Method for increasing the extraction of saccharide and glycoside components and use thereof

Abstract

The application discloses a method for increasing the extraction of saccharide components and / or glycoside components, which comprises the following steps: (1) preparing a sugar solution and / or a sugar alcohol solution; (2) soaking raw materials in the sugar solution and / or the sugar alcohol solution; and (3) heating the raw materials in the sugar solution and / or the sugar alcohol solution for 1-3 times, and obtaining an extract by filtration. The above method can significantly increase the extraction rate of saccharide components and / or glycoside components in the raw materials, thereby helping to improve the utilization rate and curative effect of the raw materials.

Description

Technical Field

[0001] This invention relates to the field of chemical component extraction, specifically to a method for increasing the leaching of sugars and glycosides and its application. Background Technology

[0002] Currently, how to extract active / effective components, such as sugars and / or glycosides, from raw materials in a green and efficient manner, while ensuring the safety and efficacy of the extracts, is an important aspect of research in the extraction of raw materials such as traditional Chinese medicine and herbal medicines. 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 resulting extracts may increase. The newly released regulations for the registration and application of new traditional Chinese medicine drugs in 2020 stipulate that if the declared indications are consistent with the traditional usage, clinical trials can be appropriately reduced or waived. For example, the "Regulations on Simplified Registration and Approval Management of Compound Preparations of Ancient Classical Formulas of Traditional Chinese Medicine" issued in 2018 clarified the scope of application for exemption from clinical trials in the development of compound preparations of ancient classic formulas, including preparation processes consistent with ancient medical records. The preparation process of most traditional Chinese medicines uses water decoction, and clinical experience and modern research show that this method is better at ensuring the safety of the extracts than organic solvent extraction. The main reason for the differences between the two extraction methods is that traditional Chinese medicine contains a wide variety of active ingredients with diverse polarities. Ethanol extraction increases the extraction rate of weakly and moderately polar components, resulting in a different composition of active ingredients in the ethanol extract compared to the decoctions used in traditional clinical practice. This may have some impact on the safety of the extract. Furthermore, the most commonly used traditional Chinese medicine extraction method, water decoction, suffers from low extraction rates and incomplete extraction. While increasing the amount of water helps improve the extraction rate of active ingredients, it increases the 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 while adhering to 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 sugar components and / or glycosides using a low-concentration aqueous solution of sugar compounds. By using a certain concentration of sugar water and / or sugar alcohol water instead of water as the extraction solvent, the extraction rate is significantly improved compared to traditional decoction methods, thereby increasing the utilization rate and efficacy of raw materials such as traditional Chinese medicine. The sugar used in this study is a commonly used excipient or edible sugar in traditional Chinese medicine, thus possessing characteristics such as safety and low price, and will not affect the safety of the extract.

[0005] Specifically, according to one aspect of the present invention, a method for increasing the leaching of carbohydrate components and / or glycoside components is provided, the method comprising the following steps:

[0006] (1) Prepare sugar solutions and / or sugar alcohol solutions;

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

[0008] (3) The raw material is extracted by heating in the sugar solution and / or the sugar alcohol solution 1-3 times, and the raw material extract is obtained by filtration.

[0009] Further, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-40 g / 100 mL. Further, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-10 g / 100 mL or 20-40 g / 100 mL. Further, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-5 g / 100 mL or 20-30 g / 100 mL. Further, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-2.5 g / 100 mL. Further, the concentration of the sugar solution and / or the sugar alcohol solution is 1-2 g / 100 mL. Furthermore, the concentration of the sugar solution and / or the sugar alcohol solution is about 0.5 g / 100 mL or about 1 g / 100 mL or about 1.5 g / 100 mL or about 2 g / 100 mL or about 20 g / 100 mL or about 30 g / 100 mL or about 40 g / 100 mL.

[0010] Further, the sugar is selected from one or more of monosaccharides, disaccharides, and trisaccharides. Further, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol, and xylitol. Further, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, sorbitol, rhamnose, ribose, xylose, and deoxyribose. Further, the disaccharide is selected from one or more of maltose, sucrose, lactose, trehalose, melibiose, gentiobiose, and kosperidose. Further, the monosaccharide is glucose and / or fructose. Further, the trisaccharide is raffinose. Further, the disaccharide is sucrose and / or maltose.

[0011] Furthermore, the sugar component is an oligosaccharide component and / or a polysaccharide component.

[0012] Furthermore, the polysaccharide component is selected from one or more of the following: astragalus polysaccharide, licorice polysaccharide, ginseng polysaccharide, and wolfberry polysaccharide.

[0013] Furthermore, the glycoside component is selected from one or more of the following: flavonoid glycosides, saponins, diterpenoid glycosides, quinone glycosides, and phenolic glycosides.

[0014] Furthermore, the flavonoid glycosides are flavonoid glycosides, isoflavone glycosides, dihydroflavonoid glycosides, dihydroflavonol glycosides, dihydroisoflavoneol glycosides, chalcone glycosides, dihydrochalcone glycosides, orange glycosides, anthocyanin glycosides, biflavonoid glycosides, and flavanol glycosides.

[0015] Furthermore, the saponin compound is a tetracyclic triterpenoid glycoside, a pentacyclic triterpenoid glycoside, and / or a steroidal glycoside.

[0016] Furthermore, the diterpenoid glycoside is selected from one or more of the following: tetracyclic diterpenoid glycosides, tricyclic diterpenoid glycosides, chain diterpenoid glycosides, monocyclic diterpenoid glycosides, and bicyclic diterpenoid glycosides.

[0017] Furthermore, the quinone glycoside is a benzoquinone glycoside, naphthoquinone glycoside, phenanthrenequinone glycoside, and / or anthraquinone glycoside.

[0018] Furthermore, the phenolic glycoside compound is a phenolic glycoside compound having a C6-C3 core structure and / or a C6-C1 core structure.

[0019] Furthermore, the steroidal glycoside compound is a spirostanol-type saponin compound, a furostanol-type saponin compound, and / or an isostanol-type saponin compound.

[0020] Furthermore, this isosspiranol-type saponin compound is a diosgenin compound.

[0021] Furthermore, this furostanol-type saponin compound is an anemarrhena saponin compound.

[0022] Furthermore, the tetracyclic triterpenoid glycoside is a lanolinane-type tetracyclic triterpenoid saponin and / or a dammarane-type tetracyclic triterpenoid saponin.

[0023] Furthermore, the pentacyclic triterpenoid glycosides are oleanane-type pentacyclic triterpenoid saponins, ursane-type pentacyclic triterpenoid saponins, and / or lupinane-type pentacyclic triterpenoid saponins.

[0024] Furthermore, the anthraquinone glycoside is a monoanthraquinone compound and / or a dianthraquinone compound.

[0025] Furthermore, the isoflavone glycoside compound is verrucoside and / or gentianin.

[0026] Further, the dihydroflavonoid glycoside compound is apigenin and / or glycyrrhizin. Further, the chalcone glycoside compound is hydroxysafflower yellow A. Further, the furostanol-type steroidal saponin compound is anemarrhenasaponin BII. Further, the lanolinane-type tetracyclic triterpenoid saponin compound is astragaloside A. Further, the dammarane-type tetracyclic triterpenoid saponin compound is ginsenoside Rg1. Further, the oleanane-type pentacyclic triterpenoid saponin compound is glycyrrhizic acid. Further, the tetracyclic diterpenoid glycoside compound is steviol glycoside and / or lebodiin. Further, the C6-C3 core-structured phenolic acid glycoside compound is hydroxysafflower yellow A. Further, the bianthraquinone nucleoside compound is sennoside A and / or sennoside B. Further, the monoanthraquinone nucleoside compound is aloin.

[0027] Furthermore, the raw material is a raw material containing sugar components and / or glycoside components.

[0028] Furthermore, the raw material is one or more traditional Chinese medicines, herbal medicines, and / or marine organisms containing sugar components and / or glycoside components.

[0029] Furthermore, the Chinese medicine and / or herbal medicine is selected from one or more of the following: ginseng, Panax notoginseng, licorice, wolfberry, stevia, astragalus, senna leaf, albizia bark, pokeweed, bupleurum, Pulsatilla chinensis, Toona sinensis, Dendrobium nobile, Achyranthes bidentata, Clematis chinensis, American ginseng, Fritillaria cirrhosa, Aesculus hippocastanum, Acanthopanax senticosus, Eleutherococcus senticosus, Gynostemma pentaphyllum, Melia azedarach, Centella asiatica, Gynostemma pentaphyllum, Ilex chinensis, Dipsacus asper, Akebia quinata, Panax notoginseng, Clematis armandii, Ziziphus jujuba leaf, hemp seed, Polygala tenuifolia, Perilla frutescens, Eclipta prostrata, Platycodon grandiflorus, Ophiopogon japonicus, Allium macrostemon, Polygonatum sibiricum, Polygonatum odoratum, Anemarrhena asphodeloides, Asparagus cochinchinensis, Ophiopogon japonicus, Paris polyphylla, Smilax glabra, Dioscorea opposita, Tribulus terrestris, Solanum nigrum, pomegranate, apricot kernel, sweet potato, Dioscorea opposita, Asparagus cochinchinensis, asparagus, Tribulus terrestris, Taro, and dragon's blood.

[0030] Furthermore, in step (2), the soaking is a room temperature soaking.

[0031] Furthermore, the soaking step includes immersing the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for 10-120 minutes.

[0032] Furthermore, the soaking step includes immersing the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for 20-60 minutes.

[0033] Furthermore, the soaking step includes immersing the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for about 30 minutes.

[0034] 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.

[0035] 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.

[0036] Furthermore, the weight-to-volume ratio of the raw material to the sugar solution and / or the sugar alcohol solution is 1:6 to 1:12.

[0037] Furthermore, in step (3), the heating extraction is a heating reflux extraction.

[0038] Furthermore, the heating time is 20 to 60 minutes.

[0039] Furthermore, the heating time is 20 to 30 minutes.

[0040] Furthermore, the heating time is approximately 20 minutes or approximately 30 minutes.

[0041] According to one aspect of the present invention, there is provided the use of the method described above in the preparation of a raw material extract or a raw material concentrate, wherein the raw material is a traditional Chinese medicine, a herbal medicine, and / or a marine organism.

[0042] Furthermore, the raw material concentrate is obtained by concentrating the raw material extract obtained in step (3) or the total raw material extract obtained in step (5) in a water bath at 20°C to 100°C.

[0043] Furthermore, the concentration was completed in a water bath at 60°C to 80°C.

[0044] Furthermore, the concentration was completed in a water bath at approximately 70°C.

[0045] According to one aspect of the present invention, there is provided the use of the method described above in the preparation of a pharmaceutical composition containing a raw material, wherein the raw material is a traditional Chinese medicine, a herbal medicine, and / or a marine organism.

[0046] According to one aspect of the present invention, there is provided the use of the method described above in the preparation of a pharmaceutical preparation or functional food or health food containing a raw material, wherein the raw material is a traditional Chinese medicine, a herbal medicine and / or a marine organism.

[0047] The beneficial effects of this invention are:

[0048] Our research has found that, compared with traditional decoction methods, the method of the present invention can significantly improve the extraction rate of sugar components and / or glycoside components in raw materials such as traditional Chinese medicine or herbal medicine. Detailed Implementation

[0049] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0050] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other. The present invention will now be described in detail with reference to the embodiments.

[0051] The present invention will be further described in detail below with reference to specific embodiments, which should not be construed as limiting the scope of protection claimed by the present invention.

[0052] As described in the background section, the composition of 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 decoction is not high. To address these issues, this invention provides a method for increasing the extraction of sugar components and / or glycosides using sugar, the method comprising the following steps:

[0053] (1) Prepare sugar solutions and / or sugar alcohol solutions;

[0054] (2) Soaking the raw materials in the sugar solution and / or the sugar alcohol solution; and

[0055] (3) The raw material is extracted by heating in the sugar solution and / or the sugar alcohol solution, and the raw material extract is obtained by filtration.

