Method for increasing extraction of licorice active ingredients using sugar

Abstract

The present application provides a method for increasing the extraction of effective components from licorice by using sugar. The method comprises the following steps: (1) preparing a sugar solution and / or a sugar alcohol solution; (2) soaking licorice in the sugar solution and / or the sugar alcohol solution; and (3) heating the licorice in the sugar solution and / or the sugar alcohol solution to extract the licorice, and obtaining licorice extract by filtration. The above method can significantly increase the extraction rate of effective components in licorice, thereby helping to improve the utilization rate and curative effect of licorice medicinal materials.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of traditional Chinese medicine extraction, and in particular to a method for increasing the extraction of effective components of licorice by using sugar. BACKGROUND

[0002] Currently, how to efficiently extract effective components of traditional Chinese medicine and ensure the safety of the extract is an important content in the field of traditional Chinese medicine extraction research. The traditional method of taking traditional Chinese medicine is mostly water decoction, and clinical experience and modern research show that this method can better ensure the effectiveness and safety of the extract than organic solvent extraction. Therefore, in the new drug registration and submission of traditional Chinese medicine released in 2020, if the indications and traditional use are consistent, the clinical trials can be appropriately reduced or exempted, such as the "Provisions on Simplifying the Registration and Approval Management of Traditional Chinese Medicine Compound Preparations of Ancient Classical Prescriptions" issued in 2018, which clearly states that the development of compound preparations of ancient classical prescriptions can be exempted from clinical trials, including preparation processes and records in ancient medical books. Modern research has found that the ethanol extraction efficiency of most components of traditional Chinese medicine is much higher than that of water decoction, but the toxicity of the extract increases. The main reason for the difference between the two extraction methods is that traditional Chinese medicine contains a wide variety of effective components with a wide range of polarity, and ethanol extraction increases the extraction rate of weakly polar and moderately polar components, resulting in differences in the composition of effective components between alcohol extracts and traditional clinical decoctions, which may have some impact on the safety of the extract. In addition, the traditional extraction method of traditional Chinese medicine, water decoction, has the problems of low extraction rate and incomplete extraction.

[0003] Therefore, how to improve the extraction rate of traditional Chinese medicine extract while adhering to the ancient method is an important research direction in the field of traditional Chinese medicine extraction research. SUMMARY

[0004] Therefore, the present application provides an efficient licorice decoction extraction method, which uses a certain concentration of sugar water and / or sugar alcohol water instead of water as the extraction solvent. Compared with the traditional decoction method, the extraction rate is significantly improved, thereby improving the utilization rate and efficacy of licorice medicinal materials. The sugar used in this study is a common auxiliary material for traditional Chinese medicine or edible sugar, and therefore has the characteristics of safety and low price, and does not affect the safety of the extract.

[0005] Specifically, according to one aspect of the present application, a method for increasing the extraction of effective components of licorice by using sugar is provided, which comprises the following steps:

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

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

[0008] (3) heating the liquor to extract the liquor from the liquor and / or the sugar alcohol solution.

[0009] Further, the liquor is raw liquor or liquor prepared by stir-frying with honey.

[0010] According to another aspect of the present application, there is provided a method for increasing the extraction of effective components from liquorice by using sugar, characterized in that the method comprises the following steps:

[0011] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0012] (2) soaking liquorice in the sugar solution and / or the sugar alcohol solution;

[0013] (3) heating the liquor to extract the liquor from the liquor and / or the sugar alcohol solution.

[0014] (4) soaking the liquorice in an aqueous solution, and heating the liquor to extract the liquor from the liquor and / or the sugar alcohol solution.

[0015] (5) combining the first liquor extract and the second liquor extract to obtain a liquor extract (i.e. a total liquor extract or a third liquor extract).

[0016] Further, the liquor is raw liquor or liquor prepared by stir-frying with honey.

[0017] According to another aspect of the present application, there is provided a method for increasing the extraction of effective components from liquorice by using sugar, characterized in that the method comprises the following steps:

[0018] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0019] (2) soaking liquorice in the sugar solution and / or the sugar alcohol solution;

[0020] (3) heating the liquor to extract the liquor from the liquor and / or the sugar alcohol solution.

[0021] (4) adding the sugar solution and / or the sugar alcohol solution to the liquorice, and heating the liquor to extract the liquor from the liquor and / or the sugar alcohol solution.

[0022] (5) combining the first liquor extract and the second liquor extract to obtain a liquor extract (i.e. a total liquor extract or a third liquor extract).

[0023] Further, the liquor is raw liquor or liquor prepared by stir-frying with honey.

[0024] Further, the sugar is selected from one or more of monosaccharides, disaccharides and trisaccharides.

[0025] Further, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol, and xylitol.

[0026] Further, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, galactose, sorbose, rhamnose, ribose, xylose, and deoxyribose.

[0027] Further, the disaccharide is selected from one or more of maltose, sucrose, lactose, and trehalose.

[0028] Further, the monosaccharide is glucose and fructose. Further, the monosaccharide is glucose.

[0029] Further, the disaccharide is sucrose and / or maltose.

