KR920005686B1 - Process for the preparation by extracting of harpagophytum - Google Patents

Abstract

A method for extracting harpagoside in Harpagophytum procumbens D.C. comprises (a) pulverizing the root of harpagophytum, and extracting it with pH 7 phosphate buffer soln. at 70 deg.C, (b) refluxextracting it with hydrocarbon solvent to remove oily material, (c) recovering the buffer part, and removing pigment-like material by use of halogen-containing hydrocarbon organic solvent, and (d) recovering the buffer part, and purifying it with ethyl acetate. In step (b), the straight chain hydrocarbon solvent is n-hexane, n-heptane or petroleum ether; and in step (c), the halogen-containing hydrocarbon solvent is CCl4, CHCl3 or CH2Cl2. Harpagoside extracts have a therapeutic effect for arthritis.

Description

Extraction Method of Harpagoside-Containing Extract

1 is an apparatus for extracting Harpagoside extract according to the present invention.

* Explanation of symbols for main parts of the drawings

1: column 2,2a, 2b: heat exchanger

3: first organic solvent chamber 4: second organic solvent chamber

5: third organic solvent chamber 6: concentration tank

7: buffer solution reservoir 8: valve

9: pH adjusting device of the buffer solution 10: pump

The present invention relates to a method for extracting an extract containing harpagoside as a main component from the roots of Harpagophytum procumbens D.C belonging to Pedaliaceae.

Harpagopitum Procumbens D.C. is a plant native to the southwestern parts of Africa, the Kalahari Desert, and the Namibian Valley of southern Africa, and has long been used as a folk medicine for the treatment of arthritis in Europe.

Thus, Harpagopitum Procumbens DC has three effects on arthritis: Harpagide, Harpagoside and Procumbide, which are pharmacologically useful ingredients for its roots or shrubs. Iridoid glucosides are included, which has been confirmed by several researchers (Ref. R. Kampf, Schweiz. Apoth.-Ztg., 114,337,1976, O. Sticher, Desch. Apoth. -Ztg., 117, 1279, 1977, M. Caprasses, J. Pharm. Belg., 35, 143, 1980).

Recently, the three iridoid glycosides have been separated and purified, and the structure thereof has been confirmed. Among them, the structure of harpagoside is represented by the following structural formula (1) including cinnamic ester.

An object of the present invention is to provide an effective method and apparatus for extracting an extract containing harpagoside represented by the structural formula (1) from harpagopitum procumbens D.C.

Hereinafter, the present invention will be described in detail.

The present invention is (a) finely pulverized toenail part of Harpagophytum procumbens DC extracted with phosphate buffer solution of pH 7 warmed to a temperature of 70 ℃, (b) the buffer thus extracted The solution is refluxed with a linear hydrocarbon solvent to remove the oil phase problem. (C) The buffer layer is recovered and the pigment material is extracted and removed with a halogen-containing hydrocarbon solvent. It is a method for extracting a harpagoside-containing extract, characterized in that the layer is recovered and purified with an ethyl acetate solvent.

In addition, the present invention includes a column (1) having a nitrogen injection pipe and a discharge pipe up and down, a first organic solvent chamber (3) containing a linear hydrocarbon-based organic solvent, a halogen-containing hydrocarbon-based organic solvent The second organic solvent chamber (4) and the third organic solvent chamber (5) containing the ethyl acetate solvent, and the buffer solution storage tank (7) are connected to each other in a circulating manner along the connection pipe, the column (1) and A pressure regulating valve 8 and a heat exchanger 2a are installed between the first organic solvent chamber 3, and a pump 10 is installed between the buffer solution reservoir 7 and the column 1. The third organic solvent (5) is an extracting device of harpagoside extract, characterized in that connected to the concentration tank (6).

Here, 2b is a heat exchanger installed in the buffer reservoir 7, 9 is a pH adjusting device of the buffer.

Hereinafter, the present invention will be described in more detail.

In the present invention, first, the well-dried Harpagopitum procumbens DC is crushed to a size of 5 cm or less in diameter, extracted with a phosphate buffer solution of pH 7, and the oily substance is removed therefrom. Hydrocarbon-based solvents such as en-nucleic acid, en-haptan, petroleum ether and the like can be used, and most preferably, an en-hexane solvent is used.

The solvent used in the extraction is recovered, reused, the buffer is separated, and then extracted with a halogen-containing hydrocarbon-based organic solvent to remove the pigment. At this time, carbon tetrachloride, chloroform, dichloroethane, etc. may be used as the organic solvent, of which carbon tetrachloride is the best.

Finally, once again, the buffer solution of the solvent is separated, the solvent is reused, and the buffer layer is purified with an ethyl acetate solvent.

The extract extracted according to the present invention contains not only harpagoside but also harpado and procumbide, and three new iridoid glucosides, 8-O- (P-coumaroyl) -harpagi, which have been recently discovered. De, 6'-O- (P-coumaroyl) -procumbide and crocumboside.

Hereinafter, the extraction method of the present invention based on the accompanying drawings.

1 is a drawing showing the extraction apparatus of the Harpago seed extract according to the present invention, finely crushed and dried the dry root of the Harpago pitum procumbens in the column 1, 80 ° C through the heat exchanger (2) The nitrogen gas warmed by to pass from the bottom of the column to the top to inactivate the sterilization of pathogenic microorganisms and various enzymes, and then the phosphate buffer solution heated to 70 ℃ by the pump 10 from the buffer resistance tank (7) Backflow from bottom to top of column (1).

At this time, the pressure adjusting valve 8 is adjusted to adjust the passage time of the column 1 of the buffer solution to take 30 minutes.

The extracted liquid is passed through a heat exchanger (2a), cooled to room temperature, and then passed through a first organic solvent chamber (3) filled with a linear hydrocarbon to extract and remove oily substances.

