阅读说明：本技术 一种联合分离多种甘草活性成分的生产工艺 (Production process for combined separation of multiple active ingredients of liquorice ) 是由 张继义 罗兴平 张丹 于 2021-08-17 设计创作，主要内容包括：本发明涉及一种联合分离多种甘草活性成分的生产工艺,该工艺包括以下步骤：⑴生产甘草酸单铵盐后的母液粉和溶剂,室温下搅拌提取,得到提取液和溶剂不溶部分；⑵提取液经过滤、回收溶解、干燥、粉碎,得到产物A；产物A中加入纯化水,经加热搅拌提取后,得到上清液和产物B；上清液经减压浓缩后加95%的乙醇加热使其溶解；然后依次加活性炭脱色、过滤、静置结晶、分离、重结晶,即得含量>90%的甘草苷；⑶溶剂不溶部分经真空干燥、常压加热,得到产物C,该产物C经干燥、粉碎,即得甘草酸含量为10%以上的脱苦甘草甜味素；⑷产物B经真空干燥、粉碎,即得含量在8～15%的甘草素和异甘草素的粗品。本发明工艺简单、成本低、安全性高,适用于工业化规模生产。(The invention relates to a production process for jointly separating multiple active ingredients of liquorice, which comprises the following steps: the method comprises the steps of stirring and extracting mother liquor powder and a solvent after production of mono-ammonium glycyrrhizinate at room temperature to obtain an extracting solution and a solvent insoluble part; filtering, recovering, dissolving, drying and crushing the extracting solution to obtain a product A; adding purified water into the product A, and heating, stirring and extracting to obtain a supernatant and a product B; concentrating the supernatant under reduced pressure, adding 95% ethanol, and heating to dissolve; then sequentially adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing to obtain liquiritin with the content of more than 90%; carrying out vacuum drying and normal-pressure heating on the solvent-insoluble part to obtain a product C, and drying and crushing the product C to obtain the debitterized liquorice monosodium glutamate with the glycyrrhizic acid content of more than 10%; and fourthly, drying and crushing the product B in vacuum to obtain crude products of liquiritigenin and isoliquiritigenin with the content of 8-15%. The method has the advantages of simple process, low cost and high safety, and is suitable for industrial mass production.)

1. A production process for jointly separating multiple active ingredients of liquorice comprises the following steps:

the method comprises the steps of respectively adding mother liquor powder and a solvent after production of mono-ammonium glycyrrhizinate into a reaction vessel, stirring and extracting for 2-3 times at room temperature to respectively obtain an extracting solution and a solvent insoluble part;

filtering, recovering, dissolving, drying and crushing the extracting solution to obtain a product A; adding purified water with the mass being 4-15 times that of the product A into the product A, heating, stirring, extracting, standing and precipitating to respectively obtain a supernatant and a product B; concentrating the supernatant under reduced pressure until the specific gravity is 1.15-1.20 at 60 ℃, adding ethanol with the volume concentration of 95%, and heating to dissolve the ethanol; then sequentially adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing to obtain liquiritin with the content of more than 90%;

thirdly, drying the solvent-insoluble part in vacuum and heating at normal pressure until the solvent is completely volatilized to obtain a product C, and drying and crushing the product C to obtain the debitterized glycyrrhizin with the glycyrrhizic acid content of more than 10%;

and fourthly, drying and crushing the product B in vacuum to obtain crude products of liquiritigenin and isoliquiritigenin with the content of 8-15%.

2. The process of claim 1, wherein the process comprises the steps of: the solvent in the step is one of ethyl acetate, ethyl formate or a mixed liquid of ethyl acetate and ethyl formate.

3. The process of claim 1, wherein the process comprises the steps of: the volume of the solvent in the step is 3-5 BV of the mother liquid powder.

4. The process of claim 1, wherein the process comprises the steps of: the heating, stirring and extracting conditions in the step II are that the temperature is 40-100 ℃, the extracting times are 2-3 times, and the extracting time is 1-2 hours each time.