[0056] The ability of sugar solutions and / or sugar alcohol solutions to improve the extraction rate of sugar components and / or glycosides in raw materials such as traditional Chinese medicine or herbal medicines is mainly related to two factors:

[0057] (1) Whether a liquid can wet a solid medicinal slice depends on its surface tension. A liquid with a low surface tension coefficient (30×10⁻⁶) will wet a solid medicinal slice. -3 The surface tension coefficient of sugar solution is approximately N / m, which can wet almost all solids; water has a higher surface tension coefficient, so it can only wet certain solids. The surface tension coefficient of sugar solution is lower than that of pure water, allowing the solution to penetrate plant cells better and release active ingredients. Therefore, it can improve the wetting of raw materials such as Chinese medicinal herbs and increase the dissolution of sugar and / or glycoside components.

[0058] (2) The sugar in the extraction solvent and the sugar and / or glycoside components in the extract both contain a large number of hydroxyl groups. Therefore, sugar can form intermolecular forces such as intermolecular hydrogen bonds with the sugar and / or glycoside components in the raw materials, such as Chinese herbal medicine slices, thus improving the extraction efficiency of sugar and / or glycoside components in the raw materials, such as Chinese herbal medicine extracts.

[0059] Therefore, compared with decoction, this method can significantly improve the extraction rate of carbohydrate and / or glycoside components in raw materials such as traditional Chinese medicine or herbal medicine.

[0060] In actual work, the number of extractions of alcohol and / or sugar alcohol solutions can be determined according to actual needs. It can be once or several times, such as 2-10 times.

[0061] In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-40 g / 100 mL.

[0062] In this invention, when concentration, proportion, equivalent, number of times or other parameters are expressed as a range, preferred range, or a range defined by a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pair of any upper or preferred value and any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range "0.1-40" is disclosed, unless otherwise stated, the range is intended to include its endpoints and all point values ​​within that range, such as 0.1, 0.2, 0.3, 0.4, 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, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 6. 0, 7.0, 8.0, 9.0, 10.0, 11.0, 12.0, 13.0, 14.0, 15.0, 16.0, 17.0, 18.0, 19.0, 20.0, 21.0, 22.0, 23.0, 24.0, 25.0, 26.0, 27.0, 28.0, 29.0, 30.0, 31.0, 32. 0, 33.0, 34.0, 35.0, 36.0, 37.0, 38.0, 39.0, 40.0, 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., and are not limited to the values ​​listed above.

[0063] In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-10 g / 100 mL or 20-40 g / 100 mL.

[0064] In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-5 g / 100 mL or 20-30 g / 100 mL.

[0065] In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is 0.1-2.5 g / 100 mL.

[0066] In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is 0.5-2 g / 100 mL.

[0067] In a preferred embodiment, the concentration of the sugar solution and / or the sugar alcohol solution is about 0.5 g / 100 mL or about 1 g / 100 mL or about 1.5 g / 100 mL or about 2 g / 100 mL or about 20 g / 100 mL or about 30 g / 100 mL or about 40 g / 100 mL.

[0068] The terms “about” or “approximately” for numerical values ​​mean ±5% of the value, but explicitly include the exact value. For example, a concentration of "about" 0.5 g / 100 mL refers to concentrations from 0.475 g / 100 mL to 0.525 g / 100 mL, but explicitly includes concentrations exactly 0.5 g / 100 mL; a concentration of "about" 1 g / 100 mL refers to concentrations from 0.95 g / 100 mL to 1.05 g / 100 mL, but explicitly includes concentrations exactly 1 g / 100 mL; a concentration of "about" 1.5 g / 100 mL refers to concentrations from 1.425 g / 100 mL to 1.575 g / 100 mL, but explicitly includes concentrations exactly 1.5 g / 100 mL; a concentration of "about" 2 g / 100 mL refers to concentrations from 1.9 g / 100 mL to 2.1 g / 100 mL, but explicitly includes concentrations exactly 2 g / 100 mL; a concentration of "about" 20 g / 100 mL refers to concentrations from 19 g / 100 mL to 19 g / 100 mL. Concentrations of g / 100 mL to 21 g / 100 mL, but explicitly include concentrations of exactly 20 g / 100 mL; a concentration of “about” 30 g / 100 mL refers to concentrations of 28.5 g / 100 mL to 31.5 g / 100 mL, but explicitly includes concentrations of exactly 30 g / 100 mL; a concentration of “about” 40 g / 100 mL refers to concentrations of 38 g / 100 mL to 42 g / 100 mL, but explicitly includes concentrations of exactly 40 g / 100 mL.

[0069] In a preferred embodiment, the sugar is selected from one or more monosaccharides, disaccharides, and trisaccharides.

[0070] In a preferred embodiment, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol, and xylitol. In practical applications, the above-mentioned sugar alcohol can be replaced with other sugar alcohols in the prior art, depending on the actual situation.

[0071] In a preferred embodiment, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, sorbose, rhamnose, ribose, xylose, and deoxyribose. In practical applications, the above monosaccharide can be replaced with other monosaccharides in the prior art, depending on the specific circumstances.

[0072] 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 disaccharide can be replaced with other disaccharides in the prior art, depending on the actual situation.

[0073] In a preferred embodiment, the monosaccharide is glucose and / or fructose.

[0074] In a preferred embodiment, the trisaccharide is raffinose.

[0075] In order to further improve the extraction rate of saponin compounds from raw materials such as traditional Chinese medicine, in a preferred embodiment, the disaccharide is sucrose and / or maltose.

[0076] In a preferred embodiment, the sugar component is an oligosaccharide component and / or a polysaccharide component.

[0077] In a preferred embodiment, the polysaccharide component is selected from one or more of the following: astragalus polysaccharide, licorice polysaccharide, ginseng polysaccharide, and wolfberry polysaccharide.

[0078] In a preferred embodiment, the glycoside component is selected from one or more of the following: flavonoid glycosides, saponins, diterpenoid glycosides, quinone glycosides, and phenolic glycosides.

[0079] In a preferred embodiment, the flavonoid glycoside compound is selected from one or more of the following: flavonoid glycoside compounds, isoflavone glycoside compounds, dihydroflavonoid glycoside compounds, dihydroflavonol glycoside compounds, dihydroisoflavoneol glycoside compounds, chalcone glycoside compounds, dihydrochalcone glycoside compounds, orange glycoside compounds, anthocyanin compounds, biflavonoid glycoside compounds, and flavanol glycoside compounds.

[0080] In a preferred embodiment, the saponin compound is a tetracyclic triterpenoid glycoside, a pentacyclic triterpenoid glycoside, and / or a steroidal glycoside.

[0081] In a preferred embodiment, the diterpenoid glycoside is selected from one or more of the following: tetracyclic diterpenoid glycosides, tricyclic diterpenoid glycosides, chain diterpenoid glycosides, monocyclic diterpenoids, and bicyclic diterpenoid glycosides.

[0082] In a preferred embodiment, the quinone glycoside is a benzoquinone glycoside, naphthoquinone glycoside, phenanthrenequinone glycoside, and / or anthraquinone glycoside.

[0083] In a preferred embodiment, the phenolic glycoside compound is a phenolic glycoside compound having a C6-C3 core structure and / or a phenolic glycoside compound having a C6-C1 core structure.

[0084] In a preferred embodiment, the steroidal glycoside compound is a spirostanol-type saponin compound, a furostanol-type saponin compound, and / or an isostanol-type saponin compound.

[0085] In a preferred embodiment, the isosspiranol-type saponin compound is a diosgenin compound.

[0086] In a preferred embodiment, the furostanol-type saponin compound is anemarrhena saponin.

[0087] In a preferred embodiment, the tetracyclic triterpenoid glycoside is a lanolinane-type tetracyclic triterpenoid saponin and / or a dammarane-type tetracyclic triterpenoid saponin.

[0088] In a preferred embodiment, the pentacyclic triterpenoid glycoside is an oleanane-type pentacyclic triterpenoid saponin, a ursane-type pentacyclic triterpenoid saponin, and / or a lupinane-type pentacyclic triterpenoid saponin.

[0089] In a preferred embodiment, the anthraquinone glycoside is a monoanthraquinone compound and / or a dianthraquinone compound.

[0090] In a preferred embodiment, the isoflavone glycoside is verrucoside glucoside and / or gentianin. In a preferred embodiment, the dihydroflavonoid glycoside is apigenin glycyrrhizin and / or glycyrrhizin. In a preferred embodiment, the chalcone glycoside is hydroxysafflower yellow A. In a preferred embodiment, the furostanol-type steroidal saponin is anemarrhenasaponin BII. In a preferred embodiment, the lanolinane-type tetracyclic triterpenoid saponin is astragaloside A. In a preferred embodiment, the dammarane-type tetracyclic triterpenoid saponin is ginsenoside Rg1. In a preferred embodiment, the oleanane-type pentacyclic triterpenoid saponin is glycyrrhizic acid. In a preferred embodiment, the tetracyclic diterpenoid glycoside is steviol glycoside and / or rebaudioside. In a preferred embodiment, the C6-C3 core-structured phenolic glycoside is hydroxysafflower yellow A. In a preferred embodiment, the dianthracene nucleoside compound is sennoside A and / or sennoside B. In a preferred embodiment, the monoanthracene nucleoside compound is aloin.

[0091] In a preferred embodiment, the raw material is a raw material containing sugar components and / or glycoside components.

[0092] In a preferred embodiment, the raw material is one or more traditional Chinese medicines, herbal medicines, and / or marine organisms containing carbohydrate components and / or glycoside components.

[0093] In a preferred embodiment, the traditional Chinese medicine and / or herbal medicine is selected from one or more of the following: ginseng, Panax notoginseng, licorice, wolfberry, stevia, astragalus, senna leaf, albizia bark, pokeweed, bupleurum, Pulsatilla chinensis, Toona sinensis, Dendrobium nobile, Achyranthes bidentata, Clematis chinensis, American ginseng, Fritillaria cirrhosa, Aesculus hippocastanum, Acanthopanax senticosus, Eleutherococcus senticosus, Gynostemma pentaphyllum, Melia azedarach, Centella asiatica, Gynostemma pentaphyllum, Ilex chinensis, Dipsacus asper, Akebia quinata, Panax notoginseng, Clematis armandii, Ziziphus jujuba leaf, hemp seed, Polygala tenuifolia, Perilla frutescens, Eclipta prostrata, Platycodon grandiflorus, Ophiopogon japonicus, Allium macrostemon, Polygonatum sibiricum, Polygonatum odoratum, Anemarrhena asphodeloides, Asparagus cochinchinensis, Ophiopogon japonicus, Paris polyphylla, Smilax glabra, Dioscorea opposita, Tribulus terrestris, Solanum nigrum, pomegranate, apricot kernel, sweet potato, Dioscorea opposita, Asparagus cochinchinensis, asparagus, Tribulus terrestris, taro, and dragon's blood.

[0094] In a preferred embodiment, in step (2), the soaking is a room temperature soaking.

[0095] In a preferred embodiment, the soaking step includes immersing the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for 10-120 min.

[0096] In a preferred embodiment, the soaking step includes immersing the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for 20-60 minutes.

[0097] In a preferred embodiment, the soaking step includes immersing the raw material in the sugar solution and / or the sugar alcohol solution at room temperature for about 30 minutes.

[0098] The terms “about” or “approximately” for numerical values ​​mean ±5% of that value, but explicitly include the exact value. For example, “about” 30 minutes means a time from 28.5 minutes to 31.5 minutes, but also explicitly includes exactly 30 minutes.

[0099] 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.

[0100] 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.

[0101] 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 1:12.

[0102] In a preferred embodiment, in step (3), the heating extraction is a heating reflux extraction.

[0103] In a preferred embodiment, the heating time is 20 min to 60 min.

