CN117327135A - Method for increasing leaching of saccharides and glycoside components and application thereof - Google Patents

Method for increasing leaching of saccharides and glycoside components and application thereof Download PDF

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CN117327135A
CN117327135A CN202210730365.4A CN202210730365A CN117327135A CN 117327135 A CN117327135 A CN 117327135A CN 202210730365 A CN202210730365 A CN 202210730365A CN 117327135 A CN117327135 A CN 117327135A
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sucrose
water
solution
compound
glycoside
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代云桃
李鹏跃
薛晓霞
赵金荣
孔沙沙
郝丽霞
陈士林
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Institute of Materia Medica of CAMS
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Priority to CN202210730365.4A priority Critical patent/CN117327135A/en
Priority to JP2023104245A priority patent/JP2024002989A/en
Publication of CN117327135A publication Critical patent/CN117327135A/en
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    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/26—Acyclic or carbocyclic radicals, substituted by hetero rings
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    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
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    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D309/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only ring hetero atom, not condensed with other rings
    • C07D309/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only ring hetero atom, not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
    • C07D309/08—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only ring hetero atom, not condensed with other rings having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D309/10—Oxygen atoms
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    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • C07H1/06—Separation; Purification
    • C07H1/08—Separation; Purification from natural products
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    • C07H15/20—Carbocyclic rings
    • C07H15/24—Condensed ring systems having three or more rings
    • C07H15/244—Anthraquinone radicals, e.g. sennosides
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    • C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
    • C07H17/06—Benzopyran radicals
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    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

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Abstract

The invention discloses a method for increasing leaching 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 the raw material in the sugar solution and/or the sugar alcohol solution; and (3) heating and extracting the raw materials in the sugar solution and/or the sugar alcohol solution for 1-3 times, and filtering to obtain an extracting solution. The method can obviously increase the extraction rate of saccharide components and/or glycoside components in the raw materials, thereby being beneficial to improving the utilization rate and the curative effect of the raw materials.

Description

Method for increasing leaching of saccharides and glycoside components and application thereof
Technical Field
The invention relates to the field of chemical component extraction, in particular to a method for increasing leaching of saccharides and glycoside components and application thereof.
Background
At present, how to extract active/effective components such as saccharide components and/or glycoside components in raw materials in a green and efficient way, and ensuring the safety and effectiveness of the extract is an important content in the extraction research field of raw materials such as traditional Chinese medicines, plant medicines and the like. Modern researches have found that the ethanol extraction efficiency of most of the components of the traditional Chinese medicine is far higher than that of water decoction, but the toxicity of the obtained extract may be increased. In registration and declaration of new traditional Chinese medicines newly released in 2020, if the declared indication is consistent with the traditional usage, clinical tests can be moderately reduced or exempted, for example, in 2018, the application range of clinical tests can be avoided in the development of the compound preparation of the ancient classical prescription, which is defined in the simplified registration approval management of the compound preparation of the ancient classical prescription, published in 2018, including the consistency of preparation process and ancient medical records. The preparation process of the traditional Chinese medicine mostly adopts a water decoction mode, and clinical experience and modern researches show that the method can ensure the safety of the extract better than an organic solvent extraction method. The main reasons for the difference between the two extraction methods are: the traditional Chinese medicine contains various active ingredients and has wide polarity range, and the extraction rate of the weak-polarity and medium-polarity ingredients is increased by ethanol extraction, so that the composition of the ethanol extract is different from that of the active ingredients in the traditional clinically used decoction, and the safety of the extract is possibly influenced to a certain extent. In addition, the most common water decoction method for traditional Chinese medicine extraction has the problems of low extraction rate, incomplete extraction and the like. Although increasing the amount of water contributes to an increase in the extraction yield of the active ingredient, the concentration time is increased and the production cost is increased.
Therefore, how to effectively improve the extraction rate of active ingredients in raw materials such as Chinese medicine extracts in a green manner on the ancient basis is an important research direction in raw materials such as Chinese medicine extraction research.
Disclosure of Invention
Based on the above, the invention provides a method for increasing the leaching of saccharide components and/or glycoside components by using low-concentration saccharide compound aqueous solution, which adopts sugar water and/or sugar alcohol water with a certain concentration as an extraction solvent instead of water, and compared with the traditional decoction method for extraction, the extraction rate is remarkably improved, thereby improving the utilization rate and the curative effect of raw materials such as traditional Chinese medicines. The sugar used in the research is common auxiliary materials of traditional Chinese medicines or edible sugar, so that the sugar has the characteristics of safety, low price and the like, and does not influence the safety of the extract.
Specifically, according to one aspect of the present invention, there is provided a method of increasing leaching of a saccharide component and/or a glycoside component, the method comprising the steps of:
(1) Preparing a sugar solution and/or a sugar alcohol solution;
(2) Soaking the raw material in the sugar solution and/or the sugar alcohol solution; and
(3) Heating and extracting the raw materials in the sugar solution and/or sugar alcohol solution for 1-3 times, and filtering to obtain raw material extract.
Further, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-40g/100mL. Further, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-10g/100mL or 20-40g/100mL. Further, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-5g/100mL or 20-30g/100mL. Further, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-2.5g/100mL. Further, the concentration of the sugar solution and/or the sugar alcohol solution is 1-2g/100mL. Further, the concentration of the sugar solution and/or the sugar alcohol solution is about 0.5g/100mL or about 1g/100mL or about 1.5g/100mL or about 2g/100mL or about 20g/100mL or about 30g/100mL or about 40g/100mL.
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, sorbose, rhamnose, ribose, xylose and deoxyribose. Further, the disaccharide is selected from one or more of maltose, sucrose, lactose, trehalose, melibiose, gentiobiose and trabiose. Further, the monosaccharide is glucose and/or fructose. Further, the trisaccharide is raffinose. Further, the disaccharide is sucrose and/or maltose.
Further, the saccharide component is an oligosaccharide component and/or a polysaccharide component.
Further, the polysaccharide component is selected from one or more of the following: astragalus polysaccharides, licorice polysaccharides, ginseng polysaccharides and wolfberry polysaccharides.
Further, the glycoside component is selected from one or more of the following: flavonoid glycoside compounds, saponins compounds, diterpenoid glycoside compounds, quinone glycoside compounds and phenolic acid glycoside compounds.
Further, the flavonoid glycoside compound is a flavonoid glycoside compound, an isoflavone glycoside compound, a flavanone glycoside compound, a flavanonol glycoside compound, a chalcone glycoside compound, a dihydrochalcone glycoside compound, an hesperidin compound, an anthocyanin compound, a biflavone glycoside compound and a flavanol glycoside compound.
Further, the saponin compounds are tetracyclic triterpene glycoside compounds, pentacyclic triterpene glycoside compounds and/or steroid glycoside compounds.
Further, the diterpenoid glycoside compound 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.
Further, the quinone glycoside compound is benzoquinone glycoside compound, naphthoquinone glycoside compound, phenanthrenequinone glycoside compound and/or anthraquinone glycoside compound.
Further, the phenolic acid glycoside compound is a phenolic acid glycoside compound with a C6-C3 parent nucleus structure and/or a phenolic acid glycoside compound with a C6-C1 parent nucleus structure.
Further, the steroid glycoside compound is a spirostanol type saponin compound, a furostanol type saponin compound and/or an isospirostanol type saponin compound.
Further, the isoparaffin type saponin compound is a dioscin type compound.
Further, the furostanol type saponin compound is a timosaponin type compound.
Further, the tetracyclic triterpene glycoside compound is lanolin alkane type tetracyclic triterpene glycoside compound and/or dammarane type tetracyclic triterpene glycoside compound.
Further, the pentacyclic triterpene glycoside compound is oleanane type pentacyclic triterpene saponin compound, ursane type pentacyclic triterpene saponin compound and/or lupin alkane type pentacyclic triterpene saponin compound.
Further, the anthraquinone glycoside compound is a monoanthracene nucleus compound and/or a dianthracene nucleus compound.
Further, the isoflavone glycoside compound is calycosin glucoside and/or formononetin.
Further, the flavanone glycoside compound is apigenin and/or liquiritin. Further, the chalcone glycoside compound is hydroxysafflor yellow A. Further, the furostanol-type steroid saponin compound is timosaponin BII. Further, the lanostane type tetracyclic triterpenoid saponin compound is astragaloside IV. Further, the dammarane type tetracyclic triterpene saponin compound is ginsenoside Rg1. Further, the oleanane type pentacyclic triterpenoid saponin compound is glycyrrhizic acid. Further, the tetracyclic diterpenoid glycoside compound is stevioside and/or rebaudioside. Further, the phenolic acid glycoside compound with a C6-C3 parent nucleus structure is hydroxysafflor yellow A. Further, the dianthrone nucleoside compound is sennoside A and/or sennoside B. Further, the monoanthracene nucleoside compound is aloin.
Further, the raw material is a raw material containing a saccharide component and/or a glycoside component.
Further, the raw material is one or more Chinese medicines, plant medicines and/or marine organisms containing saccharide components and/or glycoside components.
Further, the traditional Chinese medicine and/or the botanical is selected from one or more of the following: ginseng, notoginseng, licorice, medlar, stevia rebaudiana, astragalus, senna, cortex albiziae, pokeberry root, bupleurum, pulsatilla root, cedrela sinensis, dendrobium huoshanense, achyranthes root, clematis root, american ginseng, bolbostemma paniculatum, horse chestnut, acanthopanax root, chestnut of a senior, gynostemma pentaphylla, chinaberry, centella asiatica, radix sedge, radix dipsaci, akebia, rhizoma panacis, lysimachia foenum-graecum, clematis stem, wild jujube leaf, cannabis sativa, polygala tenuifolia, east perilla, eclipta, platycodon grandiflorum, dwarf lilyturf tuber, allium macrostemon, rhizoma polygonati, rhizoma anemarrhenae, radix asparagi, dwarf lilyturf, rhizoma paridis, glabrous greenbrier, rhizoma dioscoreae, tribulus terrestris, purple eggplant, pomegranate, almond, sweet potato, yam, asparagus, largehead atractylodes rhizome, cortex lycii radicis, purple sweet potato and resina.
Further, in step (2), the soaking is a room temperature soaking.
Further, the immersing step includes immersing the raw material in the sugar solution and/or the sugar alcohol solution at room temperature for 10 to 120 minutes.
Further, the immersing step includes immersing the raw material in the sugar solution and/or the sugar alcohol solution at room temperature for 20 to 60 minutes.
Further, the step of immersing includes immersing the raw material in the sugar solution and/or the sugar alcohol solution at room temperature for about 30 minutes.
Further, the weight-to-volume ratio of the raw material to the sugar solution and/or the sugar alcohol solution is 1:1-1:50.
Further, the weight-to-volume ratio of the raw material to the sugar solution and/or the sugar alcohol solution is 1:1-1:20.
Further, the weight-to-volume ratio of the raw material to the sugar solution and/or the sugar alcohol solution is 1:6-1:12.
Further, in step (3), the heat extraction is a heat reflux extraction.
Further, the heating time is 20min to 60min.
Further, the heating time is 20min to 30min.
Further, the heating time is about 20 minutes or about 30 minutes.
According to one aspect of the present invention there is provided a use of a method as described above for the preparation of a raw material extract or raw material concentrate, wherein the raw material is a traditional Chinese medicine, a plant drug and/or a marine organism.
Further, the raw material concentrated solution is the raw material extract solution obtained in the step (3) or the raw material total extract solution obtained in the step (5) described above, and is obtained by concentrating in a water bath at 20 to 100 ℃.
Further, the concentration is accomplished in a water bath at 60 ℃ to 80 ℃.
Further, the concentration is accomplished in a water bath at about 70 ℃.
