Comprehensive extraction process of berberine

Abstract

The invention relates to the field of traditional Chinese medicine extraction, and particularly discloses a comprehensive extraction process of berberine, which comprises the following steps: crushing fresh golden cypress to obtain golden cypress raw material powder, then taking a mixed solvent of a 10-20% methanol solution and citric acid as an entrainer, carrying out supercritical carbon dioxide extraction to obtain an extract, and concentrating the extract to obtain the berberine. Filtering to obtain cortex phellodendri filter residues and a polysaccharide extracting solution mainly containing cortex phellodendri polysaccharide; the cortex phellodendri filter residues in the step S1 are soaked with dilute sulphuric acid and extracted to obtain an extracting solution B containing berberine; adding hydrochloric acid into the extracting solution B for acidification, then adding sodium chloride for salting out, then filtering, and collecting precipitate to obtain berberine extract; the method has the characteristics that the berberine extraction period is shortened, the difficulty of viscous substances for subsequent treatment after acid extraction is solved, the extraction is more convenient and easy to operate, and meanwhile, the berberine extract with higher content is obtained through extraction.

Description

Technical Field

[0001] The present application relates to the field of plant extraction, and more specifically, to a comprehensive extraction process of berberine. Background Art

[0002] Berberine, also known as berberine, is an alkaloid extracted from plants such as Coptis chinensis and Phellodendron amurense. It is a yellow needle-shaped crystal, slightly soluble in water and ethanol, and more soluble in hot water and hot ethanol. Berberine has significant antibacterial and anti-inflammatory effects and is a traditional medicine for the treatment of digestive tract diseases such as bacterial gastroenteritis and dysentery.

[0003] The traditional water decoction method for extracting berberine has the advantages of simple operation, cheap and easy to obtain extraction solvents that are safe and non-toxic, and the extraction process does not pollute the environment. However, in addition to alkaloids, the bark of Phellodendron chinense is also rich in mucus, which is a polysaccharide substance with a similar structure to gum. The dry mucus is a white powder with strong hygroscopicity. It can expand rapidly in water and dissolve to form a viscous gum, making the Phellodendron chinense water extract more viscous and extremely difficult to filter.

[0004] Therefore, the acid water method, ethanol method, lime milk method, etc. are mainly used for the extraction of berberine. These methods extract berberine from Coptis chinensis through different solvents and acid-base conditions. The ethanol method mainly uses the property that berberine is easily soluble in hot ethanol and difficult to dissolve in cold ethanol. It uses hot ethanol to extract it, and then converts it into hydrochloride for precipitation. This method has low solvent restriction, but other components will be extracted when hot ethanol is used to extract berberine, resulting in a low berberine content in the final extract, requiring subsequent complex separation and purification processes; the commonly used extraction process is to use acidic aqueous solution for immersion extraction, but this method requires a long acid water immersion time, and after the Phellodendron chinense is immersed, a sticky substance is produced, and the high viscosity is not conducive to subsequent separation and treatment.

[0005] Therefore, it is necessary to provide a new extraction method, which can obtain a berberine extract with a higher berberine content, solve the difficulty of subsequent treatment of viscous substances after acid extraction, and shorten the extraction cycle. Summary of the invention

[0006] In order to shorten the berberine extraction cycle and solve the difficulty of subsequent treatment of sticky substances after acid extraction, the extraction is more convenient and easy to operate, and at the same time, a berberine extract with a higher content is obtained, the present application provides a comprehensive extraction process of berberine.

[0007] In the first aspect, the present application provides a comprehensive extraction process of berberine, which adopts the following technical scheme: A comprehensive extraction process of berberine comprises the following steps: S1. crushing fresh Phellodendron amurense to obtain Phellodendron amurense raw material powder, and then using a mixed solvent of 10-20% methanol solution and citric acid as an entrainer to obtain an extract through supercritical carbon dioxide extraction, and filtering to obtain Phellodendron amurense residue and a polysaccharide extract mainly containing Phellodendron amurense polysaccharide; S2, soaking the Phellodendron chinense residue in step S1 in dilute sulfuric acid to obtain an extract B containing berberine; S3. Add hydrochloric acid to the extract B obtained in step S2 to acidify it to a pH of 2-3, then add sodium chloride for salting out, and then filter and collect the precipitate to obtain a berberine extract.