[0030] Further, the trisaccharide is raffinose.

[0031] Further, the disaccharide is sucrose.

[0032] Further, in the sugar solution or the sugar alcohol solution, the concentration of the sugar or the sugar alcohol ranges from 0.1 to 30 g / 100 mL.

[0033] Further, the concentration of the sugar or the sugar alcohol ranges from 0.25 to 10 g / 100 mL.

[0034] Further, the concentration of the sugar or the sugar alcohol ranges from 0.25 to 1.25 g / 100 mL.

[0035] Further, the concentration of the sugar or the sugar alcohol ranges from 0.8 to 1.25 g / 100 mL.

[0036] Further, the concentration of the sugar or the sugar alcohol is about 1 g / 100 mL.

[0037] Further, the active ingredient comprises saponins and / or flavonoids.

[0038] Further, the saponins are triterpene saponins or steroidal saponins.

[0039] Further, the saponins are triterpene saponins.

[0040] Further, the saponins are glycyrrhizic acid.

[0041] Further, the flavonoids are dihydroflavonoids.

[0042] Further, the flavonoids are selected from one or more of apiosyl glycyrrhizin, glycyrrhizin, and glycyrrhetic acid.

[0043] Further, in step (2), the soaking is room temperature soaking.

[0044] Further, the step of soaking comprises soaking the liquorice in the sugar solution and / or the sugar alcohol solution at room temperature for 10-120 min.

[0045] Further, the step of soaking comprises soaking the liquorice in the sugar solution and / or the sugar alcohol solution at room temperature for 20-60 min.

[0046] Further, the step of soaking comprises soaking the liquorice in the sugar solution and / or the sugar alcohol solution at room temperature for about 30 min.

[0047] Further, the weight to volume ratio of the liquorice to the sugar solution and / or the sugar alcohol solution is 1:1-1:50.

[0048] Further, the weight to volume ratio of the liquorice to the sugar solution and / or the sugar alcohol solution is 1:1-1:20.

[0049] Further, the weight to volume ratio of the liquorice to the sugar solution and / or the sugar alcohol solution is 1:6-1:7.

[0050] Further, in step (3), the heating extraction is heating reflux extraction.

[0051] Further, the heating time is 20 min to 60 min.

[0052] Further, the heating time is about 30 min.

[0053] According to another aspect of the present application, there is provided a use of the above-mentioned method in the preparation of liquorice extract or liquorice concentrate.

[0054] Further, the liquorice concentrate is obtained by concentrating the liquorice extract (i.e. the total liquorice extract or the third liquorice extract) obtained in step (5) under water bath at 20°C to 100°C.

[0055] Further, in step (5), the concentrating is done under water bath at 60°C to 80°C.

[0056] Further, the concentrating is done under water bath at about 70°C.

[0057] According to another aspect of the present application, there is provided a use of the above-mentioned method in the preparation of pharmaceutical composition containing liquorice.

[0058] According to another aspect of the present application, there is provided a use of the above-mentioned method in the preparation of pharmaceutical preparation, functional food, health food and other products containing liquorice.

[0059] The beneficial effects of the present application are:

[0060] Glycyrrhiza is a commonly used traditional Chinese medicinal material, which has the effects of tonifying the spleen and replenishing qi, clearing heat and detoxifying, expelling phlegm and relieving cough, relieving acute pain, and regulating the effects of various drugs, and is commonly used for weakness of the spleen and stomach, lassitude and debilitation, palpitation and shortness of breath, cough and excessive phlegm, abdominal and limb pain, and abscesses and sores, and can relieve drug toxicity and virulence. The main chemical components of Glycyrrhiza are triterpenoid saponins and flavonoids. The most important triterpenoid saponin is glycyrrhizin, also known as glycyrrhizic acid, and glycyrrhizol, glycyrrhizin and isoglycyrrhizin are flavonoid components in Glycyrrhiza. Our research has found that, compared with the traditional water decoction method, the method of the present application can sufficiently improve the extraction rate of the effective components of Glycyrrhiza. DETAILED DESCRIPTION

[0061] The technical solutions in the embodiments of the present application will be clearly and completely described below in combination with the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0062] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict. The present application will be described in detail below in combination with the embodiments.

[0063] The present application will be further described in detail below in combination with specific embodiments, and these embodiments cannot be understood as limiting the scope of the present application.

[0064] As described in the background section, the effective component composition of the alcohol extract is different from that of the traditional clinical decoction, so the safety of the alcohol extract may be affected to a certain extent, but the extraction efficiency of the traditional water decoction is not high. In order to solve the above problems, the present application provides a method for increasing the extraction of effective components of Glycyrrhiza by using sugar, which comprises the following steps:

[0065] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0066] (2) soaking Glycyrrhiza in the sugar solution and / or the sugar alcohol solution; and

[0067] (3) heating the Glycyrrhiza in the sugar solution and / or the sugar alcohol solution to extract the Glycyrrhiza, and filtering to obtain a Glycyrrhiza extract.

