CN107723315A

CN107723315A - A kind of new method for separating giant knotweed composition and preparing resveratrol

Info

Publication number: CN107723315A
Application number: CN201610651946.3A
Authority: CN
Inventors: 梁克; 夏传递; 王惠芳; 肖虎; 焦燕玲
Original assignee: Jiangsu Bapri Biological Medicine Technology Co Ltd
Current assignee: Jiangsu Bapri Biological Medicine Technology Co Ltd
Priority date: 2016-08-11
Filing date: 2016-08-11
Publication date: 2018-02-23

Abstract

The invention belongs to natural plant component extraction separation field, and in particular to a kind of new method for separating giant knotweed composition and preparing resveratrol.Specially：Giant knotweed extracts through continuous ultrasound water, obtains giant knotweed extract solution；Extract solution directly continuously adsorbs through macroreticular resin；Solvent elutes resin, obtains polygonin mother liquor；Mother liquor concentrations remove solvent, obtain polygonin crude product；Polygonin crude product uses silica gel post separation, obtains high content polygonin；The special enzymatic conversion of high content polygonin polygonin, obtain resveratrol solid precipitate；Recrystallization, obtain resveratrol sterling.The direct ferment treatment of polygonin crude product can also be prepared the resveratrol product that content is 40~60% by this technique.For high-purity resveratrol content produced by the invention up to more than 98%, overall technology is adapted to large-scale industrial production.Compared with prior art, the inventive method has solvent safety and environmental protection, the advantages such as production cost is low, with short production cycle, transformation efficiency is high, production process loss is small.

Description

A kind of new method for separating giant knotweed composition and preparing resveratrol

Technical field

The invention belongs to natural plant component extraction separation field, and in particular to one kind separation giant knotweed natural constituent is simultaneously The method that polygonin is prepared to resveratrol with enzymatic conversion.

Background introduction

Medicinal plant giant knotweed (Polygonum cuspidatum Sieb.et Zucc.) has removing dampness through diuresis and removing jaundice, heat-clearing solution Poison, blood stasis removing analgesic, it is relieving cough and reducing sputum the effect of.Important medicinal ingredient in giant knotweed is polygonin and resveratrol etc..Resveratrol, Also known as resvertrol, it is a kind of very strong natural polyphenol of bioactivity.Modern scientific research shows that it has reduction platelet aggregation Collection, prevent and treat cardiovascular and cerebrovascular disease；The effect of whitening and anti-aging and pre- preventing tumor.Polygonin also known as polydatin, it is in vivo Resveratrol can be hydrolyzed to form by glycosidase so as to play pharmacological action.Low content composition in resveratrol and giant knotweed by In with higher medicine action activity, it is widely used, has huge in medicine, health products, cosmetic industry The market demand.Among actual production, in order to increase the yield of resveratrol, it is necessary to establish that a whole set of is simple to operate, and cost is cheap, It is easy to produce in batches the new method of resveratrol.

From extracting method angle analysis, the existing method of extraction separation polygonin and resveratrol from giant knotweed, mostly Number is organic solvent extraction, as patent CN103695479A, CN104447903A, CN03755752A, CN104130298A, CN102531851A, CN1962592 carry out soak extraction to giant knotweed using the aqueous solution of ethanol or ethanol, its solvent consumption is big, Production safety environmental protection cost is high.Partial monopoly using alkaline aqueous solution extract, as patent CN102453059A, CN102220387A, CN101759732A etc., this extracting mode add acid-base accommodation step, and operation is comparatively laborious, easily Destruction is produced to active ingredient structure.

Most of patents are all directly to be converted Rhizoma Polygoni Cuspidati, as patent CN101948370A, CN104263763A, CN101805755A, CN102838456A etc..Be converted into extracting separation after resveratrol, this method because inversion quantity is big, Need very big stacking space, complex operation, to be on the one hand difficult to the optimum pH environment for reaching enzyme, it is impossible to give full play to the work of enzyme Property, the material in another aspect giant knotweed extract solution or giant knotweed slurry can reduce the unicity that enzyme converts to polygonin, or even can be right Enzyme produces inhibitory action, so as to increase the use cost of enzyme.A small number of enzymatic conversion patents use transformation technology after extraction, such as CN103695479A, conversion of substrate is not purified in this patent, and composition is complicated, and enzyme dosage is bigger than normal, and product is various, and post-processing is difficult Degree is high.Enzymolysis product resveratrol is a kind of strong aldehydes matter of expression activitiy, with the progress of enzyme digestion reaction, this material pair Illumination, oxidant, temperature are all more sensitive, cause traditional handicraft purification efficiency not high.