According to one aspect of the present invention there is provided a use according to the above method for the preparation of a pharmaceutical composition comprising a raw material, wherein the raw material is a traditional Chinese medicine, a plant drug and/or a marine organism.
According to one aspect of the present invention there is provided a use of a method according to the above in the preparation of a pharmaceutical formulation or functional or health food comprising a raw material, wherein the raw material is a traditional Chinese medicine, a plant drug and/or a marine organism.
The invention has the beneficial effects that:
we have found that the process of the present invention can substantially increase the extraction rate of saccharide and/or glycoside components from raw materials such as Chinese medicinal materials or herbal medicines, compared with conventional water decoction methods.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely, and it is apparent that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other. The present invention will be described in detail with reference to examples.
The invention is described in further detail below in connection with specific examples which are not to be construed as limiting the scope of the invention as claimed.
As described in the background section, the composition of the alcohol extract is different from that of the effective components in the decoction for traditional clinical use, so that the safety of the alcohol extract can be influenced to a certain extent, but the extraction efficiency of the traditional water decoction is not high. In order to solve the above problems, the present invention provides a method for increasing leaching of a sugar component and/or a glycoside component by using a sugar, the method comprising the steps of:
(1) Preparing a sugar solution and/or a sugar alcohol solution;
(2) Soaking the raw material in the sugar solution and/or the sugar alcohol solution; and
(3) Extracting the raw material in the sugar solution and/or the sugar alcohol solution by heating, and filtering to obtain a raw material extracting solution.
Sugar solutions and/or sugar alcohol solutions are capable of increasing the extraction yield of saccharide components and/or glycoside components in raw materials such as traditional Chinese medicines or botanicals, mainly related to two factors:
(1) Whether a liquid can infiltrate a solid decoction piece is related to its surface tension. Surface tensionThe coefficient is smaller (30×10) -3 N/m or so), almost capable of impregnating the solid; the surface tension coefficient of water is large and it can only infiltrate some solids. The surface tension coefficient of the sugar water solution is smaller than that of pure water, and the solution can better enter plant cells to release effective components, so that the infiltration of raw materials such as traditional Chinese medicine decoction pieces can be improved, and the dissolution of saccharide components and/or glycoside components in the sugar water solution can be improved;
(2) The sugar in the extraction solvent and the sugar component and/or the glycosyl part of the glycoside component in the extract contain a large amount of hydroxyl groups, so that the sugar can form intermolecular acting forces such as intermolecular hydrogen bonds with the sugar component and/or the glycoside component in the raw materials such as traditional Chinese medicine decoction pieces, and the extraction efficiency of the sugar component and/or the glycoside component of the raw materials such as traditional Chinese medicine extract is improved;
therefore, the method can sufficiently improve the extraction rate of the saccharide component and/or glycoside component of the raw material such as the Chinese medicine or the plant medicine, compared with the water decoction mode.
In the actual working process, the extraction times of the alcohol and/or sugar alcohol solution can be determined according to actual needs, and can be 1 time or several times, for example, 2-10 times.
In a preferred embodiment, the concentration of the sugar solution and/or the sugar alcohol solution is in the range of 0.1-40g/100mL.
In the present invention, when a concentration, ratio, equivalent weight, number of times, or other parameter is expressed as a range, preferred range, or a range bounded by a list of upper preferable values and lower preferable values, this should be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "0.1-40" is disclosed, unless otherwise indicated, that range is intended to include its endpoints and all points within the range, for example 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1..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, 8.7.0, 3.8.9, 3.8, 3.9, 4.1.1, 4.1, 4.2. 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, and the like, not limited to the values recited above.
In a preferred embodiment, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-10g/100mL or 20-40g/100mL.
In a preferred embodiment, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-5g/100mL or 20-30g/100mL.
In a preferred embodiment, the concentration of the sugar solution and/or the sugar alcohol solution is in the range of 0.1-2.5g/100mL.
In a preferred embodiment, the concentration of the sugar solution and/or the sugar alcohol solution is 0.5-2g/100mL.
In a preferred embodiment, the concentration of the sugar solution and/or the sugar alcohol solution is about 0.5g/100mL or about 1g/100mL or about 1.5g/100mL or about 2g/100mL or about 20g/100mL or about 30g/100mL or about 40g/100mL.
The term "about" or "approximately" with respect to a numerical value means ± 5% of the numerical value, but explicitly includes the exact numerical value. For example, a concentration of "about" 0.5g/100mL refers to a concentration from 0.475g/100mL to 0.525g/100mL, but also specifically includes a concentration of just 0.5g/100 mL; "about" a concentration of 1g/100mL refers to a concentration from 0.95g/100mL to 1.05g/100mL, but also specifically includes a concentration of just 1g/100 mL; "about" a concentration of 1.5g/100mL refers to a concentration from 1.425g/100mL to 1.575g/100mL, but also explicitly includes concentrations of just 1.5g/100 mL; "about" a concentration of 2g/100mL refers to a concentration from 1.9g/100mL to 2.1g/100mL, but also specifically includes a concentration of just 2g/100 mL; "about" a concentration of 20g/100mL refers to a concentration from 19g/100mL to 21g/100mL, but also specifically includes a concentration of just 20g/100 mL; "about" a concentration of 30g/100mL refers to a concentration from 28.5g/100mL to 31.5g/100mL, but also specifically includes a concentration of just 30g/100 mL; "about" a concentration of 40g/100mL refers to a concentration from 38g/100mL to 42g/100mL, but also specifically includes a concentration of exactly 40g/100mL.
In a preferred embodiment, the sugar is selected from one or more of monosaccharides, disaccharides and trisaccharides.
In a preferred embodiment, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol and xylitol. In the practical application process, the sugar alcohol can be replaced by other sugar alcohols in the prior art according to the practical situation.
In a preferred embodiment, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, sorbose, rhamnose, ribose, xylose and deoxyribose. In the practical application process, the monosaccharide can be replaced by other monosaccharides according to the practical situation.
In a preferred embodiment, the disaccharide is selected from one or more of maltose, sucrose, lactose, trehalose, melibiose, gentiobiose and trabiose. In the practical application process, the disaccharide can be replaced by other disaccharides in the prior art according to practical situations.
In a preferred embodiment, the monosaccharide is glucose and/or fructose.
In a preferred embodiment, the trisaccharide is raffinose.
In order to further increase the extraction yield of the active ingredient of the saponins in the raw material such as a traditional Chinese medicine, in a preferred embodiment, the disaccharide is sucrose and/or maltose.
In a preferred embodiment, the saccharide component is an oligosaccharide component and/or a polysaccharide component.
In a preferred embodiment, the polysaccharide component is selected from one or more of the following: astragalus polysaccharides, licorice polysaccharides, ginseng polysaccharides and wolfberry polysaccharides.
In a preferred embodiment, the glycoside component is selected from one or more of the following: flavonoid glycoside compounds, saponins compounds, diterpenoid glycoside compounds, quinone glycoside compounds and phenolic acid glycoside compounds.
In a preferred embodiment, the flavonoid glycoside compound is selected from one or more of the following: flavonoid glycoside compounds, isoflavone glycoside compounds, flavanonol glycoside compounds, chalcone glycoside compounds, dihydrochalcone glycoside compounds, hesperidin compounds, anthocyanin compounds, biflavonoid glycoside compounds and flavanol glycoside compounds.
In a preferred embodiment, the saponins are tetracyclic triterpene glycosides, pentacyclic triterpene glycosides and/or steroid glycosides.
In a preferred embodiment, the diterpenoid glycosides are selected from one or more of the following: tetracyclic diterpenoid glycosides, tricyclic diterpenoid glycosides, chain diterpenoid glycosides, monocyclic diterpenoid compounds and bicyclic diterpenoid glycosides.
In a preferred embodiment, the quinone glycoside compound is benzoquinone glycoside compound, naphthoquinone glycoside compound, phenanthrenequinone glycoside compound and/or anthraquinone glycoside compound.
In a preferred embodiment, the phenolic acid glycoside compound is a phenolic acid glycoside compound having a C6-C3 parent core structure and/or a phenolic acid glycoside compound having a C6-C1 parent core structure.
In a preferred embodiment, the steroid glycoside compound is a spirostanol type saponin compound, a furostanol type saponin compound and/or an isospirostanol type saponin compound.
In a preferred embodiment, the isoparaffin type saponin compound is a dioscin type compound.
In a preferred embodiment, the furostanol-type saponin compound is a timosaponin-type compound.
In a preferred embodiment, the tetracyclic triterpene glycoside compound is a lanolate alkane type tetracyclic triterpene glycoside compound and/or a dammarane type tetracyclic triterpene glycoside compound.
In a preferred embodiment, the pentacyclic triterpene glycoside compound is an oleanane-type pentacyclic triterpene glycoside compound, an ursane-type pentacyclic triterpene glycoside compound and/or a lupin-type pentacyclic triterpene glycoside compound.
In a preferred embodiment, the anthraquinone glycoside compounds are monoanthracene nucleus compounds and/or dianthracene nucleus compounds.
In a preferred embodiment, the isoflavone glycoside compound is calycosin glucoside and/or formononetin. In a preferred embodiment, the flavanone glycoside compound is apigenin and/or liquiritin. In a preferred embodiment, the chalcone glycoside compound is hydroxysafflor yellow a. In a preferred embodiment, the furostanol-type steroid saponin compound is timosaponin BII. In a preferred embodiment, the lanostane-type tetracyclic triterpenoid saponin compound is astragaloside. In a preferred embodiment, the dammarane-type tetracyclic triterpenoid saponin compound is ginsenoside Rg1. In a preferred embodiment, the oleanane-type pentacyclic triterpene saponin compound is glycyrrhizic acid. In a preferred embodiment, the tetracyclic diterpene glycoside compound is stevioside and/or rebaudioside. In a preferred embodiment, the phenolic glycoside compound of C6-C3 parent nucleus structure is hydroxysafflor yellow A. In a preferred embodiment, the dianthroside compound is sennoside a and/or sennoside B. In a preferred embodiment, the monoanthracene nucleoside compound is aloin.
In a preferred embodiment, the raw material is a raw material containing a saccharide component and/or a glycoside component.
In a preferred embodiment, the raw material is one or more traditional Chinese medicines, botanicals and/or marine organisms containing saccharide components and/or glycoside components.
In a preferred embodiment, the traditional Chinese medicine and/or the botanical is selected from one or more of the following: ginseng, notoginseng, licorice, medlar, stevia rebaudiana, astragalus, senna, cortex albiziae, pokeberry root, bupleurum, pulsatilla root, cedrela sinensis, dendrobium huoshanense, achyranthes root, clematis root, american ginseng, bolbostemma paniculatum, horse chestnut, acanthopanax root, chestnut of a senior, gynostemma pentaphylla, chinaberry, centella asiatica, radix sedge, radix dipsaci, akebia, rhizoma panacis, lysimachia foenum-graecum, clematis stem, wild jujube leaf, cannabis sativa, polygala tenuifolia, east perilla, eclipta, platycodon grandiflorum, dwarf lilyturf tuber, allium macrostemon, rhizoma polygonati, rhizoma anemarrhenae, radix asparagi, dwarf lilyturf, rhizoma paridis, glabrous greenbrier, rhizoma dioscoreae, tribulus terrestris, purple eggplant, pomegranate, almond, sweet potato, yam, asparagus, largehead atractylodes rhizome, cortex lycii radicis, purple sweet potato and resina.
In a preferred embodiment, in step (2), the soaking is a room temperature soaking.
In a preferred embodiment, the step of soaking comprises soaking the raw material in the sugar solution and/or the sugar alcohol solution at room temperature for 10-120min.
