A method for increasing the extraction of saponin components by using a low-concentration aqueous solution of a carbohydrate compound and its application

Abstract

The present invention discloses a method for increasing the extraction of active ingredients of saponin compounds using sugar. The method comprises the following steps: (1) preparing a sugar solution and / or a sugar alcohol solution; (2) soaking a raw material in the sugar solution and / or the sugar alcohol solution; and (3) heating and extracting the raw material in the sugar solution and / or the sugar alcohol solution, and filtering to obtain an extract. The above method can significantly increase the extraction rate of the active ingredients of saponin compounds in the raw material, thereby helping to improve the utilization rate and therapeutic effect of the raw material.

Description

Technical Field

[0001] The present invention relates to the field of chemical component extraction, and in particular to a method for increasing the leaching of saponin components by utilizing a low-concentration sugar compound aqueous solution and an application thereof. Background Art

[0002] Currently, the efficient and environmentally friendly extraction of active / effective ingredients, such as saponins, from raw materials, while ensuring the safety and efficacy of the extracts, is a key focus in the field of extraction research for raw materials such as Traditional Chinese Medicine (TCM) and herbal medicines. Modern research has found that the extraction efficiency of most TCM components using ethanol is much higher than that obtained by water decoction, but this may increase the toxicity of the resulting extracts. Traditional Chinese medicine preparation processes generally utilize water decoction, and clinical experience and modern research indicate that this method offers better safety assurance than organic solvent extraction. The main reason for the differences between the two extraction methods is that TCM contains a wide variety of active ingredients with a wide polarity range. Ethanol extraction increases the extraction rate of weakly and moderately polar components, resulting in a different composition of active ingredients in the ethanol extract than in traditional clinical decoctions, potentially impacting the safety of the extract. Furthermore, the traditional water decoction method, the most commonly used for extracting TCM, suffers from low extraction yields and incomplete extraction. While increasing the amount of water used can improve the extraction rate of active ingredients, it increases concentration time and production costs.

[0003] Therefore, how to improve the extraction rate of active ingredients in raw materials such as traditional Chinese medicine extracts in a green and efficient manner based on ancient practices is an important research direction in the research of raw material extraction such as traditional Chinese medicine. Summary of the Invention

[0004] Based on this, the present invention provides a method for increasing the extraction of saponins using a low-concentration aqueous solution of a carbohydrate compound. By using a certain concentration of sugar solution and / or sugar alcohol solution instead of water as the extraction solvent, the extraction yield is significantly improved compared to traditional decoction methods, thereby enhancing the utilization and efficacy of raw materials such as traditional Chinese medicines. The sugar used in this study is a commonly used excipient in traditional Chinese medicine or edible sugar, making it safe and inexpensive, without compromising the safety of the extract.

[0005] Specifically, according to one aspect of the present invention, a method for increasing the extraction of active ingredients of saponin compounds by using sugar is provided, the method comprising the following steps:

[0006] (1) preparing sugar solution and / or sugar alcohol solution;

[0007] (2) soaking the raw material in the sugar solution and / or the sugar alcohol solution; and

[0008] (3) extracting the raw material by heating in the sugar solution and / or the sugar alcohol solution, and obtaining a raw material extract by filtering.

[0009] According to another aspect of the present invention, there is provided a method for increasing the extraction of saponin compounds by using sugar, the method comprising the following steps:

[0010] (1) preparing sugar solution and / or sugar alcohol solution;

[0011] (2) soaking the raw material in the sugar solution and / or the sugar alcohol solution;

[0012] (3) extracting the raw material by heating in the sugar solution and / or the sugar alcohol solution, and obtaining a first extract by filtering;

[0013] (4) adding the sugar solution and / or the sugar alcohol solution to the raw material, heating and extracting the raw material, and filtering to obtain a second extract; and

[0014] (5) combining the first extract and the second extract to obtain a total raw material extract.

[0015] Furthermore, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-2.5 g / 100 mL.

[0016] Furthermore, the concentration of the sugar solution and / or the sugar alcohol solution is 1-2 g / 100 mL.

[0017] Furthermore, the concentration of the sugar solution and / or the sugar alcohol solution is about 1 g / 100 mL.

[0018] Furthermore, the sugar is selected from one or more of monosaccharides, disaccharides and trisaccharides.

[0019] Furthermore, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol and xylitol.

[0020] Furthermore, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, galactose, sorbose, rhamnose, ribose, xylose and deoxyribose.

[0021] Furthermore, the disaccharide is selected from one or more of maltose, sucrose, lactose, trehalose, melibiose, gentiobiose, and kojibiose.

[0022] Furthermore, the monosaccharide is glucose and / or fructose.

[0023] Furthermore, the trisaccharide is raffinose.

[0024] Furthermore, the disaccharide is sucrose and / or maltose.

[0025] Furthermore, the saponin compound is a triterpenoid saponin compound and / or a steroidal saponin compound.

[0026] Furthermore, the steroidal saponin compound is a spirostanol saponin compound, a furostanol saponin compound and / or an isospirostanol saponin compound.

[0027] Furthermore, the triterpenoid saponin compound is a tetracyclic triterpenoid saponin compound and / or a pentacyclic triterpenoid saponin compound.

[0028] Furthermore, the isospirostanol saponin compound is a dioscin compound.

[0029] Furthermore, the furostanol saponin compound is a timosaponin compound.

[0030] Furthermore, the tetracyclic triterpenoid saponin compound is a lanolin-type tetracyclic triterpenoid saponin compound and / or a dammarane-type tetracyclic triterpenoid saponin compound.