[0104] In a preferred embodiment, the heating time is 20 to 30 minutes.

[0105] In a preferred embodiment, the heating time is about 20 minutes or about 30 minutes.

[0106] The terms “about” or “approximately” for numerical values ​​mean ±5% of that value, but explicitly include the exact value. For example, “about” 30 minutes means from 28.5 minutes to 31.5 minutes, but also explicitly includes exactly 30 minutes; “about” 20 minutes means from 19 minutes to 21 minutes, but also explicitly includes exactly 20 minutes.

[0107] According to one aspect of the present invention, there is provided the use of the method described above in the preparation of a raw material extract or a raw material concentrate, wherein the raw material is a traditional Chinese medicine, a herbal medicine, and / or a marine organism.

[0108] In a preferred embodiment, the raw material concentrate is obtained by concentrating the raw material extract obtained in step (3) or the total raw material extract obtained in step (5) in a water bath at 20°C to 100°C.

[0109] In a preferred embodiment, the concentration is performed in a water bath at 60°C to 80°C.

[0110] In a preferred embodiment, the concentration is performed in a water bath at approximately 70°C.

[0111] The terms "about" or "approximately" for numerical values ​​mean ±5% of that value, but explicitly include the exact 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 temperatures exactly 70°C.

[0112] According to one aspect of the present invention, there is provided the use of the method described above in the preparation of a pharmaceutical composition containing a raw material, wherein the raw material is a traditional Chinese medicine, a herbal medicine, and / or a marine organism.

[0113] According to one aspect of the present invention, there is provided the use of the method described above in the preparation of a pharmaceutical preparation or functional food or health food containing a raw material, wherein the raw material is a traditional Chinese medicine, a herbal medicine and / or a marine organism.

[0114] Functional foods are those that can be convincingly demonstrated to benefit one or more bodily functions, have sufficient nutritional effects to improve health or reduce disease.

[0115] Among them, health food refers to food that claims to have specific health functions or is intended to supplement vitamins and minerals. It is suitable for specific groups of people, has the function of regulating bodily functions, is not intended to treat diseases, and does not cause any acute, subacute or chronic harm to the human body.

[0116] According to one aspect of the present invention, there is provided an use of the method described above in the preparation of other products containing raw materials, wherein other products refer to all products included in the prior art, excluding pharmaceutical preparations, functional foods and health foods, containing raw materials such as traditional Chinese medicine, herbal medicine and / or biological agents, including liquid and solid forms.

[0117] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present invention will now be described in detail with reference to the embodiments.

[0118] The present invention will be further described in detail below with reference to specific embodiments, which should not be construed as limiting the scope of protection claimed in this application.

[0119] Example

[0120] Instruments and reagents

[0121] Agilent 1260 HPLC system with DAD detector (Agilent Technologies, Inc.); Agilent 1200 HPLC system with ELSD detector (Agilent Technologies, Inc.); UV-2550 UV-Vis spectrophotometer (Shimadzu Corporation); DZKW-4 electronic thermostatic water bath (Beijing Zhongxing Weiye Instrument Co., Ltd.); DK-98-ⅡA electric thermostatic water bath (Tianjin Tester Instrument Co., Ltd.); ME204 / 02 electronic balance (Mettler-Toledo Instruments (Shanghai) Co., Ltd., 0.0001 g balance); KQ-250DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); HC-3018 high-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); UniEIut C18EC solid phase extraction column (1000 mg / 6 ml) (Huapu Scientific Instruments Beijing Technology Co., Ltd.).

[0122] The reference standards, apigenin glycyrrhizin (batch number CHB180109), glycyrrhizin (batch number CHB201102), astragaloside A (batch number CHB170727), verrucoside isoflavone glucoside (batch number CHB161105), gentianin (batch number CHB150517), and D-glucose (batch number CHB190213), were all purchased from Chengdu Kloma Biotechnology Co., Ltd., with a purity > 98%; glycyrrhizic acid (batch number Z30A6B1) was purchased from Shanghai Yuanye Biotechnology Co., Ltd., with a purity > 98%; Ginsenoside Rg1 (batch number 210708), purchased from Chengdu Zhibiao Chemical Pure Biotechnology Co., Ltd., purity > 98%; Anemarrhenasaponin BII (batch number 111839-202107), purchased from the China National Institutes for Food and Drug Control, purity ≥ 95.4%; Rebaudioside (batch number 211117), Stevioside (batch number 211024), purchased from Chengdu Zhibiao Chemical Pure Biotechnology Co., Ltd., purity > 98%; Sennoside A (batch number 211203), purchased from Chengdu Zhibiao Chemical Pure Biotechnology Co., Ltd., purity HPLC > 98%; Sennoside B (batch number 220215), purchased from Chengdu Zhibiao Chemical Pure Biotechnology Co., Ltd., purity HPLC > 98%; Acetonitrile (chromatographic grade, Fisher Scientific), methanol (chromatographic grade, Fisher Scientific), phosphoric acid (analytical grade), purchased from Sinopharm Chemical Reagent Co., Ltd. Sucrose, glucose, fructose, maltose, and lactose (Kinklone (Beijing) Biotechnology Co., Ltd.); trehalose (Hebei Baiwei Biotechnology Co., Ltd.); all are pharmaceutical grade. The water is Wahaha purified water.

[0123] Example 1: Experiment on the extraction of raw licorice slices using different sugar solutions

[0124] 1.1 Preparation methods of different sugar-licorice extracts

[0125] Four portions of raw licorice root slices (Inner Mongolia) were taken from the same batch, each weighing 40 g. Seven times the volume of water (280 mL), 1% glucose solution (2.8 g glucose + 280 mL water), 1% sucrose solution (2.8 g sucrose + 280 mL water), and 1% maltose solution (2.8 g maltose + 280 mL water) were added to each portion. The mixtures were soaked for 30 min, refluxed for 30 min, and filtered through gauze while hot to obtain different sugar-licorice extracts.

[0126] 1.2 Determination of the content of apigenin, glycyrrhizin and glycyrrhizic acid (HPLC pharmacopoeia method)

[0127] 1.2.1 Preparation of the test solution

[0128] Take 0.5 mL of different sugar-licorice extracts, dilute 20 times with methanol (9.5 mL), shake well, and filter through a 0.22 μm filter membrane to obtain the final product.

[0129] 1.2.2 Preparation of reference solution

[0130] Stock solution: Accurately weigh appropriate amounts of apigenin, glycyrrhizin, and glycyrrhizic acid reference standards, and add methanol to prepare a 1 mg / mL stock solution. Store at -20 ℃.

[0131] Linear working solutions: Take appropriate amounts of apigenin and glycyrrhizin stock solution, and 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 amounts of glycyrrhizic acid stock solution, and dilute with methanol to prepare a series of linear mixed standard working solutions: 10, 20, 40, 80, 160, 200, 400 μg / mL.

[0132] 1.2.3 Content Determination Method

[0133] Chromatographic column: Thermo Acclaim 120 C18 (250 × 4.6 mm, 5 μm); mobile phase: acetonitrile (A), 0.05% phosphoric acid solution (B) as mobile phase; column temperature: 35℃; 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.

[0134] 1.3 Experimental Results

[0135] The results of the determination of the effective components in different sugar-licorice extracts are shown in Table 1.

[0136] Table 1. Results of determination of active ingredient content in different sugar-licorice extracts

[0137]

[0138] * P < 0.05 VS Water

[0139] As can be seen from Table 1, glucose, sucrose, and maltose (1 g / 100 mL concentration) can all increase the content of active ingredients in licorice to varying degrees, with sucrose and maltose having the most significant effects.

[0140] Example 2: Experiment on the extraction of raw licorice slices using different sugar solutions

[0141] 2.1 Preparation methods of different sugar-licorice extracts

[0142] Four portions (40 g each) of raw licorice root slices from the same batch (produced in Xinjiang Uygur Autonomous Region) were taken. Seven times the volume of water (280 mL), 1% sucrose solution (2.8 g sucrose + 280 mL water), 1% maltose solution (2.8 g maltose + 280 mL water), and 1% lactose solution (2.8 g lactose + 280 mL water) were added sequentially. The mixtures were soaked for 30 min, refluxed for 30 min, and filtered through gauze while hot. The residue was then further soaked in six times the volume of water, 1% sucrose solution (2.4 g sucrose + 240 mL water), 1% maltose solution (2.4 g maltose + 240 mL water), and 1% lactose solution (2.4 g lactose + 240 mL water), refluxed for 20 min, and filtered while hot. The two filtrates were combined to obtain different sugar-licorice extracts.

[0143] 2.2 Determination of the content of apigenin, glycyrrhizin and glycyrrhizic acid (HPLC pharmacopoeia method)

[0144] 2.2.1 Preparation of the test solution

[0145] The preparation method is the same as that for the test solution in Example 1.

[0146] 2.2.2 Preparation of the reference solution

[0147] The preparation method is the same as that of the reference solution in Example 1.

[0148] 2.2.3 Content determination method

[0149] The content determination method is the same as that in Example 1.

[0150] 2.3 Experimental Results

[0151] The results of the determination of the effective components in different sugar-licorice extracts are shown in Table 2.

[0152] Table 2. Results of determination of active ingredient content in different sugar-licorice extracts

[0153]

[0154] * P < 0.05 VS Water

[0155] As can be seen from Table 2, both sucrose and maltose (1 g / 100 mL concentration) can increase the content of effective components in licorice to varying degrees, with sucrose having the most significant effect.

[0156] The results from Tables 1 and 2 show that sucrose can significantly improve the extraction rate of the main active components in licorice, including flavonoids (apigenin, glycyrrhizin) and saponins (glycyrrhizic acid).

[0157] Example 3: Experiment on the extraction of raw Astragalus membranaceus slices using different sugar solutions

[0158] 3.1 Preparation methods of different sugar-Astragalus extracts (two extractions with different 1% sugars)

[0159] Five portions (50 g each) of raw Astragalus membranaceus slices from the same batch (originating from Hunyuan, Shanxi) were taken. Each portion was soaked in one of the following solutions: 7 times the volume of water (350 ml), 1% glucose solution (3.5 g glucose diluted to 350 ml), 1% fructose solution (3.5 g fructose diluted to 350 ml), 1% sucrose solution (3.5 g sucrose diluted to 350 ml), and 1% trehalose solution (3.5 g trehalose diluted to 350 ml). The solutions were then refluxed for 30 minutes, filtered through gauze while still hot, and the residue was further separated. Add 6 times the volume of water (300ml), 1% glucose solution (3.0g glucose diluted with water to 300ml), 1% fructose solution (3.0g fructose diluted with water to 300ml), 1% sucrose solution (3.0g sucrose diluted with water to 300ml), and 1% trehalose solution (3.0g trehalose diluted with water to 300ml). Reflux for 20 minutes, filter while hot, and combine the two filtrates to obtain different sugar-Astragalus extracts.

[0160] 3.2 Determination of the contents of astragaloside A, verbascoside, and gentianoside

[0161] 3.2.1 Preparation of the test solution

[0162] Preparation of saponin component test solution: Take 5 ml of different sugar-Astragalus extract, add 20 ml of 60% methanol, mix by sonication, add 10 ml of ammonia water, mix well, centrifuge at 3600 r / min for 10 min, pass the supernatant through a Huapu UniEIut C18 EC column (1000 mg / 6 ml), elute with 10 ml of purified water, then elute with 5 ml of methanol, collect the methanol eluent, centrifuge at 8000 r / min for 5 min, take the supernatant, filter through a 0.45 μm microporous membrane, and obtain the solution.

[0163] Preparation of flavonoid sample solution: Take 300 μl of different sugar-Astragalus extract, add 700 μl of water, place in a 1.5 ml centrifuge tube, centrifuge at 12000 r / min for 5 min, take the supernatant, filter through a 0.45 µm microporous membrane to obtain the sample solution.