[0068] The sugar solution and / or the sugar alcohol solution can improve the extraction rate of the effective components in Glycyrrhiza, which is mainly related to two factors:

[0069] (1) Whether the liquid can infiltrate the solid decoction piece is related to its surface tension. The smaller the surface tension coefficient (30×10 -3N / m or so, and can almost soak into solid; water has a larger surface tension coefficient, and can only soak into some solid. The surface tension coefficient of sugar water solution is smaller than that of pure water, and the solution can better enter into plant cells to release effective components, thus improving the soaking of licorice decoction pieces and improving the dissolution of effective components;

[0070] (2) Sugar can form hydrogen bonds and other forces with active components in licorice decoction pieces, thus improving the extraction efficiency of licorice extract components;

[0071] Therefore, compared with the water decoction method, the method can sufficiently improve the extraction rate of licorice effective components.

[0072] In actual work, the number of times of extraction of the alcohol and / or sugar alcohol solution can be determined according to actual needs, and can be 1 time or several times, for example, 2-10 times.

[0073] In a preferred embodiment, the licorice is raw licorice or fried licorice.

[0074] According to another aspect of the present application, a method for increasing the extraction of licorice effective components by using sugar is provided, and the method comprises the following steps:

[0075] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0076] (2) soaking licorice in the sugar solution and / or the sugar alcohol solution;

[0077] (3) heating and extracting the licorice in the sugar solution and / or the sugar alcohol solution, and obtaining a first licorice extract liquid through filtration;

[0078] (4) soaking the licorice in an aqueous solution, heating and extracting the licorice in the aqueous solution, and obtaining a second licorice extract liquid through filtration; and

[0079] (5) combining the first licorice extract liquid and the second licorice extract liquid to obtain a licorice extract liquid (i.e., a total licorice extract liquid or a third licorice extract liquid).

[0080] In a preferred embodiment, the licorice is raw licorice or fried licorice.

[0081] According to another aspect of the present application, a method for increasing the extraction of licorice effective components by using sugar is provided, and the method comprises the following steps:

[0082] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0083] (2) soaking licorice in the sugar solution and / or the sugar alcohol solution;

[0084] (3) heating the liquor to extract the liquoram, and filtering to obtain a first liquoram extract;

[0085] (4) adding the liquor to the sugar and / or sugar alcohol solution, heating to extract the liquoram, and obtaining a second liquoram extract; and

[0086] (5) combining the first and second liquoram extracts to obtain a liquoram extract (i.e., a total liquoram extract or a third liquoram extract).

[0087] In a preferred embodiment, the liquoram is raw liquoram or roasted liquoram.

[0088] The term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, a concentration of "about" 0.5 g crude drug / mL means a concentration of from 0.475 g crude drug / mL to 0.525 g crude drug / mL, but also expressly includes a concentration of exactly 0.5 g crude drug / mL.

[0089] In a preferred embodiment, the sugar is selected from one or more of monosaccharides, disaccharides, and trisaccharides.

[0090] In a preferred embodiment, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol, and xylitol. In actual practice, the sugar alcohol can be replaced by other sugar alcohols known in the art.

[0091] 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 practice, the monosaccharide can be replaced by other monosaccharides known in the art.

[0092] In a preferred embodiment, the disaccharide is selected from one or more of maltose, sucrose, lactose, and trehalose. In actual practice, the disaccharide can be replaced by other disaccharides known in the art.

[0093] In a preferred embodiment, the monosaccharide is glucose and fructose.

[0094] In a preferred embodiment, the monosaccharide is glucose.

[0095] In a preferred embodiment, the disaccharide is sucrose and / or maltose.

[0096] In a preferred embodiment, the trisaccharide is raffinose.

[0097] To further improve the extraction rate of the active ingredients in licorice, in a preferred embodiment, the disaccharide is sucrose.

[0098] In a preferred embodiment, the concentration of the sugar or sugar alcohol in the sugar solution or sugar alcohol solution ranges from 0.1 to 30 g / 100 mL.

[0099] In a preferred embodiment, the concentration of the sugar or sugar alcohol ranges from 0.25 to 10 g / 100 mL.

[0100] In a preferred embodiment, the concentration of the sugar or sugar alcohol ranges from 0.25 to 1.25 g / 100 mL.

[0101] To further improve the extraction rate of the active ingredients in licorice, in a preferred embodiment, the concentration of the sugar or sugar alcohol ranges from 0.8 to 1.25 g / 100 mL.

[0102] Further optimizing the above concentration range, in a preferred embodiment, the concentration of the sugar or sugar alcohol is about 1 g / 100 mL.

[0103] The term "about" or "approximately" in reference to a numerical value means ± 5% of the numerical value, but expressly 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 expressly includes a concentration of exactly 1 g / 100 mL.

[0104] In a preferred embodiment, the active ingredients include saponins and / or flavonoids.

[0105] In a preferred embodiment, the saponins are triterpene saponins or steroidal saponins.

[0106] In a preferred embodiment, the saponins are triterpene saponins.

[0107] In practical applications, the saponins include other saponins identified in the art from licorice in addition to triterpene saponins or steroidal saponins.

[0108] In a preferred embodiment, the saponins are glycyrrhizic acid.