[0031] Furthermore, the pentacyclic triterpenoid saponin compound is an oleanane-type pentacyclic triterpenoid saponin compound, an ursane-type pentacyclic triterpenoid saponin compound and / or a lupane-type pentacyclic triterpenoid saponin compound.

[0032] Furthermore, the furostanol-type steroidal saponin compound is anemarrhena saponin BII.

[0033] Furthermore, the lanolin-type tetracyclic triterpenoid saponin compound is astragaloside IV.

[0034] Furthermore, the dammarane-type tetracyclic triterpenoid saponin compound is ginsenoside Rg1.

[0035] Furthermore, the oleanane-type pentacyclic triterpenoid saponin compound is glycyrrhizic acid.

[0036] Furthermore, the raw material is a raw material containing saponin compounds.

[0037] Furthermore, the raw materials are one or more traditional Chinese medicines, botanicals and / or marine organisms containing saponin compounds.

[0038] Furthermore, the Chinese medicine and / or herbal medicine is selected from one or more of the following: ginseng, Panax notoginseng, licorice, astragalus, julibrissin bark, pokeweed, bupleurum, whitehead, toona sinensis, Huoshan dendrobium, ox knee, clematis, American ginseng, fritillaria, horse chestnut, Acanthopanax senticosus, chestnut, Gynostemma pentaphyllum, chinaberry, Centella asiatica, red hairy notoginseng, holly, dipsaccharum officinale, akebia, japonica, angelica officinalis, clematis, jujube leaf, cannabis, polygala, eastern perilla, Eclipta prostrata, platycodon, ophiopogon, scallion, polygonatum, polygonatum, anemarrhena, asparagus, ophiopogon, parsley, parsley, smilax, veratrum, yam, tribulus terrestris, eggplant, pomegranate, almond, sweet potato, yam, asparagus, trillium, perilla bark, purple taro and dragon's blood.

[0039] Furthermore, in step (2), the soaking is performed at room temperature.

[0040] Furthermore, the soaking step comprises soaking the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for 10-120 minutes.

[0041] Furthermore, the soaking step comprises soaking the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for 20-60 minutes.

[0042] Furthermore, the soaking step comprises soaking the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for about 30 minutes.

[0043] Furthermore, the weight-to-volume ratio of the raw material to the sugar solution and / or the sugar alcohol solution is 1:1 to 1:50.

[0044] Furthermore, the weight-to-volume ratio of the raw material to the sugar solution and / or the sugar alcohol solution is 1:1 to 1:20.

[0045] Furthermore, the weight-to-volume ratio of the raw material to the sugar solution and / or the sugar alcohol solution is 1:6 to 1:7.

[0046] Furthermore, in step (3), the heating extraction is heating reflux extraction.

[0047] Furthermore, the heating time is 20 min to 60 min.

[0048] Furthermore, the heating time is about 30 minutes.

[0049] According to one aspect of the present invention, there is provided a use of the above method in preparing a raw material extract or a raw material concentrate, wherein the raw material is a traditional Chinese medicine, a botanical medicine and / or a marine organism.

[0050] According to one aspect of the present invention, there is provided a use of the above method in preparing a pharmaceutical composition containing a raw material, wherein the raw material is a traditional Chinese medicine, a botanical medicine and / or a marine organism.

[0051] According to one aspect of the present invention, there is provided a use of the above method in preparing a pharmaceutical preparation or functional food or health food containing raw materials, wherein the raw materials are traditional Chinese medicines, botanicals and / or marine organisms.

[0052] Beneficial effects of the present invention:

[0053] Our research has found that compared with the traditional water decoction method, the method of the present invention can fully improve the extraction rate of the active ingredients of saponin compounds in raw materials such as traditional Chinese medicine. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0055] Figure 1 The diagram below shows an example of saponin compounds in the raw materials, where A is astragaloside IV, B is ginsenoside Rg1, C is glycyrrhizic acid, and D is anemarrhena saponin BII. DETAILED DESCRIPTION

[0056] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work shall fall within the scope of protection of the present invention.

[0057] It should be noted that, in the absence of conflict, the embodiments and features of the embodiments of the present invention can be combined with each other. The present invention will be described in detail below with reference to the embodiments.

[0058] The present invention is further described in detail below with reference to specific examples. These examples should not be construed as limiting the scope of protection claimed in the present invention.

[0059] As described in the background technology section, the composition of the active ingredients in alcohol extracts differs from that in traditional clinical decoctions, which may affect the safety of alcohol extracts. However, the extraction efficiency of traditional water decoctions is not high. To address the above issues, the present invention provides a method for increasing the extraction of the active ingredients of saponin compounds using sugar, comprising the following steps:

[0060] (1) preparing sugar solution and / or sugar alcohol solution;

[0061] (2) soaking the raw material in the sugar solution and / or the sugar alcohol solution; and

[0062] (3) extracting the raw material by heating in the sugar solution and / or the sugar alcohol solution, and obtaining a raw material extract by filtering.

[0063] The ability of sugar solutions and / or sugar alcohol solutions to improve the extraction rate of active ingredients of saponins in raw materials such as traditional Chinese medicine is mainly related to two factors:

[0064] (1) Whether a liquid can soak into a solid medicinal piece is related to its surface tension. The smaller the surface tension coefficient (30×10 -3N / m), which can almost wet solids; water has a large surface tension coefficient and can only wet certain solids. The surface tension coefficient of a sugar solution is lower than that of pure water, allowing the solution to better penetrate plant cells and release active ingredients. This can improve the wetting of raw materials such as Chinese herbal medicine slices and increase the dissolution of saponin compounds.

