (like Uncaria gambier Roxb.), the Rosaceae (like tormentic, Fruit of Nanking Cherry, potentilla anserina, list hawthorn), polygonaceae (holding like rheum officinale, adder-wort 、 pivot), Theaceae (like camellia), Hippocastanaceae (like red horse-chestnut, california horse-chestnut) wherein.Catechin has tangible removing interior free yl, anticancer, anti-inflammatory, mutation, anti-ageing and improve biological activitys such as liver function, can be used for body-care and meat preservation etc.At present; The process for extracting of catechin perhaps is that raw material makes through further processing with the tea-polyphenol for from tealeaves, to extract, and the production of tea-polyphenol is comparatively universal at home; Because the restriction of extraction process; Often foreign matter content is high in the product, some also contain be detrimental to health such as organic solvent residual and pesticide residue such as chloroforms, so the market development has received great restriction.

Torrid zone medicinal plant cap stamen wood (Mitragyna rotundifolia Kuntze.) is that Rubiaceae (Rubiaceae) cap stamen wood belongs to (Mitragyna L.) plant.Among the people of many uses, can be used for generating heat, the treatment of hernia, myalgia, and can expelling parasite.The research of cap stamen fogfruit mainly concentrates on the indoles alkaloid composition; Pharmacological research is also quite clear and definite; Mainly concentrate on antitumor, cardiovascular disorder and anti-microbial activity aspect, at present, from Rubiaceae cap stamen fogfruit cap stamen wood, extracting catechin does not also have report.

Summary of the invention

It is abundant to the purpose of this invention is to provide a kind of raw material sources, a large amount of methods that prepare the compound catechin of extracting from tropical medicinal plant cap stamen wood of organic solvent-free and pesticide residue.

To achieve these goals, technical scheme of the present invention has adopted a kind of method of extracting catechin, may further comprise the steps:

(1) gets raw material Rubiaceae cap stamen fogfruit cap stamen wood, after drying, pulverizing, at room temperature carry out cold soaking, obtain extract medicinal extract behind the reclaim under reduced pressure reagent, medicinal extract is suspended in water, use sherwood oil, ETHYLE ACETATE and n-butanol extraction successively with solvent;

(2) acetic acid ethyl ester extract is mixed an appearance back dress post with 60～100 order silica gel according to weight ratio at 1: 30, sherwood oil and acetone by volume (9: 1-10: 1) wash-out, thin layer detects, and merges identical point and carries out decompression and solvent recovery, gets the catechin bullion;

(3) add acetone in the bullion, heating adds acetone more gradually up to dissolving crude product, adds the ether of acetone volume (1/10-1/9) then, through placing, separates out crystal, crosses the pure article of catechin that filter.

Described raw material Rubiaceae cap stamen fogfruit is a cap stamen wood.

Used raw material is preferably cap stamen wood skin or cap stamen wood leaf.

Described solvent is that concentration is 70% aqueous acetone solution or in water, methyl alcohol, ethanol and the ETHYLE ACETATE any one.

Cold soaking in the step (1) is specially cold soaking 2～4 times, each 6～9 days.

The add-on of acetone is 2～5 times of volumes of catechin bullion weight in the step (3).

Method of the present invention is the extraction that raw material carries out catechin with Rubiaceae cap stamen fogfruit cap stamen wood, and plant cap stamen wood distributes extensively at occurring in nature, has expanded the value of exploiting and utilizing of this medicinal plant of cap stamen wood greatly.Process for extracting of the present invention is simple, and raw material sources are abundant, and cost is low, and the compound catechu that obtains that extracts have wide biological activity, and extract and obtain catechin monomers, the recyclable utilization of the solvent of using in the leaching process, extracted amount is big, but suitability for industrialized production.Adopt method extraction efficiency of the present invention high, yield reaches 50%～90%.

Embodiment

Embodiment 1

The process for extracting of the catechin of present embodiment may further comprise the steps:

(1) get exsiccant cap stamen wood skin (3kg) and pulverize after, with 70% aqueous acetone solution cold soaking 3 times at room temperature, each 7 days, obtain extract medicinal extract behind the reclaim under reduced pressure reagent, medicinal extract is suspended in water, use sherwood oil, ETHYLE ACETATE and n-butanol extraction successively;

(2) acetic acid ethyl ester extract 35g mixes a kind back dress post with 60～100 order silica gel according to weight ratio at 1: 30,9: 1 by volume wash-outs of sherwood oil and acetone, and thin layer detects, and merges identical point and carries out decompression and solvent recovery, gets the catechin bullion;

(3) in bullion, add proper amount of acetone, heating adds acetone more gradually and just dissolves up to bullion; The ether that adds acetone volume 1/10 is then placed, and separates out crystal, crosses and filters the pure article 20g of catechin.The yield that this kind method is extracted catechin is 85%.