In a preferred embodiment, the step of soaking comprises soaking the raw material in the sugar solution and/or the sugar alcohol solution at room temperature for 20-60min.
In a preferred embodiment, the step of soaking comprises soaking the feedstock in the sugar solution and/or the sugar alcohol solution at room temperature for about 30 minutes.
The term "about" or "approximately" with respect to a numerical value means ± 5% of the numerical value, but explicitly includes the exact numerical value. For example, a time of "about" 30 minutes refers to a time from 28.5 minutes to 31.5 minutes, but also explicitly includes a time of just 30 minutes.
In a preferred embodiment, the weight to volume ratio of the starting material to the sugar solution and/or the sugar alcohol solution is from 1:1 to 1:50.
In a preferred embodiment, the weight to volume ratio of the starting material to the sugar solution and/or the sugar alcohol solution is from 1:1 to 1:20.
In a preferred embodiment, the weight to volume ratio of the starting material to the sugar solution and/or the sugar alcohol solution is 1:6 to 1:12.
In a preferred embodiment, in step (3), the heat extraction is a heat reflux extraction.
In a preferred embodiment, the heating time is from 20min to 60min.
In a preferred embodiment, the heating time is from 20 minutes to 30 minutes.
In a preferred embodiment, the heating time is about 20 minutes or about 30 minutes.
The term "about" or "approximately" with respect to a numerical value means ± 5% of the numerical value, but explicitly includes the exact numerical value. For example, a time of "about" 30 minutes refers to a time from 28.5 minutes to 31.5 minutes, but also explicitly includes a time of just 30 minutes; the term "about" 20 minutes refers to a time from 19 minutes to 21 minutes, but also explicitly includes a time of just 20 minutes.
According to one aspect of the present invention there is provided a use of a method as described above for the preparation of a raw material extract or raw material concentrate, wherein the raw material is a traditional Chinese medicine, a plant drug and/or a marine organism.
In a preferred embodiment, the raw material concentrate is the raw material extract obtained in the above step (3) or the raw material total extract obtained in the step (5) is concentrated in a water bath at 20 to 100 ℃.
In a preferred embodiment, the concentration is accomplished in a water bath at 60 to 80 ℃.
In a preferred embodiment, the concentration is accomplished in a water bath at about 70 ℃.
The term "about" or "approximately" with respect to a numerical value means ± 5% of the numerical value, but explicitly includes the exact numerical value. For example, a temperature of "about" 70 ℃ refers to a ratio of from 66.5 ℃ to 73.5 ℃, but also specifically includes temperatures of just 70 ℃.
According to one aspect of the present invention there is provided a use according to the above method for the preparation of a pharmaceutical composition comprising a raw material, wherein the raw material is a traditional Chinese medicine, a plant drug and/or a marine organism.
According to one aspect of the present invention there is provided a use of a method according to the above in the preparation of a pharmaceutical formulation or functional or health food comprising a raw material, wherein the raw material is a traditional Chinese medicine, a plant drug and/or a marine organism.
Among these, functional foods are foods that can convincingly prove beneficial to a certain function or functions of the body, have sufficient nutritional effects to improve health or reduce illness.
Wherein, the health food is food which is claimed to have specific health care function or aims to supplement vitamins and minerals, namely is suitable for specific people to eat, has the function of regulating organism, does not aim to treat diseases, and does not generate any acute, subacute or chronic harm to human bodies.
According to one aspect of the present invention there is provided the use of a method according to the above in the preparation of other products containing raw materials, wherein other products refer to all prior art included products containing raw materials such as traditional Chinese medicine, botanicals and/or biologicals, except pharmaceutical formulations, functional foods and health foods, including liquid and solid forms and the like.
It should be noted that, in the case of no conflict, the embodiments and features in the embodiments may be combined with each other. The present invention will be described in detail with reference to examples.
The invention is described in further detail below in connection with specific embodiments, which should not be construed as limiting the scope of the invention as claimed.
Examples
Instrument and reagent
Agilent 1260 type high performance liquid chromatograph equipped with DAD detector (Agilent, usa); agilent model 1200 high performance liquid chromatograph equipped with ELSD detector (Agilent, usa); UV-2550 type ultraviolet-visible spectrophotometer (shimadzu corporation); DZKW-4 type electronic thermostat water bath (Beijing Zhongxing Wei industry instruments Co., ltd.); DK-98-IIA electric thermostatic water bath (Test instruments Co., tianjin); ME204/02 electronic balance (Metrele Tolyduo instruments (Shanghai) Co., ltd., one ten million balance); KQ-250DE type digital controlled ultrasonic cleaner (Kunshan ultrasonic instruments Co., ltd.); HC-3018 high-speed centrifuge (Anhui middle Ke Zhongjia scientific instruments Co., ltd.); a solid phase extraction column UniEIut C18 EC column (1000 mg/6 ml) (Hua Puke Instrument Beijing technologies Co., ltd.).
The reference substances of apigenin (lot number CHB 180109), glycyrrhizin (lot number CHB 201102), astragaloside IV (lot number CHB 170727), calycosin glucoside (lot number CHB 161105), formononetin (lot number CHB 150517) and D-glucose (lot number CHB 190213) are all purchased from Chengdu Crohn biotechnology Co., ltd, and the purity is >98%; glycyrrhizic acid (lot number Z30A6B 1) was purchased from shanghai source leaf biotechnology limited with purity >98%; ginsenoside Rg1 (lot number 210708), purchased from Chengdu plant labeling pure biotechnology Co., ltd, purity >98%; timosaponin BII (lot No. 111839-202107) purchased from China food and drug assay institute with purity of 95.4% or more; rebaudioside (lot 211117), stevioside (lot 211024), purchased from adzuki plant labeled pure biotechnology limited, purity >98%; sennoside A (lot number 211203), purchased from Chengdu plant labeled pure biotechnology Co., ltd, purity HPLC >98%; sennoside B (lot number 220215), purchased from Chengdu plant labeling pure biotechnology Co., ltd, purity HPLC >98%; acetonitrile (chromatographic purity, fisher Co.), methanol (chromatographic purity, fisher Co.) and phosphoric acid (analytical purity, available from Guogu Chemicals Co., ltd.). Sucrose, glucose, fructose, maltose, lactose (gold cloning (Beijing) biotechnology Co., ltd.); trehalose (Hebei hundred biotech Co., ltd.); are all of pharmaceutical grade. The water is child ha purified water.
Example one experiment for extracting Licorice decoction pieces with different sugar solutions
1.1 preparation method of different sugar-Licorice root extract
4 parts of raw licorice decoction pieces (inner Mongolia) in the same batch are taken, and each part is 40g. 7 times of water (280 mL of water), 1% glucose aqueous solution (2.8 g glucose+280 mL of water), 1% sucrose aqueous solution (2.8 g sucrose+280 mL of water) and 1% maltose aqueous solution (2.8 g maltose+280 mL of water) were added respectively, immersed for 30min, refluxed for 30min, and filtered with gauze while hot to obtain different sugar-licorice extracts.
1.2 determination of the content of the apigenin, the glycyrrhizin and the glycyrrhizic acid (HPLC pharmacopoeia method)
1.2.1 preparation of sample solutions
Taking 0.5mL of different sugar-licorice extract, adding methanol (9.5 mL) for dilution by 20 times, shaking uniformly, and filtering with 0.22 μm filter membrane to obtain the final product.
1.2.2 preparation of control solution
Stock solution: taking appropriate amounts of apigenin, glycyrrhizin and glycyrrhizic acid reference substances, precisely weighing, and adding methanol to prepare 1mg/mL stock solution. Stored in a-20 ℃ refrigerator.
Linear working solution: taking a proper amount of apigenin and glycyrrhizin stock solution, and adding methanol to dilute into a series of linear mixed labeling working solutions: 2.5,5, 10, 20, 40, 50, 100 μg/mL; taking a proper amount of glycyrrhizic acid stock solution, and adding methanol to dilute into a series of linear mixed labeling working solutions: 10 20, 40, 80, 160, 200, 400 μg/mL.
1.2.3 content determination method
Chromatographic column: thermo Acclaim 120C 18 (250X 4.6mm,5 μm); mobile phase: acetonitrile (a), 0.05% phosphoric acid solution (B) as mobile phase, column temperature: 35 ℃, detection wavelength is 237nm, sample injection amount: 10 μl, flow rate: gradient elution at 1 ml/min: 0-8min,19% A;8-35min,19% -50% of A;35-36min,50% -100% A;36-40min,100% -19% A;40-50min,19% A.
1.3 experimental results
The results of measuring the content of the active ingredients in the different sugar-licorice extracts are shown in Table 1.
TABLE 1 determination of the content of active principles in different sugar-Glycyrrhiza extracts
* P <0.05VS water
As can be seen from Table 1, glucose, sucrose and maltose (1 g/100mL concentration) can raise the content of the effective components in licorice to different degrees, wherein the effects of sucrose and maltose are most obvious.
Example two experiment for extracting Licorice decoction pieces with different sugar solutions
2.1 preparation method of different sugar-Glycyrrhrizae radix extractive solutions
4 parts of raw licorice decoction pieces (produced by Xinjiang) in the same batch are taken, and each part is 40g. 7 times of water (280 mL of water), 1% sucrose aqueous solution (2.8 g sucrose+280 mL of water), 1% maltose aqueous solution (2.8 g maltose+280 mL of water), 1% lactose aqueous solution (2.8 g lactose+280 mL of water) were sequentially added, immersed for 30min, refluxed for 30min, filtered with gauze while hot, 6 times of water was added to the residue, 1% sucrose aqueous solution (2.4 g sucrose+240 mL of water), 1% maltose aqueous solution (2.4 g maltose+240 mL of water), 1% lactose aqueous solution (2.4 g lactose+240 mL of water), refluxed for 20min, filtered while hot, and 2 times of filtrates were combined to obtain different sugar-licorice extracts.
2.2 determination of the content of the apigenin, the glycyrrhizin and the glycyrrhizic acid (HPLC pharmacopoeia method)
2.2.1 preparation of sample solutions
The same procedure was used for the preparation of the sample solution of example one.
2.2.2 preparation of control solution
The same procedure was used for the preparation of the control solution of example one.
2.2.3 content determination method
The same method as in example one was used for the content measurement.
2.3 experimental results
The results of measuring the content of the active ingredients in the different sugar-licorice extracts are shown in Table 2.
TABLE 2 determination of the active ingredient content in different sugar-Glycyrrhiza extracts
* P <0.05VS water
As can be seen from Table 2, sucrose and maltose (1 g/100mL concentration) can raise the content of the effective components in licorice to different degrees, wherein the sucrose effect is most obvious.
From a combination of the results shown in tables 1 and 2, sucrose can significantly improve the extraction rate of main active ingredients in licorice, including flavonoids (apioside, liquiritin) and saponins (glycyrrhizic acid).
Example three experiment for extracting raw astragalus decoction pieces by different sugar solutions
3.1 preparation method of different sugar-radix astragali extract (different 1% sugar extraction twice)
5 parts of raw astragalus decoction pieces (from a pistachio source) in the same batch are taken, and each part is 50g. 7 times of water (350 ml of water), 1% glucose aqueous solution (3.5 g glucose added water fixed volume at 350 ml), 1% fructose aqueous solution (3.5 g fructose added water fixed volume at 350 ml), 1% sucrose aqueous solution (3.5 g sucrose added water fixed volume at 350 ml), 1% trehalose aqueous solution (3.5 g trehalose added water fixed volume at 350 ml), soaking for 30min, refluxing for 30min, filtering with gauze while hot, adding 6 times of water (300 ml of water) to the residues, 1% glucose aqueous solution (3.0 g glucose added water fixed volume at 300 ml), 1% fructose aqueous solution (3.0 g fructose added water fixed volume at 300 ml), 1% sucrose aqueous solution (3.0 g sucrose added water fixed volume at 300 ml), 1% trehalose aqueous solution (3.0 g trehalose added water fixed volume at 300 ml), refluxing for 20min, filtering while hot, combining 2 times of filtrate to obtain different sugar-astragalus extracts.
