CN101007797B

CN101007797B - A method for extracting dihydromyricetin from Hovenia dulcis

Info

Publication number: CN101007797B
Application number: CN200710026288A
Authority: CN
Inventors: 张德志; 申向荣
Original assignee: Guangdong Pharmaceutical University
Current assignee: Guangdong Pharmaceutical University
Priority date: 2007-01-12
Filing date: 2007-01-12
Publication date: 2010-05-19
Anticipated expiration: 2027-01-12
Also published as: CN101007797A

Abstract

The invention discloses a method for extracting dihydromyricetin from Hovenia dulcis. The extraction method of the present invention firstly obtains the extract that is rich in dihydromyricetin by pretreating medicinal materials, then further purifies by using resin, and finally recrystallizes to obtain high-purity dihydromyricetin (purity > 99%), which overcomes the The cumbersome operation process of the previous extraction method has the advantages of simple operation, low toxicity, low cost, strong controllability, and large-scale production.

Description

A kind of method of from trifoliate orange seed, extracting dibydro myricetrin

Technical field

The present invention relates to dibydro myricetrin, particularly relate to a kind of method of from trifoliate orange seed, extracting dibydro myricetrin.

Background technology

Had medicinal history in several thousand as liver-protecting sobering up good medicine trifoliate orange seed commonly used among the people (Rhamnaceae trifoliate orange Dulcis belongs to the mature seed of HoveniaThunb. plant Semen Hoveniae (Hovenia dulcis Thunb.), trifoliate orange Dulcis (Hovenia acerba Lindl.) and comospore trifoliate orange Dulcis (Hovenia trichocarpa Chun et Tsiang)), the whole world has 3 kinds, 2 mutation mainly are distributed in China, Korea, Japan and India.China natural resources is very abundant, and except that northeast, the Inner Mongol, Xinjiang, Ningxia, Qinghai and Taiwan, all there is distribution each provinces and regions.Beginning is stated from Tang Materia Medica, is used as medicine with fruit, seed, wooden juice, wooden skin, is now recorded by " The People's Republic of China's the Sanitation Ministry medicine standard Chinese medicinal materials ".Flavonoid compound is the effective constituent of trifoliate orange seed, pharmacological evaluation according to Japanese scholar, confirmed that the dibydro myricetrin that extracts has shown fabulous liver-protecting activity from the trifoliate orange seed fruit that Japan produces, its activity is higher than present European countries and controls hepatopathy medicine silybin (Hase K commonly used, OhsugiM, Xiong Q, et al.Hepatoprotective effect of Hovenia dulcis THUNB.on experimental liver injuries induced by carbon tetrachioride orD-galactosamine/lipopolysaccharide[J] .Biol.Pharm.Bull., 1997,20 (4): 381-385.).In addition, modern science also confirms, dibydro myricetrin has liver protecting, anti-inflammatory, analgesia, reducing blood-fat, SOD activity improving, very strong antioxygenation and good pharmacological effect such as antibacterial, antiviral are expected to be developed as a kind of novel healthcare products and antioxidant from natural food and country's one, two kind new medicines.Its chemical name is 3,5,7,3 ', 4 ', 5 '-hexahydroxy-, 2,3 flavanones, and English name Dihydromyricetin (being abbreviated as DMY) or (+)-ampelopsin, its molecular structural formula is shown in (I).

Yet the main plant source of current dibydro myricetrin is the ampelopsis vine tea, and its extraction process is loaded down with trivial details, and toxicity is big.

Summary of the invention

The objective of the invention is to overcome the deficiency that exists on the existing dibydro myricetrin extracting method, a kind of simple to operate, method of extracting dibydro myricetrin from trifoliate orange seed that toxicity is little is provided.

The present invention extracts the method for dibydro myricetrin from the trifoliate orange seed medicinal material, its step is as follows:

1. with clean trifoliate orange seed pulverizing medicinal materials, cross 80～100 purposes sieve, extract, filter, get filter residue with low polar organic solvent;

2. the filter residue with above-mentioned gained carries out refluxing extraction with alcohol solution;

3. resin absorption is separated: the extracting solution of step (2) gained is concentrated, make it pass through macroporous resin or polyamide column, wash with water earlier and remove partial impurities, use 5%～10% (V/V) alcohol solution wash-out to remove partial impurities again, use the alcohol solution of 20%～30% (V/V) to carry out wash-out at last; When being eluted to 6～7 BV, stop wash-out, merge the elutriant of this section;

4. concentrate recrystallization: the elutriant of step 3 gained is concentrated, come the thermosol recrystallization with alcohol solution again, leave standstill, filtering for crystallizing, drying can make dibydro myricetrin, and purity reaches more than 99%.

The trifoliate orange seed medicinal material of above-mentioned steps 1 comprises that Rhamnaceae trifoliate orange Dulcis belongs to mature fruit or the dry seed of Hovenia Thunb. plant Semen Hoveniae (Hovenia dulcis Thunb.), trifoliate orange Dulcis (Hovenia acerba Lindl.) and comospore trifoliate orange Dulcis (Hovenia trichocarpa Chun et Tsiang).

The low polar organic solvent of above-mentioned steps 1 is aliphatic ether or 6# gasoline; Described aliphatic ether is ether, sherwood oil or normal hexane.

When above-mentioned steps 2 described alcohol solutions extracted, its alcohol solution was the aqueous solution of methyl alcohol, ethanol, propyl alcohol, butanols or amylalcohol, and the volume of organic solvent specific concentration is 50%～100% (V/V) in its mixed solvent.