[0008] By adopting the above technical scheme, when extracting berberine from Phellodendron chinense, the viscous liquid in the acid soaking process is mainly the polysaccharide contained in Phellodendron chinense, which undergoes structural transformation under acidic conditions and exhibits viscous characteristics. Therefore, in this application, fresh Phellodendron chinense is first extracted with supercritical carbon dioxide, and a mixed solvent with a low concentration of methanol solution and weak acid citric acid is added as an entrainer. Phellodendron chinense polysaccharide has more hydroxyl groups, and berberine is an isoquinoline alkaloid substance containing nitrogen elements and benzene rings. A large number of hydroxyl groups in Phellodendron chinense polysaccharide can form hydrogen bonds with water, and its epoxy atoms and The glycosidic oxygen atoms connected to the sugar ring can also form hydrogen bonds with water, so that in the mixed solvent of low concentration methanol solution and citric acid, the low concentration methanol solvent has a stronger effect on the dissolution enhancement of Phellodendron chinense polysaccharide in supercritical carbon dioxide, while citric acid has a greater effect on the solubility of Phellodendron chinense polysaccharide and less effect on berberine. Therefore, in the entrainer system in the present application, supercritical carbon dioxide is used to selectively extract the polysaccharides in Phellodendron chinense, and the obtained extract contains more Phellodendron chinense polysaccharide, while berberine is mainly present in the Phellodendron chinense filter residue, thereby achieving the separation of berberine and Phellodendron chinense polysaccharide.

[0009] The obtained Phellodendron chinense residue is then soaked in dilute sulfuric acid, and berberine exists in the form of sulfate, so as to realize the extraction of berberine in the Phellodendron chinense residue. Since the Phellodendron chinense residue has been subjected to high-pressure treatment during the previous supercritical extraction with carbon dioxide and treatment with a low-concentration methanol solution, it is easier to dissolve berberine when the Phellodendron chinense residue is soaked in dilute sulfuric acid, thus shortening the acid leaching time. Then, hydrochloric acid is added for acidification and sodium chloride is used for salting out to realize the precipitation of berberine, and crude berberine is obtained by filtration. Finally, the extraction method provided in the present application greatly shortens the berberine extraction process to be completed within 24 hours. At the same time, it realizes the removal of sticky substances in Phellodendron chinense, improves the purity of berberine, and obtains a higher content of berberine in the berberine extract. Moreover, in the present application, the removal of Phellodendron chinense polysaccharide is realized before the acid leaching treatment, thus solving the problem that the sticky substances produced in the current acid extraction make subsequent filtration and other operations difficult, and the extraction is more convenient and easy to operate.

[0010] Optionally, in step S1, the specific operation of supercritical carbon dioxide extraction is: First, the extraction is carried out at an extraction temperature of 30-40° C. and an extraction pressure of 25-30 MPa for 1-2 hours, and then the extraction is carried out at an extraction temperature of 25-30° C. and an extraction pressure of 10-15 MPa for 1-2 hours to obtain an extract.

[0011] By adopting the above technical solution, the supercritical carbon dioxide extraction conditions are controlled to be carried out under low temperature conditions, which can reduce the temperature-induced changes in the solubility of berberine and phellodendron polysaccharides in the system, thereby increasing the difficulty of subsequent separation.

[0012] Optionally, in step S1, the amount of entrainer added is 5-8wt% of the mass of the Phellodendron chinense raw material powder, and the mass ratio of the methanol solution to the citric acid added is 1:(0.2-0.3).

[0013] By adopting the above technical solution and selecting the above ratio of addition, more Phellodendron amurense polysaccharides are extracted during supercritical extraction of Phellodendron amurense, while more berberine is retained in Phellodendron amurense.

[0014] Optionally, the Phellodendron chinense residue in step S1 is first soaked in dilute sulfuric acid for 8-10 hours, and then a gelatin solution is added and filtered to obtain a filtrate containing tannins and an extract B containing berberine, wherein the mass concentration of the dilute sulfuric acid is 0.1-0.3%, and the amount of dilute sulfuric acid added is 10-15 times the mass of the amount of the Phellodendron chinense residue added, the mass concentration of the gelatin solution is 20-30wt%, and the amount of gelatin added is until no precipitation is produced.

[0015] By adopting the above-mentioned technical scheme, the technical scheme in the present application is adopted, and the Phellodendron chinense raw material is pretreated after supercritical carbon dioxide extraction. On the one hand, the viscous liquid can be removed, and it is also more conducive to the dissolution of berberine in subsequent treatment. Therefore, in the present application, dilute sulfuric acid with lower concentration and less addition amount is used, and more berberine can be extracted in a shorter time.