3.2 determination of Astragaloside IV, calycosin glucoside and formononetin content
3.2.1 preparation of sample solutions
Preparation of a sample solution of saponin components: taking 5ml of different sugar-astragalus extract, adding 20ml of 60% methanol, carrying out ultrasonic mixing, adding 10ml of ammonia water, uniformly mixing, centrifuging for 10min at 3600r/min, passing the supernatant through a UniEIut C18 EC column (1000 mg/6 ml), eluting with 10ml of purified water, eluting with 5ml of methanol, collecting methanol eluent, centrifuging for 5min at 8000r/min, taking the supernatant, and passing through a 0.45 μm microporous filter membrane.
Preparation of flavonoid component sample solution: taking 300 μl of different sugar-radix astragali extractive solutions, adding 700 μl of water, placing into 1.5ml centrifuge tube, centrifuging at 12000r/min for 5min, collecting supernatant, and filtering with 0.45 μm microporous membrane.
3.2.2 preparation of control solution
Taking appropriate amounts of astragaloside IV, calycosin glucoside and formononetin reference substances, precisely weighing, and adding methanol to prepare 1mg/ml stock solution. Stock solutions were diluted to different series of concentrations and working curves were established.
The linear equation of astragaloside is lnY =1.3271 lnx+9.9314, r=1.0000; the linear equation of calycosin glucoside is y=35129x+14, r=0.9998; the linear equation for formononetin is y=10024x+33, r=0.9998.
3.2.3 content determination method
The method for measuring the content of the saponins comprises the following steps: analysis was performed using an Agilent type 1200 liquid chromatograph (equipped with ELSD detector), column: thermo Acclaim 120C 18 (250X 4.6mm,5 μm); mobile phase: 0.1% formic acid water (A) to 0.1% formic acid acetonitrile (B), gradient elution: 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), column temperature: the flow rate is 1ml/min at room temperature, the temperature of the drift tube is 100 ℃, the flow rate of carrier gas is 2.5l/min, and the sample injection amount is 20 mu l.
The method for measuring the content of flavonoid components comprises the following steps: analysis was performed using an Agilent 1260 type high performance liquid chromatograph (equipped with DAD detector), column: thermo Acclaim 120C 18 (250X 4.6mm,5 μm); mobile phase: gradient elution of 0.1% aqueous formic acid (a) with 0.1% acetonitrile formate (B): 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; the flow rate is 1ml/min; the detection wavelength is 260nm; column temperature 25 ℃; the sample was introduced in an amount of 10. Mu.l.
3.3 experimental results
The results of measuring the content of the active ingredients in the different sugar-astragalus extracts are shown in table 3.
TABLE 3 determination of the content of active ingredients in different sugar-Astragalus extracts
* P <0.05VS astragalus water extract
As can be seen from Table 3, sucrose, glucose, fructose and trehalose (1 g/100ml concentration) can increase the content of the active ingredients in astragalus to different degrees, wherein the effects of sucrose and trehalose are most obvious, but sucrose is more economical than trehalose.
Example four experiment for extracting raw astragalus decoction pieces with different sugar solutions
4.1 preparation method of different sugar-radix astragali extract (different 1% sugar is extracted once)
The same batch of raw astragalus decoction pieces (from a pistachio) is taken in 4 parts, and each part is 50g. 7 times of water (350 ml of water) and 1% glucose aqueous solution (3.5 g glucose added with water to constant volume of 350 ml) and 1% sucrose aqueous solution (3.5 g sucrose added with water to constant volume of 350 ml) are respectively added, and 1% maltose aqueous solution (3.5 g maltose added with water to constant volume of 350 ml) are soaked for 30min, refluxed for 30min and filtered by gauze while hot to obtain different sugar-astragalus extract.
4.2 determination of Astragaloside IV, calycosin glucoside and formononetin content
4.2.1 preparation of sample solutions
The same method as in example three was used to prepare the test solutions.
4.2.2 preparation of control solution
The same procedure was used for the preparation of the control solution of example three.
4.2.3 content determination method
The same method as in example three was used for content measurement.
4.3 experimental results
The results of measuring the content of the active ingredients in the different sugar-astragalus extracts are shown in table 4.
TABLE 4 determination of the content of active principles in different sugar-Astragalus extracts
* P <0.05VS astragalus water extract
As can be seen from Table 4, glucose, sucrose and maltose (1 g/100ml concentration) can raise the content of the effective components in astragalus root to different degrees, wherein the effect of sucrose is most obvious.
Example five different sugar solutions extraction of raw radix astragali decoction pieces experiment
5.1 preparation method of different sugar-radix astragali extract (different 1% sugar extraction twice)
The same batch of raw astragalus decoction pieces (from Shanxi Lingchuan) is taken in 4 parts, and each part is 50g. 7 times of water (350 ml of water), 1% sucrose aqueous solution (3.5 g sucrose water fixed volume at 350 ml), 1% maltose aqueous solution (3.5 g maltose water fixed volume at 350 ml), 1% lactose aqueous solution (3.5 g lactose water fixed volume at 350 ml) are respectively added, soaked for 30min, refluxed for 30min, filtered with gauze while hot, the residues are respectively added with 6 times of water (300 ml of water), 1% sucrose aqueous solution (3.0 g sucrose water fixed volume at 300 ml), 1% maltose aqueous solution (3.0 g maltose water fixed volume at 300 ml), 1% lactose aqueous solution (3.0 g lactose water fixed volume at 300 ml), refluxed for 20min, filtered while hot, and combined with 2 times of filtrate to obtain different sugar-astragalus extracts.
5.2 determination of Astragaloside IV, calycosin glucoside and formononetin content
5.2.1 preparation of sample solution
The same method as in example three was used to prepare the test solutions.
5.2.2 preparation of control solution
The same procedure was used for the preparation of the control solution of example three.
5.2.3 content determination method
The same method as in example three was used for content measurement.
5.3 experimental results
The results of measuring the content of the active ingredients in the different sugar-astragalus extracts are shown in table 5.
TABLE 5 determination of the active ingredient content in different sugar-Astragalus extracts
* P <0.05VS astragalus water extract
As can be seen from Table 5, sucrose, maltose and lactose (1 g/100ml concentration) can increase the content of the effective components in astragalus root to different degrees, wherein the extraction rate of three components can be increased by sucrose, and the effect is most obvious.
From the results of tables 3 to 5, it is apparent that sucrose can significantly improve the extraction rate of the main active ingredients in astragalus, including flavonoids (calycosin glucoside, formononetin) and saponins (astragaloside IV).
Example six extraction of raw radix astragali decoction pieces from sucrose solutions with different concentrations
6.1 preparation of sucrose-Astragalus root extract with different concentrations (0-10% sucrose extraction with different concentrations)
7 parts of raw astragalus decoction pieces (from mountain cinquefoil) in the same batch are taken, and each part is 50g. 7 times of water (350 ml of water), 0.5% sucrose aqueous solution (1.8 g sucrose water fixed volume: 350 ml), 1.25% sucrose aqueous solution (3.5 g sucrose water fixed volume: 350 ml), 1.5% sucrose aqueous solution (4.4 g sucrose water fixed volume: 350 ml), 1.3% sucrose aqueous solution (5.3 g sucrose water fixed volume: 350 ml), 2% sucrose aqueous solution (7 g sucrose water fixed volume: 350 ml), 10% sucrose aqueous solution (35 g sucrose water fixed volume: 350 ml), soaking for 30min, refluxing for 30min, filtering with gauze while hot, adding 6 times of water (300 ml of water) to the dregs, 0.5% sucrose aqueous solution (1.5 g sucrose water fixed volume: 300 ml), 1.25% sucrose aqueous solution (3.8 g sucrose water fixed volume: 300 ml), 1.5% sucrose aqueous solution (4.5 g sucrose water fixed volume: 300 ml), 2% sucrose aqueous solution (2.5 g sucrose water fixed volume: 300 ml), filtering while refluxing for 30min, mixing the aqueous solutions (2-300 ml) and filtering when the aqueous solution is not hot.
6.2 determination of Astragaloside IV, calycosin glucoside and formononetin content
6.2.1 preparation of sample solution
The same method as in example three was used to prepare the test solutions.
6.2.2 preparation of control solution
The same procedure was used for the preparation of the control solution of example three.
6.2.3 content determination method
The same method as in example three was used for content measurement.
6.3 experimental results
The determination results of the content of the active ingredients in the astragalus extract obtained by extracting the raw astragalus decoction pieces by using the sucrose solutions with different concentrations are shown in table 6.
TABLE 6 determination of the active ingredient content in Astragalus extract obtained with sucrose at different concentrations
* P <0.05VS astragalus water extract
As can be seen from Table 6, the concentration of sucrose can be increased to different degrees in the range of 0.25g/100ml to 1.25g/100ml, wherein the effect is most obvious when the concentration of sucrose is in the range of 1g/100ml to 1.25g/100 ml. In addition, the concentration of sucrose can also increase the content of active ingredients in the astragalus membranaceus when the concentration is 10g/100 ml.
Seventh experiment for extracting ginsenoside Rg1 from Notoginseng radix with sugar solutions of different concentrations
7.1 preparation method of sucrose-pseudo-ginseng extract with different concentrations
The same batch of radix Notoginseng decoction pieces (origin: yunnan; manufacturer: tianjin Shengsheng Utility Co., ltd.; lot number: X20102703A) was taken in 4 parts, 50g each. 7 times of water (350 ml of water), 0.5% sucrose aqueous solution (1.75 g sucrose water constant volume at 350 ml), 1% sucrose aqueous solution (3.5 g sucrose water constant volume at 350 ml), 1.5% sucrose aqueous solution (5.25 g sucrose water constant volume at 350 ml), soaking for 30min, refluxing for 30min, filtering while hot with gauze, adding 6 times of water (300 ml of water) to the residues, 0.5% sucrose aqueous solution (1.5 g sucrose water constant volume at 300 ml), 1% sucrose aqueous solution (3.0 g sucrose water constant volume at 300 ml), 1.5% sucrose aqueous solution (4.5 g sucrose water constant volume at 300 ml), refluxing for 20min, filtering while hot, and combining 2 times of filtrates to obtain sucrose-pseudo-ginseng extract with different concentrations.
7.2 determination of ginsenoside Rg1 content
7.2.1 preparation of sample solution
Precisely measuring 2mL of different Notoginseng radix extractive solutions, respectively adding methanol to constant volume to 10mL volumetric flask, mixing, and 13000 r.min -1 Centrifuging for 5min, collecting supernatant, and filtering with 0.22 μm microporous membrane.
7.2.2 preparation of control solution
Taking a proper amount of ginsenoside Rg1 reference substance, precisely weighing, and adding methanol to prepare a stock solution of 1 mg/ml. Stock solutions were diluted to different series of concentrations and working curves were established.
The linear equation of ginsenoside Rg1 is: y= 3246.4X-4.5152, r 2 ＝0.9999。
7.2.3 content determination method
Analysis was performed using an Agilent type 1200 liquid chromatograph, column: roc C18 (250×4.6mm,5 μm); mobile phase: water (a) -acetonitrile (B), gradient elution: 0-12 min,19% -19% B; 12-60 min,19% -36% B; column temperature: the flow rate is 1ml/min at 35 ℃, the sample injection amount is 10 μl, and the detection wavelength is 203nm.
7.3 experimental results
The effect of sucrose at different concentrations on the extraction rate of index components in Notoginseng radix is shown in Table 7.