[0164] 3.2.2 Preparation of reference solution

[0165] Accurately weigh appropriate amounts of astragaloside A, verbascoside, and gentianoside reference standards, and prepare a 1 mg / ml stock solution with methanol. Dilute the stock solution to different concentrations and establish a working curve.

[0166] The linear equation for astragaloside A is lnY=1.3271lnX+9.9314, r=1.0000; the linear equation for verrucoside isoflavone glucoside is Y=35129X+14, r=0.9998; and the linear equation for gentianoside is Y=10024X+33, r=0.9998.

[0167] 3.2.3 Content determination method

[0168] Method for determining the content of saponins: Analysis was performed using an Agilent 1200 liquid chromatograph (equipped with an ELSD detector). The chromatographic column was a Thermo Acclaim 120 C18 (250×4.6 mm, 5 μm). The mobile phase was 0.1% formic acid water (A) ~ 0.1% formic acid acetonitrile (B). 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℃, the carrier gas flow rate was 2.5 l / min, and the injection volume was 20 μl.

[0169] The method for determining the content of flavonoid components was as follows: Analysis was performed using an Agilent 1260 high-performance liquid chromatograph (equipped with a DAD detector). The chromatographic column was a Thermo Acclaim 120 C18 (250 × 4.6 mm, 5 μm). The mobile phase consisted of 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile (B). Gradient elution was used: 0–8 min, 5%–20% B; 8–15 min, 20%–25% B; 15–20 min, 25% B; 20–30 min, 25%–40% B; 30–40 min, 40%–60% B; flow rate was 1 ml / min; detection wavelength was 260 nm; column temperature was 25 °C; and injection volume was 10 μl.

[0170] 3.3 Experimental Results

[0171] The results of the determination of the effective components in different sugar-Astragalus extracts are shown in Table 3.

[0172] Table 3. Results of determination of effective components in different sugar-Astragalus extracts

[0173]

[0174] * P < 0.05 VS Astragalus membranaceus aqueous extract

[0175] As can be seen from Table 3, sucrose, glucose, fructose, and trehalose (1g / 100ml concentration) can all increase the content of effective components in Astragalus membranaceus to varying degrees. Among them, sucrose and trehalose have the most obvious effects, but sucrose is more economical than trehalose.

[0176] Example 4: Experiment on the extraction of raw Astragalus membranaceus slices using different sugar solutions

[0177] 4.1 Preparation methods of different sugar-Astragalus extracts (one extraction with different 1% sugars)

[0178] Four portions of raw Astragalus membranaceus slices (from Hunyuan, Shanxi) were taken, each weighing 50 g. Seven times the volume of water (350 ml), a 1% glucose solution (3.5 g glucose diluted with water to 350 ml), a 1% sucrose solution (3.5 g sucrose diluted with water to 350 ml), and a 1% maltose solution (3.5 g maltose diluted with water to 350 ml) were added to each portion. The mixtures were soaked for 30 minutes, refluxed for 30 minutes, and filtered through gauze while hot to obtain different sugar-Astragalus membranaceus extracts.

[0179] 4.2 Determination of the contents of astragaloside A, verbascoside, and gentianoside

[0180] 4.2.1 Preparation of the test solution

[0181] The preparation method is the same as that for the test solution in Example 3.

[0182] 4.2.2 Preparation of reference solution

[0183] The preparation method is the same as that of the reference solution in Example 3.

[0184] 4.2.3 Content Determination Method

[0185] The content determination method is the same as that in Example 3.

[0186] 4.3 Experimental Results

[0187] The results of the determination of the effective components in different sugar-Astragalus extracts are shown in Table 4.

[0188] Table 4. Results of determination of effective component content in different sugar-Astragalus extracts

[0189]

[0190] * P < 0.05 VS Astragalus membranaceus aqueous extract

[0191] As can be seen from Table 4, glucose, sucrose, and maltose (1g / 100ml concentration) can all increase the content of effective components in Astragalus membranaceus to varying degrees, with sucrose showing the most significant effect.

[0192] Example 5: Experiment on the extraction of raw Astragalus membranaceus slices using different sugar solutions

[0193] 5.1 Preparation methods of different sugar-Astragalus extracts (two extractions with different 1% sugars)

[0194] Four portions of raw Astragalus membranaceus slices (from Lingchuan, Shanxi Province) were taken, each weighing 50 g. Seven times the volume of water (350 ml), 1% sucrose solution (3.5 g sucrose diluted to 350 ml), 1% maltose solution (3.5 g maltose diluted to 350 ml), and 1% lactose solution (3.5 g lactose diluted to 350 ml) were added to each portion and soaked for 30 minutes. The mixture was then refluxed for 30 minutes and filtered through gauze while hot. The residue was then further treated with six times the volume of water (300 ml), 1% sucrose solution (3.0 g sucrose diluted to 300 ml), 1% maltose solution (3.0 g maltose diluted to 300 ml), and 1% lactose solution (3.0 g lactose diluted to 300 ml), refluxed for 20 minutes, and filtered while hot. The two filtrates were combined to obtain different sugar-Astragalus membranaceus extracts.

[0195] 5.2 Determination of the contents of astragaloside A, verbascoside, and gentianoside

[0196] 5.2.1 Preparation of the test solution

[0197] The preparation method is the same as that for the test solution in Example 3.

[0198] 5.2.2 Preparation of reference solution

[0199] The preparation method is the same as that of the reference solution in Example 3.

[0200] 5.2.3 Content Determination Method

[0201] The content determination method is the same as that in Example 3.

[0202] 5.3 Experimental Results

[0203] The results of the determination of the effective components in different sugar-Astragalus extracts are shown in Table 5.

[0204] Table 5. Results of determination of effective components in different sugar-Astragalus extracts

[0205]

[0206] * P < 0.05 VS Astragalus membranaceus aqueous extract

[0207] As can be seen from Table 5, sucrose, maltose, and lactose (1g / 100ml concentration) can all increase the content of effective components in Astragalus membranaceus to varying degrees. Among them, sucrose can increase the extraction rate of all three components at the same time, and its effect is the most obvious.

[0208] The results in Tables 3-5 show that sucrose can significantly improve the extraction rate of the main active components in Astragalus membranaceus, including flavonoids (verrucoside and gentianin) and saponins (astragaloside A).

[0209] Example 6: Extraction of raw Astragalus membranaceus slices from sucrose solutions of different concentrations

[0210] 6.1 Preparation of sucrose-Astragalus extracts at different concentrations (0-10% sucrose extracts)

[0211] Seven portions (50 g each) of raw Astragalus membranaceus slices from the same batch (originating from Lingchuan, Shanxi) were taken. Seven times the volume of water (350 ml) was added to each portion, along with the following solutions: 0.5% sucrose solution (1.8 g sucrose diluted to 350 ml), 1% sucrose solution (3.5 g sucrose diluted to 350 ml), 1.25% sucrose solution (4.4 g sucrose diluted to 350 ml), 1.5% sucrose solution (5.3 g sucrose diluted to 350 ml), 2% sucrose solution (7 g sucrose diluted to 350 ml), and 10% sucrose solution (35 g sucrose diluted to 350 ml). The solutions were soaked for 30 minutes, refluxed for 30 minutes, and filtered while hot through gauze. The residue was then further treated with 6... Double the volume of water (300ml), 0.5% sucrose aqueous solution (1.5g sucrose diluted with water to 300ml), 1% sucrose aqueous solution (3.0g sucrose diluted with water to 300ml), 1.25% sucrose aqueous solution (3.8g sucrose diluted with water to 300ml), 1.5% sucrose aqueous solution (4.5g sucrose diluted with water to 300ml), 2% sucrose aqueous solution (6.0g sucrose diluted with water to 300ml), 10% sucrose aqueous solution (30g sucrose diluted with water to 300ml), reflux for 20min, filter while hot, combine the two filtrates to obtain sucrose-Astragalus extracts of different concentrations.

[0212] 6.2 Determination of the contents of astragaloside A, verbascoside, and gentianoside

[0213] 6.2.1 Preparation of the test solution

[0214] The preparation method is the same as that for the test solution in Example 3.

[0215] 6.2.2 Preparation of reference solution

[0216] The preparation method is the same as that of the reference solution in Example 3.

[0217] 6.2.3 Content Determination Method

[0218] The content determination method is the same as that in Example 3.

[0219] 6.3 Experimental Results

[0220] The results of the determination of the effective components in the extracts of Astragalus membranaceus obtained by extracting raw Astragalus membranaceus slices with sucrose solutions of different concentrations are shown in Table 6.

[0221] Table 6. Determination of the content of active ingredients in Astragalus extracts obtained with different concentrations of sucrose.

[0222]

[0223] * P < 0.05 VS Astragalus membranaceus aqueous extract

[0224] As shown in Table 6, sucrose concentrations ranging from 0.25 g / 100 ml to 1.25 g / 100 ml all increased the content of active ingredients in Astragalus membranaceus to varying degrees, with the most significant effect observed in concentrations between 1 g / 100 ml and 1.25 g / 100 ml. Furthermore, a sucrose concentration of 10 g / 100 ml also increased the content of active ingredients in Astragalus membranaceus.

[0225] Example 7: Experiment on the extraction of ginsenoside Rg1 from Panax notoginseng using sugar solutions of different concentrations

[0226] 7.1 Preparation methods of sucrose-Panax notoginseng extracts of different concentrations

[0227] Four portions of raw Panax notoginseng slices (origin: Yunnan; manufacturer: Tianjin Shengshi Pharmaceutical Co., Ltd.; batch number: X20102703A) were taken, each portion weighing 50 g. Add 7 times the volume of water (350 ml), 0.5% sucrose aqueous solution (1.75 g sucrose diluted to 350 ml), 1% sucrose aqueous solution (3.5 g sucrose diluted to 350 ml), and 1.5% sucrose aqueous solution (5.25 g sucrose diluted to 350 ml) to the residue respectively. Soak for 30 min, reflux for 30 min, and filter with gauze while hot. Add 6 times the volume of water (300 ml), 0.5% sucrose aqueous solution (1.5 g sucrose diluted to 300 ml), 1% sucrose aqueous solution (3.0 g sucrose diluted to 300 ml), and 1.5% sucrose aqueous solution (4.5 g sucrose diluted to 300 ml) to the residue respectively. Reflux for 20 min, filter while hot, and combine the two filtrates to obtain sucrose-Panax notoginseng extracts of different concentrations.

[0228] 7.2 Determination of Ginsenoside Rg1 Content

[0229] 7.2.1 Preparation of the test solution

[0230] Accurately measure 2 mL of different Panax notoginseng extracts, add methanol to each and bring to a final volume of 10 mL in a volumetric flask, mix well, and incubate at 13000 rpm. -1 Centrifuge for 5 minutes, collect the supernatant, and filter it through a 0.22 μm microporous membrane to obtain the final product.

[0231] 7.2.2 Preparation of reference solution

[0232] Accurately weigh an appropriate amount of ginsenoside Rg1 reference standard and prepare a 1 mg / ml stock solution with methanol. Dilute the stock solution to different concentrations and establish a working curve.

[0233] The linear equation for ginsenoside Rg1 is: Y = 3246.4X - 4.5152, R2 = 0.9999.

[0234] 7.2.3 Content Determination Method

[0235] Analysis was performed using an Agilent 1200 liquid chromatograph. The chromatographic column was a Roc C18 (250×4.6 mm, 5 μm); the mobile phase was water (A) to acetonitrile (B), with gradient elution: 0–12 min, 19% → 19%B; 12–60 min, 19% → 36%B; the column temperature was 35℃, the flow rate was 1 ml / min, the injection volume was 10 μl, and the detection wavelength was 203 nm.

[0236] 7.3 Experimental Results

[0237] The effects of different sucrose concentrations on the extraction rates of key components in Panax notoginseng are shown in Table 7.

[0238] Table 7. Effects of different sucrose concentrations on the extraction rates of key components in Panax notoginseng.