[0016] In addition, since there will be more tannin components when berberine is extracted with dilute sulfuric acid, which will affect the berberine content in the final berberine extract, gelatin is added after the Phellodendron chinense residue is acid-leached during the acid extraction process in this application. The gelatin and the tannins in the acid extraction solution form an insoluble precipitate, and then the tannins in the Phellodendron chinense extract are removed by filtration to obtain a berberine extract with higher purity.

[0017] Optionally, the Phellodendron chinense residue in step S1 is soaked in dilute sulfuric acid for 2-3h, and then sodium chloride is added for salting out, the amount of sodium chloride added is 5-10wt% of the Phellodendron chinense residue, and then filtered to obtain a crude berberine extract A, and dilute sulfuric acid is added to the filtrate and continued to be soaked for 2-3h, the amount of dilute sulfuric acid added is 0.05-0.1wt% of the mass of the Phellodendron chinense residue, and the mass concentration of the supplemented dilute sulfuric acid is 0.01-0.05%, to obtain a crude berberine extract and a crude berberine extract B, and the obtained berberine crude extract A and berberine crude extract B are combined with the crude berberine extract, and then a gelatin solution is added to filter to obtain a filtrate containing tannins and an extract B containing berberine.

[0018] By adopting the above technical scheme, in the present application, when the Phellodendron chinense residue is extracted in dilute sulfuric acid, sodium chloride is added for salting out after a period of extraction, so that the solubility of the extracted berberine is reduced and precipitated, and then the crude berberine extract A is obtained by filtration, and a small amount of dilute sulfuric acid is added to the filtrate for continued soaking, so that the reduction of the berberine concentration in the filtrate is more conducive to the continued leaching of berberine from Phellodendron chinense, and the addition of a small amount of dilute sulfuric acid allows the soaking process to be carried out in two stages, which is conducive to the leaching of berberine, can further shorten the acid leaching time of berberine, shorten the extraction cycle, and extract more berberine, so that the extract has a higher content of berberine and more berberine is extracted.

[0019] Optionally, the polysaccharide extract in step S1 is further subjected to the following separation treatment: Adding a long-chain quaternary ammonium salt to a polysaccharide extract, filtering after standing to obtain an acidic polysaccharide residue and a filtrate, mixing the acidic polysaccharide residue with an inorganic alkali alcohol solution to obtain free acidic polysaccharide, then adding ethanol and filtering, washing the obtained precipitate with water, crystallizing and drying to obtain an acidic polysaccharide; Adding lime milk to the filtrate to adjust the pH to weak alkalinity, leaving it to stand and then filtering it to obtain a polysaccharide precipitate containing neutral polysaccharides and alkaline polysaccharides and an extract A containing berberine; Then in step S3, the obtained extract B is combined with the extract A and then hydrochloric acid is added for acidification.

[0020] By adopting the above technical scheme, the study found that Phellodendron polysaccharide has anti-inflammatory, immunomodulatory, antioxidant and other activities, and has a wide range of medical uses and clinical applications, especially the acidic Phellodendron pollen acidic polysaccharide in the acidic Phellodendron polysaccharide shows strong antioxidant capacity and protective effect against radiation-induced oxidative stress, and has stronger antioxidant properties and biological activity. Although a small number of studies have found that adding lime milk to the Phellodendron extract can form a water-insoluble precipitate with the polysaccharide to remove impurities and improve the purity of berberine, it has caused a waste of resources for Phellodendron polysaccharide.

[0021] Therefore, in the present application, long-chain quaternary ammonium salts are added to the separated polysaccharide extract, and the long-chain quaternary ammonium salts and acidic polysaccharides form a precipitate. After standing, the filter residue obtained by filtration is acidic polysaccharides, while the filtrate mainly contains neutral polysaccharides and alkaline polysaccharides. Lime milk is added, and the lime milk and polysaccharides form a precipitate, thereby realizing the separation of polysaccharides and a small amount of berberine in the polysaccharide extract. In this way, a small amount of berberine extract A and extract B are obtained, which are subsequently treated together to increase the berberine yield. The acidic polysaccharide filter residue is mixed with an inorganic alkali alcohol solution to destroy the interaction between the acidic polysaccharide and the long-chain quaternary ammonium salt, so that the acidic polysaccharide is freed. Then ethanol is added, and the long-chain quaternary ammonium salt is soluble in ethanol, while the acidic polysaccharide is insoluble in cold ethanol, so that the acidic polysaccharide is precipitated, thereby realizing the separation and extraction of the acidic polysaccharide in the polysaccharide extract, and finally realizing the extraction of berberine and acidic polysaccharides in Phellodendron chinense.