[0109] In a preferred embodiment, the flavonoids are dihydroflavonoids. In practical applications, the flavonoids include other flavonoids identified in the art from licorice in addition to dihydroflavonoids.

[0110] In a preferred embodiment, the flavonoid compound is selected from one or more of apiosyl glycyrrhizin, glycyrrhizin, and glycyrrhetic acid.

[0111] In a preferred embodiment, the soaking in step (2) is room temperature soaking.

[0112] In a preferred embodiment, the step of soaking comprises soaking the liquorice in the sugar solution and / or the sugar alcohol solution at room temperature for 10-120 min.

[0113] In a preferred embodiment, the step of soaking comprises soaking the liquorice in the sugar solution and / or the sugar alcohol solution at room temperature for 20-60 min.

[0114] In a preferred embodiment, the step of soaking comprises soaking the liquorice in the sugar solution and / or the sugar alcohol solution at room temperature for about 30 min.

[0115] The term "about" or "approximately" in relation to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, a time of "about" 30 min means a time from 28.5 min to 31.5 min, but also expressly includes a time of exactly 30 min.

[0116] In a preferred embodiment, the weight to volume ratio of the liquorice to the sugar solution and / or the sugar alcohol solution is 1 : 1 to 1 : 50.

[0117] In a preferred embodiment, the weight to volume ratio of the liquorice to the sugar solution and / or the sugar alcohol solution is 1 : 1 to 1 : 20.

[0118] In a preferred embodiment, the weight to volume ratio of the liquorice to the sugar solution and / or the sugar alcohol solution is 1 : 6 to 1 : 7.

[0119] In a preferred embodiment, the heating extraction in step (3) is heating reflux extraction.

[0120] In a preferred embodiment, the time of heating is 20 min to 60 min.

[0121] In a preferred embodiment, the time of heating is about 30 min.

[0122] The term "about" or "approximately" in relation to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, a time of "about" 30 min means a time from 28.5 min to 31.5 min, but also expressly includes a time of exactly 30 min.

[0123] According to another aspect of the present application, there is provided a use of the above-mentioned method in the preparation of licorice extract or licorice concentrate.

[0124] In a preferred embodiment, the licorice concentrate is the licorice extract (i.e. the total licorice extract or the third licorice extract) obtained in step (5) which is concentrated at 20°C to 100°C water bath.

[0125] In a preferred embodiment, the concentration in step (5) is done at 60°C to 80°C water bath.

[0126] In a preferred embodiment, the concentration is done at about 70°C water bath.

[0127] The term "about" or "approximately" in relation to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, a temperature of "about" 70°C means a range of 66.5°C to 73.5°C, but also expressly includes a temperature of exactly 70°C.

[0128] According to another aspect of the present application, there is provided a use of the above-mentioned method in the preparation of pharmaceutical composition containing licorice.

[0129] According to another aspect of the present application, there is provided a use of the above-mentioned method in the preparation of pharmaceutical preparation, functional food, health food and other products containing licorice.

[0130] The functional food is a food which can convincingly prove to have benefits to one or more functions of the body, has sufficient nutritional effect to improve health condition or can reduce the risk of disease.

[0131] The health food refers to a food which claims to have specific health functions or is intended for supplementing vitamins and minerals, i.e. is suitable for specific population, has the function of regulating the body, is not intended for treating diseases and does not cause any acute, subacute or chronic harm to the human body.

[0132] The other products refer to all products other than pharmaceutical preparation, functional food and health food which are included in the prior art, including liquid and solid forms, etc.

[0133] Example 1: Experiment of extracting raw licorice decoction pieces with different sugar solutions

[0134] 1.1 Preparation method of different sugar-licorice extract

[0135] The same batch of Radix Glycyrrhizae decoction pieces (Chifeng, Inner Mongolia Autonomous Region) were taken 5 (R, R + 1% glucose, R + 1% fructose, R + 1% sucrose, R + 1% trehalose), 50g each. Respectively, 7 times the amount of water (350mL water), 1% glucose aqueous solution (3.5g glucose + 350mL water), 1% fructose aqueous solution (3.5g fructose + 350mL water), 1% sucrose aqueous solution (3.5g sucrose + 350mL water), 1% trehalose aqueous solution (3.5g trehalose + 350mL water), soaked for 30min, refluxed for 30min, hot filtered with gauze, the residue was added with 6 times the amount of water (300mL water) and refluxed for 20min, hot filtered, and the two filtrates were combined to obtain different sugar-glycyrrhiza extract.

[0136] 1.2.1 Instrument and reagent

[0137] 1.2.1 Instrument and reagent

[0138] Agilent 1260 type high performance liquid chromatograph with DAD detector (American Agilent Company); DZKW-4 type electronic constant temperature water bath (Beijing Zhongxing Weiyeye Instrument Co., Ltd.); DK-98-IIA electric heating constant temperature water bath (Tianjin Test Instrument Co., Ltd.); ME204 / 02 type electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd., one hundred thousandth balance); KQ-250DE type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).