The content of the invention

The present invention is easily destroyed for producing composition existing for resveratrol technique at this stage, extraction process very complicated； Organic solvent environmentally friendly cost safe to use is high；Alkali extraction and acid precipitation is complex for operation step, is easily destroyed the problems such as active ingredient, invention A kind of High-efficient Water ultrasonic extraction purifies polygonin, then digests the new method that conversion prepares resveratrol, and having reached reduces cost, fast Rate produces the purpose of resveratrol.

A kind of that the method that conversion prepares resveratrol is extracted from giant knotweed, detailed process is：Giant Knotweed Rhizome is through continuous ultrasound Water extracts, and obtains giant knotweed aqueous extract；After extract solution is separated off precipitation, directly continuously adsorbed through macroreticular resin；Solvent elutes Resin, obtain polygonin mother liquor；Mother liquor concentrations remove solvent, obtain polygonin crude product；Polygonin crude product uses silica gel post separation, Elution, concentration, obtain high content polygonin；High content polygonin uses the special enzymatic conversion of polygonin, obtains white black false hellebore Alcohol solid precipitate；Resveratrol solid precipitate recrystallizes, and obtains the sterling of Resveratrol content 98%.

The direct ferment treatment of polygonin crude product can be prepared the resveratrol product that content is 40~60% by this technique.

The present invention prepares giant knotweed natural constituent, joint enzymatic conversion using water ultrasound assisted extraction technique and chromatography technology Technique prepares resveratrol, has following advantage：First, this patent uses water ultrasound assisted extraction technique, utilizes " the cavitation of ultrasonic wave Effect ", mechanical effect and fuel factor, efficiently, sufficiently extract giant knotweed natural constituent, extraction process safety and environmental protection, cost It is cheap；Secondly, giant knotweed extract solution directly can be enriched with through macroporous resin adsorption, avoid the heating concentration process of extract solution, contracting The short extraction process time to polygonin active ingredient, the destruction for preventing high temperature, oxidized zone from coming；Again, by preliminary pure Polygonin after change is converted in controllable environmental system, and enzyme can be carried out under optimum reaction conditionses for single substrate Enzyme digestion reaction, the usage amount of enzyme saved, improved the conversion ratio of substrate；Finally, special polygonin specific enzyme can be short (a few hours) complete conversion in time, shorten the script resveratrol production cycle of 3-5 days.

Compared with prior art, the inventive method has solvent safety and environmental protection, and production cost is low, the cycle is short, transformation efficiency The advantages such as height, yield height.Among actual industrial production, the method for ultrasonic water extraction giant knotweed composition is not by solvent and material pre-treatment Limitation, it is possible to achieve short cycle, scale, sustainable production.

Embodiment

Example 1

2 kilograms of crushing of Rhizoma Polygoni Cuspidati are taken, add 35 DEG C of water 40L immersions, ultrasonic device extracts 2 hours, then is obtained by filtering Obtain giant knotweed extract solution.Giant knotweed extract solution directly uses D101 macroporous resin adsorptions, then elutes resin with ethyl acetate, obtains acetic acid Ethyl ester eluent, is concentrated under reduced pressure, and obtains polygonin crude product 140g, Determination of Polydatin 28.2%.Polygonin crude product uses silica gel Post separation, ethyl acetate elution, retains polygonin efflux, is concentrated under reduced pressure and obtains high content polygonin 50g, and Determination of Polydatin is 80.5%.High content polygonin is dissolved in 16L buffer solutions (pH=4.7), adds polygonin specific enzyme 15g, 40 DEG C of reactions 12 Hour.Resveratrol caused by enzymolysis is by recrystallization, the white solid powder 21g of acquisition content 98%.