[0022] Optionally, when the polysaccharide extract is separated, the inorganic alkali alcohol solution is an ethanol solution and a sodium hydroxide solution with a mass concentration of 8-15wt%, and the addition of the sodium hydroxide solution makes the pH value 7.5-8.

[0023] By adopting the above technical scheme, the acidic polysaccharide residue is mixed with an inorganic alkali alcohol solution. The addition of the alcohol solution reduces the polarity of the solution, thereby changing the interaction between the polysaccharide and the quaternary ammonium salt. The addition of the inorganic base causes the acidic polysaccharide to lose protons, ionizes the acidic polysaccharide, and promotes the release of the acidic polysaccharide, thereby achieving the separation and extraction of the acidic polysaccharide in the polysaccharide extract.

[0024] Optionally, the long-chain quaternary ammonium salt is cetyltrimethylammonium bromide or cetyltrimethylammonium base.

[0025] Optionally, the amount of sodium chloride added in step S3 is 3-5 wt % of the liquid mixture after adding hydrochloric acid.

[0026] Optionally, the precipitate collected after filtration in step S3 is further subjected to heat dissolution, filtration, crystallization and drying treatments to obtain a berberine extract.

[0027] By adopting the above technical scheme, in the present application, the berberine precipitate obtained after salting out is first thermally dissolved and then filtered and then crystallized and dried. The impurities are further removed by cooling and crystallizing after thermal dissolution, thereby improving the purity of berberine.

[0028] Optionally, the thermal dissolution is carried out using water or ethanol, and when water is used, the dissolution is carried out at 80-90°C, and when ethanol is used, the dissolution is carried out at 65-75°C.

[0029] In summary, this application has the following beneficial effects: 1. The extraction method provided in the present application greatly shortens the berberine extraction process to within 24 hours. At the same time, it realizes the removal of tannins and sticky substances in Phellodendron amurense, improves the purity of berberine, and obtains a berberine extract with a higher content of berberine. In addition, the present application realizes the removal of Phellodendron amurense polysaccharides before acid leaching, which solves the problem that sticky substances are produced in the current acid extraction, making subsequent filtration and other operations difficult to carry out, and the extraction is more convenient and easy to operate; 2. In the present application, when the Phellodendron chinense residue is leached in dilute sulfuric acid, sodium chloride is added for salting out after leaching for a period of time, so that the solubility of the leached berberine is reduced and precipitated, and then the crude berberine extract A is obtained by filtration, and a small amount of dilute sulfuric acid is added to the filtrate for continued immersion, so that the concentration of berberine in the filtrate is reduced, which is more conducive to the continued leaching of berberine from the Phellodendron chinense. In combination with the addition of a small amount of dilute sulfuric acid, the immersion process is divided into two stages, which is conducive to the leaching of berberine, and can further shorten the acid leaching time of berberine, shorten the extraction cycle, and more berberine is leached, so that the extract has a higher content of berberine and more berberine is extracted; 3. In the present application, fresh Phellodendron chinense is first extracted with supercritical carbon dioxide, and a mixed solvent of low-concentration methanol solution and weak acid citric acid is added as an entrainer, so that the supercritical carbon dioxide selectively extracts polysaccharides in Phellodendron chinense. The obtained extract contains more Phellodendron chinense polysaccharides, and berberine is mainly present in the Phellodendron chinense filter residue, thereby realizing the separation of berberine and Phellodendron chinense polysaccharides. Then, the obtained Phellodendron chinense filter residue is soaked in dilute sulfuric acid, and berberine exists in the form of sulfate, thereby realizing the extraction of berberine in the Phellodendron chinense filter residue, shortening the acid leaching time, and then adding hydrochloric acid for acidification and cooperating with sodium chloride salting-out to realize the precipitation of berberine, and filtering to obtain crude berberine. During the process, gelatin is also added after the Phellodendron chinense filter residue is acidified, and the gelatin and tannin in the acid leaching solution form an insoluble precipitate. Then, the tannin in the Phellodendron chinense extract is removed by filtration to obtain a berberine extract with higher purity. DETAILED DESCRIPTION

[0030] The present application is further described in detail below in conjunction with the examples. It is particularly noted that if no specific conditions are specified in the following examples, the experiments are carried out according to conventional conditions or the conditions recommended by the manufacturer. Unless otherwise specified, the raw materials used in the following examples can be obtained from common commercial sources.