[0139] Reference CHB180109, CHB201102 and CHB201103 were purchased from Chengdu Keluoma Biological Technology Co., Ltd., with purity > 98%; Z30A6B1 was purchased from Shanghai Yuanye Biological Technology Co., Ltd., with purity > 98%; acetonitrile (chromatographic pure, Fisher Company), methanol (chromatographic pure, Fisher Company), phosphoric acid was analytical pure, purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd. Sucrose, glucose and fructose (Jikelong (Beijing) Biotechnology Co., Ltd.); trehalose (Hebei Bawei Biological Technology Co., Ltd.); all are pharmaceutical grade. Water is Wahaha pure water.

[0140] 1.2.2 Preparation of test solution

[0141] Take 0.5mL of different sugar-glycyrrhiza extract, dilute 20 times with methanol (9.5mL), shake well, and pass through a 0.22μm filter membrane to obtain the test solution.

[0142] 1.2.3 Preparation of reference solution

[0143] Stock solution: The stock solution of 1 mg / mL was prepared by accurately weighing the appropriate amount of the reference substances of glycosyl-liquiritin, liquiritin, glycyrrhizin and glycyrrhizic acid, and dissolving them in methanol. The stock solution was stored in a refrigerator at -20°C.

[0144] Linear working solution: The stock solution of glycosyl-liquiritin, liquiritin and glycyrrhizin was diluted with methanol to prepare a series of linear mixed standard working solutions: 2.5, 5, 10, 20, 40, 50, 100 μg / mL; the stock solution of glycyrrhizic acid was diluted with methanol to prepare a series of linear mixed standard working solutions: 10, 20, 40, 80, 160, 200, 400 μg / mL.

[0145] 1.2.4 Content determination method

[0146] Chromatographic column: Thermo Acclaim 120 C18 (250 x 4.6 mm, 5 μm); mobile phase: acetonitrile (A) and 0.05% phosphoric acid solution (B), column temperature: 35°C, detection wavelength: 237 nm, injection volume: 10 μL, flow rate: 1 ml / min, gradient elution: 0-8 min, 19% A; 8-35 min, 19%-50% A; 35-36 min, 50%-100% A; 36-40 min, 100%-19% A; 40-50 min, 19% A.

[0147] 1.3 Experimental results

[0148] The content determination results of the effective ingredients in different sugar-liquorice extract solutions are shown in Table 1.

[0149] Table 1 Content determination results of effective ingredients in different sugar-liquorice extract solutions

[0150]

[0151] *P <0.05 VS water

[0152] As can be seen from Table 1, sucrose, glucose and fructose (1 g / 100 mL concentration) can all increase the content of effective ingredients in liquorice to different extents, and the effect of sucrose is the most obvious.

[0153] Example Two: Experiment of different sugar solutions extracting raw liquorice decoction pieces

[0154] 2.1 Preparation method of different sugar-liquorice extract solutions

[0155] The same batch of Radix Glycyrrhizae decoction pieces (Inner Mongolia Autonomous Region) were taken 4 times (R, R + 1% glucose, R + 1% sucrose, R + 1% maltose), 40 g each. Respectively, 7 times the amount of water (280 mL water), 1% glucose aqueous solution (2.8 g glucose + 280 mL water), 1% sucrose aqueous solution (2.8 g sucrose + 280 mL water), 1% maltose aqueous solution (2.8 g maltose + 280 mL water) was added, soaked for 30 min, refluxed for 30 min, and then filtered with gauze while hot to obtain different sugar-glycyrrhiza extract.

[0156] 2.2.2 Preparation of test solution

[0157] 2.2.1 Instruments and reagents

[0158] Agilent 1260 type high performance liquid chromatograph with DAD detector (American Agilent Company); DZKW-4 type electronic constant temperature water bath (Beijing Zhongxing Weiyeye Instrument Co., Ltd.); DK-98-IIA electric heating constant temperature water bath (Tianjin Test Instrument Co., Ltd.); ME204 / 02 type electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd., one hundred thousandth balance); KQ-250DE type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).

[0159] Reference substances CHB180109, CHB201102 and CHB201103 were purchased from Chengdu Keluoma Biological Technology Co., Ltd., with a purity of >98%; Z30A6B1 was purchased from Shanghai Yuanye Biological Technology Co., Ltd., with a purity of >98%; acetonitrile (chromatographic pure, Fisher Company), methanol (chromatographic pure, Fisher Company), phosphoric acid were analytical pure, purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd. Water was Wahaha pure water.

[0160] 2.2.2 Preparation of test solution

[0161] Take 0.5 mL of different sugar-glycyrrhiza extract, dilute 20 times with methanol (9.5 mL), shake well, and pass through a 0.22 μm filter membrane to obtain the test solution.

[0162] 2.2.3 Preparation of reference solution

[0163] Stock solution: Take appropriate amounts of CHB180109, CHB201102, CHB201103 and Z30A6B1 reference substances, accurately weigh and prepare 1 mg / mL stock solution with methanol. Store in a -20°C refrigerator.

[0164] Linear working solution: take glycyrrhizic acid stock solution, add methanol to dilute into a series of linear mixed standard working solutions: 10, 20, 40, 80, 160, 200, 400 μg / mL.