Embodiment 2

(1) get cap stamen wood leaf 1kg through dry, pulverize after, with ethyl acetate solution cold soaking 3 times at room temperature, each 7 days, obtain extract medicinal extract behind the reclaim under reduced pressure reagent, medicinal extract is suspended in water, use sherwood oil, ETHYLE ACETATE and n-butanol extraction successively;

(2) acetic acid ethyl ester extract 15g mixes a kind back dress post with 60～100 order silica gel according to weight ratio at 1: 30,10: 1 by volume wash-outs of sherwood oil and acetone, and thin layer detects, and merges identical point and carries out decompression and solvent recovery, gets the catechin bullion;

(3) in bullion, add proper amount of acetone, heating adds acetone more gradually and just dissolves up to bullion; The ether that adds acetone volume 1/9 is then placed, and separates out crystal, crosses and filters the pure article 8g of catechin.The yield that this kind method is extracted catechin is 90%.

Adopt the structure of the catechin that method of the present invention extracts to identify as follows:

[proterties] yellow powder

[evaluation] molecular formula: C ₁₅H ₁₄O ₆, molecular weight: 290.272

The instrument material: fusing point is measured with the micro-fusing point appearance of XRC-1 type that scientific instrument factory in Sichuan produces, and TM is not proofreaied and correct; Optically-active is measured with the SPEA-300 appearance; Infrared at Bio-Rad FTS-135 infrared measurement; Ultraviolet is measured on UV-210A ultraviolet spectroscopy appearance; 1H, 13C NMR collection of illustrative plates is by the Brukeram-400MHz nmr determination.TMS is interior mark; The two dimensional NMR collection of illustrative plates is by the DRX-500MHz nmr determination; Mass spectrum is measured by VG.AUTO Spec-3000 type mass spectrograph.

Experimental result: Mp:240; Optical Rotation: [α] _D ²⁰39.7 (MeOH), utilize EI MS, 1H NMR and 13C NMR test, normal data is following: LC-MS m/z:290,152,139,123,69.1H?NMR(400M?Hz，DMSO)δ：4.72(1H，d，J＝8.0Hz，H-2)，3.98(1H，m，H-3)，2.50(2H，m，H-4)，5.70(1H，d，J＝2.0Hz，H-6)，5.88(1H，d，J＝2.0Hz，H-8)，6.64(1H，d，J＝2.0Hz，H-2′，5′)，6.87(1H，dd，J＝2.0，8.0Hz，H-6′)。13C?NMR(100MHz，DMSO)δ：28.2(C-4)，64.9(C-3)，78.1(C-2)，95.1(C-6)，94.1(C-8)，98.5(C-4a)，114.9(C-2′，6′)，144.4(C-5′)，130.8(C-1′)，144.6(C-3′)，117.9(C-4′)，151.8(C-5)，156.3(C-7)，155.5(C-8a)。

[inspection] utilizes TLC, is hyperfluorescence at UV 254nm; With the sulfuric acid developer, 110 ℃ following 5 minutes, present yellow

[assay] utilizes HPLC, chromatographic condition: the YWG-C18 10 μ m (chromatographic column of 250mm * 4.6mm).Moving phase: methyl alcohol: water: phosphoric acid (55: 44.6: 0.4), flow velocity: 1.0mL/min detects wavelength 255nm, RT value 8.9min.

It should be noted last that: above embodiment is only in order to explanation; And unrestricted technical scheme of the present invention; Although the present invention is specified with reference to the foregoing description; Those of ordinary skill in the art is to be understood that: still can make amendment or be equal to replacement the present invention, and not break away from any modification or the local replacement of the spirit and scope of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.

Claims

1. method of extracting catechin is characterized in that: may further comprise the steps:

(2) acetic acid ethyl ester extract is mixed a kind back dress post with 60～100 order silica gel according to weight ratio 1:30, sherwood oil and acetone is 9:1-10:1 wash-out by volume, and thin layer detects, and merges identical point and carries out decompression and solvent recovery, gets the catechin bullion;

(3) add acetone in the bullion, heating adds acetone more gradually up to dissolving crude product, adds the ether of acetone volume 1:10-1:9 then, through placing, separates out crystal, crosses the pure article of catechin that filter;

Wherein, described solvent is that concentration is 70% aqueous acetone solution or ETHYLE ACETATE.

2. the method for extraction catechin according to claim 1 is characterized in that: used raw material is preferably cap stamen wood skin or cap stamen wood leaf.

3. according to the method for each described extraction catechin in the claim 1～2, it is characterized in that: the cold soaking in the step (1) is specially cold soaking 2～4 times, each 6～9 days.

4. according to the method for each described extraction catechin in the claim 1～2, it is characterized in that: the add-on of acetone is 2～5 times of volumes of catechin bullion weight in the step (3).