5. The process of claim 1, wherein the process comprises the steps of: in the step II, the addition amount of the ethanol is 0.2-6 BV of the supernatant.

Technical Field

The invention relates to the field of food and medicine, in particular to a production process for jointly separating multiple active ingredients of liquorice.

Background

Licorice (Glycyrrhiza uralensis Fisch) is a plant belonging to genus Glycyrrhiza of family Leguminosae, and has 29 varieties of 6 varieties in the whole world, wherein 18 varieties of 3 varieties exist in China and are widely distributed in northwest, north China, northeast China and other areas, and meanwhile, Glycyrrhiza uralensis, Glycyrrhiza inflata and Glycyrrhiza glabra are one of the basic plants of medicinal material licorice, and are widely applied to industries such as medicine, food, cosmetics and the like.

The main active components of Glycyrrhrizae radix extract include glycyrrhizic acid and its multiple analogues of glycyrrhizin, Glycyrrhrizae radix total flavonoids such as liquiritin, liquiritigenin, isoliquiritigenin, glabridin, etc., and Glycyrrhrizae radix polysaccharide.

The Liquiritin (Liquiritin) is an important monomer active component in flavonoid compounds and has strong pharmacological activity, researches show that the Liquiritin has obvious antidepressant effect, neuroprotective effect on nerve cell injury and protective effect on cardiac system and liver toxicity, in addition, the Liquiritin has various activities of antivirus and anti-inflammation, recent research results of big research teams show that the Liquiritin can inhibit the replication of novel coronavirus, and the Liquiritin is expected to be developed into a new medicine for resisting SARS-CoV-2. Liquiritigenin (liquiritigenin) is a flavanone monomer compound existing in Glycyrrhrizae radix, and has physiological effects of resisting cancer, ulcer, inflammation and protecting liver. Isoglycyrrhiza is a "strong analog" of the organ transplant rejection drug "rapamycin" in its natural products. Glycyrrhizic acid has antiinflammatory and antiviral effects, and also has pharmacological effects of resisting bacteria, resisting cancer, resisting allergy, resisting blood coagulation activity, and removing local excessive fat. Glycyrrhizic acid can also be used as raw material to prepare glycyrrhizin, which is widely used in food field.

Qinghai lake pharmaceutical industry Co., Ltd applies for a patent (application number: 201610229840.4) named as "a method for separating and purifying liquiritin", which adopts 2 times of extraction and enrichment of extract, silica gel column chromatography separation and crystallization to obtain high-purity liquiritin. However, the extracting solution used in the process is methanol, chloroform, dichloromethane and the like, and the organic solvents such as benzene, toluene and the like used in the purification of a chromatographic column have strong toxicity, and the use of the organic solvents can cause solvent residue to influence the quality of liquiritin; the process uses silica gel column for separation and purification, and the silica gel is not easy to regenerate, so that the use times of the silica gel are limited, and the silica gel cannot be repeatedly used, so that the production cost is high; the flash point of the ethyl acetate used for chromatographic separation is low, and potential safety hazards exist; only one product of liquiritin is used, the glycyrrhizin and the total flavone in the ammoniation mother liquor cream are not comprehensively utilized, the production cost is high, the waste is caused, and the environment is polluted.

A patent (application number: 201110287907.7) named as a method for extracting liquiritin from liquorice is applied by Tianjin peak natural product development limited company, and the patent process comprises the steps of crushing liquorice, extracting with deionized water, filtering to obtain liquiritin crude extract, and then separating, purifying and recrystallizing the crude extract on a mixed resin column to obtain the liquiritin. However, the process only extracts and separates the liquiritin from the liquorice, discards other important functional components in the liquorice, such as glycyrrhizic acid and general flavone, and has no comprehensive utilization, only theoretical significance and no market prospect.