[0065] (2) Sugar can form hydrogen bonds and other interactions with active ingredients such as saponin compounds in raw materials such as Chinese herbal medicine slices, thereby improving the extraction efficiency of saponin components in raw materials such as Chinese herbal medicine extracts;

[0066] Therefore, compared with the water decoction method, this method can fully improve the extraction rate of the active ingredients of saponin compounds in raw materials such as traditional Chinese medicine.

[0067] In actual working process, the number of extractions of alcohol and / or sugar alcohol solution can be determined according to actual needs, which can be 1 time or several times, for example, 2-10 times.

[0068] According to another aspect of the present invention, there is provided a method for increasing the extraction of saponin compounds by using sugar, the method comprising the following steps:

[0069] (1) preparing sugar solution and / or sugar alcohol solution;

[0070] (2) soaking the raw material in the sugar solution and / or the sugar alcohol solution;

[0071] (3) extracting the raw material by heating in the sugar solution and / or the sugar alcohol solution, and obtaining a first extract by filtering;

[0072] (4) adding the sugar solution and / or the sugar alcohol solution to the raw material, heating and extracting the raw material, and filtering to obtain a second extract; and

[0073] (5) combining the first extract and the second extract to obtain a total raw material extract. In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-2.5 g / 100 mL.

[0074] In the present invention, when concentration, ratio, equivalent, number or other parameters are expressed as a range, a preferred range, or a range defined by a series of upper preferred values ​​and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any upper range limit or preferred value with any lower range limit or preferred value, regardless of whether the range is disclosed separately. .4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 0.61, 0.62, 0.63, 0.64, 0.65, 1.21, 1.22, 1.23, 1.24, 1.25, 1.26, 1.27, 1.28, 1.29, etc., without limitation to the values ​​listed above, unless otherwise stated.

[0075] In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is 1-2 g / 100 mL.

[0076] In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is about 1 g / 100 mL.

[0077] The term "about" or "approximately" with respect to a numerical value means ±5% of the numerical value, but explicitly includes the exact numerical value. For example, a concentration of "about" 1 g / 100 mL means a concentration from 0.95 g / 100 mL to 1.05 g / 100 mL, but also explicitly includes a concentration of exactly 1 g / 100 mL.

[0078] In a preferred embodiment, the sugar is selected from one or more of monosaccharides, disaccharides and trisaccharides.

[0079] In a preferred embodiment, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol and xylitol. In actual application, the above sugar alcohols can be replaced with other sugar alcohols in the prior art according to actual conditions.

[0080] In a preferred embodiment, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, galactose, sorbose, rhamnose, ribose, xylose, and deoxyribose. In actual application, the above monosaccharides can be replaced with other monosaccharides in the prior art according to actual conditions.

[0081] In a preferred embodiment, the disaccharide is selected from one or more of maltose, sucrose, lactose, trehalose, melibiose, gentiobiose and kojibiose. In practical applications, the above disaccharides can be replaced with other disaccharides in the prior art according to actual conditions.

[0082] In a preferred embodiment, the monosaccharide is glucose and / or fructose.

[0083] In a preferred embodiment, the trisaccharide is raffinose.

[0084] In order to further improve the extraction rate of the active ingredients of saponin compounds in raw materials such as traditional Chinese medicine, in a preferred embodiment, the disaccharide is sucrose and / or maltose.

[0085] In a preferred embodiment, the saponin compound is a triterpenoid saponin compound and / or a steroidal saponin compound.

[0086] In a preferred embodiment, the steroidal saponin compound is a spirostanol saponin compound, a furostanol saponin compound and / or an isospirostanol saponin compound.

[0087] In a preferred embodiment, the triterpenoid saponin compound is a tetracyclic triterpenoid saponin compound and / or a pentacyclic triterpenoid saponin compound.

[0088] In a preferred embodiment, the isospirostanol saponin compound is a dioscin compound.

[0089] In a preferred embodiment, the furostanol saponin compound is a timosaponin compound.

[0090] In a preferred embodiment, the tetracyclic triterpenoid saponin compound is a lanolin-type tetracyclic triterpenoid saponin compound and / or a dammarane-type tetracyclic triterpenoid saponin compound.

[0091] In a preferred embodiment, the pentacyclic triterpenoid saponin compound is an oleanane-type pentacyclic triterpenoid saponin compound, an ursane-type pentacyclic triterpenoid saponin compound and / or a lupane-type pentacyclic triterpenoid saponin compound.

[0092] In a preferred embodiment, the furostanol-type steroidal saponin compound is anemarrhena saponin BII.

[0093] In a preferred embodiment, the lanolin-type tetracyclic triterpenoid saponin compound is astragaloside IV.

[0094] In a preferred embodiment, the dammarane-type tetracyclic triterpenoid saponin compound is ginsenoside Rg1.

[0095] In a preferred embodiment, the oleanane-type pentacyclic triterpenoid saponin compound is glycyrrhizic acid.

[0096] In a preferred embodiment, the raw material is a raw material containing saponin compounds.

[0097] In a preferred embodiment, the raw material is one or more traditional Chinese medicines, botanicals and / or marine organisms containing saponin compounds.