[0031] In the following examples, percentages refer to mass percentages unless otherwise specified.

[0032] Example 1 A comprehensive extraction process of berberine comprises the following steps: S1. crushing fresh Phellodendron amurense (Phellodendron amurense rhizome) to obtain Phellodendron amurense raw material powder, and then using a mixed solvent of 15% mass concentration of methanol solution and citric acid as an entrainer, extracting by supercritical carbon dioxide to obtain an extract, and filtering to obtain Phellodendron amurense residue and a polysaccharide extract mainly containing Phellodendron amurense polysaccharide; Among them, the amount of entrainer added in step S1 is 6wt% of the mass of the cortex phellodendri raw material powder, and the added mass ratio of methanol solution to citric acid is 1:0.25. The specific operation of supercritical carbon dioxide extraction is: First, the extraction was performed at an extraction temperature of 35°C and an extraction pressure of 28 MPa for 1.5 h, and then the extraction was performed at an extraction temperature of 28°C and an extraction pressure of 12 MPa for 1.5 h to obtain an extract; S2, soaking the Phellodendron chinense residue in step S1 in dilute sulfuric acid for 9 hours, wherein the mass concentration of the dilute sulfuric acid is 0.2%, and the amount of the dilute sulfuric acid added is 12 times the mass of the amount of the Phellodendron chinense residue added, adding a gelatin solution for filtration, wherein the mass concentration of the gelatin solution is 25wt%, and the amount of gelatin added is until no precipitation is produced, to obtain a filtrate containing tannins and an extract B containing berberine; S3. Add hydrochloric acid to the extract B obtained in step S2 to acidify it to a pH of 2.5, then add sodium chloride for salting out, wherein the amount of sodium chloride added is 4wt% of the liquid mixture after the addition of hydrochloric acid, then filter and collect the precipitate, and then sequentially subject the collected precipitate to thermal dissolution, filtration, crystallization and drying to obtain a berberine extract, wherein the thermal dissolution is to dissolve the precipitate in hot ethanol at 70°C.

[0033] Example 2 A comprehensive extraction process of berberine comprises the following steps: S1. crushing fresh Phellodendron amurense (Phellodendron amurense rhizome) to obtain Phellodendron amurense raw material powder, taking the same amount of Phellodendron amurense raw material powder as in Example 1 for treatment, and then using a mixed solvent of 10% methanol solution and citric acid as an entrainer, extracting by supercritical carbon dioxide to obtain an extract, filtering to obtain Phellodendron amurense filter residue and a polysaccharide extract mainly containing Phellodendron amurense polysaccharide; Among them, the amount of entrainer added in step S1 is 5wt% of the mass of the cortex phellodendri raw material powder, and the added mass ratio of methanol solution to citric acid is 1:0.2. The specific operation of supercritical carbon dioxide extraction is: First, extracting at an extraction temperature of 30°C and an extraction pressure of 25 MPa for 2 h, and then extracting at an extraction temperature of 25°C and an extraction pressure of 10 MPa for 2 h to obtain an extract; S2, soaking the Phellodendron amurense residue in step S1 in dilute sulfuric acid for 8 hours, wherein the mass concentration of the dilute sulfuric acid is 0.3%, and the amount of dilute sulfuric acid added is 15 times the mass of the amount of the Phellodendron amurense residue added, adding a gelatin solution for filtration, wherein the mass concentration of the gelatin solution is 20wt%, and the amount of gelatin added is until no precipitation is produced, to obtain a filtrate containing tannins and an extract B containing berberine; S3. Add hydrochloric acid to the extract B obtained in step S2 to acidify it to a pH of 2, then add sodium chloride for salting out, wherein the amount of sodium chloride added is 3wt% of the liquid mixture after the addition of hydrochloric acid, then filter and collect the precipitate, and then sequentially subject the collected precipitate to thermal dissolution, filtration, crystallization and drying to obtain a berberine extract, wherein the thermal dissolution is to dissolve the precipitate in hot ethanol at 65°C.