A patent named 'liquiritin and a preparation method thereof' is applied by Jiangsu Tiancheng pharmaceutical industry Co., Ltd (application number: 201010579521.9). the patent process comprises the steps of firstly ammoniating glycyrrhizic acid powder, extracting to obtain liquiritin, then separating, purifying and crystallizing by using a polyamide column to obtain the liquiritin. The process only extracts and separates liquiritin from liquorice, and discards other important functional components in the liquorice, such as glycyrrhizic acid and total flavonoids, which are not comprehensively utilized. Also has high production cost and no market competitiveness.

The university of Chinese academy of sciences applies for a patent (application number: 200710011041.0) named as a preparation method of liquiritin, the technology of the patent extracts and deposits liquiritin by alcohol, separates the liquiritin by a molecular membrane separator, macroporous resin and a high-efficiency industrial chromatographic column in sequence, and freezes and dries to obtain the liquiritin, and the technology is complex, has high requirements on equipment, large operation difficulty and long time consumption, and is not suitable for industrial mass production.

Gansu pan-plant medicine industry Co., Ltd applies for a process for the combined separation of high purity glycyrrhizin, bitter-removed glycyrrhizin and licorice total flavonoids from a glycyrrhiza monoammonium salt mother liquor paste (application No. 202011128456.8), the process comprises eluting the glycyrrhiza monoammonium salt mother liquor paste with different types of resins to obtain the glycyrrhizin and co-produce glycyrrhizin and licoflavone, although the raw materials can be fully and effectively utilized, the flavonoid components such as glycyrrhizin and isoliquiritigenin which coexist with the glycyrrhizin are not classified and collected, the macroporous resin elution solvent consumption is large, and the operation is complicated and is not convenient for large-scale production.

In conclusion, the prior art has the defects of complex process, high requirement on equipment, high operation difficulty, use of various organic solvents which easily cause potential safety hazards, incomplete and effective utilization of raw materials, low additional value, high production cost and the like.

Disclosure of Invention

The invention aims to solve the technical problem of providing a production process for jointly separating multiple licorice active ingredients with low cost and high safety.

In order to solve the problems, the production process for jointly separating multiple active ingredients of liquorice comprises the following steps:

the method comprises the steps of respectively adding mother liquor powder and a solvent after production of mono-ammonium glycyrrhizinate into a reaction vessel, stirring and extracting for 2-3 times at room temperature to respectively obtain an extracting solution and a solvent insoluble part;

filtering, recovering, dissolving, drying and crushing the extracting solution to obtain a product A; adding purified water with the mass being 4-15 times that of the product A into the product A, heating, stirring, extracting, standing and precipitating to respectively obtain a supernatant and a product B; concentrating the supernatant under reduced pressure until the specific gravity is 1.15-1.20 at 60 ℃, adding ethanol with the volume concentration of 95%, and heating to dissolve the ethanol; then sequentially adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing to obtain liquiritin with the content of more than 90%;

thirdly, drying the solvent-insoluble part in vacuum and heating at normal pressure until the solvent is completely volatilized to obtain a product C, and drying and crushing the product C to obtain the debitterized glycyrrhizin with the glycyrrhizic acid content of more than 10%;

and fourthly, drying and crushing the product B in vacuum to obtain crude products of liquiritigenin and isoliquiritigenin with the content of 8-15%.

The solvent in the step is one of ethyl acetate, ethyl formate or a mixed liquid of ethyl acetate and ethyl formate.

The volume of the solvent in the step is 3-5 BV of the mother liquid powder.

The heating, stirring and extracting conditions in the step II are that the temperature is 40-100 ℃, the extracting times are 2-3 times, and the extracting time is 1-2 hours each time.

In the step II, the addition amount of the ethanol is 0.2-6 BV of the supernatant.