[0098] In a preferred embodiment, the Chinese medicine and / or herbal medicine is selected from one or more of the following: ginseng, Panax notoginseng, licorice, astragalus, julibrissin bark, pokeweed, bupleurum, whitehead, toona sinensis, Huoshan dendrobium, ox knee, clematis, American ginseng, fritillaria, horse chestnut, acanthopanax senticosus, tianshi chestnut, gynostemma pentaphyllum, chinaberry, centella asiatica, red hair seven, holly, dipsaccharum officinale, akebia, japonica, angelica officinalis, clematis, jujube leaf, cannabis, polygala, eastern perilla, Eclipta prostrata, platycodon, ophiopogon, scallion, polygonatum, polygonatum, anemarrhena, asparagus, ophiopogon, parsley, parsley, smilax, veratrum, yam, tribulus terrestris, purple flower eggplant, pomegranate, almond, sweet potato, yam, asparagus, trillium, perilla bark, purple taro and dragon's blood.

[0099] In a preferred embodiment, in step (2), the soaking is performed at room temperature.

[0100] In a preferred embodiment, the soaking step comprises soaking the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for 10-120 minutes.

[0101] In a preferred embodiment, the soaking step comprises soaking the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for 20-60 minutes.

[0102] In a preferred embodiment, the soaking step comprises soaking the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for about 30 minutes.

[0103] The term "about" or "approximately" with respect to a numerical value means ±5% of the numerical value, but explicitly includes the exact numerical value. For example, a time of "about" 30 minutes means a time from 28.5 minutes to 31.5 minutes, but also explicitly includes a time of exactly 30 minutes.

[0104] In a preferred embodiment, the weight-to-volume ratio of the raw material to the sugar solution and / or the sugar alcohol solution is 1:1 to 1:50.

[0105] In a preferred embodiment, the weight-to-volume ratio of the raw material to the sugar solution and / or the sugar alcohol solution is 1:1 to 1:20.

[0106] In a preferred embodiment, the weight-to-volume ratio of the raw material to the sugar solution and / or the sugar alcohol solution is 1:6 to 1:7.

[0107] In a preferred embodiment, in step (3), the heating extraction is heating reflux extraction.

[0108] In a preferred embodiment, the heating time is 20 min to 60 min.

[0109] In a preferred embodiment, the heating time is about 30 minutes.

[0110] The term "about" or "approximately" with respect to a numerical value means ±5% of the numerical value, but explicitly includes the exact numerical value. For example, a time of "about" 30 minutes means a time from 28.5 minutes to 31.5 minutes, but also explicitly includes a time of exactly 30 minutes.

[0111] According to one aspect of the present invention, there is provided a use of the above method in preparing a raw material extract or a raw material concentrate, wherein the raw material is a traditional Chinese medicine, a botanical medicine and / or a marine organism.

[0112] In a preferred embodiment, the raw material concentrated solution is obtained by concentrating the raw material extract obtained in the above step (3) or the raw material total extract obtained in step (5) in a water bath at 20° C. to 100° C.

[0113] In a preferred embodiment, the concentration is performed in a water bath at 60°C to 80°C.

[0114] In a preferred embodiment, the concentration is accomplished in a water bath at about 70°C.

[0115] The term "about" or "approximately" with respect to a numerical value means ±5% of the numerical value, but explicitly includes the exact numerical value. For example, a temperature of "about" 70°C refers to a range from 66.5°C to 73.5°C, but also explicitly includes a temperature of exactly 70°C.

[0116] According to one aspect of the present invention, there is provided a use of the above method in preparing a pharmaceutical composition containing a raw material, wherein the raw material is a traditional Chinese medicine, a botanical medicine and / or a marine organism.

[0117] According to one aspect of the present invention, there is provided a use of the above method in preparing a pharmaceutical preparation or functional food or health food containing raw materials, wherein the raw materials are traditional Chinese medicines, botanicals and / or marine organisms.

[0118] Functional foods are foods that can be convincingly proven to be beneficial to one or more functions of the body and have sufficient nutritional effects to improve health or reduce illness.

[0119] Among them, health food refers to food that claims to have specific health functions or is intended to supplement vitamins and minerals. That is, it is suitable for consumption by specific groups of people, has the function of regulating body functions, is not intended to treat diseases, and does not cause any acute, subacute or chronic harm to the human body.

[0120] According to one aspect of the present invention, there is provided a use of the above-mentioned method in preparing other products containing raw materials, wherein other products refer to all products included in the prior art containing raw materials such as traditional Chinese medicine, botanicals and / or biological preparations, except for pharmaceutical preparations, functional foods and health foods, including liquid and solid forms.

[0121] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present invention will be described in detail below with reference to the embodiments.

[0122] The present invention is further described in detail below with reference to specific examples. These examples should not be construed as limiting the scope of protection claimed in this application.

[0123] Example

[0124] Example 1 Experiment on extracting raw licorice slices with different sugar solutions

[0125] 1.1 Preparation methods of different sugar-licorice extracts

[0126] Four portions of the same batch of raw licorice slices (produced in Xinjiang Uygur Autonomous Region) were taken and numbered, each 40 g. Seven times the amount of water (280 mL of water), 1% sucrose aqueous solution (2.8 g of sucrose + 280 mL of water), 1% maltose aqueous solution (2.8 g of maltose + 280 mL of water), and 1% lactose aqueous solution (2.8 g of lactose + 280 mL of water) were added in sequence and soaked for 30 min, refluxed for 30 min, and filtered with gauze while hot. The residues were then added with six times the amount of water (240 mL of water), 1% sucrose aqueous solution (2.4 g of sucrose + 240 mL of water), 1% maltose aqueous solution (2.4 g of maltose + 240 mL of water), and 1% lactose aqueous solution (2.4 g of lactose + 240 mL of water), refluxed for 20 min, filtered while hot, and the two filtrates were combined to obtain different sugar-licorice extracts.

