KR19980066516A - Method of extracting active ingredient including saponin contained in plant by enzyme treatment - Google Patents

Abstract

A method of extracting active ingredients containing saponins from herbs and plants, including ginseng, which is contained in plants by immersing the leachate in water, boiling and cooling to 40-60 ℃ to fully react with degrading enzymes that do not destroy saponins. Extraction of saponin or active ingredient as a solvent in the state of weakening the binding strength of chemical bond form or adsorption form with other high molecular substance such as polysaccharide pectin or protein molecule, and extracting only active ingredient including saponin or pure saponin in high yield .

Description

Method of extracting active ingredient including saponin contained in plant by enzyme treatment

It is an object of the present invention to provide an extraction method for increasing the extraction efficiency of saponin-containing active ingredients by enzymatic treatment of herbs and plants including ginseng containing saponin as an active ingredient.

The present invention relates to a technical field for extracting saponin and other active ingredients from herbals and plants including ginseng. The method of extracting an active ingredient including saponins from conventional plants is used to leach water or leach it with water or ethyl alcohol, methyl alcohol. And other organic solvents, for example, 40-70% ethanol leaching method (Chinese pharmacy magazine published in 1992: 25 (5), 281-293), methyl alcohol leaching (Shibate et al., Japanese Patent Publication No. 91109 published in 1978), but the biggest drawback of the solvent leaching method is that no matter which solvent is used, more than 60% of the saponins or active ingredients in ginseng, herbal medicine, and plants are used. It can't be extracted and the rest is left in the residue, which wastes valuable medicine and nutritional ingredients.

Saponins or active ingredients originally contained in ginseng, herbal medicine, and plants are strongly bound to polysaccharide pectin, protein molecules, which are large molecules of other substances in chemical bond form or adsorption, and thus, saponins contained in plants by conventional leaching or extraction methods. However, since there is a limit to the extraction of the active ingredient, it will be used industrially by increasing the extraction efficiency by weakening the binding force of the chemical bond form or adsorption form.

1 HPLC diagram of ginseng saponin by conventional methanol extraction method

Figure 2 HPLC diagram of ginseng saponin by conventional ethanol extraction method

Fig. 3 Saponin HPLC diagram of ethyl ginseng by ethyl alcohol extraction method after ginseng treatment according to the present invention

Figure 4 HPLC diagram of saponin by ethanol extraction method after treatment of ginseng according to the present invention yeast leaching complexase

The present invention overcomes the inefficient drawbacks of solvent leaching or extraction described above, and reacts saponins with endo-glucose degrading enzymes that do not destroy or decompose saponins from herbs or plants containing ginseng. By separating the binding of molecular material by water, it is made into a free saponin form, and the saponins bound by adsorption or other forms are also easily extracted by decomposition. The enzyme used in the invention

(1) starch degrading enzymes: α-amylase, β-amylase, gloco amylase

Iso-amylase

(2) endo-cellulase

(3) pectinase

(4) mold and monoclonal solid or liquid yeast complex enzymes, natural yeast-leaved complex enzymes, plant seeds or plant seed germination complexes, and one or more of the above enzymes in combination

(5) Degradative enzymes containing heat treatment or enzyme inhibitors if enzymes that destroy saponins are present in the leachate and the complex enzyme

The above enzymes can be used alone or in combination to treat medicinal herbs containing phosphoric acid and saponin as the main active ingredients, plants with high saponin content, and then filtered to obtain leaching liquids with high saponin content, or conventional solvent leaching of water, ethanol and methanol. Pure saponins can be extracted to yield 1.5-2 times higher yield than conventional non-enzymatic solvent extraction.

TLC (Thin Layer Chromatography: plato was analyzed by Kieselgel 60F-254: Merk) method and HPLC (High Performance Liquid Chromatography) method. In addition, the saponin extraction amount was greatly increased, and it can be seen that the extraction method by the present method was successful, and the graph by the analysis method of HPLC is shown in FIGS. 1 to 4.

Here, FIG. 1 is a HPLC diagram of ginseng saponin by conventional methanol extraction method, FIG. 2 is a HPLC diagram of ginseng saponin by conventional ethyl alcohol extraction method, and FIG. 3 is a saponin HPLC diagram by ethyl alcohol extraction method after treating ginseng polysaccharide enzyme. 4 is saponin HPLC diagram of ethyl ginseng extracted with ethyl alcohol after yeast leaching complex enzyme treatment.

1 and 2 are diagrams analyzing the extract by the conventional saponin extraction method and FIGS. 3 and 4 are diagrams analyzing the saponin extract according to the present invention.

As mentioned above, in order to make the method and effect by this invention clearer, it shows in the following Example.

