CN1240695C

CN1240695C - Technical method for extracting salvianolic acid B from radix salivae miltiorrhizae

Info

Publication number: CN1240695C
Application number: CN 03151476
Authority: CN
Inventors: 张立国; 倪力军; 高秀蛟
Original assignee: East China University of Science and Technology
Current assignee: East China University of Science and Technology
Priority date: 2003-09-29
Filing date: 2003-09-29
Publication date: 2006-02-08
Anticipated expiration: 2023-09-29
Also published as: CN1528756A

Abstract

The present invention relates to a set of new technology which is suitable for separating and purifying salvianolic acid B. The present invention is the organic combination of several kinds of methods from danshen root for extracting salvianolic acid B. The new technology comprises the steps of extracting by water, flocculating by chitosan, alcohol sinking, extracting respectively by petroleum ether and ethyl acetate. Finally, the present invention uses silica gel column chromatography to obtain the salvianolic acid B with higher purity. The new technology method of the present invention has low cost, is suitable for industrial production, and can extract the salvianolic acid B with higher purity.

Description

A kind of processing method of from the red sage root, extracting salvianolic acid B

Technical field

The present invention relates to a kind of new processing method of from the red sage root, extracting salvianolic acid B.

Background technology

Red sage root beginning is stated from Shennong's Herbal and classifies as top gradely, is one of medicine commonly used in China's traditional medicine, and long clinical application history is arranged; Show that according to modern pharmacological research pressure differential self has very strong antioxygenation, can remove intravital superoxide anion and hydroxyl radical free radical, suppress peroxidatic reaction of lipid, microcirculation improvement; The clinical treatment heart, cerebrovascular disease and the associated all kinds of diseases of being mainly used in of the red sage root, the field has a wide range of applications.

Salvianolic acid B in the red sage root water soluble ingredient is one of main effective constituent, content height and strong drug action.The separation and purification route of effective constituent salvianolic acid B is mainly in the red sage root of existing bibliographical information: extract---removal of impurities---column chromatography; These extraction and separation method key distinctions exist:

1, the difference of extracting method has water to carry, methods such as different concentration ethanol extraction with aqueous solution;

2, the difference of impurity-removing method has alcohol precipitation, and macroporous resin is crossed in flocculation, extraction;

Column chromatography adopts the preparative chromatography post to separate, but volume containing the sample can only obtain the product of trace less.Because the structure and the physicochemical property of pressure differential self are very close, all have pyrocatechol and Ar-CH=CH-COOR structure.The molecular structure of salvianolic acid B is:

Because the common ground of its structure will be isolated very difficulty of highly purified salvianolic acid B, also do not report so far and separate its pure product that obtain, the highest also the having only about 70% of content of danshinolic acid B in the existing literature report, major part is about 60%, even lower.

Summary of the invention

The purpose of this patent invention is the defective that overcomes prior art, develop a kind of simple, be suitable for suitability for industrialized production, and the novel process of effective constituent salvianolic acid B in the low extraction purifying red sage root of cost.

Technical conceive of the present invention be sample:

Several extracting and purifying method of integrated use of the present invention carry out organic assembling with it, have developed the novel process of the suitable separation and purification salvianolic acid B of a cover.Elder generation's water extracts, and uses flocculate with chitosan then, then carries out alcohol precipitation, extracts respectively with sherwood oil and ethyl acetate again, adopts silica gel column chromatography to obtain the salvianolic acid B of higher degree at last.

The present invention is achieved through the following technical solutions:

1, with the red sage root with 3-15 water doubly, the water of 6 times of amounts preferably, water is carried 1-5 hour, best 2 hours; Again with 1-10 water doubly, the water of 4 times of amounts preferably, water is carried 1-5 hour, and best 1.5 hours, extracting temperature was 30-100 degree centigrade, is preferably 65 degrees centigrade;

2, merge extracted twice liquid, add the chitosan of 0.5%-5%, be preferably 2% chitosan aqueous solution 100ml, flocculation time is 0.5-5 hour, is preferably 1 hour, and flocculation temperature is 0-40 degree centigrade, is preferably 0 degree centigrade;

3, filter, filtrate is concentrated into the doubling dose of crude drug quality, the aqueous ethanolic solution volumetric concentration is adjusted to 50%-90%, be preferably 75%, carried out alcohol precipitation 0.5-10 hour, be preferably 2 hours; The alcohol precipitation temperature is 0-40 degree centigrade, is preferably 0 degree centigrade;

4, refilter the concentrated ethanol of removing, the water constant volume is used twice of equal-volume petroleum ether extraction to 2 times of amounts of crude drug quality; Use the salt acid for adjusting pH value to 1-4 water, be preferably 2, use twice of equal-volume ethyl acetate extraction again;

5, with ethyl acetate phase evaporate to dryness, cross 100-120 order silica gel column chromatography (eluent is ethyl acetate, ethanol), obtaining purity is the phenolic acid B product of 50%-96%.