[0127] 1.2 Determination of glycyrrhizic acid content

[0128] 1.2.1 Instruments and reagents

[0129] An Agilent 1260 high-performance liquid chromatograph equipped with a DAD detector (Agilent Technologies, Inc., USA); a DZKW-4 electronic constant-temperature water bath (Beijing Zhongxing Weiye Instrument Co., Ltd.); a DK-98-ⅡA electric constant-temperature water bath (Tianjin Test Instrument Co., Ltd.); an ME204 / 02 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd., 1 / 100,000 balance); and a KQ-250DE digitally controlled ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).

[0130] The reference substance, glycyrrhizic acid (Z30A6B1), was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with a purity of >98%. Acetonitrile (chromatographic grade, Fisher), methanol (chromatographic grade, Fisher), and analytical grade phosphoric acid were purchased from Sinopharm Chemical Reagent Co., Ltd. Maltose, lactose, and sucrose were purchased from Golden Clone (Beijing) Biotechnology Co., Ltd., and Golden Clone (Beijing Biotechnology Co., Ltd.) were all pharmaceutical grade. Water was Wahaha purified water.

[0131] 1.2.2 Preparation of test solution

[0132] Take 0.5 mL of different sugar-licorice extracts, add methanol (9.5 mL) to dilute 20 times, shake well, and pass through a 0.22 μm filter membrane to obtain the product.

[0133] 1.2.3 Preparation of reference solution

[0134] Stock solution: Accurately weigh an appropriate amount of glycyrrhizic acid reference substance and add methanol to prepare a 1 mg / mL stock solution. Store in a -20°C refrigerator.

[0135] Linear working solution: Take an appropriate amount of glycyrrhizic acid stock solution and dilute it with methanol to form a series of linear mixed standard working solutions: 10, 20, 40, 80, 160, 200, 400 μg / mL.

[0136] 1.2.4 Content determination method

[0137] Chromatographic column: Thermo Acclaim 120C18 (250×4.6mm, 5μm); mobile phase: acetonitrile (A), 0.05% phosphoric acid solution (B) as mobile phase, column temperature: 35°C, detection wavelength: 237nm, injection volume: 10μL, flow rate: 1ml / min, gradient elution: 0-8min, 19%A; 8-35min, 19%-50%A; 35-36min, 50%-100%A; 36-40min, 100%-19%A; 40-50min, 19%A.

[0138] 1.3 Experimental Results

[0139] The results of determination of the active ingredient contents in different sugar-licorice extracts are shown in Table 1.

[0140] Table 1 Determination results of active ingredient content in different sugar-licorice extracts

[0141]

[0142] *P<0.05 vs water

[0143] As can be seen from Table 1, both sucrose and maltose (1 g / 100 mL concentration) can increase the content of the active ingredient in licorice - saponins (glycyrrhizic acid) to varying degrees, among which sucrose has the most obvious effect.

[0144] Example 2 Experiment on Extracting Raw Astragalus Root Pieces with Different Sugar Solutions

[0145] 2.1 Preparation methods of different sugar-astragalus extracts

[0146] Four portions of 50 g each of the same batch of raw Astragalus root slices (from Lingchuan, Shanxi) were taken. Seven times the amount of water (350 ml of water), 1% sucrose aqueous solution (3.5 g of sucrose was added to 350 ml of water), 1% maltose aqueous solution (3.5 g of maltose was added to 350 ml of water), and 1% lactose aqueous solution (3.5 g of lactose was added to 350 ml of water) were added, respectively, and soaked for 30 minutes. The mixture was refluxed for 30 minutes and filtered with gauze while hot. The residue was then added with six times the amount of water (300 ml of water), 1% sucrose aqueous solution (3.0 g of sucrose was added to 300 ml of water), 1% maltose aqueous solution (3.0 g of maltose was added to 300 ml of water), and 1% lactose aqueous solution (3.0 g of lactose was added to 300 ml of water). The mixture was refluxed for 20 minutes and filtered while hot. The two filtrates were combined to obtain different sugar-Astragalus root extracts.

[0147] 2.2 Determination of Astragaloside IV Content

[0148] 2.2.1 Instruments and reagents

[0149] Agilent 1200 high-performance liquid chromatograph equipped with an ELSD detector (Agilent Technologies, Inc., USA); DZKW-4 electronic constant-temperature water bath (Beijing Zhongxing Weiye Instrument Co., Ltd.); DK-98-ⅡA electric constant-temperature water bath (Tianjin Test Instrument Co., Ltd.); ME204 / 02 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd., 1 / 100,000 balance); KQ-250DE digitally controlled ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); HC-3018 high-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); and solid-phase extraction UniEIut C18EC column (1000 mg / 6 ml) (Beijing Huapu Science and Technology Co., Ltd.).

[0150] The reference substance, astragaloside IV (batch number CHB170727), was purchased from Chengdu Kroma Biotechnology Co., Ltd. with an HPLC purity of ≥98%. Acetonitrile (chromatographic grade, Thermo Fisher Scientific), methanol (chromatographic grade, Thermo Fisher Scientific), and formic acid were of analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd. Maltose, lactose, and sucrose were purchased from Golden Clone (Beijing) Biotechnology Co., Ltd., and were pharmaceutical grade. Water was Wahaha purified water.