[0034] Example 3 A comprehensive extraction process of berberine comprises the following steps: S1. crushing fresh Phellodendron amurense (Phellodendron amurense rhizome) to obtain Phellodendron amurense raw material powder, treating the same amount of Phellodendron amurense raw material powder as in Example 1, and then extracting the extract by supercritical carbon dioxide with a mixed solvent of 20% methanol solution and citric acid as an entrainer, and filtering to obtain Phellodendron amurense filter residue and a polysaccharide extract mainly containing Phellodendron amurense polysaccharide; Among them, the amount of entrainer added in step S1 is 8wt% of the mass of the cortex phellodendri raw material powder, and the added mass ratio of methanol solution to citric acid is 1:0.3. The specific operation of supercritical carbon dioxide extraction is: First, extracting at an extraction temperature of 40°C and an extraction pressure of 30 MPa for 1 hour, and then extracting at an extraction temperature of 30°C and an extraction pressure of 15 MPa for 1 hour to obtain an extract; S2, soaking the Phellodendron chinense residue in step S1 in dilute sulfuric acid for 10 hours, wherein the mass concentration of the dilute sulfuric acid is 0.1%, and the amount of the dilute sulfuric acid added is 10 times the mass of the amount of the Phellodendron chinense residue added, adding a gelatin solution for filtration, wherein the mass concentration of the gelatin solution is 30wt%, and the amount of gelatin added is until no precipitation is produced, to obtain a filtrate containing tannins and an extract B containing berberine; S3. Add hydrochloric acid to the extract B obtained in step S2 to acidify it to a pH of 3, then add sodium chloride for salting out, wherein the amount of sodium chloride added is 5wt% of the liquid mixture after the addition of hydrochloric acid, then filter and collect the precipitate, and then sequentially subject the collected precipitate to thermal dissolution, filtration, crystallization and drying to obtain a berberine extract, wherein the thermal dissolution is to dissolve the precipitate in hot ethanol at 75°C.

[0035] Example 4 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that the specific operation of step S2 is: The Phellodendron chinense residue in step S1 is soaked in dilute sulfuric acid for 2.5 hours, and then sodium chloride is added for salting out, the amount of sodium chloride added is 8wt% of the Phellodendron chinense residue, and then filtered to obtain berberine crude extract A, dilute sulfuric acid is added to the filtrate and continued to be soaked for 2.5 hours, the amount of dilute sulfuric acid added is 0.08wt% of the mass of the Phellodendron chinense residue, and the mass concentration of the supplemented dilute sulfuric acid is 0.02%, to obtain berberine crude extract A and berberine crude extract B, the obtained berberine crude extract A and berberine crude extract B are combined with the berberine crude extract, and then gelatin solution is added for filtration to obtain a filtrate containing tannins and an extract B containing berberine.

[0036] Example 5 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that the specific operation of step S2 is: The Phellodendron chinense residue in step S1 is soaked in dilute sulfuric acid for 2 hours, and then sodium chloride is added for salting out, the amount of sodium chloride added is 5wt% of the Phellodendron chinense residue, and then filtered to obtain berberine crude extract A, dilute sulfuric acid is added to the filtrate and the immersion is continued for 3 hours, the amount of dilute sulfuric acid added is 0.05wt% of the mass of the Phellodendron chinense residue, and the mass concentration of the supplemented dilute sulfuric acid is 0.01%, to obtain berberine crude extract A and berberine crude extract B, the obtained berberine crude extract A and berberine crude extract B are combined with the berberine crude extract, and then a gelatin solution is added for filtration to obtain a filtrate containing tannins and an extract B containing berberine.

[0037] Example 6 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that the specific operation of step S2 is: The Phellodendron chinense residue in step S1 is soaked in dilute sulfuric acid for 3 hours, and then sodium chloride is added for salting out, the amount of sodium chloride added is 10wt% of the Phellodendron chinense residue, and then filtered to obtain berberine crude extract A, dilute sulfuric acid is added to the filtrate and continued to be soaked for 2 hours, the amount of dilute sulfuric acid added is 0.1wt% of the mass of the Phellodendron chinense residue, and the mass concentration of the supplemented dilute sulfuric acid is 0.05%, to obtain berberine crude extract A and berberine crude extract B, the obtained berberine crude extract A and berberine crude extract B are combined with the berberine crude extract, and then gelatin solution is added for filtration to obtain a filtrate containing tannins and an extract B containing berberine.