Example 2

20 kilograms of crushing of Rhizoma Polygoni Cuspidati are taken, add 60 DEG C of water 200L immersions, ultrasonic device is extracted 1 hour, obtained by filtering Obtain giant knotweed extract solution.Giant knotweed extract solution directly using DM130 macroporous resin adsorptions to saturation, then with acetone water elution tree, acquisition Acetone eluant, it is concentrated under reduced pressure, obtains polygonin crude product 1208g, Determination of Polydatin 25.4%.Polygonin crude product uses silica gel Post separation, ethyl acetate elution, retains polygonin efflux, is concentrated under reduced pressure and obtains high content polygonin 395g, Determination of Polydatin For 76.5%.High content polygonin is dissolved in 100L buffer solutions (pH=5.2), adds polygonin specific enzyme 30g, 50 DEG C of reactions 16 hours.Resveratrol caused by enzymolysis is by recrystallization, the white solid powder 150g of acquisition content 98%.

Example 3

4 kilograms of crushing of Rhizoma Polygoni Cuspidati are taken, 55 DEG C of water 90L ultrasonic devices is added and extracts 3 hours, then by filtering, obtain tiger Cane extract solution.Giant knotweed extract solution is directly using HP-20 joint D101 macroporous resin adsorptions, then resin is washed with ethyl acetate It is de-, ethyl acetate eluent is obtained, is concentrated under reduced pressure, obtains polygonin crude product 300g, Determination of Polydatin 25.1%.Polygonin is thick Product are directly dissolved in 30L buffer solutions (pH=4.0), add polygonin specific enzyme 30g, 40 DEG C are reacted 6 hours.Caused by enzymolysis Resveratrol solid precipitate is by recrystallization, the red solid powder 73g of acquisition content 58%.

Claims

1. a kind of new method for separating giant knotweed composition and preparing resveratrol, detailed process：Giant Knotweed Rhizome extracts through continuous ultrasound water, Obtain giant knotweed extract solution；After extract solution is separated off precipitation, directly continuously adsorbed through macroreticular resin；Solvent elutes resin, obtains Polygonin mother liquor；Mother liquor concentrations remove solvent, obtain polygonin crude product；Polygonin crude product uses silica gel post separation, and eluent is washed It is de-, concentration, obtain high content polygonin；High content polygonin uses the special enzymatic conversion of polygonin, obtains the analysis of resveratrol solid Go out thing；Resveratrol solid precipitate recrystallizes, and obtains resveratrol sterling.

2. a kind of new method for separating giant knotweed composition and preparing resveratrol according to claim 1, its ultrasonic water extraction bar Part is characterised by that the mass volume ratio of Giant Knotweed Rhizome and water is 1: 5~60；Ultrasonic extraction temperature is 20~80 DEG C.

3. a kind of new method for separating giant knotweed composition and preparing resveratrol according to claim 1, its macroreticular resin are continuous Adsorpting characteristic is the resin combination or mixing for using skeleton for styrene, preferably one kind in D101, DM130, HP-20, AB-8 It is or several.

4. a kind of new method for separating giant knotweed composition and preparing resveratrol according to claim 1, its solvent elution resin It is characterised by the preferred water of eluant, eluent, ethyl acetate, methanol, ethanol, the one or more combination in acetone or mixing.

5. a kind of new method for separating giant knotweed composition and preparing resveratrol according to claim 1, its polygonin crude product are adopted With silica gel post separation, elution, eluent is characterised by that the silicagel column after absorption is used in ethyl acetate, ethanol, acetone One or more combination or mixing elution.

6. a kind of new method for separating giant knotweed composition and preparing resveratrol according to claim 1, its polygonin specific enzyme It is characterized in a kind of enzymatic mixture with beta-glucosidase key hydrolysis, average enzymatic activity is excellent in 15000~40000 units Select the one or more of mixture of cellulase, beta-glucosidase, 1,4 beta-glucanase.

7. a kind of new method for separating giant knotweed composition and preparing resveratrol according to claim 1, it is special using polygonin It is to digest pH of buffer=3.0~6.5,36~54 DEG C of hydrolysis temperature, polygonin specific enzyme and height with enzymatic conversion process feature The mass ratio of content polygonin is 1: 0.5~50.

8. a kind of separation giant knotweed composition described in claim 1 prepares the new method of resveratrol, its production technology can also adopt With the direct ferment treatment of polygonin crude product, the resveratrol that content is 40~60% is prepared.