TABLE 7 Effect of sucrose at different concentrations on extraction yield of index Components in Notoginseng radix
* P <0.05vs pseudo-ginseng water extract
As can be seen from Table 7, sucrose (1 g/100ml-1.5g/100ml concentration) can raise the content of the active ingredient-saponins (ginsenoside Rg 1) in Notoginseng radix to different degrees.
EXAMPLE eight experiment for extracting timosaponin BII from rhizoma anemarrhenae decoction pieces by sucrose solutions with different concentrations
8.1 preparation method of sucrose-rhizoma anemarrhenae extract with different concentrations
5 parts of raw rhizoma anemarrhenae decoction pieces in the same batch are taken, and 50g of each part is taken. 7 times of water (350 mL of water), 0.5% sucrose aqueous solution (1.75 g sucrose water fixed volume: 350 mL), 1% sucrose aqueous solution (3.5 g sucrose water fixed volume: 350 mL), 1.5% sucrose aqueous solution (5.25 g sucrose water fixed volume: 350 mL), 2.0% sucrose aqueous solution (7.0 g sucrose water fixed volume: 350 mL) were respectively added, immersed for 30 minutes, refluxed for 30 minutes, filtered with gauze while hot, the dregs were respectively added with 6 times of water (300 mL of water), 0.5% sucrose aqueous solution (1.5 g sucrose water fixed volume: 300 mL), 1% sucrose aqueous solution (3.0 g sucrose water fixed volume: 300 mL), 1.5% sucrose aqueous solution (4.5 g sucrose water fixed volume: 300 mL), 2.0% sucrose aqueous solution (6 g sucrose water fixed volume: 300 mL), refluxed for 20 minutes, filtered while hot, combined with 2 times of filtrate, fixed volume: 500mL, thus obtaining the rhizoma anemarrhenae extract with different concentrations.
8.2 timosaponin BII content determination
8.2.1 preparation of sample solutions
Precisely measuring 2mL of different rhizoma anemarrhenae extractive solutions, respectively adding methanol to constant volume to 10mL volumetric flask, mixing, and 13000 r.min -1 Centrifuging for 5min, collecting supernatant, and filtering with 0.22 μm microporous membrane.
8.2.2 preparation of control solution
Taking a proper amount of timosaponin BII reference substance, precisely weighing, and adding 30% acetonitrile to prepare a stock solution with the mass concentration of 1 mg/ml. Stock solutions were diluted to different series of concentrations and working curves were established.
The linear equation for timosaponin BII is: lnY =1.8921lnx+10.274, r 2 ＝0.9977。
8.2.3 content determination method
Analysis was performed using an Agilent type 1200 liquid chromatograph, column: inertsil ODS-3-C18 (250X 4.6mm,5 μm); mobile phase: water (A) to acetonitrile (B), 25% acetonitrile isocratic elution; column temperature: the flow rate is 0.8ml/min at 35 ℃ and the sample injection amount is 10 μl. Drift tube temperature: 104 ℃, carrier gas flow rate: 2.8 L.min -1 。
8.3 experimental results
The effect of sucrose at different concentrations on extraction rate of index components in rhizoma anemarrhenae is shown in Table 8.
TABLE 8 Effect of sucrose at different concentrations on extraction yield of index Components in Anemarrhena asphodeloides
* P <0.05vs rhizoma anemarrhenae water extract
As can be seen from Table 8, sucrose (1.0 g/100ml-2.0g/100ml concentration) can raise the content of the active ingredient-saponins (timosaponin BII) in rhizoma anemarrhenae to different degrees.
Experiment for extracting sennoside B and sennoside A from senna with sucrose solutions of different concentrations
9.1 preparation method of sucrose-senna leaf extract with different concentrations
The senna leaves of the same batch (Guangxi Yulin, batch number: 200101). 8 parts of each 20g of the solution were taken, 12 times of water (240 mL of water), 0.5% aqueous sucrose solution (1.2 g of sucrose was dissolved in 240mL of water), 0.75% aqueous sucrose solution (1.8 g of sucrose was dissolved in 240mL of water), 1% aqueous sucrose solution (2.4 g of sucrose was dissolved in 240mL of water), 1.5% aqueous sucrose solution (3.6 g of sucrose was dissolved in 240mL of water), 2% aqueous sucrose solution (4.8 g of sucrose was dissolved in 240mL of water), 3% aqueous sucrose solution (7.2 g of sucrose was dissolved in 240mL of water), 5% aqueous sucrose solution (12 g of sucrose was dissolved in 240mL of water), immersing for 30 minutes, refluxing for 30 minutes, filtering with 3 layers of gauze while hot, adding 10 times of water (200 mL of water) to the dregs, 0.5% aqueous sucrose solution (1 g of sucrose was dissolved in 200mL of water), 1.5% aqueous sucrose solution (1.5 g of sucrose was dissolved in 200mL of water), 1% aqueous sucrose solution (2 g of sucrose was dissolved in 200mL of water), 3% aqueous sucrose solution (200 mL of 2% aqueous solution was dissolved in 200mL of distilled water), and aqueous solution (200 mL of aqueous solution was filtered, and aqueous solution was added to 200mL of aqueous solution was filtered.
9.2 determination of sennoside B and sennoside A content
9.2.1 preparation of sample solutions
Precisely measuring 1mL of senna leaf extract with different concentrations, metering 5mL of the senna leaf extract to a volume of 5mL by using a volumetric flask, diluting the sample by 5 times, uniformly mixing, and carrying out 15000 r.min -1 Centrifuging for 5min, collecting supernatant, and sieving with 0.2And (3) a microporous filter membrane with the diameter of 2 mu m.
9.2.2 preparation of control solution
Taking appropriate amounts of sennoside B and sennoside A as reference substances, adding 0.1% NaHCO 3 Preparing mother solution with mass concentration of 1mg/ml, diluting with methanol to give reference stock solution of 0.2mg/ml, 0.1mg/ml, 0.05mg/ml, 0.025mg/ml, and 0.0125mg/ml, and shaking. The linear equation of sennoside B is y=8988.5x+2.5708, r 2 =0.9999; the linear equation of sennoside a is y=8564.2x+9.9917, r 2 ＝0.9999。
9.2.3 content determination method
Analysis was performed using an Agilent 1260 type liquid chromatograph (equipped with an ultraviolet detector):
chromatographic column: c18 (250 mm. Times.4.6 mm,5 μm). The sample loading was 10ul. The detection wavelength is 340nm, the flow rate is 1ml/min, and the column temperature is 30 ℃. The mobile phase A is 0.1% phosphoric acid water, the B phase is acetonitrile, and the gradient flow is carried out (0-10 min,79% A, 10-15 min,79% to 75% A, 15-16 min,75% A, 16-17 min,75% to 79% A).
9.3 experimental results
The effect of varying concentrations of sucrose on the extraction of the index ingredient from senna is shown in table 9.
TABLE 9 Effect of sucrose at various concentrations on extraction yield of index ingredients from senna leaves
* P <0.05vs senna leaf water extract
As can be seen from Table 9, the concentration of sucrose can significantly increase the content of the index ingredient in the senna leaf in the concentration range of 2g/100mL to 5g/100 mL.
Example experiment for extracting aloin from aloe by using sucrose solutions with different concentrations and sugar solutions with the same concentration
Method for preparing 10.1.1 sucrose-aloe extract with different concentrations
The aloe vera (purchased from Beijing Tongren Tang.) is prepared by taking 6 parts of aloe vera (20 g) and adding 12 times of water (240 mL water), 0.5% sucrose aqueous solution (1.2 g sucrose is dissolved in 240mL water), 1% sucrose aqueous solution (2.4 g sucrose is dissolved in 240mL water), 1.5% sucrose aqueous solution (3.6 g sucrose is dissolved in 240mL water), 2% sucrose aqueous solution (4.8 g sucrose is dissolved in 240mL water) and 3% sucrose aqueous solution (7.2 g sucrose is dissolved in 240mL water) respectively, soaking for 30min, refluxing for 30min, filtering with 1 layer of terylene while hot, and fixing volume to 240 mL.
10.1.2 preparation method of different sugar-aloe extracts
The aloe beverage is prepared by taking 4 parts of aloe (purchased from Beijing Tongrentang) in the same batch, adding 12 times of water (240 mL of water), 0.5% sucrose aqueous solution (1.2 g sucrose is dissolved in 240mL of water), 0.5% maltose aqueous solution (1.2 g maltose is dissolved in 240mL of water), 0.5% trehalose aqueous solution (1.2 g trehalose is dissolved in 240mL of water) into each 20g of aloe, refluxing for 30min, filtering with 1 layer of terylene while hot, and fixing volume to 240 mL.
10.2 Aloesin content determination
10.2.1 preparation of sample solutions
Precisely measuring 0.2mL of aloe extract with different concentrations, metering to 10mL with volumetric flask, diluting the sample by 50 times, mixing, and 15000 r.min -1 Centrifuging for 5min, collecting supernatant, and filtering with 0.22 μm microporous membrane.
Preparation of 10.2.2 control solution
Adding methanol into appropriate amount of aloin reference substance to obtain mother solution with mass concentration of 1mg/ml, diluting with methanol to obtain reference substance stock solution of 0.2mg/ml, 0.1mg/ml, 0.05mg/ml, 0.3mg/ml, 0.15mg/ml, and 0.075mg/ml, and shaking.
The linear equation of the aloin is y= 7459.1x-25.173R 2 ＝0.9991
10.2.3 content determination method
Analysis was performed using an Agilent 1260 type liquid chromatograph equipped with an ultraviolet detector,
chromatographic column: c18 (250 mm. Times.4.6 mm,5 μm). The sample loading was 5ul. The detection wavelength is 355nm, the flow rate is 1ml/min, the mobile phase A is water, the mobile phase B is acetonitrile, and the isocratic flow is carried out (0-15 min,75% A;).
10.3 experimental results
The effect of sucrose at various concentrations on the extraction rate of index components in aloe is shown in Table 10.
TABLE 10 Effect of sucrose at different concentrations on extraction yield of index ingredients in aloe
* P <0.05vs aloe water extract
The effect of different disaccharides on the extraction of index components from aloe is shown in table 11.
TABLE 11 Effect of different disaccharides on extraction yield of index Components in Aloe
* P <0.05vs aloe water extract
As can be seen from the descriptions in tables 10 and 11, sucrose concentration at 0.5g/100mL significantly increased the content of the index component (i.e., aloin) in aloe.
Eleven experiments for extracting stevioside and rebaudioside in stevia rebaudiana with sucrose solutions of different concentrations and different sugar solutions of the same concentration
11.1 preparation method of sucrose-stevia Rebaudiana extract with different concentrations
5 parts of stevia rebaudiana in the same batch are taken, and each part is 20g. Adding 12 times of water (240 ml water), 0.5% sucrose aqueous solution (1.2 g sucrose is dissolved in 240ml water), 1% sucrose aqueous solution (2.4 g sucrose is dissolved in 240ml water), 1.5% sucrose aqueous solution (3.6 g sucrose is dissolved in 240ml water), 2% sucrose aqueous solution (4.8 g sucrose is dissolved in 240ml water), soaking for 30min, refluxing for 30min, filtering with 3 layers of gauze while hot; the medicinal materials are respectively added with 10 times of water (200 mL of water), 0.5 percent (1 g of sucrose is dissolved in 200mL of water), 1 percent of sucrose aqueous solution (2 g of sucrose is dissolved in 200mL of water), 1.5 percent of sucrose aqueous solution (3 g of sucrose is dissolved in 200mL of water) and 2 percent (4 g of sucrose is dissolved in 200mL of water), reflux is carried out for 20min, 3 layers of gauze are filtered while the medicinal materials are hot, 2 times of filtrate are combined, the volume is fixed to 400mL, and sucrose-stevia rebaudiana extract with different concentrations is obtained.