[0239]

[0240] * P < 0.05 vs Panax notoginseng aqueous extract

[0241] As can be seen from Table 7, sucrose (concentrations of 1 g / 100 ml to 1.5 g / 100 ml) can all increase the content of saponins (ginsenoside Rg1), the effective component in Panax notoginseng, to varying degrees.

[0242] Example 8: Experiment on the extraction of anemarrhena saponin BII from Anemarrhena asphodeloides slices using sucrose solutions of different concentrations

[0243] 8.1 Preparation methods of sucrose-Anemarrhena asphodeloides extracts of different concentrations

[0244] Five portions of raw Anemarrhena asphodeloides slices were taken from the same batch, each portion weighing 50 g. Add 7 times the volume of water (350 ml), 0.5% sucrose solution (1.75 g sucrose diluted to 350 ml), 1% sucrose solution (3.5 g sucrose diluted to 350 ml), 1.5% sucrose solution (5.25 g sucrose diluted to 350 ml), and 2.0% sucrose solution (7.0 g sucrose diluted to 350 ml) to the residue respectively. Soak for 30 min, reflux for 30 min, and filter with gauze while hot. Add 6 times the volume of water (300 ml), 0.5% sucrose solution (1.5 g sucrose diluted to 300 ml), 1% sucrose solution (3.0 g sucrose diluted to 300 ml), 1.5% sucrose solution (4.5 g sucrose diluted to 300 ml), and 2.0% sucrose solution (6 g sucrose diluted to 300 ml) to the residue respectively, reflux for 20 min. After 1 minute, filter while hot, combine the two filtrates, and bring the volume to 500 mL to obtain sucrose-Anemarrhena extracts of different concentrations.

[0245] 8.2 Determination of Anemarrhena saponin BⅡ content

[0246] 8.2.1 Preparation of the test solution

[0247] Accurately measure 2 mL of different Anemarrhena asphodeloides extracts, add methanol to each and bring to a final volume of 10 mL in a volumetric flask, mix well, and incubate at 13000 rpm. -1 Centrifuge for 5 min, collect the supernatant, and filter it through a 0.22 μm microporous membrane to obtain the final product.

[0248] 8.2.2 Preparation of reference solution

[0249] Accurately weigh an appropriate amount of Anemarrhena saponin BⅡ reference standard, and prepare a stock solution with a mass concentration of 1 mg / mL by adding 30% acetonitrile. Dilute the stock solution to different concentrations to establish a working curve.

[0250] The linear equation for Anemarrhena saponin BII is: lnY = 1.8921lnX + 10.274, R 2 = 0.9977.

[0251] 8.2.3 Content Determination Method

[0252] Analysis was performed using an Agilent 1200 high-performance liquid chromatograph. The column was an Inertsil ODS-3-C18 (250 × 4.6 mm, 5 μm). The mobile phase was water (A) to acetonitrile (B), with isocratic elution of 25% acetonitrile. The column temperature was 35℃, the flow rate was 0.8 ml / min, and the injection volume was 10 μl. The drift tube temperature was 104℃, and the carrier gas flow rate was 2.8 L·min. -1 .

[0253] 8.3 Experimental Results

[0254] The effects of different sucrose concentrations on the extraction rates of indicator components in Anemarrhena asphodeloides are shown in Table 8.

[0255] Table 8. Effects of different sucrose concentrations on the extraction rate of indicator components in Anemarrhena asphodeloides.

[0256]

[0257] * P < 0.05 vs Anemarrhena asphodeloides aqueous extract

[0258] As can be seen from Table 8, sucrose (concentrations of 1.0 g / 100 ml to 2.0 g / 100 ml) can all increase the content of saponins (anemarrhena saponin BⅡ), the effective component in Anemarrhena asphodeloides, to varying degrees.

[0259] Example 9: Extraction of sennoside B and sennoside A from senna leaves using sucrose solutions of different concentrations

[0260] 9.1 Preparation methods of sucrose-senna leaf extracts at different concentrations

[0261] The same batch of senna leaves (originating from Yulin, Guangxi, batch number: 200101) was used. Eight portions, each 20 g, were prepared and soaked in the following solutions: 12 times the volume of water (240 ml), 0.5% sucrose solution (1.2 g sucrose dissolved in 240 ml), 0.75% sucrose solution (1.8 g sucrose dissolved in 240 ml), 1% sucrose solution (2.4 g sucrose dissolved in 240 ml), 1.5% sucrose solution (3.6 g sucrose dissolved in 240 ml), 2% sucrose solution (4.8 g sucrose dissolved in 240 ml), 3% sucrose solution (7.2 g sucrose dissolved in 240 ml), and 5% sucrose solution (12 g sucrose dissolved in 240 ml). Each solution was soaked for 30 minutes and then refluxed for 30 minutes. The extract was filtered through three layers of gauze while still hot. The residue was then mixed with 10 times the amount of water (200 ml), 0.5% sucrose solution (1 g sucrose dissolved in 200 ml water), 0.75% sucrose solution (1.5 g sucrose dissolved in 200 ml water), 1% sucrose solution (2 g sucrose dissolved in 200 ml water), 1.5% sucrose solution (3 g sucrose dissolved in 200 ml water), 2% sucrose solution (4 g sucrose dissolved in 200 ml water), 3% sucrose solution (6 g sucrose dissolved in 200 ml water), and 5% sucrose solution (10 g sucrose dissolved in 200 ml water). The mixture was refluxed for 20 min, filtered through three layers of gauze while still hot, and the two filtrates were combined and brought to a final volume of 400 mL to obtain sucrose-senna leaf extracts of different concentrations.

[0262] 9.2 Determination of sennoside B and sennoside A content

[0263] 9.2.1 Preparation of the test solution

[0264] Accurately measure 1 mL of senna leaf extract at different concentrations, and dilute to 5 mL in a volumetric flask. Dilute the sample 5 times, mix well, and incubate at 15000 rpm. -1 Centrifuge for 5 minutes, collect the supernatant, and filter it through a 0.22 μm microporous membrane to obtain the final product.

[0265] 9.2.2 Preparation of reference solution

[0266] Take appropriate amounts of sennoside B and sennoside A reference standards, add 0.1% NaHCO3 to prepare a stock solution with a mass concentration of 1 mg / ml, and then dilute with methanol to prepare reference standard stock solutions with concentrations of 0.2 mg / ml, 0.1 mg / ml, 0.05 mg / ml, 0.025 mg / ml, and 0.0125 mg / ml. Shake well to obtain the final solution. The linear equation for sennoside B is y = 8988.5x + 2.5708, R0. 2 = 0.9999; the linear equation for sennoside A is y = 8564.2x + 9.9917, R0 2 = 0.9999.

[0267] 9.2.3 Content Determination Method

[0268] Analysis was performed using an Agilent 1260 liquid chromatograph (equipped with a UV detector):

[0269] Column: C18 (250 mm × 4.6 mm, 5 μm). Injection volume: 10 μL. Detection wavelength: 340 nm, flow rate: 1 mL / min, column temperature: 30 °C. Mobile phase A: 0.1% phosphoric acid aqueous solution; mobile phase B: acetonitrile, gradient desorption (0–10 min, 79% A; 10–15 min, 79% → 75% A; 15–16 min, 75% A; 16–17 min, 75% → 79% A).

[0270] 9.3 Experimental Results

[0271] The effects of different concentrations of sucrose on the extraction rates of key components in senna leaves are shown in Table 9.

[0272] Table 9. Effects of different sucrose concentrations on the extraction rates of key components in senna leaves.

[0273]

[0274] * P < 0.05 vs senna leaf aqueous extract

[0275] As can be seen from Table 9, sucrose concentrations ranging from 2 g / 100 mL to 5 g / 100 mL can significantly increase the content of indicator components in senna leaves.

[0276] Example 10: Experiment on the extraction of aloin from aloe vera using sucrose solutions of different concentrations and sugar solutions of the same concentration but different concentrations.

[0277] 10.1.1 Preparation methods of sucrose-aloe extracts of different concentrations

[0278] Take 6 portions of aloe vera (purchased from Beijing Tongrentang), each 20 g, and add 12 times the amount of water (240 ml), 0.5% sucrose solution (1.2 g sucrose dissolved in 240 ml water), 1% sucrose solution (2.4 g sucrose dissolved in 240 ml water), 1.5% sucrose solution (3.6 g sucrose dissolved in 240 ml water), 2% sucrose solution (4.8 g sucrose dissolved in 240 ml water), and 3% sucrose solution (7.2 g sucrose dissolved in 240 ml water) to each portion. Soak for 30 min, reflux for 30 min, filter while hot through a single layer of polyester, and bring the volume to 240 mL.

[0279] 10.1.2 Preparation methods of different sugar-aloe vera extracts

[0280] Four portions of aloe vera (purchased from Beijing Tongrentang) were taken, each weighing 20 g. The aloe vera was soaked in 12 times the volume of water (240 ml), 0.5% sucrose solution (1.2 g sucrose dissolved in 240 ml), 0.5% maltose solution (1.2 g maltose dissolved in 240 ml), and 0.5% trehalose solution (1.2 g trehalose dissolved in 240 ml) for 30 min, then refluxed for 30 min. The aloe vera was then filtered through a single layer of polyester while still hot and brought to a final volume of 240 mL.

[0281] 10.2 Determination of Aloe Vera Glycoside Content

[0282] 10.2.1 Preparation of the test solution

[0283] Accurately measure 0.2 mL of aloe vera extract at different concentrations, and dilute to 10 mL in a volumetric flask. Dilute the sample 50 times, mix well, and incubate at 15000 rpm. -1 Centrifuge for 5 minutes, collect the supernatant, and filter it through a 0.22 μm microporous membrane to obtain the final product.

[0284] 10.2.2 Preparation of reference solution

[0285] Take an appropriate amount of aloin reference standard, add methanol to prepare a stock solution with a mass concentration of 1 mg / ml, and then dilute with methanol to prepare reference standard stock solutions with concentrations of 0.2 mg / ml, 0.1 mg / ml, 0.05 mg / ml, 0.3 mg / ml, 0.15 mg / ml, and 0.075 mg / ml. Shake well to obtain the final product.

[0286] The linear equation for aloin is y = 7459.1x - 25.173 R. 2 = 0.9991

[0287] 10.2.3 Content Determination Method

[0288] Analysis was performed using an Agilent 1260 liquid chromatograph (equipped with a UV detector).

[0289] Chromatographic column: C18 (250 mm × 4.6 mm, 5 μm). Injection volume: 5 μL. Detection wavelength: 355 nm. Flow rate: 1 mL / min. Mobile phase A: water. Mobile phase B: acetonitrile. Isocratic desorption (0–15 min, 75% A).

[0290] 10.3 Experimental Results

[0291] The effects of different concentrations of sucrose on the extraction rates of key components in aloe vera are shown in Table 10.

[0292] Table 10 Effect of different sucrose concentrations on the extraction rate of index components in aloe vera

[0293]

[0294] * P < 0.05 vs Aloe Vera Water Extract

[0295] The effects of different types of disaccharides on the extraction rate of indicator components in aloe vera are shown in Table 11.

[0296] Table 11 Effects of different types of disaccharides on the extraction rate of index components in aloe vera

[0297]

[0298] * P < 0.05 vs Aloe Vera Water Extract

[0299] As can be seen from the records in Tables 10 and 11, a sucrose concentration of 0.5 g / 100 mL can significantly increase the content of the indicator component (i.e., aloin) in aloe vera.

[0300] Example 11: Extraction of steviol glycosides and rebaudioside from stevia using sucrose solutions of different concentrations and different sugar solutions of the same concentration.