Compared with the prior art, the invention has the following advantages:

1. the resource utilization rate is high, and the added value of the product is high:

the invention adopts a scientific and feasible method to separate and prepare high-purity liquiritin (content 90%), flavonoid-removed glycyrrhizin (glycyrrhizic acid content is more than 10%) and crude liquiritigenin and isoliquiritigenin (content is 8-15%) from ammonium glycyrrhetate mother liquor powder through the steps of dissolution and combined separation of different solvents, crystallization, recrystallization and the like. Compared with the prior art, the obtained liquiritin has high purity, and simultaneously, three products of flavonoid-removed glycyrrhizin, crude liquiritigenin and isoliquiritigenin are co-produced, so that the raw materials can be fully and effectively utilized.

2. The production safety is high:

in the invention, only two organic solvents of edible solvents of ethyl acetate, methyl acetate and ethanol are used, other organic solvents with low flammable and explosive flash points are not used, other organic solvent residues are avoided, and the potential safety hazard in the production process can be reduced.

3. The production cost is low:

the solvents used in the invention can be recycled, thereby reducing the emission and the production cost.

4. Is suitable for large-scale industrial production:

the method has the advantages of simple process, simple and convenient operation, easy realization of standardization and automation, and is suitable for industrial large-scale production.

Detailed Description

A production process for jointly separating multiple active ingredients of liquorice comprises the following steps:

the method comprises the steps of adding mother liquor powder and a solvent after production of mono-ammonium glycyrrhizinate into a reaction vessel, stirring and extracting for 2-3 times at room temperature, and obtaining an extracting solution and a solvent insoluble part respectively.

Wherein: the solvent is one of ethyl acetate, ethyl formate or a mixture of the ethyl acetate and the ethyl formate. The volume of the solvent is 3-5 BV of the mother liquor powder.

Filtering, recovering, dissolving, drying and crushing the extracting solution to obtain a product A; and adding purified water with the mass being 4-15 times of that of the product A into the product A, heating and stirring the product A at the temperature of 40-100 ℃, and extracting the product A for 2-3 times, wherein each time of extraction lasts for 1-2 hours. Stirring, extracting, standing and precipitating to respectively obtain a supernatant and a product B. Product B is the water insoluble fraction.

Concentrating the supernatant under reduced pressure until the specific gravity is 1.15-1.20 (60 ℃), adding 0.2-6 BV of ethanol with the volume concentration of 95%, and heating to dissolve the ethanol; then sequentially adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing to obtain the liquiritin with the content of more than 90%.

And thirdly, drying the solvent-insoluble part in vacuum, heating under normal pressure until the solvent is volatilized completely to obtain a product C, and drying and crushing the product C to obtain the debitterized glycyrrhizin (namely the flavonoid-removed glycyrrhizin) with the glycyrrhizic acid content of more than 10%.

And fourthly, drying and crushing the product B in vacuum to obtain crude products of liquiritigenin and isoliquiritigenin with the content of 8-15%. The crude product can be used as raw material for preparing high-content glycyrrhizin and isoliquiritigenin.

Example 1 a process for the combined separation of multiple active ingredients of licorice, comprising the steps of:

firstly, adding 10kg of mother liquor powder after producing mono-ammonium glycyrrhizinate into a jacketed kettle, adding 50L of ethyl acetate each time, stirring and extracting for 3 times at room temperature to respectively obtain an extracting solution and an ethyl acetate insoluble substance.

Filtering, recovering, dissolving, drying and crushing the extracting solution to obtain 3.5kg of a product A; and adding the product A into a jacketed kettle, adding 12L of purified water each time, heating and stirring at 100 ℃, and extracting for 3 times, wherein the extraction time is 1-2 hours each time. Stirring, extracting, standing and precipitating to respectively obtain a supernatant and a product B. Product B is the water insoluble fraction.

Concentrating the supernatant under reduced pressure to obtain fluid extract with a specific gravity of 1.15 (60 deg.C) 2.5kg, adding 5BV ethanol with a volume concentration of 95%, and heating to dissolve; then sequentially adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing to obtain 306g of white solid liquiritin with the content of 92.7 percent and the weight yield of 3.06 percent.