11.2 preparation method of different sugar-stevia Rebaudiana extract
5 parts of stevia rebaudiana in the same batch are taken, and each part is 20g. Respectively adding 12 times of water (240 ml water), 1.5% sucrose aqueous solution (3.6 g sucrose is dissolved in 240ml water), 1.5% maltose aqueous solution (3.6 g maltose is dissolved in 240ml water), 1.5% trehalose aqueous solution (3.6 g trehalose is dissolved in 240ml water), 1.5% lactose aqueous solution (3.6 g lactose is dissolved in 240ml water), soaking for 30min, refluxing for 30min, filtering with 3 layers gauze while hot; the medicinal materials are respectively added with 10 times of water (200 mL of water), 1.5% sucrose aqueous solution (3 g sucrose is dissolved in 200mL of water), 1.5% maltose aqueous solution (3 g maltose is dissolved in 200mL of water), 1.5% trehalose aqueous solution (3 g trehalose is dissolved in 200mL of water), 1.5% lactose aqueous solution (3 g lactose is dissolved in 200mL of water), reflux is carried out for 20min, 3 layers of gauze while the medicinal materials are hot are filtered, 2 times of filtrate are combined, the volume is fixed to 400mL, and different sugar-stevia rebaudiana extracts with the same concentration are obtained.
11.3 determination of index component content
Preparation of 11.3.1 sample solution
Precisely measuring 0.5mL of stevia rebaudiana Bertoni extract with different concentrations, adding methanol to constant volume to 10mL, diluting the sample by 20 times, mixing well, and 14000 r.min -1 Centrifuging for 5min, collecting supernatant, and filtering with 0.22 μm microporous membrane.
Preparation of 11.3.2 control solution
Rebaudioside d: adding methanol into a proper amount of reference substance to prepare a mother solution with the mass concentration of 1mg/ml, diluting with methanol to prepare reference substance stock solutions of 0.4mg/ml, 0.3mg/ml, 0.2mg/ml, 0.15mg/ml, 0.1mg/ml and 0.05mg/ml, and shaking uniformly to obtain the final product.
Stevioside: adding methanol into a proper amount of reference substance to prepare a mother solution with the mass concentration of 1mg/ml, diluting with methanol to prepare reference substance stock solutions of 0.3mg/ml, 0.2mg/ml, 0.15mg/ml, 0.1mg/ml, 0.05mg/ml and 0.025mg/ml, and shaking uniformly to obtain the final product.
The linear equation for rebaudioside is: y=999.64x+1.6452, r 2 ＝0.9995
The linear equation for stevioside is: y=y=1184.2x+0.987R 2 ＝0.9998
11.3.3 content determination method
Analysis was performed using an Agilent 1260 type liquid chromatograph (equipped with an ultraviolet detector) with a column size of C18 (250 mm. Times.4.6 mm,5 μm) and an amount of sample introduced of 5ul. The detection wavelength is 210nm, the flow rate is 0.6ml/min, and the column temperature is set at 35 ℃. The mobile phase is 0.1% phosphoric acid water A-acetonitrile B, 30% acetonitrile is eluted at the flow equality degree, and the time is 25min.
11.4 experimental results
The effect of varying concentrations of sucrose on the extraction of the index component from stevia is shown in Table 12.
TABLE 12 Effect of sucrose at different concentrations on extraction yield of index ingredient in stevia
* Stevia rebaudiana water extract with P <0.05vs
As can be seen from Table 12, sucrose concentrations ranging from 0.5g/100mL to 1.5g/100mL are capable of increasing the levels of the indicated components in stevia to varying degrees, with sucrose concentrations at 1.5g/100mL being most effective.
The effect of different classes of disaccharides on the extraction of index components in stevia is shown in Table 13.
TABLE 13 Effect of different disaccharides on the extraction yield of index ingredients in stevia rebaudiana
* Stevia rebaudiana water extract with P <0.05vs
As can be seen from Table 13, different disaccharides can increase the level of the index ingredient in stevia to varying degrees at 1.5g/100mL concentrations.
Example twelve experiments on extraction of hydroxy safflor yellow A from safflower with sucrose solutions of different concentrations and sugar solutions of the same concentration
12.1 preparation method of sucrose-safflower extract with different concentrations
7 parts of safflower (Tibet, beijing Tongrentang Co., ltd.) was taken in the same batch, 20g of each was added with 12 times of water (240 mL of water), 0.5% sucrose aqueous solution (1.2 g sucrose was dissolved in 240mL of water), 1.5% sucrose aqueous solution (2.4 g sucrose was dissolved in 240mL of water), 1.5% sucrose aqueous solution (3.6 g sucrose was dissolved in 240mL of water), 2.5% sucrose aqueous solution (6 g sucrose was dissolved in 240mL of water), 5% sucrose aqueous solution (12 g sucrose was dissolved in 240mL of water), immersed for 30 minutes, refluxed for 30 minutes, filtered with 3 layers of gauze while hot, and the dregs were again added with 10 times of water (200 mL of water), 0.5% sucrose aqueous solution (1 g sucrose was dissolved in 200mL of water), 1% sucrose aqueous solution (2 g sucrose was dissolved in 200mL of water), 1.5% sucrose aqueous solution (3 g sucrose was dissolved in 200mL of water), 2.8 g sucrose was dissolved in 200mL of 2% sucrose aqueous solution (4 g sucrose was dissolved in 200mL of water), and the filtrate was obtained by mixing the two layers of sucrose aqueous solutions (200 mL of 2g sucrose aqueous solution, 2% aqueous solution and refluxing for 200mL of water, and filtering.
12.2 preparation method of different sugar-safflower extracts
3 parts of safflower (Tibet, beijing Tongrentang Co., ltd.) of the same batch are taken, 20g of each part is added with 12 times of water (240 mL of water), 0.5% sucrose aqueous solution (1.2 g of sucrose is dissolved in 240mL of water), 0.5% maltose aqueous solution (1.2 g of maltose is dissolved in 240mL of water) is soaked for 30min, refluxed for 30min, filtered by 3 layers of gauze while hot, 10 times of water (200 mL of water) is added to dregs, 0.5% sucrose aqueous solution (1 g of sucrose is dissolved in 200mL of water), 0.5% maltose aqueous solution (1 g of maltose is dissolved in 200mL of water) is added, refluxed for 20min, 3 layers of gauze are filtered while hot, and 2 times of filtrate are combined and the volume is fixed to 400 mL.
12.3 determination of index component content
Preparation of 12.3.1 sample solution
Precisely measuring 1mL of safflower extract with different concentrations, adding 1mL of methanol, diluting the sample for 2 times, mixing well, 15000 r.min -1 Centrifuging for 5min, collecting supernatant, and filtering with 0.22 μm microporous membrane.
Preparation of 12.3.2 control solution
Adding methanol into appropriate amount of hydroxysafflor yellow A reference substance to obtain mother liquor with mass concentration of 1mg/ml, diluting with methanol to obtain reference substance stock solution of 0.2mg/ml, 0.1mg/ml, 0.05mg/ml, 0.025mg/ml and 0.0125mg/ml, and shaking.
The linear equation of hydroxysafflor yellow A is y=13131x-26.496, R 2 ＝0.9993。
12.3.3 content determination method
Analysis was performed using an Agilent 1260 type liquid chromatograph (equipped with an ultraviolet detector), column: c18 (250 mm. Times.4.6 mm,5 μm). The sample loading was 5ul. The detection wavelength was 403nm and the flow rate was 1ml/min. The mobile phase A is 0.5% phosphoric acid water, the B phase is acetonitrile, and the gradient flow is carried out (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, 60-65 min,31% -90% B, 65-67 min,90% -5% B).
12.4 experimental results
The effect of sucrose at various concentrations on the extraction rate of index components in safflower is shown in Table 14.
TABLE 14 Effect of sucrose at different concentrations on extraction yield of index ingredients in safflower
* P <0.05 vs. safflower water extract
As can be seen from Table 14, the concentration of sucrose increased to varying degrees in the range of 0.5g/100mL to 2.5g/100mL, with the effects being most pronounced at sucrose concentrations of either 0.5g/100mL or 2g/100 mL.
The effect of different disaccharides on the extraction of the index component from safflower is shown in Table 15.
TABLE 15 Effect of different disaccharides on the extraction yield of index ingredients in safflower
* P <0.05 vs. safflower water extract
As can be seen from Table 15, sucrose concentration at 0.5g/100mL significantly increased the content of the index ingredient in safflower.
Embodiment thirteen different sugar-Astragalus polysaccharide extraction experiments
13.1 preparation method of different sugar-astragalus extract
5 parts of astragalus membranaceus decoction pieces in the same batch are taken, 50g of each part is added with 7 times of water (350 ml of water), 1% sucrose aqueous solution (3.5 g of sucrose is added with water to a volume of 350 ml), 1% glucose aqueous solution (3.5 g of glucose is added with water to a volume of 350 ml), 1% fructose aqueous solution (3.5 g of fructose is added with water to a volume of 350 ml), 1% lactose aqueous solution (3.5 g of lactose is added with water to a volume of 350 ml), 1% maltose aqueous solution (3.5 g of maltose is added with water to a volume of 350 ml), soaking is performed for 30min, reflux is performed for 30min, 6 times of water (300 ml of water) is added to the residues, 1% sucrose aqueous solution (3.0 g of sucrose is added with water to a volume of 300 ml), 1% glucose aqueous solution (3.0 g of glucose is added with water to a volume of 300 ml), 1% fructose aqueous solution (3.0 g of fructose is added with water to a volume of 300 ml), 1% lactose aqueous solution (3.0 g of trehalose is added with water to a volume of 300 ml), 1% lactose aqueous solution (3.0 g of lactose is added with water to a volume of 300 ml), and filtrate is obtained after reflux is filtered with water for 2 min.
13.2 determination of Total polysaccharide content
13.2.1 sample solution preparation method
Precisely measuring 1mL of radix astragali water extract, placing in a 15mL centrifuge tube, adding 4mL of absolute ethanol, mixing, soaking for 12h, 70000 r.min -1 Centrifuging for 20min, removing supernatant, dissolving precipitate in water, standing in 25ml measuring flask, and shaking.
13.2.2 preparation method of control solution
Taking proper amount of anhydrous glucose reference substance, precisely weighing, and adding water to obtain 0.132mg of anhydrous glucose solution per 1 ml.
13.2.3 selection of measurement wavelength
Precisely measuring 0.6ml of sample solution, placing the sample solution into 10ml of test tubes with plug scales, adding water to 2.0ml respectively, shaking uniformly, precisely adding 1ml of 5% phenol solution respectively, swirling for 20s, swirling for 5ml, swirling for 20s, carrying out water bath heat preservation at 80 ℃ for 20min, taking out, rapidly cooling in an ice water bath for 10min, taking out, and scanning in a wavelength range of 400-800 nm. There is a maximum absorption peak at 490nm, and thus, the total polysaccharide content in the aqueous extract of Astragalus membranaceus was determined using 490 nm.
Drawing of 13.2.4 standard curve
Precisely measuring 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml and 1ml of reference substance solution, respectively placing into 10ml test tubes with plug scales, respectively adding water to 2.0ml, shaking uniformly, respectively precisely adding 1ml of 5% phenol solution, vortexing for 20s, concentrated sulfuric acid for 5ml, vortexing for 20s, taking out after water bath heat preservation for 20min at 80 ℃, rapidly cooling for 10min in an ice water bath, and taking out, and taking the corresponding reagent as blank. Absorbance was measured at 490nm by uv-vis spectrophotometry (general 0401). And drawing a standard curve by taking absorbance as an ordinate and concentration as an abscissa to obtain a linear equation.