[0301] 11.1 Preparation methods of sucrose-stevia extracts at different concentrations

[0302] Five portions of stevia from the same batch were taken, each portion weighing 20 g. Add 12 times the volume of water (240 ml), 0.5% sucrose aqueous solution (1.2 g sucrose dissolved in 240 ml water), 1% sucrose aqueous solution (2.4 g sucrose dissolved in 240 ml water), 1.5% sucrose aqueous solution (3.6 g sucrose dissolved in 240 ml water), and 2% sucrose aqueous solution (4.8 g sucrose dissolved in 240 ml water) to the herbs respectively, soak for 30 min, reflux for 30 min, and filter while hot through 3 layers of gauze; then add 10 times the volume of water (200 ml water), 0.5% (1 g sucrose dissolved in 200 ml water), 1% sucrose aqueous solution (2 g sucrose dissolved in 200 ml water), 1.5% sucrose aqueous solution (3 g sucrose dissolved in 200 ml water), and 2% (4 g sucrose dissolved in 200 ml water) to the herbs respectively, reflux for 20 min, filter while hot through 3 layers of gauze, combine the two filtrates, and make up to 400 mL to obtain sucrose-stevia extracts of different concentrations.

[0303] 11.2 Preparation methods of different sugar-stevia extracts

[0304] Five portions of stevia from the same batch were taken, each portion weighing 20 g. Add 12 times the volume of water (240ml), 1.5% sucrose solution (3.6g sucrose dissolved in 240ml water), 1.5% maltose solution (3.6g maltose dissolved in 240ml water), 1.5% trehalose solution (3.6g trehalose dissolved in 240ml water), and 1.5% lactose solution (3.6g lactose dissolved in 240ml water) to the herbs respectively, soak for 30 minutes, reflux for 30 minutes, and filter while hot through three layers of gauze. Then add 10 times the volume of water (200ml water), 1.5% sucrose solution (3g sucrose dissolved in 200ml water), 1.5% maltose solution (3g maltose dissolved in 200ml water), 1.5% trehalose solution (3g trehalose dissolved in 200ml water), and 1.5% lactose solution (3g lactose dissolved in 200ml water) to the herbs respectively, reflux for 20 minutes. The solution was filtered through three layers of gauze while still hot. The two filtrates were combined and brought to a final volume of 400 mL to obtain stevia extracts of the same concentration but different sugar concentrations.

[0305] 11.3 Determination of the content of indicator components

[0306] 11.3.1 Preparation of the test solution

[0307] Accurately measure 0.5 mL of stevia extract at different concentrations, add methanol to bring the volume to 10 mL, dilute the sample 20 times, mix well, and incubate at 14000 rpm. -1Centrifuge for 5 minutes, collect the supernatant, and filter it through a 0.22 μm microporous membrane to obtain the final product.

[0308] 11.3.2 Preparation of reference solution

[0309] Lebodiin: Take an appropriate amount of reference standard, add methanol to prepare a stock solution with a mass concentration of 1 mg / ml, and then dilute with methanol to prepare reference standard stock solutions of 0.4 mg / ml, 0.3 mg / ml, 0.2 mg / ml, 0.15 mg / ml, 0.1 mg / ml and 0.05 mg / ml, and shake well to obtain the solution.

[0310] Stevioside: Take an appropriate amount of reference standard, add methanol to prepare a stock solution with a mass concentration of 1 mg / ml, and then dilute with methanol to prepare reference standard stock solutions of 0.3 mg / ml, 0.2 mg / ml, 0.15 mg / ml, 0.1 mg / ml, 0.05 mg / ml and 0.025 mg / ml, and shake well to obtain the solution.

[0311] The linear equation for lebodiin is: y = 999.64x + 1.6452, R0 2 = 0.9995

[0312] The linear equation for steviol glycoside is: y = y = 1184.2x + 0.987 R 2 = 0.9998

[0313] 11.3.3 Content Determination Method

[0314] Analysis was performed using an Agilent 1260 liquid chromatograph (equipped with a UV detector).

[0315] The chromatographic column was a C18 (250 mm × 4.6 mm, 5 μm), with a UV detector and an injection volume of 5 μL. The detection wavelength was 210 nm, the flow rate was 0.6 mL / min, and the column temperature was set at 35 °C. The mobile phase was 0.1% phosphoric acid solution A-acetonitrile B, with isocratic elution of 30% acetonitrile for 25 min.

[0316] 11.4 Experimental Results

[0317] The effects of different sucrose concentrations on the extraction rates of key components in stevia are shown in Table 12.

[0318] Table 12 Effects of different sucrose concentrations on the extraction rates of key components in stevia.

[0319]

[0320] * P < 0.05 vs Stevia aqueous extract

[0321] As can be seen from Table 12, sucrose concentrations ranging from 0.5 g / 100 mL to 1.5 g / 100 mL can increase the content of indicator components in stevia to varying degrees, with the most significant effect observed at a sucrose concentration of 1.5 g / 100 mL.

[0322] The effects of different types of disaccharides on the extraction rates of key components in stevia are shown in Table 13.

[0323] Table 13 Effects of different types of disaccharides on the extraction rate of index components in stevia

[0324]

[0325] * P < 0.05 vs Stevia aqueous extract

[0326] As can be seen from Table 13, different types of disaccharides can all increase the content of indicator components in stevia to varying degrees at a concentration of 1.5 g / 100 mL.

[0327] Example 12: Extraction of hydroxysafflower yellow A from safflower using sucrose solutions of different concentrations and sugar solutions of the same concentration but different concentrations.

[0328] 12.1 Preparation methods of sucrose-safflower extracts of different concentrations

[0329] Seven portions (20 g each) of safflower from the same batch (Tibet Autonomous Region, Beijing Tongrentang Co., Ltd.) were taken and soaked in the following solutions respectively: 12 times the volume of water (240 ml), 0.5% sucrose solution (1.2 g sucrose dissolved in 240 ml), 1% sucrose solution (2.4 g sucrose dissolved in 240 ml), 1.5% sucrose solution (3.6 g sucrose dissolved in 240 ml), 2% sucrose solution (4.8 g sucrose dissolved in 240 ml), 2.5% sucrose solution (6 g sucrose dissolved in 240 ml), and 5% sucrose solution (12 g sucrose dissolved in 240 ml). Each solution was soaked for 30 minutes and then refluxed for 30 minutes. Filter the solution while hot through three layers of gauze. Add 10 times the amount of water (200ml) to the residue, then add the following solutions: 0.5% sucrose solution (1g sucrose dissolved in 200ml water), 1% sucrose solution (2g sucrose dissolved in 200ml water), 1.5% sucrose solution (3g sucrose dissolved in 200ml water), 2% sucrose solution (4g sucrose dissolved in 200ml water), 2.5% sucrose solution (5g sucrose dissolved in 200ml water), and 5% sucrose solution (10g sucrose dissolved in 200ml water). Reflux for 20 minutes, then filter while hot through three layers of gauze. Combine the two filtrates and bring the volume to 400 mL.

[0330] 12.2 Preparation methods of different sugar-safflower extracts

[0331] Take three portions of safflower from the same batch (Tibet Autonomous Region, Beijing Tongrentang Co., Ltd.), each 20 g. Add 12 times the amount of water (240 ml), 0.5% sucrose solution (1.2 g sucrose dissolved in 240 ml), and 0.5% maltose solution (1.2 g maltose dissolved in 240 ml) to each portion and soak for 30 min. Reflux for 30 min and filter while hot through three layers of gauze. Add 10 times the amount of water (200 ml), 0.5% sucrose solution (1 g sucrose dissolved in 200 ml), and 0.5% maltose solution (1 g maltose dissolved in 200 ml) to each portion and reflux for 20 min. Filter while hot through three layers of gauze. Combine the two filtrates and bring the volume to 400 mL.

[0332] 12.3 Determination of the content of indicator components

[0333] 12.3.1 Preparation of the test solution

[0334] Accurately measure 1 mL of safflower extract at different concentrations, add 1 mL of methanol, dilute the sample by 2 times, mix well, and incubate at 15000 rpm. -1 Centrifuge for 5 minutes, collect the supernatant, and filter it through a 0.22 μm microporous membrane to obtain the final product.

[0335] 12.3.2 Preparation of reference solution

[0336] Take an appropriate amount of hydroxysaffron yellow A reference standard, add methanol to prepare a stock solution with a mass concentration of 1 mg / ml, and then dilute with methanol to prepare reference standard stock solutions with concentrations of 0.2 mg / ml, 0.1 mg / ml, 0.05 mg / ml, 0.025 mg / ml, and 0.0125 mg / ml. Shake well to obtain the final product.

[0337] The linear equation for hydroxysaffron yellow A is y = 13131x - 26.496, R0 2 = 0.9993.

[0338] 12.3.3 Content Determination Method

[0339] Analysis was performed using an Agilent 1260 liquid chromatograph (equipped with a UV detector). The column was C18 (250 mm × 4.6 mm, 5 μm). The injection volume was 5 μL. The detection wavelength was 403 nm, and the flow rate was 1 mL / min. Mobile phase A was 0.5% phosphoric acid aqueous solution, and phase B was acetonitrile, with a gradient desorption process (0–10 min, 5% → 11% B; 10–16 min, 11% → 14% B; 16–23 min, 14% B; 23–30 min, 14% → 20% B; 30–60 min, 20% → 31% B; 30–60 min, 20% → 31% B; 60–65 min, 31% → 90% B; 65–67 min, 90% → 5% B).

[0340] 12.4 Experimental Results

[0341] The effects of different sucrose concentrations on the extraction rates of key components in safflower are shown in Table 14.

[0342] Table 14 Effects of different sucrose concentrations on the extraction rate of indicator components in safflower

[0343]

[0344] * P < 0.05 vs safflower aqueous extract

[0345] As can be seen from Table 14, sucrose concentrations ranging from 0.5 g / 100 mL to 2.5 g / 100 mL can increase the content of indicator components in safflower to varying degrees, with the most significant effects observed at sucrose concentrations of 0.5 g / 100 mL or 2 g / 100 mL.

[0346] The effects of different types of disaccharides on the extraction rate of index components in safflower are shown in Table 15.

[0347] Table 15 Effects of different types of disaccharides on the extraction rate of index components in safflower

[0348]

[0349] * P < 0.05 vs safflower aqueous extract

[0350] As can be seen from Table 15, a sucrose concentration of 0.5 g / 100 mL can significantly increase the content of indicator components in safflower.

[0351] Example 13: Extraction Experiment of Different Sugars-Astragalus Polysaccharides

[0352] 13.1 Preparation methods of different sugar-Astragalus extracts

[0353] Five portions of Astragalus membranaceus slices from the same batch, each weighing 50g, were added to the following solutions: 7 times the volume of water (350ml), 1% sucrose solution (3.5g sucrose diluted to 350ml), 1% glucose solution (3.5g glucose diluted to 350ml), 1% fructose solution (3.5g fructose diluted to 350ml), 1% trehalose solution (3.5g trehalose diluted to 350ml), 1% lactose solution (3.5g lactose diluted to 350ml), and 1% maltose solution (3.5g maltose diluted to 350ml). Soak the herbs in 50ml of water for 30 minutes, reflux for 30 minutes, and filter while hot through gauze. Add 6 times the amount of water (300ml of water), 1% sucrose solution (3.0g sucrose diluted with water to 300ml), 1% glucose solution (3.0g glucose diluted with water to 300ml), 1% fructose solution (3.0g fructose diluted with water to 300ml), 1% trehalose solution (3.0g trehalose diluted with water to 300ml), 1% lactose solution (3.0g lactose diluted with water to 300ml), and 1% maltose solution (3g maltose diluted with water to 300ml) to the reflux solution for 20 minutes, filter while hot, and combine the two filtrates to obtain different sugar-Astragalus extracts.

[0354] 13.2 Determination of total polysaccharide content

[0355] 13.2.1 Preparation method of test solution

[0356] Accurately measure 1 ml of Astragalus membranaceus aqueous extract and place it in a 15 mL centrifuge tube. Add 4 mL of anhydrous ethanol, mix well, and incubate for 12 h at 7000 r·min. -1 Centrifuge for 20 min, discard the supernatant, dissolve the precipitate in water, dilute to a volumetric flask with water, and shake well to obtain the final product.