[0165] 2.2.4 Content determination method

[0166] Column: Thermo Acclaim 120 C18 (250 x 4.6 mm, 5 μm); mobile phase: acetonitrile (A), 0.05% phosphoric acid solution (B) as mobile phase, column temperature: 35°C, detection wavelength: 237 nm, injection volume: 10 μL, flow rate: 1 mL / min, gradient elution: 0-8 min, 19% A; 8-35 min, 19%-50% A; 35-36 min, 50%-100% A; 36-40 min, 100%-19% A; 40-50 min, 19% A.

[0167] The content determination results of effective components in different sugar-glycyrrhiza extract solutions are shown in Table 2.

[0168] Table 2 Content determination results of effective components in different sugar-glycyrrhiza extract solutions

[0169]

[0170] *P < 0.05 VS water

[0171] As can be seen from Table 2, glucose, sucrose and maltose (1 g / 100 mL concentration) can all increase the content of effective components in glycyrrhiza to different extents, and the effects of sucrose and maltose are the most obvious.

[0172] Example Three: Experiment of extracting raw glycyrrhiza slices with different sugar solutions

[0173] 3.1 Preparation method of different sugar-glycyrrhiza extract solutions

[0174] The same batch of raw licorice pieces (produced in Xinjiang) was taken 4 times, numbered (R, R+1% sucrose, R+1% maltose, R+1% lactose), each 40 g. In turn, 7 times the amount of water (280 mL water), 1% sucrose aqueous solution (2.8 g sucrose + 280 mL water), 1% maltose aqueous solution (2.8 g maltose + 280 mL water), 1% lactose aqueous solution (2.8 g lactose + 280 mL water) were added to soak for 30 min, reflux for 30 min, filter with gauze while hot, and then add 6 times the amount of water (240 mL water), 1% sucrose aqueous solution (2.4 g sucrose + 240 mL water), 1% maltose aqueous solution (2.4 g maltose + 240 mL water), 1% lactose aqueous solution (2.4 g lactose + 240 mL water) to the residue, respectively, and reflux for 20 min, filter while hot, and combine the two filtrates to obtain different sugar-licorice extract solutions.

[0175] 3.2 Apigenin, Glycyrrhizin, Glycyrrhizic Acid and Glycyrrhizic Acid Content Determination (HPLC Pharmacopoeia Method)

[0176] 3.2.1 Instruments and reagents

[0177] Agilent 1260 type high performance liquid chromatograph with DAD detector (American Agilent Company); DZKW-4 type electronic constant temperature water bath (Beijing Zhongxing Weiyeye Instrument Co., Ltd.); DK-98-IIA electric heating constant temperature water bath (Tianjin Test Instrument Co., Ltd.); ME204 / 02 type electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd., one hundred thousandth balance); KQ-250DE type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).

[0178] The reference substances apigenin (CHB180109), glycyrrhizin (CHB201102) and glycyrrhizin (CHB201103) were purchased from Chengdu Keluoma Biological Technology Co., Ltd., with a purity of >98%; glycyrrhizic acid (Z30A6B1) was purchased from Shanghai Yuanye Biological Technology Co., Ltd., with a purity of >98%; acetonitrile (chromatographic pure, Fisher Company), methanol (chromatographic pure, Fisher Company), phosphoric acid were analytical pure, purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd. Maltose, lactose (Jikelong (Beijing) Biotechnology Co., Ltd.); pharmaceutical grade. Water is Wahaha pure water.

[0179] 3.2.2 Preparation of test solution

[0180] Take 0.5 mL of different sugar-licorice extract solution, dilute 20 times with methanol (9.5 mL), shake well, and pass through a 0.22 μm filter membrane to obtain the test solution.

[0181] 3.2.3 Preparation of reference solution

[0182] Stock solution: Take appropriate amount of apigenin- 20-β-D-glucopyranosyl- 1-β-D-glucopyranosyl- 1-β-D-glucopyranoside, glycyrrhizin, glycyrrhizinic acid and the same amount of methanol to prepare a stock solution of 1 mg / mL. Stored in a refrigerator at -20°C.

[0183] Linear working solution: Take appropriate amount of apigenin- 20-β-D-glucopyranosyl- 1-β-D-glucopyranosyl- 1-β-D-glucopyranoside, glycyrrhizin, glycyrrhizin stock solution, dilute with methanol to prepare a series of linear mixed standard working solutions: 2.5, 5, 10, 20, 40, 50, 100 μg / mL; Take appropriate amount of glycyrrhizin stock solution, dilute with methanol to prepare a series of linear mixed standard working solutions: 10, 20, 40, 80, 160, 200, 400 μg / mL.

[0184] 3.2.4 Content determination method

[0185] Chromatographic column: Thermo Acclaim 120 C18 (250 x 4.6 mm, 5 μm); mobile phase: acetonitrile (A), 0.05% phosphoric acid solution (B) as mobile phase, column temperature: 35°C, detection wavelength: 237 nm, injection volume: 10 μL, flow rate: 1 ml / min, gradient elution: 0-8 min, 19% A; 8-35 min, 19%-50% A; 35-36 min, 50%-100% A; 36-40 min, 100%-19% A; 40-50 min, 19% A.