13.3 experimental results
The results of the total polysaccharide content measurements in the different sugar-astragalus extracts are shown in table 16.
TABLE 16 determination of total polysaccharide content in different sugar-Astragalus root extracts
* P <0.05VS astragalus water extract
As can be seen from Table 16, sucrose, glucose, fructose, trehalose, lactose, maltose (1 g/100mL concentration) all increased the polysaccharide content of Astragalus mongholicus to varying degrees, with the sucrose effect being most pronounced.
Fourteen experiments on extraction of sucrose-Astragalus polysaccharides with different concentrations
14.1 preparation method of sucrose-radix astragali extract with different concentrations
7 parts of raw astragalus decoction pieces in the same batch are taken, and 50g of each part is taken. 7 times of water (350 ml of water), 1% sucrose aqueous solution (3.5 g sucrose to volume: 350 ml), 2% sucrose aqueous solution (7 g sucrose to volume: 350 ml), 10% sucrose aqueous solution (35 g sucrose to volume: 350 ml), 20% sucrose aqueous solution (70 g sucrose to volume: 350 ml), 30% sucrose aqueous solution (105 g sucrose to volume: 350 ml), 40% sucrose aqueous solution (140 g sucrose to volume: 350 ml), soaking for 30min, refluxing for 30min, filtering while hot, adding 6 times of water (300 ml of water) to the residues, 1% sucrose aqueous solution (3.0 g sucrose to volume: 300 ml), 2% sucrose aqueous solution (6.0 g sucrose to volume: 300 ml), 10% sucrose aqueous solution (30 g sucrose to volume: 300 ml), 20% sucrose aqueous solution (60 g sucrose to volume: 300 ml), 30% sucrose aqueous solution (90 g to volume: 300 ml), 40% sucrose to volume: 300 ml), filtering while hot, mixing the above filtrates, and filtering while the filtrate.
14.2 determination of Total polysaccharide content
The same as in the thirteenth embodiment.
14.3 experimental results
The measurement results of the total polysaccharide content in the astragalus extract obtained by different concentrations of sucrose are shown in table 17.
TABLE 17 determination of total polysaccharide content in Astragalus extract obtained with sucrose at different concentrations
* P <0.05VS astragalus water extract
As can be seen from Table 17, sucrose solutions (1-40 g/100 mL) with different concentrations can raise the content of the effective component (astragalus polysaccharide) in astragalus to different degrees, wherein the effect is most obvious when 30g/100mL of sucrose solution is used.
EXAMPLE fifteen experiments on sucrose-Glycyrrhiza polysaccharide extraction at different concentrations
15.1 preparation method of sucrose-Glycyrrhrizae radix extractive solutions with different concentrations
7 parts of raw licorice decoction pieces (produced by Xinjiang) in the same batch are taken, and each part is 50g. 7 times of water (350 ml water), 1% sucrose aqueous solution (3.5 g sucrose+350 ml water), 2% sucrose aqueous solution (7.0 g sucrose+350 ml water), 10% sucrose aqueous solution (35 g sucrose+350 ml water), 20% sucrose aqueous solution (70 g sucrose+350 ml water), 30% sucrose aqueous solution (105 g sucrose+350 ml water), 40% sucrose aqueous solution (140 g sucrose+350 ml water), soaking for 30 minutes, refluxing for 30 minutes, filtering with gauze while hot, adding 6 times of water (300 ml water) to the residues, 1% sucrose aqueous solution (3.0 g sucrose+300 ml water), 2% sucrose aqueous solution (6.0 g sucrose+300 ml water), 10% sucrose aqueous solution (30 g sucrose+300 ml water), 20% sucrose aqueous solution (60 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), refluxing for 20 minutes, filtering while hot, and combining 2 times to obtain filtrate.
15.2 determination of Total polysaccharide content
Preparation of 15.2.1 sample solution
Precisely weighing 2ml of Glycyrrhrizae radix water extract, adding 8ml of ethanol, mixing, soaking for 12 hr, 5000 r.min -1 Centrifuging for 20min, removing supernatant, dissolving the precipitate in water, fixing volume in 50ml measuring flask, and shaking.
Preparation of 15.2.2 control solution
Taking proper amount of anhydrous glucose reference substance, precisely weighing, and adding water to obtain solution (10 ml) containing 0.34mg of anhydrous glucose per 1 ml.
15.2.3 selection of measurement wavelength
Precisely measuring 1.0ml of sample solution, placing into 10ml test tubes with plug scales, adding water to 2.0ml, shaking uniformly, slowly dripping 8ml of 0.2% anthrone-sulfuric acid solution, mixing uniformly, placing into boiling water bath, preserving heat for 10 minutes, taking out, immediately placing into ice water bath, cooling for 10 minutes, taking out, and scanning within a wavelength range of 400-800 nm. There was a maximum absorption peak at 593nm, and therefore, the total polysaccharide content in the aqueous licorice extract was determined using 593 nm.
Drawing of 15.2.4 standard curve
Precisely measuring 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml and 1ml of reference substance solution, respectively placing into 10ml test tubes with plug scales, respectively adding water to 2.0ml, shaking uniformly, slowly dripping 8ml of 0.2% anthrone-sulfuric acid solution, mixing uniformly, placing into boiling water bath, preserving heat for 10 minutes, taking out, immediately placing into ice water bath, cooling for 10 minutes, taking out, and taking the corresponding reagent as blank. Absorbance was measured at a wavelength of 593nm by uv-vis spectrophotometry (general rule 0401). And drawing a standard curve by taking absorbance as an ordinate and concentration as an abscissa to obtain a linear equation.
15.3 experimental results
The results of measuring the total polysaccharide content in the different sugar-licorice aqueous extracts are shown in Table 18.
TABLE 18 determination of total polysaccharide content in Glycyrrhiza concentrated solutions obtained with sucrose at different concentrations
* P <0.05VS licorice water extract
As can be seen from Table 18, sucrose solutions (1-40 g/100 mL) with different concentrations can raise the content of effective components (Glycyrrhiza polysaccharide) in Glycyrrhiza uralensis to different degrees, wherein the effect is most obvious with 20g/100mL sucrose solution.
Example sixteen different sugar-Glycyrrhiza polysaccharide extraction experiments
16.1 preparation method of different sugar-Glycyrrhrizae radix extractive solutions
3 parts of raw licorice decoction pieces (produced by Xinjiang) in the same batch are taken, and each part is 50g. 7 times of water (350 ml of water), 20% maltose aqueous solution (70 g maltose+350 ml of water), 20% sucrose aqueous solution (70 g lactose+350 ml of water) are respectively added for soaking for 30min, refluxing for 30min, filtering with gauze while hot, adding 6 times of water (300 ml of water) into the residues, 20% maltose aqueous solution (60 g maltose+300 ml of water), 20% sucrose aqueous solution (60 g lactose+300 ml of water), refluxing for 20min, filtering while hot, and combining 2 times of filtrates. Obtaining the product.
16.2 determination of Total polysaccharide content
The same as in the fifteenth embodiment.
16.3 experimental results
The results of measuring the total polysaccharide content in the different sugar-licorice aqueous extracts are shown in Table 19.
TABLE 19 determination of total polysaccharide content in different sugar-Glycyrrhiza aqueous extracts
* P <0.05VS licorice water extract
As can be seen from Table 19, sucrose and maltose (20 g/100mL concentration) can raise the content of the active ingredient (glycyrrhizin) in licorice to different degrees, wherein the effect of maltose is most obvious.
Example seventeen experiments on extraction of sucrose-ginseng polysaccharide at different concentrations
17.1 preparation method of sucrose-Ginseng radix extractive solutions with different concentrations
6 parts of ginseng decoction pieces (Jilin Fusong) of the same batch are taken, and 40g of ginseng decoction pieces are taken for each part. Respectively adding 7 times of water, 1%, 10%, 20%, 25% and 30% sucrose aqueous solution, soaking for 30min, refluxing for 30min, filtering with gauze while hot, respectively adding 6 times of water, 1%, 10%, 20%, 25% and 30% sucrose aqueous solution into the residue, refluxing for 20min, filtering with two layers of gauze while hot, mixing 2 times of filtrates, and adding water to 800 mL.
17.2 determination of Total polysaccharide content
17.2.1 preparation method of sample solution
Accurately measuring 1mL of Ginseng radix extractive solutions containing sucrose with different concentrations, respectively adding 4mL of absolute ethanol, mixing, soaking for 12 hr, 70000 r.min -1 Centrifuging for 20min, removing supernatant, dissolving the precipitate in water, fixing volume in 10mL measuring flask, and shaking.
17.2.2 preparation method of control solution
Taking a proper amount of anhydrous glucose reference substance, precisely weighing, and adding water to prepare a solution containing 0.132mg of anhydrous glucose per 1 mL.
17.2.3 Total polysaccharide assay
Precisely measuring 0.4, 0.3 and 0.2mL of ginseng extract sample solutions with different concentrations of sucrose, placing the sample solutions into 10mL of test tubes with stopper scales, adding water to 2.0mL of each sample solution, shaking uniformly, precisely adding 1mL of 5% phenol solution, swirling 20s, 5mL of concentrated sulfuric acid, swirling 20s, taking out after water bath heat preservation at 80 ℃ for 20min, rapidly cooling for 10min in an ice water bath, and taking corresponding reagent (water) as a blank. Absorbance was measured at wavelength 489 nm.
Drawing of 17.2.4 standard curve
Precisely measuring 0.1mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL and 1mL of reference substance solution, respectively placing the reference substance solution and the reference substance solution into 10mL test tubes with plug scales, respectively adding water to 2.0mL, shaking uniformly, respectively precisely adding 1mL of 5% phenol solution, vortexing for 20s, concentrating sulfuric acid for 5mL, vortexing for 20s, carrying out water bath heat preservation at 80 ℃ for 20min, taking out, rapidly cooling in an ice water bath for 10min, and taking out, wherein the corresponding reagent is blank. Absorbance was measured at 489nm by uv-vis spectrophotometry (generally 0401). And drawing a standard curve by taking absorbance as an ordinate and concentration as an abscissa.
17.3 experimental results
The effect of sucrose at various concentrations on the extraction yield of ginseng polysaccharide is shown in Table 20.
TABLE 20 Effect of sucrose at different concentrations on extraction yield of ginseng polysaccharide
* P <0.05VS ginseng water extract
As can be seen from Table 20, sucrose solutions (10-40 g/100 mL) of different concentrations can significantly increase the content of the active ingredient in ginseng to different degrees.
Example eighteen different sugar-ginseng polysaccharide extraction experiments
18.1 preparation method of disaccharide-Ginseng radix extractive solutions of different types
3 parts of ginseng decoction pieces (Jilin Fusong) of the same batch are taken, and 40g of ginseng decoction pieces are taken for each part. 7 times of water, 30% sucrose and 30% maltose aqueous solution were added respectively, and immersed for 30 minutes, followed by the same procedure as in example seventeen.
18.2 determination of Total polysaccharide content
The same as in example seventeenth.
18.3 experimental results
The effect of different disaccharides on the total polysaccharide extraction yield of ginseng is shown in Table 21.
TABLE 21 Effect of disaccharides of different classes on the extraction yield of Total polysaccharide from Ginseng radix
* P <0.05VS ginseng water extract
As can be seen from Table 21, sucrose and maltose (30 g/100mL concentration) can increase the content of the active ingredient (ginseng polysaccharide) in ginseng to different degrees, wherein the effect of sucrose is most obvious.