[0151] 2.2.2 Preparation of test solution

[0152] Take 5 ml of different sugar-astragalus extracts, add 20 ml of 60% methanol, ultrasonically mix, add 10 ml of ammonia water, mix well, and centrifuge at 3600 r / min for 10 min. The supernatant is passed through a Huapu UniEIut C18 EC column (1000 mg / 6 ml), eluted with 10 ml of pure water, and then eluted with 5 ml of methanol. The methanol eluate is collected and centrifuged at 8000 r / min for 5 min. The supernatant is taken and passed through a 0.45 μm microporous filter membrane to obtain the product.

[0153] 2.2.3 Preparation of reference solution

[0154] Take an appropriate amount of Astragaloside IV reference substance, accurately weigh it, and add methanol to prepare a 1 mg / ml stock solution. Dilute the stock solution into different concentration series and establish a working curve.

[0155] The linear equation of astragaloside IV is lnY=1.3271lnX+9.9314, r=1.0000.

[0156] 2.2.4 Content determination method

[0157] Saponin component (astragaloside IV) content determination method: Agilent 1200 liquid chromatograph (equipped with ELSD detector) was used for analysis. The chromatographic column was Thermo Acclaim 120C18 (250×4.6 mm, 5 μm). The mobile phase was 0.1% formic acid in water (A) to 0.1% formic acid in acetonitrile (B). The gradient elution was as follows: 0-5 min, 5%-10% B; 5-10 min, 10%-32% B; 10-30 min, 32%-45% B; 30-35 min, 45%-95% B; 35-40 min, 95%-20% B. The column temperature was room temperature. The flow rate was 1 ml / min, the drift tube temperature was 100°C, the carrier gas flow rate was 2.5 l / min, and the injection volume was 20 μl.

[0158] 2.3 Experimental Results

[0159] The results of the determination of the active ingredient contents in different sugar-astragalus extracts are shown in Table 2.

[0160] Table 2 Determination results of effective ingredient content in different sugar-astragalus extracts

[0161]

[0162] *P<0.05 vs water

[0163] As can be seen from Table 2, sucrose, maltose and lactose (1g / 100ml concentration) can all increase the content of the active ingredient in Astragalus - saponins (astragaloside IV) to varying degrees, among which maltose has the most obvious effect.

[0164] Example 3: Experiment on Extraction of Ginsenoside Rg1 from Panax notoginseng Using Sugar Solutions of Different Concentrations

[0165] 3.1 Preparation of ginsenoside Rg1 from Panax notoginseng by extraction with different sugar concentrations

[0166] Take 5 portions of the same batch of raw Panax notoginseng slices (origin: Yunnan; manufacturer: Tianjin Shengshi Pharmaceutical Co., Ltd.; batch number: X20102703A), each 50g. Add 7 times the amount of water (350ml water), 0.5% sucrose aqueous solution (1.75g ​​sucrose is added to 350ml with water), 1% sucrose aqueous solution (3.5g sucrose is added to 350ml with water), and 1.5% sucrose aqueous solution (5.25g sucrose is added to 350ml with water), soak for 30min, reflux for 30min, filter with gauze while hot, and add 6 times the amount of water (300ml water), 0.5% sucrose aqueous solution (1.5g sucrose is added to 300ml with water), 1% sucrose aqueous solution (3.0g sucrose is added to 300ml with water), and 1.5% sucrose aqueous solution (4.5g sucrose is added to 300ml with water), reflux for 20min, filter while hot, and combine the two filtrates to obtain different sugar-Sanqi extracts.

[0167] 3.2 Determination of ginsenoside Rg1 content

[0168] 3.2.1 Instruments and reagents

[0169] An Agilent 1260 high-performance liquid chromatograph equipped with a DAD detector (Agilent Technologies, Inc., USA); a DZKW-4 electronic constant-temperature water bath (Beijing Zhongxing Weiye Instrument Co., Ltd.); a DK-98-ⅡA electric constant-temperature water bath (Tianjin Test Instrument Co., Ltd.); an ME204 / 02 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd., 1 / 100,000 balance); a KQ-250DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); and an HC-3018 high-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.).

[0170] The reference substance, ginsenoside Rg1 (batch number 10703-201731), had a purity of ≥93.6% as determined by HPLC. Acetonitrile (chromatographically pure, Thermo Fisher Scientific Inc.), methanol (chromatographically pure, Thermo Fisher Scientific Inc.), and formic acid were all analytically pure and purchased from Sinopharm Chemical Reagent Co., Ltd. Sucrose was pharmaceutical grade (Golden Clone Beijing Biotechnology Co., Ltd.). Water was Wahaha purified water.

[0171] 3.2.2 Preparation of test solution

[0172] Accurately measure 2 mL of different Panax notoginseng extracts, add methanol to the 10 mL volumetric flask, mix well, and spin at 13000 r·min. -1 Centrifuge for 5 minutes, take the supernatant, and filter it through a 0.22 μm microporous membrane.

[0173] 3.2.3 Preparation of reference solution

[0174] Take an appropriate amount of ginsenoside Rg1 reference substance, accurately weigh it, and add methanol to prepare a 1 mg / ml stock solution. Dilute the stock solution into different concentration series and establish a working curve.

[0175] The linear equation of ginsenoside Rg1 is: Y = 3246.4X - 4.5152R 2 =0.9999.