And thirdly, drying the ethyl acetate insoluble substance in vacuum, heating the ethyl acetate insoluble substance at normal pressure until the solvent in the ethyl acetate insoluble substance is volatilized completely to obtain a product C, and drying and crushing the product C to obtain 6.5kg of the debitterized glycyrrhizin, wherein the glycyrrhizic acid content is 12.42 percent, and the weight yield is 65.0 percent.

And fourthly, drying and crushing the product B in vacuum to obtain 2.8kg of crude product containing the liquiritigenin and the isoliquiritigenin. Wherein: the content of liquiritigenin is 11.20%, the content of isoliquiritigenin is 11.6%, and the weight yield is 28.0%.

Embodiment 2 a process for the combined separation of multiple active ingredients of licorice, comprising the steps of:

firstly, adding 10kg of mother liquor powder after producing mono-ammonium glycyrrhizinate into a jacketed kettle, adding 30L of ethyl acetate each time, stirring and extracting for 3 times at room temperature to respectively obtain an extracting solution and an ethyl acetate insoluble substance.

Filtering, recovering, dissolving, drying and crushing the extracting solution to obtain 2.8kg of a product A; and adding the product A into a jacketed kettle, adding 0.4L of purified water each time, heating and stirring at 60 ℃, and extracting for 3 times, wherein the extraction time is 1-2 hours each time. Stirring, extracting, standing and precipitating to respectively obtain a supernatant and a product B. Product B is the water insoluble fraction.

Concentrating the supernatant under reduced pressure to obtain fluid extract with a specific gravity of 1.20 (60 deg.C) 2kg, adding 3BV ethanol with a volume concentration of 95%, and heating to dissolve; then sequentially adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing to obtain 280g of white solid liquiritin, wherein the content is 96.5 percent, and the weight yield is 2.80 percent.

And thirdly, drying the ethyl acetate insoluble substance in vacuum, heating the ethyl acetate insoluble substance at normal pressure until the solvent in the ethyl acetate insoluble substance is volatilized completely to obtain a product C, and drying and crushing the product C to obtain 7.5kg of the debitterized glycyrrhizin, wherein the glycyrrhizic acid content is 11.38%, and the weight yield is 75.0%.

And fourthly, drying and crushing the product B in vacuum to obtain 2.24kg of crude product containing the liquiritigenin and the isoliquiritigenin. Wherein: the content of liquiritigenin is 12.60%, the content of isoliquiritigenin is 10.8%, and the weight yield is 22.4%.

Example 3 a process for the combined separation of multiple active ingredients of licorice, comprising the steps of:

firstly, adding 10kg of mother liquor powder after producing mono-ammonium glycyrrhizinate into a jacketed kettle, adding 40L of ethyl acetate each time, stirring and extracting for 2 times at room temperature to respectively obtain an extracting solution and an ethyl acetate insoluble substance.

Filtering, recovering, dissolving, drying and crushing the extracting solution to obtain 2.2kg of a product A; adding the product A into a jacketed kettle, adding 8.8L of purified water each time, heating and stirring at 40 ℃, and extracting for 1 time, wherein the extraction time is 1-2 hours each time. Stirring, extracting, standing and precipitating to respectively obtain a supernatant and a product B. Product B is the water insoluble fraction.

Concentrating the supernatant under reduced pressure to obtain fluid extract 1.5kg with specific gravity of 1.18 (60 deg.C), adding 1L ethanol with volume concentration of 95%, and heating to dissolve; then adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing in sequence to obtain 180g of white solid liquiritin, wherein the content is 91.56%, and the weight yield is 1.80%.

And thirdly, drying the ethyl acetate insoluble substance in vacuum, heating the ethyl acetate insoluble substance at normal pressure until the solvent in the ethyl acetate insoluble substance is volatilized completely to obtain a product C, and drying and crushing the product C to obtain 8.2kg of the debitterized glycyrrhizin, wherein the glycyrrhizic acid content is 10.38 percent, and the weight yield is 82.0 percent.