Example nineteen experiments on extraction of sucrose-Lycium barbarum polysaccharides at different concentrations
19.1 preparation method of extractive solution of fructus Lycii
6 parts of medlar (Ningxia) in the same batch are taken, and 40g of medlar is taken as each part. 7 times of water (280 mL of water), 1% sucrose aqueous solution (2.8 g sucrose+280 m of water), 10% sucrose aqueous solution (28 g sucrose+280 m of water), 20% sucrose aqueous solution (56 g sucrose+280 m of water), 30% sucrose aqueous solution (84 g sucrose+280 m of water), 40% sucrose aqueous solution (112 g sucrose+280 m of water) and 30% maltose aqueous solution (84 g maltose+280 m of water) are respectively added, the mixture is soaked for 30 minutes, the mixture is refluxed for 30 minutes, the mixture is filtered by gauze while the mixture is still hot, 6 times of water, 1% sucrose aqueous solution, 10% sucrose aqueous solution, 20% sucrose aqueous solution, 30% sucrose aqueous solution, 40% sucrose aqueous solution and 30% maltose aqueous solution are respectively added, the mixture is refluxed for 20 minutes, and two layers of gauze while the mixture is hot are filtered, 2 times of filtrates are combined, and the water is added to a volume of 800mL.
19.2 determination of Total polysaccharide content
19.2.1 sample solution preparation method
The same procedure was used for the preparation of the sample solutions as in example seventeen.
Preparation of 19.2.2 control solution
The same procedure was used for the preparation of the control solution as in example seventeen.
19.2.3 Total polysaccharide assay
Precisely measuring 0.4, 0.3 and 0.3mL of medlar extract sample solutions with different concentrations of sucrose, placing the medlar extract sample solutions into 10mL of test tubes with stopper scales, respectively adding water to 2.0mL, shaking uniformly, respectively precisely adding 1mL of 5% phenol solution, swirling 20s, 5mL of concentrated sulfuric acid, swirling 20s, carrying out water bath heat preservation at 80 ℃ for 20min, taking out, rapidly cooling in an ice water bath for 10min, and taking corresponding reagent (water) as blank. Absorbance was measured at wavelength 489 nm.
Drawing of 19.2.4 standard curve
The same method as in the seventeenth embodiment was used for the standard curve drawing.
19.3 experimental results
The effect of varying concentrations of sucrose on the extraction of Lycium barbarum polysaccharide is shown in Table 22.
TABLE 22 Effect of sucrose at different concentrations on Lycium barbarum polysaccharide extraction yield
* P <0.05VS medlar water extract
As can be seen from Table 22, sucrose solutions of different concentrations (10-40 g/100 mL) are capable of increasing the content of the active ingredient (Lycium barbarum polysaccharide) in Lycium barbarum to different degrees, wherein the effect is most obvious when 40g/100mL of sucrose solution is used.
Example twenty different sugar-Lycium Barbarum polysaccharide extraction experiments
20.1 preparation methods of disaccharide-Lycium barbarum extracts of different classes
The medlar decoction pieces (producing area: ningxia; manufacturer: tianjin Shengshun Utility Co., ltd.; lot number: X20122702A) of the same batch were taken 3 parts, 40g each. 7 times of water, 30% sucrose and 30% maltose aqueous solution were added respectively, and immersed for 30 minutes, followed by the same procedure as in example nineteenth.
20.2 determination of Total polysaccharide content
The same as in example nineteenth.
20.3 experimental results
The effect of different classes of disaccharides on the extraction yield of Lycium barbarum polysaccharides is shown in Table 23.
TABLE 23 influence of disaccharides of different classes on the extraction yield of Lycium barbarum polysaccharides
* P <0.05VS medlar water extract
As can be seen from Table 23, sucrose and maltose (30 g/100mL concentration) can increase the content of the active ingredient (Lycium barbarum polysaccharide) in Lycium barbarum to different degrees.
The foregoing is merely exemplary embodiments of the present invention, and specific structures and features that are well known in the art are not described in detail herein. It should be noted that modifications and improvements can be made by those skilled in the art without departing from the scope of the invention, which is also to be considered as the scope of the invention, and which does not affect the effect of the invention and the utility of the patent. The protection scope of the present invention is subject to the content of the claims, and the description of the specific embodiments and the like in the specification can be used for explaining the content of the claims.
The foregoing has outlined rather broadly the more detailed description of embodiments of the invention in order that the detailed description of the principles and embodiments of the invention may be implemented in conjunction with the detailed description of embodiments of the invention that follows. Meanwhile, based on the idea of the present invention, those skilled in the art can make changes or modifications on the specific embodiments and application scope of the present invention, which belong to the protection scope of the present invention. In view of the foregoing, this description should not be construed as limiting the invention.

Claims (8)

1. A method of increasing leaching of saccharide components and/or glycoside components, the method comprising the steps of:
(1) Preparing a sugar solution and/or a sugar alcohol solution;
(2) Soaking a raw material in the sugar solution and/or the sugar alcohol solution; and
(3) Heating and extracting the raw materials in the sugar solution and/or the sugar alcohol solution for 1-3 times, and filtering to obtain a raw material extracting solution.
2. The method according to claim 1, characterized in that the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-40g/100mL;
preferably, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-10g/100mL or 20-40g/100mL;
preferably, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-5g/100mL or 20-30g/100mL;
preferably, the concentration of the sugar solution and/or the sugar alcohol solution is 0.1-2.5g/100mL;
preferably, the concentration of the sugar solution and/or the sugar alcohol solution is 1-2g/100mL;
preferably, the concentration of the sugar solution and/or the sugar alcohol solution is about 0.5g/100mL or about 1g/100mL or about 1.5g/100mL or about 2g/100mL or about 20g/100mL or about 30g/100mL or about 40g/100mL;
preferably, the sugar is selected from one or more of monosaccharides, disaccharides and trisaccharides;
Preferably, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol and xylitol;
preferably, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, sorbose, rhamnose, ribose, xylose and deoxyribose;
preferably, the disaccharide is selected from one or more of maltose, sucrose, lactose, trehalose, melibiose, gentiobiose and trabiose;
particularly preferably, the monosaccharide is glucose and/or fructose; particularly preferably, the trisaccharide is raffinose;
most preferably, the disaccharide is sucrose and/or maltose.
3. The method according to claim 1, wherein the saccharide fraction is an oligosaccharide fraction and/or a polysaccharide fraction;
preferably, the polysaccharide component is selected from one or more of the following: astragalus polysaccharides, licorice polysaccharides, ginseng polysaccharides and wolfberry polysaccharides;
preferably, the glycoside component is selected from one or more of the following: flavonoid glycoside compounds, saponins compounds, diterpenoid glycoside compounds, quinone glycoside compounds and phenolic acid glycoside compounds;
more preferably, the flavonoid glycoside compound is selected from one or more of the following: flavonoid glycoside compounds, isoflavone glycoside compounds, flavanonol glycoside compounds, chalcone glycoside compounds, dihydrochalcone glycoside compounds, hesperidin compounds, anthocyanin compounds, biflavone glycoside compounds and flavanol glycoside compounds;
More preferably, the saponins compound is a tetracyclic triterpene glycoside compound, a pentacyclic triterpene glycoside compound and/or a steroid glycoside compound;
more preferably, the diterpenoid glycosides are selected from one or more of the following: tetracyclic diterpenoid glycosides, tricyclic diterpenoid glycosides, chain diterpenoid glycosides, monocyclic diterpenoid glycosides and bicyclic diterpenoid glycosides;
more preferably, the quinone glycoside compound is benzoquinone glycoside compound, naphthoquinone glycoside compound, phenanthrenequinone glycoside compound and/or anthraquinone glycoside compound;
more preferably, the phenolic acid glycoside compound is a phenolic acid glycoside compound with a C6-C3 parent nucleus structure and/or a phenolic acid glycoside compound with a C6-C1 parent nucleus structure;
more preferably, the steroid glycoside compound is a spirostanol type saponin compound, a furostanol type saponin compound and/or an isospirostanol type saponin compound;
preferably, the isospirostanol type saponin compound is a dioscin type compound; preferably, the furostanol type saponin compound is a timosaponin type compound;
preferably, the tetracyclic triterpenoid saponins are lanolin alkane type tetracyclic triterpenoid saponins and/or dammarane type tetracyclic triterpenoid saponins; preferably, the pentacyclic triterpenoid saponins are oleanane-type pentacyclic triterpenoid saponins, ursane-type pentacyclic triterpenoid saponins and/or lupin-type pentacyclic triterpenoid saponins;
Preferably, the anthraquinone glycoside compound is a monoanthracene nucleus compound and/or a dianthracene nucleus compound;
particularly preferably, the isoflavone glycoside compound is calycosin glucoside and/or formononetin; particularly preferably, the flavanone glycoside compound is apigenin and/or liquiritin; the chalcone glycoside compound is hydroxysafflor yellow A;
particularly preferably, the furostanol-type steroid saponin compound is timosaponin BII; particularly preferably, the lanolin alkane type tetracyclic triterpene saponin compound is astragaloside IV; particularly preferably, the dammarane type tetracyclic triterpene saponin compound is ginsenoside Rg1; particularly preferably, the oleanane-type pentacyclic triterpene saponin compound is glycyrrhizic acid;
particularly preferably, the tetracyclic diterpenoid glycosides are stevioside and/or rebaudioside;
particularly preferably, the phenolic acid glycoside compound with a C6-C3 parent nucleus structure is hydroxysafflor yellow A;
particularly preferably, the dianthrone nucleoside compound is sennoside A and/or sennoside B; the monoanthracene nucleoside compound is aloin.
4. The method according to claim 1, wherein the raw material is a raw material containing a saccharide component and/or a glycoside component;
Preferably, the raw materials are one or more traditional Chinese medicines, plant medicines and/or marine organisms containing saccharide components and/or glycoside components.
5. The method according to claim 1, wherein, in step (2),
the soaking is performed at room temperature;
preferably, the step of soaking comprises soaking the raw material in the sugar solution and/or the sugar alcohol solution at room temperature for 10-120min;
still preferably, the step of soaking includes soaking the raw material in the sugar solution and/or the sugar alcohol solution at room temperature for 20 to 60 minutes;
more preferably, the step of soaking comprises soaking the feedstock in the sugar solution and/or the sugar alcohol solution at room temperature for about 30 minutes;
particularly preferably, 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;
still more preferably, 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;
particularly preferably, the weight-to-volume ratio of the raw material to the sugar solution and/or the sugar alcohol solution is 1:6-1:12;
still more preferably, in step (3), the heat extraction is heat reflux extraction;
Particularly preferably, the heating time is from 20min to 60min;
particularly preferably, the heating time is from 20min to 30min;
most preferably, the heating is for a time of about 20 minutes or about 30 minutes.
6. Use of the method according to any one of claims 1 to 5 for the preparation of a raw material extract or a raw material concentrate, wherein the raw material is a traditional Chinese medicine, a plant drug and/or a marine organism.
7. Use of the method according to any one of claims 1 to 5 for the preparation of a pharmaceutical composition comprising raw materials, wherein the raw materials are traditional Chinese medicines, botanicals and/or marine organisms.
8. Use of the method according to any one of claims 1 to 5 for the preparation of a pharmaceutical formulation or a functional or health food comprising raw materials, wherein the raw materials are traditional Chinese medicines, botanicals and/or marine organisms.
CN202210730365.4A 2022-06-24 2022-06-24 Method for increasing leaching of saccharides and glycoside components and application thereof Pending CN117327135A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116850217A (en) * 2022-06-27 2023-10-10 中国中医科学院中药研究所 Method for improving leaching rate of compound containing phenolic hydroxyl group by utilizing aqueous sugar solution

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116850217A (en) * 2022-06-27 2023-10-10 中国中医科学院中药研究所 Method for improving leaching rate of compound containing phenolic hydroxyl group by utilizing aqueous sugar solution

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