[0186] 3.3 Experimental results

[0187] The results of determination of the content of effective components in different sugar-glycyrrhiza extract solutions are shown in Table 3.

[0188] Table 3 Determination results of effective component content in different sugar-glycyrrhiza extract solutions

[0189]

[0190]

[0191] *P <0.05 VS water

[0192] As can be seen from Table 3, sucrose and maltose (1 g / 100 mL concentration) can increase the content of effective components in glycyrrhiza to varying degrees, of which sucrose has the most obvious effect.

[0193] According to the results of Table 1 and Table 2, sucrose can significantly increase the extraction rate of main active components in glycyrrhiza, including flavonoids (apigenin- 20-β-D-glucopyranosyl- 1-β-D-glucopyranosyl- 1-β-D-glucopyranoside, glycyrrhizin, glycyrrhizin) and saponin components (glycyrrhizinic acid).

[0194] Example Four: Extraction of raw glycyrrhiza decoction pieces with different concentrations of sucrose solution

[0195] 4.1 Preparation of different concentrations of sucrose-glycyrrhiza extract solutions

[0196] The same batch of licorice decoction pieces (Chifeng, Inner Mongolia Autonomous Region) were taken 8 times (R, R + 0.25% sucrose, R + 0.5% sucrose, R + 1% sucrose, R + 1.25% sucrose, R + 1.5% sucrose, R + 2% sucrose, R + 10% sucrose), 50g each. Respectively, 7 times the amount of water (350mL water), 0.25% sucrose solution (0.9g sucrose + 350mL water), 0.5% sucrose solution (1.8g sucrose + 350mL water), 1% sucrose solution (3.5g sucrose + 350mL water), 1.25% sucrose solution (4.4g sucrose + 350mL water), 1.5% sucrose solution (5.3g sucrose + 350mL water), 2% sucrose solution (7g sucrose + 350mL water), 10% sucrose solution (35g sucrose + 350mL water) were added, soaked for 30min, refluxed for 30min, filtered with gauze while hot, the residue was added with 6 times the amount of water (300mL water) respectively, refluxed for 20min, filtered while hot, and the two filtrates were combined to obtain licorice extract with different concentrations of sucrose.

[0197] 4.2 Content determination of chikusetsu licorice glycoside, licorice glycoside, glycyrrhizin and glycyrrhizic acid (HPLC pharmacopoeia method)

[0198] 4.2.1 Instruments and reagents

[0199] Agilent 1260 type high performance liquid chromatograph with DAD detector (Agilent, USA); DZKW-4 type electronic constant temperature water bath (Beijing Zhongxing Weiyeye Instrument Co., Ltd.); DK-98-IIA electric heating constant temperature water bath (Tianjin Test Instrument Co., Ltd.); ME204 / 02 type electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd., 1 / 10000 balance); KQ-250DE type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).

[0200] Reference substances chikusetsu licorice glycoside (CHB180109), licorice glycoside (CHB201102) and glycyrrhizin (CHB201103) were purchased from Chengdu Keloma Biological Technology Co., Ltd., with purity >98%; glycyrrhizic acid (Z30A6B1) was purchased from Shanghai Yuanye Biological Technology Co., Ltd., with purity >98%; acetonitrile (chromatographic pure, Fisher Company), methanol (chromatographic pure, Fisher Company), phosphoric acid were analytical pure, purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd. Water was Wahaha pure water.

[0201] 4.2.2 Preparation of test solution

[0202] Take 0.5mL of licorice extract, dilute 20 times with methanol (9.5mL), shake well, and filter through a 0.22μm filter membrane to obtain the test solution.

[0203] 4.2.3 Preparation of control solution

[0204] Stock solution: Take appropriate amount of control samples of glycyrrhizin, glycyrrhizin, glycyurol and glycyrrhizic acid, accurately weigh and prepare 1 mg / mL stock solution with methanol. Store in -20°C refrigerator.

[0205] Linear working solution: Take appropriate amount of stock solution of glycyrrhizin, glycyrrhizin and glycyurol, dilute with methanol to prepare a series of linear mixed standard working solutions: 2.5, 5, 10, 20, 40, 50, 100 μg / mL; Take appropriate amount of glycyrrhizic acid stock solution, dilute with methanol to prepare a series of linear mixed standard working solutions: 10, 20, 40, 80, 160, 200, 400 μg / mL.

[0206] 4.2.4 Content determination method

[0207] Chromatographic column: Thermo Acclaim 120 C18 (250 x 4.6 mm, 5 μm); mobile phase: acetonitrile (A), 0.05% phosphoric acid solution (B) as mobile phase, column temperature: 35°C, detection wavelength: 237 nm, injection volume: 10 μL, flow rate: 1 mL / min, gradient elution: 0-8 min, 19% A; 8-35 min, 19%-50% A; 35-36 min, 50%-100% A; 36-40 min, 100%-19% A; 40-50 min, 19% A.

[0208] 4.3 Experimental results

[0209] The content determination results of effective components in glycyrrhiza extract obtained by extracting raw licorice slices with different concentrations of sucrose solution are shown in Table 4.