Example (1)

100 g of 40 mesh ginseng powder was added to 200-1500 ml of distilled water, mixed, cooled to 60 ° C. for 20 minutes, and saccharifying endo-cellulase (Endo-Cellulase EC 3, 2, 1, 4). Jin Fengxie: The third Japan China Joint Symposium on Enzyme Engineering Sep. 19-20, 1994, Kyoto.p17) was mixed and reacted at 40-60 ℃ for 2-4 hours at 95%, ethyl alcohol. The mixture was filtered and filtered to a concentration of 40-50% (v / v), and the filtrate was extracted several times at 70-80 ° C. with 40-50% ethyl alcohol. After distillation under reduced pressure to 1 / 10-1 / 20 volume, 3-5-degreased with the same volume of ether (C ₂ H ₅ -OC ₂ H ₅ ), the aqueous layer was collected and unsaturated butanol (CH ₃ CH ₂ CH ₂ CH ₂ —OH) 50 ml of mixture was mixed with saponins, and the butanol layers were combined several times.

If treated by the above-described method without enzyme treatment can be obtained 3-6g saponin of the same purity in 100g ginseng powder, and 1-3g saponin can be obtained by enzymatic treatment of the remaining ginseng residue.

The saponins obtained by the enzyme treatment and the non-enzyme treatment extracted by this method were almost similar to the difference in purity as measured by Vanillin color development method using the Re saponin standard method. Almost the same saponin was obtained.

Example (2)

100 g of 40 mesh ginseng powder, 200-2000 ml of distilled water, mixed at 100 ° C. for 20 minutes, cooled to 50 ° C., and then reacted with starch enzyme for 2-3 hours. Distilled, degreased, leached with butanol and distilled under reduced pressure to obtain a dried product.

This was again dissolved in 80 ml of methanol and filtered. The filtrate was slowly added to cold 1000 ml of Acetone, and the precipitate was collected and dried to obtain 5-8 g of slightly yellow saponin.

If you do not go through the enzyme treatment process as described above to obtain 2.5-4g saponin from 100g ginseng, the above ginseng residue is treated by the enzyme treatment method and then treated with the same method can be obtained saponin 1-2.5g.

Example (3)

100 g of 40 mesh ginseng powder was mixed with 200-1500 ml of distilled water and heated at 100 ° C. for 20 minutes. In this complex enzyme, beta-glucosidase, alpha, beta-Xylosidase, alpha, beta-Arabinosidase, α, β-Rhamosidase, etc. If there is a degrading enzyme, it should be removed by heating or adding an inhibitor) and reacted for 2-5 hours, and then extracting saponin by the method of Example (1) or (2) described above. Up to 150% increase over the method.

The extracted saponins were almost the same as the saponin purity obtained by the method of vanillin development (中草藥, 13 (8), 19-23, 1982). Since there is little difference with the obtained saponin, it can be seen that the amount of saponin extracted is greatly increased.

As described above, it can be seen that the enzymatic treatment of ginseng powder greatly increases the extraction amount. If saponins are extracted in the same manner as in Example (1) (2) (3) instead of ginseng extract, The amount of saponin is increased by 20-100%, and saponin extract is higher than non-enzyme treatment when saponin is extracted from herbs such as sasamin, macmundong, emperor, gangyong, and soybeans containing saponin. It can be seen that the increase is increased by 20-100%. In addition, 40-70% of ginseng itself becomes a soluble substance, so that the enzyme treatment solution can be directly used for manufacturing other ginseng products, and other saponin as the main active ingredient Or, in the case of plants, the saponin content in the filtrate is greatly increased when the filtrate is treated after the enzyme treatment, and the content of the soluble substance, which is another active ingredient, is also greatly increased. The filtrate can be used directly for the manufacture of other products.

By treating the herbal medicines and plants containing ginseng with decomposition enzymes, they weaken the binding force of saponins or other active ingredients that are strongly bound to molecules of other substances in the plant, making it easier to extract and greatly improve the extraction efficiency. It is a way to extract valuable herbs or nutrients left in the extraction residue with a high yield.

Claims (3)

Method of extracting the active ingredient containing saponin by solvent leaching after sufficiently reacting Chinese herbal medicines and plants including ginseng immersed in water and cooled to 50-60 ℃ and the addition of the enzyme. The method of extracting saponin according to claim 1, wherein the degrading enzyme is an enzyme as follows. (1) starch enzymes: alpha-amylase, beta-amylase, glucoamylase, isoamylase (2) endo-cellulases (3) pectinase (4) Degrading enzymes mixed with one or more enzymes of solid or liquid yeasts of mold and monocot, complex enzymes leaching natural yeast, or complex enzyme abnormalities leaching plant seeds or seed germination The method of extracting saponin according to claim (1), wherein the degrading enzyme is a degrading enzyme from which an exo-glucose is removed by heat treatment or an inhibitor when the degrading enzyme includes an exo-sugar-binding enzyme.