With method of the present invention extracting effective components phenolic acid B from the red sage root, technical have following remarkable advantage and a progress:

(1) adopts suitable flocculation agent chitosan, macromolecular substance such as protein are flocculated out;

(2) not only water-soluble but also be dissolved in the alcoholic acid characteristics according to salvianolic acid B, adopt the heavy most of carbohydrate content and other the various macromolecular substance that extract of going of suitable alcohol concn;

(3) adopt petroleum ether extraction, remove lipid material, further separation is simplified;

(4) salvianolic acid B is the slightly acidic compound, is ionic compound in the aqueous solution, so its aqueous solution is regulated the pH value, salvianolic acid B changes into the molecule-type compound, extracts with ethyl acetate, removes other compositions that are insoluble to ethyl acetate;

(5) adopt silica gel column chromatography, select ethyl acetate, sherwood oil and ethanol to carry out purifying, obtain the salvianolic acid B of required purity as eluent according to the molecular structure characteristics of salvianolic acid B;

(6) this process using extraction chromatography coupling makes that the silica gel volume containing the sample is big, can prepare salvianolic acid B in a large number;

(7) can provide the salvianolic acid B product (50%---96%) of various content;

Description of drawings:

Fig. 1, Fig. 2, Fig. 3 are the HPLC test results, and Fig. 4 is the IR spectrogram, and Fig. 5 is H ^EThe NMR spectrum, Fig. 6 is the MS spectrum.

Below in conjunction with embodiment and accompanying drawing the present invention is further specified.

Embodiment 1

Content of danshinolic acid B is 80% product

Salvia miltiorrhiza raw material 200 grams add the water of 6 times of amounts and carry 2 hours, carry 1.5 hours with the water of 4 times of amounts again, t=65 degree centigrade; Merge extracted twice liquid, the chitosan 100ml flocculation of adding 2% 1 hour, t=0 degree centigrade; Filter, filtrate decompression is concentrated into the doubling dose of raw medicinal herbs quality, add dehydrated alcohol 1200ml sedimentation 2 hours, t=0 degree centigrade; Filter, concentrate and remove ethanol, the water constant volume is to 2 times of amounts of crude drug quality, with twice of equal-volume petroleum ether extraction, water is transferred pH to 2, with ethyl acetate extraction twice, with ethyl acetate phase evaporate to dryness, get total liposoluble ingredient, faint yellow, the HPLC test result is seen accompanying drawing 1, retention time 8.697min, be salvianolic acid B, content is 80%.

Embodiment 2

Content of danshinolic acid B is 90% product

It with content of danshinolic acid B 80% pale yellow powder, admix proper silica gel, pack in the post, use ethyl acetate: sherwood oil (1: 1) mixing solutions 1000ml wash-out, lose elutriant, use the 400ml ethanol elution again, elutriant is concentrated into dried, pale powder, the HPLC test result is seen accompanying drawing 2, retention time 8.272min is a salvianolic acid B, and content is 90%.

Embodiment 3

Content of danshinolic acid B is 96% product

It with content of danshinolic acid B 80% pale yellow powder, admix proper silica gel, in the post of packing into, use ethyl acetate: sherwood oil (1: 1) mixing solutions 2000ml wash-out, lose elutriant, use the 400ml ethanol elution again, elutriant is concentrated into dried, blackish green powder, the HPLC test result is seen Fig. 3, retention time 8.328min is salvianolic acid B, and content is 96%.

HPLC, H ^ENMR, IR, MS analyze:

IR, H ^ENMR, MS check and analysis are salvianolic acid B, product is carried out HPLC detect, and the results are shown in accompanying drawing 4,5,6.

Claims

1, a kind of processing method of extracting salvianolic acid B from the red sage root is characterized in that, comprises the steps:

(1) with the water of the red sage root with 3-15 times, water is carried 1-5 hour; With 1-10 water doubly, water is carried 1-5 hour again, and extracting temperature is 30-100 degree centigrade;

(2) merge extracted twice liquid, add the chitosan 100ml of 0.5%-5%, flocculation time is 0.5-5 hour, and flocculation temperature is 0-40 degree centigrade;

(3) filter, filtrate is concentrated into the doubling dose of crude drug, regulating the ethanol volumetric concentration is 50%-90%, carries out alcohol precipitation 0.5-10 hour; The alcohol precipitation temperature is 0-40 degree centigrade;

(4) refilter the concentrated ethanol of removing, the water constant volume is used twice of equal-volume petroleum ether extraction to 2 times of amounts of crude drug; Use the salt acid for adjusting pH value to 1-4 water, use twice of equal-volume ethyl acetate extraction again;

(5) with ethyl acetate phase evaporate to dryness, cross 100-120 order silica gel column chromatography, be eluent with ethyl acetate, ethanol, obtaining purity is the phenolic acid B product of 50%-96%.

2, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 1 is characterized in that, when use water extraction, for the first time with the water of 6 times of amounts, the second time is with the water of 4 times of amounts.

3, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 2 is characterized in that, to carry the time be 2 hours to water for the first time, is extracted as 1.5 hours for the second time.

4, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 3 is characterized in that, extracting temperature is 65 degrees centigrade.

5, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 1 is characterized in that, uses 2% flocculate with chitosan.

6, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 5 is characterized in that flocculation time is 1 hour.

7, the processing method of stating as claim 6 of extracting salvianolic acid B from the red sage root is characterized in that flocculation temperature is 0 degree centigrade.

8, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 1 is characterized in that, regulating the ethanol volumetric concentration during alcohol precipitation is 75%.

9, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 8 is characterized in that the alcohol precipitation time is 2 hours.

10, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 9 is characterized in that the alcohol precipitation temperature is 0 degree centigrade.

11, the processing method of extracting salvianolic acid B from the red sage root as claimed in claim 1 is characterized in that, in step (4), carries out equal-volume petroleum ether extraction twice, and water is regulated pH to 2, twice of equal-volume ethyl acetate extraction with hydrochloric acid.