[0210] Table 4 Content determination results of effective components in glycyrrhiza extract obtained by different concentrations of sucrose

[0211]

[0212] *P <0.05 VS water

[0213] As can be seen from Table 4, sucrose concentrations in the range of 0.25 g / 100 mL to 2 g / 100 mL can increase the content of effective components in licorice to varying degrees, and the effect is most obvious when the concentration of sucrose is in the range of 1 g / 100 mL to 1.25 g / 100 mL.

[0214] The above is only the embodiment of the present application, and the common knowledge of the specific structure and characteristics in the scheme is not described in detail. It should be pointed out that for those skilled in the art, without departing from the present application, a number of modifications and improvements can be made, which should be considered as the protection scope of the present application, which will not affect the effect and practicality of the patent. The protection scope of the present application should be subject to the content of its claims, and the specific implementation mode in the specification can be used to explain the content of the claims.

[0215] The above describes the embodiments of the present application in detail, and the principles and implementation modes of the present application are described by applying specific examples. The above embodiment is only used to help understand the method and core idea of the present application. Meanwhile, the changes or deformations made by those skilled in the art on the basis of the specific implementation mode and application range of the present application all belong to the protection scope of the present application. In summary, the content of the present application should not be understood as a limitation of the present application.

Claims

1. A method for increasing the extraction of licorice active ingredients by using sugar, characterized by, The method comprises the following steps: (1) preparing a sugar solution with a concentration of about 1 g / 100 mL; (2) soaking liquorice in the sugar solution; and (3) heating extraction of the liquorice in the sugar solution once, and obtaining liquorice extract liquid through filtration; wherein the sugar is one or more of glucose, sucrose and maltose; wherein the effective components are glycyrrhizinate, glycyrrhizin and glycyrrhizic acid.

2. The method of claim 1, wherein, The liquorice is raw liquorice or roasted liquorice.

3. A method for increasing the extraction of licorice active ingredients using sugar, characterized by, The method comprises the following steps: (1) preparing a sugar solution with a concentration of about 1 g / 100 mL; (2) soaking liquorice in the sugar solution; (3) heating extraction of the liquorice in the sugar solution, and obtaining first liquorice extract liquid through filtration; (4) soaking the liquorice in an aqueous solution, heating extraction of the liquorice in the aqueous solution, and obtaining second liquorice extract liquid through filtration; and (5) combining the first liquorice extract liquid and the second liquorice extract liquid to obtain liquorice extract liquid; wherein the sugar is sucrose; wherein the effective components are glycyrrhizinate, glycyrrhizin and glycyrrhizic acid.

4. The method of claim 3, wherein, The liquorice is raw liquorice or roasted liquorice.

5. A method for increasing the extraction of licorice active ingredients by using sugar, characterized by, The method comprises the following steps: (1) preparing a sugar solution with a concentration of about 1 g / 100 mL; (2) soaking liquorice in the sugar solution; (3) heating extraction of the liquorice in the sugar solution, and obtaining first liquorice extract liquid through filtration; (4) adding the sugar solution to the liquorice, heating extraction of the liquorice, and obtaining second liquorice extract liquid; and (5) combining the first liquorice extract liquid and the second liquorice extract liquid to obtain liquorice extract liquid; wherein the sugar is sucrose; wherein the effective components are glycyrrhizinate, glycyrrhizin, glycyrrhizin and glycyrrhizic acid.

6. The method of claim 5, wherein, The liquorice is raw liquorice or roasted liquorice.

7. The method according to any one of claims 1 to 6, characterized in that, In step (2), the soaking is room temperature soaking.

8. The method of claim 7, wherein, The step of soaking comprises soaking the liquorice in the sugar solution at room temperature for 10-120 min.

9. The method of claim 7, wherein, The step of soaking comprises soaking the liquorice in the sugar solution at room temperature for 20-60 min.

10. The method of claim 7, wherein, The step of soaking comprises soaking the liquorice in the sugar solution at room temperature for about 30 min.

11. The method according to any one of claims 1 to 6, characterized in that, The weight-to-volume ratio of the liquorice to the sugar solution is 1:1-1:

50.

12. The method according to any one of claims 1 to 6, characterized in that, The weight-to-volume ratio of the liquorice to the sugar solution is 1:1-1:

20.

13. The method according to any one of claims 1 to 6, characterized in that, The weight-to-volume ratio of the liquorice to the sugar solution is 1:6-1:

7.

14. The method of any one of claims 1 to 6, wherein, In step (3), the heating extraction is heating reflux extraction.

15. The method of claim 14, wherein, The heating time is 20 min to 60 min.

16. The method of claim 14, wherein, The heating time is about 30 min.

17. Use of the method according to any one of claims 1 to 16 in the preparation of liquorice extract liquid or liquorice concentrated liquid.

18. Use of the method according to any one of claims 1 to 16 in the preparation of a pharmaceutical composition containing liquorice.

19. Use of the method according to any one of claims 1 to 16 in the preparation of a pharmaceutical preparation or functional food or health food containing liquorice.

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