The buffer layer separated into the lower part is sprayed upward from the lower part of the second organic solvent chamber 4 containing the organic solvent containing hydrogen halide to remove the pigment material.

Subsequently, the buffer layer separated from the upper layer was sprayed and purified in the third organic solvent chamber 5 containing ethyl acetate, and then transferred to the phosphate buffer storage tank 7 to readjust the buffer solution at pH 7 to perform the above operation. Will repeat.

The above operation is continued for 6 hours and all 12 times are repeated. Finally, the extract from the column is transferred to a concentration tank (6), concentrated under reduced pressure to form a viscous liquid, which is transferred to a drying panel and dried in a vacuum dryer to obtain a powder with a light brown bitter taste. Get This extract powder contains more than 3% of Harpagoside, which is very suitable for the manufacture of pharmaceuticals.

On the other hand, in order to purify the extract prepared according to the present invention for injection, the extract is added to a mixed solvent of water and ethyl acetate, it is allowed to stand alone, and then the two layers are separated.

Subsequently, the aqueous layer was eluted with charcoal through column chromatography, and the residue obtained by concentrating the resultant was passed through column chromatography filled with silica gel, and then extracted with methanol: chloroform = 1: 9 mixed solvent. Distillation under reduced pressure gives pure harpagoside. The target product thus obtained is suitable for injection.

Hereinafter, examples of the present invention will be described.

Example 1

Harpagopitum Procumbens D.C. on column (1). 285 kg of finely crushed and finely filled, the nitrogen gas is heated to 80 ℃ in the heat exchanger (2) and passed through the bottom of the column (1) for 30 minutes.

The buffer solution of the phosphate buffer reservoir 7 is warmed to 70 ° C. and then passed from the bottom of the column 1 to the top through the transfer pump 10. At this time, adjust the pressure control valve (8) so that it takes 30 minutes to pass through the 3000l buffer.

The buffer solution passed through is cooled to room temperature in the heat exchanger (2a) and sprayed from top to bottom in 1500 l of N-hexane in the first organic solvent chamber (3) to extract and remove the oil component in the extract, and the buffer layer separated at the bottom is Subsequently, it is sprayed from the lower part to the upper part in 1500 L of carbon tetrachloride solvent of the 2nd container plum 4, and a pigment | dye material is melt | dissolved in carbon tetrachloride.

The buffer separated at the upper part of the second organic solvent chamber 4 is sprayed from the upper part to the lower part to 1500 l of the ethyl acetate layer of the third organic solvent chamber 5 to be purified, and then sent to the buffer storage tank 7 to pH 7 The above operation is repeated 12 times by adjusting the buffer solution.

The extraction process is continued for a total of 6 hours, and the final flow through is transferred to a reduced pressure condenser in unit 6 and concentrated.

The concentrated solution was transferred to a drying panel in a viscous state, and dried in a vacuum dryer at 50 ° C. for 24 hours to obtain 100 kg of a light brown bitter or odorous powder.

The powder contained more than 3.2% of Harpagoside, and was tested by thin layer chromatography and showed the same spots as the standard product.

Example 2

100 kg of the extract extract obtained in Example 1 was extracted four times with 600 l of methanol, and the resultant residue was diluted with 500 l of water and extracted three times with 400 l of ethyl acetate.

The aqueous layer is taken and passed through column chromatography filled with 60 kg of charcoal and adsorbed.

First, pass through 1000l of water to wash the column, then pass through 3000l of 1: 1 methanol: water solution, elute and concentrate to obtain 1.5kg of residue. Then elute by passing through a water: methanol solution of 3: 7 to obtain 0.5kg of residue. Finally, eluting with 6000 liters of methanol yields a methanol solution containing mainly harpagoside.

The solution was concentrated to contain 2.4 kg of residue, which was dissolved in 1500 l of methanol and passed through a column containing 120 kg of silica gel to adsorb. Subsequently, the mixture was eluted by passing through a mixed solution of methanol-chloroform 1: 9, and concentrated under reduced pressure to obtain 1.4 kg of harpagoside.

HPLC analysis was consistent with the purity of the standard and was suitable for the preparation of injectable drugs.

Claims (6)

(A) Finely grind the toenails of Harpagophytum procumbens DC, extract it with a phosphate buffer solution of pH 7, heated to 70 ° C, and (B) extract the buffer solution After refluxing with a chain hydrocarbon solvent to remove the oily substance, (C) recovering the buffer layer, extracting and removing the pigment material with a halogen-containing hydrocarbon solvent, and (D) again removing the buffer layer. A method for extracting harpagoside-containing extracts, which is recovered and purified by an ethyl acetate solvent. The method according to claim 1, wherein (a) the pulverized Harpagophytum procumbens D.C of step (A) is sterilized with nitrogen gas warmed to 80 ° C. prior to extraction with phosphate buffer solution. The method of claim 1, wherein (b) N-nucleic acid, N-haptan, petroleum ether are used as the solvent of the linear hydrocarbon-based process. 2. The method according to claim 1, wherein carbon tetrachloride, chloroform, and dichloroethane are used as the halogen-containing hydrocarbon-based organic solvent in the step (c). The method according to claim 1, wherein (a) (b) (c) and (d) are repeated 12 times in succession, and when (d) is moved from step (a) to step (a), it is readjusted to pH7 buffer solution. How to. The harpagoside-containing extract obtained in claim 1 is concentrated, extracted with a mixed solvent of water and ethyl acetate, the aqueous layer is separated and eluted with methanol in charcoal column chromatography, and then methanol: chloroform = silica gel column chromatography. A method for producing an injectable harpagoside extract, which is eluted with a mixed solvent of 1: 9, and the eluate is depressurized and concentrated.

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