[0176] 3.2.4 Content determination method

[0177] An Agilent 1200 liquid chromatograph was used for analysis. The chromatographic column was Roc C18 (250×4.6 mm, 5 μm). The mobile phase was water (A) to acetonitrile (B). The gradient elution was as follows: 19% to 19% B in 0 to 12 min and 19% to 36% B in 12 to 60 min. The column temperature was 35°C. The flow rate was 1 ml / min. The injection volume was 10 μl. The detection wavelength was 203 nm.

[0178] 3.3 Experimental Results

[0179] The effects of different concentrations of sucrose on the extraction rate of index components in Panax notoginseng are shown in Table 3.

[0180] Table 3 Effects of different sucrose concentrations on the extraction rate of index components in Panax notoginseng

[0181]

[0182] *P<0.05 vs Panax notoginseng aqueous extract

[0183] As can be seen from Table 3, sucrose (concentration of 1g / 100ml-1.5g / 100ml) can increase the content of the active ingredient in Panax notoginseng - saponin components (ginsenoside Rg1) to varying degrees.

[0184] Example 4 Experiment on Extracting Anemarrhena Saponin BⅡ from Anemarrhena Decoction Pieces with Sugar Solutions of Different Concentrations

[0185] 4.1 Preparation of Anemarrhena Saponin BⅡ from Anemarrhena Rhizoma by Extraction of Sugars at Different Concentrations

[0186] Take 5 portions of the same batch of raw Anemarrhena slices, 50g each. Add 7 times the amount of water (350ml water), 0.5% sucrose aqueous solution (1.75g ​​sucrose is added to 350ml with water), 1% sucrose aqueous solution (3.5g sucrose is added to 350ml with water), 1.5% sucrose aqueous solution (5.25g sucrose is added to 350ml with water), 2.0% sucrose aqueous solution (7.0g sucrose is added to 350ml with water) and soak for 30min, reflux for 30min, filter with gauze while hot, and add 6 times the amount of water (350ml water), 1% sucrose aqueous solution (350ml with water), 1.5% sucrose aqueous solution (5.25g sucrose is added to 350ml with water), 2.0% sucrose aqueous solution (7.0g sucrose is added to 350ml with water) and soak for 30min. times the amount of water (300ml water), 0.5% sucrose aqueous solution (1.5g sucrose is added to 300ml with water), 1% sucrose aqueous solution (3.0g sucrose is added to 300ml with water), 1.5% sucrose aqueous solution (4.5g sucrose is added to 300ml with water), 2.0% sucrose aqueous solution (6g sucrose is added to 300ml with water), reflux for 20min, filter while hot, combine the two filtrates, and adjust the volume to 500mL to obtain different sugar-Anemarrhena extracts.

[0187] 4.2 Determination of Anemarrhena Saponin BⅡ Content

[0188] 4.2.1 Instruments and reagents

[0189] An Agilent 1200 high-performance liquid chromatograph equipped with an ELSD detector was used (Agilent Technologies, Inc., USA); an ME204 / 02 electronic balance was purchased from Mettler-Toledo Instruments (Shanghai) Co., Ltd., and a HC-3018 high-speed centrifuge was used (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.).

[0190] Anemarrhena saponin BⅡ (batch number 111839-201706, purity 94.5% by HPLC) with a purity of ≥94.5% by HPLC. Acetonitrile (chromatographically pure, Thermo Fisher Scientific), methanol (chromatographically pure, Thermo Fisher Scientific), and formic acid were all analytically pure and purchased from Sinopharm Chemical Reagent Co., Ltd. Sucrose was pharmaceutical grade (Gold Clonal Beijing Biotechnology Co., Ltd.). Water was Wahaha purified water.

[0191] 4.2.2 Preparation of test solution

[0192] Accurately measure 2 mL of different Anemarrhena extracts, add methanol to the 10 mL volumetric flask, mix well, and spin at 13000 r·min. -1 Centrifuge for 5 minutes, take the supernatant, and filter it through a 0.22 μm microporous membrane.

[0193] 4.2.3 Preparation of reference solution

[0194] Take an appropriate amount of Anemarrhena saponin BⅡ reference substance, accurately weigh it, and add 30% acetonitrile to prepare a stock solution with a mass concentration of 1 mg / ml. Dilute the stock solution into different concentration series and establish a working curve.

[0195] The linear equation of Anemarrhena saponin BⅡ is: lnY=1.8921lnX+10.274, R 2 =0.9977.

[0196] 4.2.4 Content determination method

[0197] Analysis was performed using an Agilent 1200 liquid chromatograph, using an Inertsil ODS-3-C18 column (250 × 4.6 mm, 5 μm); mobile phase: water (A) to acetonitrile (B), isocratic elution with 25% acetonitrile; column temperature: 35°C, flow rate: 0.8 ml / min, injection volume: 10 μl; drift tube temperature: 104°C, carrier gas flow rate: 2.8 L min -1 .

[0198] 4.3 Experimental Results

[0199] The effects of different concentrations of sucrose on the extraction rate of index components in Anemarrhena asphodeloides are shown in Table 4.

[0200] Table 4 Effects of different sucrose concentrations on the extraction rate of index components in Anemarrhena asphodeloides

[0201]

[0202] *P<0.05 vs Anemarrhena asphodeloides aqueous extract

[0203] As can be seen from Table 4, sucrose (concentrations of 1g / 100ml-2.0g / 100ml) can increase the content of the effective ingredient in Anemarrhena asphodeloides - saponin components (Anemarrhena saponin BⅡ) to varying degrees.