And fourthly, drying and crushing the product B in vacuum to obtain 1.85kg of crude product containing the liquiritigenin and the isoliquiritigenin. Wherein: the content of liquiritigenin is 10.60%, the content of isoliquiritigenin is 11.23%, and the weight yield is 18.5%.

Example 4 a process for the combined separation of multiple active ingredients of licorice, comprising the steps of:

adding 10kg of mother liquor powder after producing mono-ammonium glycyrrhizinate into a jacketed kettle, adding 60L of ethyl formate each time, stirring and extracting for 2 times at room temperature to obtain an extracting solution and an ethyl formate insoluble substance respectively.

Filtering, recovering, dissolving, drying and crushing the extracting solution to obtain 2.6kg of a product A; and adding the product A into a jacketed kettle, adding 10.4L of purified water each time, heating and stirring at 80 ℃, and extracting for 2 times, wherein the extraction time is 1-2 hours each time. Stirring, extracting, standing and precipitating to respectively obtain a supernatant and a product B. Product B is the water insoluble fraction.

Concentrating the supernatant under reduced pressure to obtain fluid extract 1.8kg with specific gravity of 1.18 (60 deg.C), adding 0.36L ethanol with volume concentration of 95%, and heating to dissolve; then sequentially adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing to obtain 160g of white solid liquiritin with the content of 92.22% and the weight yield of 1.60%.

And thirdly, drying the ethyl formate insoluble substance in vacuum, heating the ethyl formate insoluble substance at normal pressure until the solvent in the ethyl formate insoluble substance is volatilized completely to obtain a product C, and drying and crushing the product C to obtain 7.2kg of the debitterized glycyrrhizin, wherein the glycyrrhizic acid content is 10.68 percent, and the weight yield is 72.0 percent.

And fourthly, drying and crushing the product B in vacuum to obtain 1.95kg of crude product containing the liquiritigenin and the isoliquiritigenin. Wherein: the content of liquiritigenin is 12.60%, the content of isoliquiritigenin is 11.58%, and the weight yield is 19.5%.

Example 5 a process for the combined isolation of multiple active ingredients from licorice, comprising the steps of:

adding 10kg of mother liquor powder after producing mono-ammonium glycyrrhizinate into an interlayer pot, wherein ethyl formate is added each time: ethyl acetate =1:1, and the mixture was stirred and extracted at room temperature for 3 times to obtain an extract and an insoluble ethyl formate-ethyl acetate mixture, respectively.

Filtering, recovering, dissolving, drying and crushing the extracting solution to obtain 3.2kg of a product A; and adding the product A into a jacketed kettle, adding 10.4L of purified water each time, heating and stirring at 80 ℃, and extracting for 2 times, wherein the extraction time is 1-2 hours each time. Stirring, extracting, standing and precipitating to respectively obtain a supernatant and a product B. Product B is the water insoluble fraction.

Concentrating the supernatant under reduced pressure to obtain fluid extract with a specific gravity of 1.17 (60 deg.C) 2.1kg, adding 0.36L ethanol with volume concentration of 95%, and heating to dissolve; then adding activated carbon for decolorization, filtering, standing for crystallization, separating and recrystallizing in sequence to obtain 188g of white solid liquiritin with the content of 93.50 percent and the weight yield of 1.88 percent.

And thirdly, drying the ethyl formate-ethyl acetate mixed solvent insoluble substance in vacuum, heating the mixture at normal pressure until the solvent in the mixture is volatilized completely to obtain a product C, and drying and crushing the product C to obtain 7.7kg of the debitterized glycyrrhizin, wherein the glycyrrhizic acid content is 13.68 percent, and the weight yield is 77.0 percent.

And fourthly, drying and crushing the product B in vacuum to obtain 1.65kg of crude product containing the liquiritigenin and the isoliquiritigenin. Wherein: the content of the liquiritigenin is 10.60 percent, the content of the isoliquiritigenin is 12.58 percent, and the weight yield is 16.5 percent.