The consumption that above-mentioned steps 2 is extracted solvent is a 6-10 times of medicinal material amount, and extraction time is 2-4 time, and extracting method is that extraction, supersound extraction or diafiltration extraction are extracted, flooded in thermal backflow.

Above-mentioned steps 3 used macroporous resin column are raw material with vinylbenzene, and divinylbenzene, methacrylonitrile or methyl acrylate are the macroporous resin column that linking agent is formed.Macroporous resin is meant the macroporous resin of D-101 type, HPD-500 type, HP-20 type, XAD-4 type, AB-8 type, SP-825 type, NKA type, SIP-1100 type or XAD-2 type.

The alcohol solution of the recrystallization solvent of above-mentioned steps 4 is the aqueous solution of its methyl alcohol, ethanol, propyl alcohol, butanols or amylalcohol, and the volume of organic solvent specific concentration is 0%～50% (V/V) in its mixed solvent.

The drying of above-mentioned steps 4 is constant pressure and dry, reduced vacuum drying, spraying drying or lyophilize.

Compared with prior art; the present invention has following beneficial effect: extracting method of the present invention is at first by the pre-treatment medicinal material; obtain being rich in the extracting solution of dibydro myricetrin, by adopting resin to be further purified, last recrystallization obtains highly purified dibydro myricetrin (purity＞99%) again; overcome the troublesome operation process of extracting method in the past; have simple to operately, toxicity is little, and cost is low; controllability is strong, but advantages such as large-scale production.

Description of drawings

Fig. 1 is dibydro myricetrin (DMY) extraction separation route map;

Fig. 2 is the hydrogen spectrogram of DMY.

Embodiment

Embodiment 1

1. get the dry seed (1kg) of Semen Hoveniae, remove impurity,, cross 80-100 purpose sieve its pulverizing;

2. be 60～90 sherwood oil refluxing extraction 2 hours with 8 times of amount boiling ranges, and then be 60～90 sherwood oil refluxing extraction 2 times with the boiling range of 6 times of amounts, each 1.5 hours, filter, filter residue, united extraction liquid, recovery solvent.

3. the filter residue of above-mentioned gained is again with 8 times of amount alcohol reflux of 70% 2 hours, and then with 70% alcohol reflux 2 times, each 1.5 hours;

4. extracting solution concentrating under reduced pressure, last AB-8 type macroporous resin column slowly goes up sample, it is closely colourless that elder generation's water is eluted to outflow, uses 5 times of post retention volume of 10% ethanol elution instead, uses 30% ethanol elution at last, when being eluted to 6 times of post retention volume, stop to collect, with the partially recycled solvent of 30% ethanol elution;

5. the concentrated solution of above-mentioned gained, the ethanol with 20% carries out thermosol, leaves standstill 12 hours at 0-5 ℃, separates out the needle crystal of white, the suction filtration crystallization, spraying drying, dibydro myricetrin 4.0g, its hydrogen spectrogram is seen Fig. 2.

Claims

1. a method for extracting dihydromyricetin from Hovenia dulcis, the steps are as follows:

(1) Pulverize Hovenia dulcis medicinal material, pass through a sieve of 80 to 100 meshes, extract with fatty ether, n-hexane or 6# gasoline, and filter to obtain filter residue;

(2) Carry out reflux extraction with the above-mentioned obtained filter residue with alcoholic aqueous solution;

(3) Resin adsorption separation: Concentrate the extract obtained in step (2), make it pass through a macroporous resin column or polyamide resin, first wash with water to remove some impurities, and then use an aqueous alcohol solution with a volume ratio of 5 to 10:100 Remove some impurities by eluting, and finally elute with an aqueous alcohol solution with a volume ratio of 20 to 30:100; when the elution reaches 6 to 7 times the column retention volume, stop the elution and combine the eluents in this section;

(4) Concentration and recrystallization: Concentrate the eluent obtained in step (3), then heat-dissolve and recrystallize it with an aqueous alcohol solution, let stand, filter the crystals, and dry to obtain dihydromyricetin.

2. according to the described method of claim 1, it is characterized in that, the Hovenia dulcis medicinal material described in step (1) is the mature fruit or dried seed.

3. by the described method of claim 1, it is characterized in that, described aliphatic ether is ether or sherwood oil.

4. by the described method of claim 1, it is characterized in that, the aqueous alcohol solution described in step (2) is the aqueous solution of methyl alcohol, ethanol, propanol, butanol or pentanol, and the volume ratio of alcohol and water is 50～100 : 100.

5. according to the described method of claim 1, it is characterized in that, the consumption of the described alcoholic aqueous solution of step (2) is 6～10 times medical material amount, and extraction times is 2～4 times; Described extraction method is changed into by reflux extraction Heat reflux extraction, maceration extraction, ultrasonic extraction or percolation extraction.

6. by the described method of claim 1, it is characterized in that, the described macroporous resin post of step (3) is to be raw material with styrene, and divinylbenzene, methacrylonitrile or methyl acrylate are crosslinking agent to form Macroporous resin column.

7. Press the described method of claim 7, it is characterized in that, described macroporous resin refers to D-101 type, HPD-500 type, HP-20 type, XAD-4 type, AB-8 type, SP- 825 type, NKA type, SIP-1100 type or XAD-2 type macroporous resin.

8. according to the described method of claim 1, it is characterized in that, the described alcohol aqueous solution of step (4) is the aqueous solution of methyl alcohol, ethanol, propanol, butanol or pentanol, and the volume ratio of alcohol and water is 0～50 : 100.

9. The method according to claim 1, characterized in that the drying described in step (4) is normal pressure drying, vacuum drying under reduced pressure, spray drying or freeze drying.