[0204] The above is only an embodiment of the present invention, and the common knowledge such as the specific structure and characteristics of the scheme is not described in detail here. It should be pointed out that for those skilled in the art, several variations and improvements can be made without departing from the present invention, and these should also be regarded as the scope of protection of the present invention. These will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by the present invention shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

[0205] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only intended to help understand the method of the present invention and its core concept. At the same time, changes or modifications made by those skilled in the art based on the concept of the present invention, the specific implementation methods of the present invention, and the scope of application are all within the scope of protection of the present invention. In summary, the contents of this specification should not be understood as limiting the present invention.

Claims

1. A method for increasing the extraction of saponin compounds by using sugar, characterized in that: The method comprises the following steps: (1) Prepare a sugar solution with a concentration of 1-1.5 g / 100 mL; (2) soaking the raw materials in the sugar solution; (3) extracting the raw material by heating in the sugar solution, and obtaining a first extract by filtering; (4) adding the sugar solution to the raw material, heating and extracting the raw material, and filtering to obtain a second extract; and (5) combining the first extract and the second extract to obtain a total raw material extract; Wherein, the sugar is sucrose and / or maltose; Wherein, the saponin compound is glycyrrhizic acid, astragaloside IV, ginsenoside Rg1 and / or timosaponin BII; Wherein, the raw material is a raw material containing the saponin compound.

2. The method according to claim 1, characterized in that The raw materials are one or more traditional Chinese medicines containing the saponin compounds.

3. The method according to claim 1, characterized in that The raw materials are one or more botanical medicines containing the saponin compounds.

4. The method according to claim 1, wherein The raw materials are one or more marine organisms containing the saponin compounds.

5. The method according to claim 2, characterized in that The Chinese medicine is selected from one or more of the following: ginseng, Panax notoginseng, liquorice, astragalus and anemarrhena.

6. The method according to claim 1, characterized in that In step (2), the soaking is done at room temperature.

7. The method according to claim 6, characterized in that The soaking step comprises soaking the raw material in the sugar solution at room temperature for 10-120 minutes.

8. The method according to claim 7, characterized in that The soaking step comprises soaking the raw material in the sugar solution at room temperature for 20-60 minutes.

9. The method according to claim 8, characterized in that The soaking step comprises soaking the raw material in the sugar solution at room temperature for 28.5-31.5 minutes.

10. The method according to claim 1, characterized in that The weight-to-volume ratio of the raw materials to the sugar solution is 1:1 to 1:

50.

11. The method according to claim 10, characterized in that The weight-to-volume ratio of the raw materials to the sugar solution is 1:1 to 1:

20.

12. The method according to claim 11, characterized in that The weight-to-volume ratio of the raw materials to the sugar solution is 1:6 to 1:

7.

13. The method according to claim 1, wherein In step (3), the heating extraction is heating reflux extraction.

14. The method according to claim 13, characterized in that The heating time is 20 min to 60 min.

15. The method according to claim 14, characterized in that The heating time is 28.5-31.5 minutes.

16. Use of the method according to any one of claims 1, 2, 5 to 15 in preparing a raw material extract or a raw material concentrate, wherein the raw material is a traditional Chinese medicine.

17. Use of the method according to any one of claims 1, 3, 6 to 15 in preparing a raw material extract or a raw material concentrate, wherein the raw material is a botanical drug.

18. Use of the method according to any one of claims 1, 4, 6 to 15 in preparing a raw material extract or a raw material concentrate, wherein the raw material is marine organisms.

19. Use of the method according to any one of claims 1, 2, 5 to 15 in preparing a pharmaceutical composition containing a raw material, wherein the raw material is a traditional Chinese medicine.

20. Use of the method according to any one of claims 1, 3, 6 to 15 in the preparation of a pharmaceutical composition containing a raw material, wherein the raw material is a botanical drug.

21. Use of the method according to any one of claims 1, 4, 6 to 15 in the preparation of a pharmaceutical composition containing a raw material, wherein the raw material is a marine organism.

22. Use of the method according to any one of claims 1, 2, 5 to 15 in the preparation of a pharmaceutical preparation containing a raw material, wherein the raw material is a traditional Chinese medicine.

23. Use of the method according to any one of claims 1, 2, 5 to 15 in preparing functional foods containing raw materials, wherein the raw materials are traditional Chinese medicines.

24. Use of the method according to any one of claims 1, 2, 5 to 15 in preparing a health food containing raw materials, wherein the raw materials are traditional Chinese medicines.

25. Use of the method according to any one of claims 1, 3, 6 to 15 for the preparation of a pharmaceutical preparation containing a raw material, wherein the raw material is a botanical drug.

26. Use of the method according to any one of claims 1, 3, 6 to 15 in the preparation of a functional food containing a raw material, wherein the raw material is a botanical drug.

27. Use of the method according to any one of claims 1, 3, 6 to 15 in the preparation of a health food containing a raw material, wherein the raw material is a botanical drug.

28. Use of the method according to any one of claims 1, 4, 6 to 15 in the preparation of a pharmaceutical preparation containing a raw material, wherein the raw material is a marine organism.

29. Use of the method according to any one of claims 1, 4, 6 to 15 in the preparation of a functional food containing a raw material, wherein the raw material is a marine organism.

30. Use of the method according to any one of claims 1, 4, 6 to 15 in the preparation of a health food containing a raw material, wherein the raw material is a marine organism.

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