[0357] 13.2.2 Preparation method of reference solution

[0358] Take an appropriate amount of anhydrous glucose reference standard, accurately weigh it, and add water to prepare a solution containing 0.132 mg of anhydrous glucose per 1 ml.

[0359] 13.2.3 Selection of Measurement Wavelength

[0360] Accurately measure 0.6 ml of the test solution and place it in a 10 ml stoppered graduated test tube. Add water to each tube to a final volume of 2.0 ml, shake well, and then accurately add 1 ml of 5% phenol solution to each tube. Vortex for 20 seconds, then add 5 ml of concentrated sulfuric acid and vortex for 20 seconds. Incubate at 80°C for 20 minutes, then remove and quickly cool in an ice-water bath for 10 minutes. Scan the sample in the wavelength range of 400–800 nm. A maximum absorption peak is observed at 490 nm; therefore, 490 nm is selected as the wavelength for determining the total polysaccharide content in the Astragalus membranaceus aqueous extract.

[0361] 13.2.4 Plotting the Standard Curve

[0362] Accurately measure 0.1 ml, 0.2 ml, 0.4 ml, 0.6 ml, 0.8 ml, and 1 ml of the reference solution into separate 10 ml stoppered graduated test tubes. Add water to each tube to a final volume of 2.0 ml, shake well, and then accurately add 1 ml of 5% phenol solution to each. Vortex for 20 seconds. Add 5 ml of concentrated sulfuric acid and vortex for 20 seconds. Incubate in an 80°C water bath for 20 minutes, then remove and quickly cool in an ice-water bath for 10 minutes. Use the corresponding reagents as blanks. Measure the absorbance at 490 nm using UV-Vis spectrophotometry (General Rule 0401). Plot a standard curve with absorbance on the ordinate and concentration on the abscissa to obtain the linear equation.

[0363] 13.3 Experimental Results

[0364] The results of the determination of total polysaccharide content in different sugar-Astragalus extracts are shown in Table 16.

[0365] Table 16. Results of determination of total polysaccharide content in different sugar-Astragalus extracts

[0366]

[0367] *P < 0.05 VS Astragalus membranaceus aqueous extract

[0368] As can be seen from Table 16, sucrose, glucose, fructose, trehalose, lactose, and maltose (1 g / 100 mL concentration) can all increase the polysaccharide content in Astragalus membranaceus to varying degrees, with sucrose having the most significant effect.

[0369] Example 14: Extraction Experiment of Astragalus Polysaccharides at Different Concentrations

[0370] 14.1 Preparation methods of sucrose-Astragalus extracts of different concentrations

[0371] Seven portions of raw Astragalus membranaceus slices, each weighing 50 g, were taken from the same batch. Seven times the volume of water (350 ml) was added to each portion, along with the following solutions: 1% sucrose solution (3.5 g sucrose diluted to 350 ml), 2% sucrose solution (7 g sucrose diluted to 350 ml), 10% sucrose solution (35 g sucrose diluted to 350 ml), 20% sucrose solution (70 g sucrose diluted to 350 ml), 30% sucrose solution (105 g sucrose diluted to 350 ml), and 40% sucrose solution (140 g sucrose diluted to 350 ml). Each solution was soaked for 30 minutes, then refluxed for 30 minutes. The mixture was then filtered through gauze while still hot. The residue was then further processed separately. Add 6 times the volume of water (300ml), 1% sucrose aqueous solution (3.0g sucrose diluted with water to 300ml), 2% sucrose aqueous solution (6.0g sucrose diluted with water to 300ml), 10% sucrose aqueous solution (30g sucrose diluted with water to 300ml), 20% sucrose aqueous solution (60g sucrose diluted with water to 300ml), 30% sucrose aqueous solution (90g sucrose diluted with water to 300ml), and 40% sucrose aqueous solution (120g sucrose diluted with water to 300ml). Reflux for 20 minutes, filter while hot, and combine the two filtrates to obtain sucrose-Astragalus extracts of different concentrations.

[0372] 14.2 Determination of total polysaccharide content

[0373] Same as Example Thirteen.

[0374] 14.3 Experimental Results

[0375] The results of the determination of total polysaccharide content in Astragalus extract obtained with different concentrations of sucrose are shown in Table 17.

[0376] Table 17. Results of total polysaccharide content determination in Astragalus extracts obtained with different concentrations of sucrose.

[0377]

[0378] * P < 0.05 VS Astragalus membranaceus aqueous extract

[0379] As can be seen from Table 17, different concentrations of sucrose solutions (1-40 g / 100 mL) can all increase the content of the effective components (astragalus polysaccharides) in Astragalus to varying degrees, with the 30 g / 100 mL sucrose solution showing the most significant effect.

[0380] Example 15: Extraction Experiment of Sucrose-Glycyrrhiza Polysaccharide at Different Concentrations

[0381] 15.1 Preparation methods of sucrose-licorice extracts of different concentrations

[0382] Seven portions of raw licorice slices (produced in Xinjiang Uygur Autonomous Region) were taken from the same batch, each portion weighing 50 g. Add 7 times the amount of water (350ml), 1% sucrose solution (3.5g sucrose + 350ml water), 2% sucrose solution (7.0g sucrose + 350ml water), 10% sucrose solution (35g sucrose + 350ml water), 20% sucrose solution (70g sucrose + 350ml water), 30% sucrose solution (105g sucrose + 350ml water), and 40% sucrose solution (140g sucrose + 350ml water) to the dregs respectively. Soak for 30 minutes, reflux for 30 minutes, filter with gauze while hot, and then add 6 times the amount of water (300ml water), 1% sucrose solution (3.0g sucrose + 300ml water), 2% sucrose solution (6.0g sucrose + 300ml water), 10% sucrose solution (30g sucrose + 300ml water), and 20% sucrose solution (60g sucrose + 300ml water) to the dregs respectively. 90 g sucrose + 300 ml water), 30% sucrose aqueous solution (90 g sucrose + 300 ml water), 40% sucrose aqueous solution (120 g sucrose + 300 ml water), reflux for 20 min, filter while hot, combine the two filtrates to obtain the final product.

[0383] 15.2 Determination of total polysaccharide content

[0384] 15.2.1 Preparation of the test solution

[0385] Accurately measure 2 ml of licorice water extract, add 8 ml of ethanol, mix well, and soak for 12 hours at 5000 rpm. -1 Centrifuge for 20 minutes, discard the supernatant, dissolve the precipitate in water, dilute to a volumetric flask with water, and shake well to obtain the final product.

[0386] 15.2.2 Preparation of reference solution

[0387] Take an appropriate amount of anhydrous glucose reference standard, accurately weigh it, and add water to prepare a solution (10 ml) containing 0.34 mg of anhydrous glucose per ml.

[0388] 15.2.3 Selection of Measurement Wavelength

[0389] Accurately measure 1.0 ml of the test solution and place it in a 10 ml stoppered graduated test tube. Add water to each tube to a final volume of 2.0 ml, shake well, and slowly add 8 ml of 0.2% anthrone-sulfuric acid solution. Mix well, incubate in a boiling water bath for 10 minutes, remove, and immediately cool in an ice-water bath for 10 minutes. Scan the sample in the wavelength range of 400–800 nm. A maximum absorption peak is observed at 593 nm; therefore, 593 nm is selected as the wavelength for determining the total polysaccharide content in licorice aqueous extract.

[0390] 15.2.4 Plotting the Standard Curve

[0391] Accurately measure 0.1 ml, 0.2 ml, 0.4 ml, 0.6 ml, 0.8 ml, and 1 ml of the reference solution into separate 10 ml stoppered graduated test tubes. Add water to each tube to a final volume of 2.0 ml, shake well, and slowly add 8 ml of 0.2% anthrone-sulfuric acid solution. Mix well, incubate in a boiling water bath for 10 minutes, remove, and immediately cool in an ice-water bath for 10 minutes. Use the corresponding reagents as blanks. Measure the absorbance at 593 nm using ultraviolet-visible spectrophotometry (General Rule 0401). Plot a standard curve with absorbance on the ordinate and concentration on the abscissa to obtain the linear equation.

[0392] 15.3 Experimental Results

[0393] The results of the determination of total polysaccharide content in different sugar-licorice aqueous extracts are shown in Table 18.

[0394] Table 18. Results of determination of total polysaccharide content in licorice concentrates obtained with different concentrations of sucrose.

[0395]

[0396] * P < 0.05 VS Licorice aqueous extract

[0397] As can be seen from Table 18, different concentrations of sucrose solutions (1-40 g / 100 mL) can all increase the content of active ingredients (glycyrrhiza polysaccharides) in licorice to varying degrees, with the 20 g / 100 mL sucrose solution showing the most significant effect.

[0398] Example 16: Extraction Experiment of Different Sugars-Licorice Polysaccharides

[0399] 16.1 Preparation methods of different sugar-licorice extracts

[0400] Take three portions (50g each) of raw licorice root slices from the same batch (produced in Xinjiang Uygur Autonomous Region). Add 7 times the volume of water (350ml), a 20% maltose solution (70g maltose + 350ml water), and a 20% sucrose solution (70g lactose + 350ml water) to each portion, respectively. Soak for 30 minutes, reflux for 30 minutes, and filter while hot through gauze. Add 6 times the volume of water (300ml water), a 20% maltose solution (60g maltose + 300ml water), and a 20% sucrose solution (60g lactose + 300ml water) to each portion of the residue, respectively. Reflux for 20 minutes, filter while hot, and combine the two filtrates. The desired result is obtained.

[0401] 16.2 Determination of total polysaccharide content

[0402] Same as Example 15.

[0403] 16.3 Experimental Results

[0404] The results of the determination of total polysaccharide content in different sugar-licorice aqueous extracts are shown in Table 19.

[0405] Table 19 Results of determination of total polysaccharide content in different sugar-licorice aqueous extracts

[0406]

[0407] * P < 0.05 VS Licorice aqueous extract

[0408] As can be seen from Table 19, both sucrose and maltose (20 g / 100 mL concentration) can increase the content of active ingredients (licorice polysaccharides) in licorice to varying degrees, with maltose showing the most significant effect.

[0409] Example 17: Extraction Experiment of Sucrose-Ginseng Polysaccharide at Different Concentrations

[0410] 17.1 Preparation methods of sucrose-ginseng extracts of different concentrations

[0411] Take 6 portions of ginseng slices from the same batch (Fusong, Jilin), each 40 g. Add 7 times the amount of water and 1%, 10%, 20%, 25%, and 30% sucrose aqueous solutions to each portion, respectively. Soak for 30 min, reflux for 30 min, and filter while hot through gauze. Add 6 times the amount of water and 1%, 10%, 20%, 25%, and 30% sucrose aqueous solutions to the residue, respectively, reflux for 20 min, and filter while hot through two layers of gauze. Combine the two filtrates, add water to make up to 800 mL, and the product is ready.

[0412] 17.2 Determination of total polysaccharide content

[0413] 17.2.1 Preparation method of test solution

[0414] Accurately measure 1 mL of ginseng extract with different sucrose concentrations, add 4 mL of anhydrous ethanol to each, mix well, and soak for 12 h at 7000 r·min. -1 Centrifuge for 20 min, discard the supernatant, dissolve the precipitate in water, dilute to 10 mL in a volumetric flask, and shake well to obtain the final product.

[0415] 17.2.2 Preparation method of reference solution

[0416] Take an appropriate amount of anhydrous glucose reference standard, accurately weigh it, and add water to prepare a solution containing 0.132 mg of anhydrous glucose per 1 mL.

[0417] 17.2.3 Determination of total polysaccharide content

[0418] Accurately measure 0.4, 0.4, 0.3, 0.3, 0.3, and 0.2 mL of ginseng extract test solutions with different sucrose concentrations and place them in 10 mL stoppered graduated test tubes. Add water to each tube to a final volume of 2.0 mL, shake well, and then accurately add 1 mL of 5% phenol solution to each tube. Vortex for 20 s. Add 5 mL of concentrated sulfuric acid and vortex for 20 s. Incubate in an 80°C water bath for 20 min, then remove and quickly cool in an ice-water bath for 10 min. Use the corresponding reagent (water) as a blank. Measure the absorbance at a wavelength of 489 nm.

[0419] 17.2.4 Plotting the Standard Curve

[0420] Accurately measure 0.1 mL, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1 mL of the reference solution into separate 10 mL stoppered graduated test tubes. Add water to each tube to a final volume of 2.0 mL, shake well, and then accurately add 1 mL of 5% phenol solution to each. Vortex for 20 s. Add 5 mL of concentrated sulfuric acid and vortex for 20 s. Incubate in an 80°C water bath for 20 min, then remove and immediately cool in an ice-water bath for 10 min. Use the corresponding reagents as blanks. Measure the absorbance at 489 nm using UV-Vis spectrophotometry (General Rule 0401). Plot a standard curve with absorbance on the ordinate and concentration on the abscissa.

[0421] 17.3 Experimental Results

[0422] The effects of different sucrose concentrations on the extraction rate of ginseng polysaccharides are shown in Table 20.

[0423] Table 20 Effect of different sucrose concentrations on the extraction rate of ginseng polysaccharides

[0424]

[0425] * P < 0.05 VS Ginseng aqueous extract

[0426] As can be seen from Table 20, sucrose solutions of different concentrations (10-40 g / 100 mL) can significantly increase the content of active ingredients in ginseng to varying degrees.

[0427] Example 18: Extraction Experiment of Different Sugars-Ginseng Polysaccharides

[0428] 18.1 Preparation methods of different types of disaccharide-ginseng extracts

[0429] Take 3 portions of ginseng slices from the same batch (Fusong, Jilin), each 40 g. Add 7 times the amount of water, 30% sucrose, and 30% maltose aqueous solution to each portion, and soak for 30 minutes. The subsequent operation is the same as in Example 17.

[0430] 18.2 Determination of total polysaccharide content

[0431] Same as Example 17.

[0432] 18.3 Experimental Results

[0433] The effects of different types of disaccharides on the extraction rate of total ginseng polysaccharides are shown in Table 21.

[0434] Table 21 Effects of different types of disaccharides on the extraction rate of total ginseng polysaccharides

[0435]

[0436] * P < 0.05 VS Ginseng aqueous extract

[0437] As can be seen from Table 21, both sucrose and maltose (30 g / 100 mL concentration) can increase the content of active ingredients (ginseng polysaccharides) in ginseng to varying degrees, with sucrose having the most significant effect.

[0438] Example 19: Extraction Experiment of Sucrose-Lycium barbarum Polysaccharides at Different Concentrations

[0439] 19.1 Preparation method of wolfberry extract

[0440] Six portions of the same batch of goji berries (originating in Ningxia) were taken, each portion containing 40 g. Add 7 times the volume of water (280 ml water), 1% sucrose solution (2.8 g sucrose + 280 ml water), 10% sucrose solution (28 g sucrose + 280 ml water), 20% sucrose solution (56 g sucrose + 280 ml water), 30% sucrose solution (84 g sucrose + 280 ml water), 40% sucrose solution (112 g sucrose + 280 ml water), and 30% maltose solution (84 g maltose + 280 ml water) to the residue, soak for 30 min, reflux for 30 min, filter while hot through gauze, and then add 6 times the volume of water, 1% sucrose solution, 10% sucrose solution, 20% sucrose solution, 30% sucrose solution, 40% sucrose solution, and 30% maltose solution to the residue, reflux for 20 min, filter while hot through two layers of gauze, combine the two filtrates, and add water to make up to 800 mL.

[0441] 19.2 Determination of total polysaccharide content

[0442] 19.2.1 Preparation method of test solution

[0443] The preparation method is the same as that of the test solution in Example 17.

[0444] 19.2.2 Preparation of reference solution

[0445] The preparation method is the same as that for the reference solution in Example 17.

[0446] 19.2.3 Determination of total polysaccharide content

[0447] Accurately measure 0.4, 0.4, 0.4, 0.3, and 0.3 mL of wolfberry extract with different sucrose concentrations into 10 mL stoppered graduated test tubes. Add water to each tube to a final volume of 2.0 mL, shake well, and then accurately add 1 mL of 5% phenol solution. Vortex for 20 s. Add 5 mL of concentrated sulfuric acid and vortex for 20 s. Incubate in an 80°C water bath for 20 min, then remove and quickly cool in an ice-water bath for 10 min. Use the corresponding reagent (water) as a blank. Measure the absorbance at a wavelength of 489 nm.

[0448] 19.2.4 Plotting the Standard Curve

[0449] The method for plotting the standard curve is the same as in Example 17.

[0450] 19.3 Experimental Results

[0451] The effects of different sucrose concentrations on the extraction rate of Lycium barbarum polysaccharides are shown in Table 22.

[0452] Table 22 Effect of different sucrose concentrations on the extraction rate of Lycium barbarum polysaccharides

[0453]

[0454] * P < 0.05 VS Lycium barbarum aqueous extract

[0455] As can be seen from Table 22, sucrose solutions of different concentrations (10-40 g / 100 mL) can all increase the content of active ingredients (goji polysaccharides) in goji berries to varying degrees, with the 40 g / 100 mL sucrose solution showing the most significant effect.

[0456] Example 20: Extraction Experiment of Different Sugars-Lycium barbarum Polysaccharides

[0457] 20.1 Preparation methods of different types of disaccharide-lycium barbarum extract

[0458] Take three portions of the same batch of wolfberry slices (origin: Ningxia; manufacturer: Tianjin Shengshi Pharmaceutical Co., Ltd.; batch number: X20122702A), each weighing 40 g. Add 7 times the amount of water, 30% sucrose, and 30% maltose aqueous solution to each portion, and soak for 30 minutes. The subsequent procedures are the same as in Example 19.

[0459] 20.2 Determination of total polysaccharide content

[0460] Same as Example 19.

[0461] 20.3 Experimental Results

[0462] The effects of different types of disaccharides on the extraction rate of Lycium barbarum polysaccharides are shown in Table 23.

[0463] Table 23 Effects of different types of disaccharides on the extraction rate of Lycium barbarum polysaccharides

[0464]

[0465] * P < 0.05 VS Lycium barbarum aqueous extract

[0466] As can be seen from Table 23, both sucrose and maltose (30 g / 100 mL concentration) can increase the content of effective components (goji polysaccharides) in goji berries to varying degrees.

[0467] The above descriptions are merely embodiments of the present invention, and common knowledge regarding specific structures and characteristics is not elaborated upon here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the present invention or the practicality of the patent. The scope of protection claimed by the present invention should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

[0468] The embodiments of the present invention have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the present invention. The descriptions of the embodiments above are only for the purpose of helping to understand the method and core ideas of the present invention. Furthermore, any changes or modifications made by those skilled in the art based on the ideas of the present invention, its specific implementation methods, and its application scope, are all within the scope of protection of the present invention. Therefore, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. A method for increasing the leaching of carbohydrate components and / or glycoside components, characterized in that, The method includes the following steps: (1) Prepare a sucrose solution of 0.95~1.05 g / 100 mL or 10~30 g / 100 mL; (2) Soaking the raw materials in the 0.95~1.05 g / 100 mL or 10~30 g / 100 mL sucrose solution; and (3) The raw material is heated and extracted 1-3 times in the sucrose solution of 0.95~1.05 g / 100 mL or 10~30 g / 100 mL, and the raw material extract is obtained by filtration; The concentration of the sucrose solution extracted from the sugar components is increased to 10-30 g / 100 mL, and the sugar components are astragalus polysaccharide, licorice polysaccharide, ginseng polysaccharide or wolfberry polysaccharide; The concentration of the sucrose solution extracted from the glycosides is increased to 0.95~1.05 g / 100 mL. The glycosides are verbascoside, gentianoside, astragaloside A, apigenin glycyrrhizin, glycyrrhizin, glycyrrhizic acid, ginsenoside Rg1, anemarrhenaside BII, lebodiin, or hydroxysafflower yellow A.

2. The method according to claim 1, characterized in that, The raw materials are those containing sugar components and / or glycoside components.

3. The method according to claim 2, characterized in that, The raw materials are one or more traditional Chinese medicines containing sugar components and / or glycoside components.

4. The method according to claim 2, characterized in that, The raw material is one or more herbal medicines containing sugar components and / or glycoside components.

5. The method according to claim 2, characterized in that, The raw material is one or more marine organisms containing sugars and / or glycosides.

6. The method according to claim 3, characterized in that, The Chinese medicinal herbs are selected from one or more of the following: Astragalus membranaceus, Glycyrrhiza uralensis, Ginseng, Panax notoginseng, Lycium barbarum, Anemarrhena asphodeloides, Stevia rebaudiana, and Carthamus tinctorius.

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

8. The method according to claim 1, characterized in that, In step (2), the soaking step includes soaking the raw material in the 0.95~1.05 g / 100 mL or 10~30 g / 100 mL sucrose solution at room temperature for 10-120 min.

9. The method according to claim 1, characterized in that, In step (2), the soaking step includes soaking the raw material in the 0.95~1.05 g / 100 mL or 10~30 g / 100 mL sucrose solution at room temperature for 20-60 min.

10. The method according to claim 1, characterized in that, In step (2), the soaking step includes soaking the raw material in the 0.95~1.05 g / 100 mL or 10~30 g / 100 mL sucrose solution at room temperature for 28.5~31.5 min.

11. The method according to claim 1, characterized in that, In step (2), the weight-to-volume ratio of the raw material to the 0.95~1.05g / 100mL or 10~30g / 100mL sucrose solution is 1:1~1:

50.

12. The method according to claim 1, characterized in that, In step (2), the weight-to-volume ratio of the raw material to the 0.95~1.05g / 100mL or 10~30g / 100mL sucrose solution is 1:1~1:

20.

13. The method according to claim 1, characterized in that, In step (2), the weight-to-volume ratio of the raw material to the 0.95~1.05g / 100mL or 10~30g / 100mL sucrose solution is 1:6~1:

12.

14. The method according to claim 1, characterized in that, In step (3), the heating extraction is a heating reflux extraction.

15. The method according to claim 1, characterized in that, In step (3), the heating time is 20 min to 60 min.

16. The method according to claim 1, characterized in that, In step (3), the heating time is 20 min to 30 min.

17. The method according to claim 1, characterized in that, In step (3), the heating time is 19~21 min or 28.5~31.5 min.

18. Use of the method according to any one of claims 1 to 3, 6 to 17 in the preparation of raw material extract, or raw material concentrate, or pharmaceutical composition containing the raw material, or pharmaceutical preparation or functional food or health food containing the raw material, wherein the raw material is Astragalus membranaceus, Glycyrrhiza uralensis, Ginseng, Lycium barbarum, Panax notoginseng, Anemarrhena asphodeloides, Stevia rebaudiana, or Carthamus tinctorius.

19. The use of the method according to any one of claims 1, 2, 4, 7 to 17 in the preparation of a raw material extract, or a raw material concentrate, or a pharmaceutical composition containing the raw material, or a pharmaceutical preparation or functional food or health food containing the raw material, wherein the raw material is one or more herbal medicines containing sugar components and / or glycoside components.

20. The use of the method according to any one of claims 1, 2, 5, 7 to 17 in the preparation of a raw material extract, or a raw material concentrate, or a pharmaceutical composition containing the raw material, or a pharmaceutical preparation or functional food or health food containing the raw material, wherein the raw material is one or more marine organisms containing carbohydrate components and / or glycoside components.

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