[0038] Example 7 A comprehensive extraction process of berberine is carried out according to the method in Example 4, except that dilute sulfuric acid is not added when the filtrate continues to be immersed in dilute sulfuric acid in step S2.

[0039] Example 8 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that the polysaccharide extract in step S1 is further subjected to the following separation treatment: Adding a long-chain quaternary ammonium salt, specifically hexadecyltrimethylammonium bromide, to a polysaccharide extract, letting it stand and then filtering to obtain an acidic polysaccharide residue and a filtrate, mixing the acidic polysaccharide residue with an inorganic alkali alcohol solution to obtain free acidic polysaccharide, wherein the amount of the inorganic alkali alcohol solution added is 2.5 times the mass of the acidic polysaccharide residue, and the inorganic alkali alcohol solution is a mixed solution of an ethanol solution with a mass concentration of 10wt% and a sodium hydroxide solution, wherein the addition of the sodium hydroxide solution makes the pH value of the inorganic alkali alcohol 7.5, then adding ethanol and filtering, wherein the amount of ethanol added is 2.5 times the mass of the acidic polysaccharide residue, washing the obtained precipitate with water, crystallizing and drying, and obtaining an acidic polysaccharide; Adding lime milk to the obtained filtrate to adjust the pH to 7.5, letting it stand and then filtering it to obtain a polysaccharide precipitate containing neutral polysaccharides and alkaline polysaccharides and an extract A containing berberine; Then, in step S3, the extract B obtained in step S2 is combined with the extract A obtained above and then hydrochloric acid is added for acidification.

[0040] Example 9 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that the polysaccharide extract in step S1 is further subjected to the following separation treatment: Adding a long-chain quaternary ammonium salt, specifically hexadecyltrimethylammonium bromide, to a polysaccharide extract, letting it stand and then filtering to obtain an acidic polysaccharide residue and a filtrate, mixing the acidic polysaccharide residue with an inorganic alkali alcohol solution to obtain free acidic polysaccharide, wherein the amount of the inorganic alkali alcohol solution added is twice the mass multiple of the acidic polysaccharide residue, and the inorganic alkali alcohol solution is a mixed solution of an ethanol solution with a mass concentration of 8wt% and a sodium hydroxide solution, wherein the addition of the sodium hydroxide solution makes the pH value of the inorganic alkali alcohol to be 7.5, then adding ethanol and filtering, wherein the amount of ethanol added is twice the mass multiple of the acidic polysaccharide residue, washing the obtained precipitate with water, crystallizing and drying, and obtaining an acidic polysaccharide; Adding lime milk to the obtained filtrate to adjust the pH to 8, letting it stand and then filtering it to obtain a polysaccharide precipitate containing neutral polysaccharides and alkaline polysaccharides and an extract A containing berberine; Then, in step S3, the extract B obtained in step S2 is combined with the extract A obtained above and then hydrochloric acid is added for acidification.

[0041] Example 10 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that the polysaccharide extract in step S1 is further subjected to the following separation treatment: A long-chain quaternary ammonium salt, specifically hexadecyltrimethylammonium bromide, is added to a polysaccharide extract, and the mixture is allowed to stand and then filtered to obtain an acidic polysaccharide residue and a filtrate. The acidic polysaccharide residue is mixed with an inorganic alkali alcohol solution to obtain free acidic polysaccharide, wherein the amount of the inorganic alkali alcohol solution added is 3 times the mass of the acidic polysaccharide residue, and the inorganic alkali alcohol solution is a mixed solution of an ethanol solution with a mass concentration of 15wt% and a sodium hydroxide solution. The addition of the sodium hydroxide solution makes the pH value of the inorganic alkali alcohol 8, and then ethanol is added and filtered, wherein the amount of ethanol added is 3 times the mass of the acidic polysaccharide residue. The obtained precipitate is washed with water, crystallized and dried to obtain an acidic polysaccharide; lime milk is added to the obtained filtrate to adjust the pH to weak alkalinity, and the mixture is allowed to stand and then filtered to obtain a polysaccharide precipitate containing neutral polysaccharides and alkaline polysaccharides and an extract A containing berberine; Then, in step S3, the extract B obtained in step S2 is combined with the extract A obtained above and then hydrochloric acid is added for acidification.

[0042] Comparative Example 1 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that step S1 is not carried out, and the Phellodendron chinense raw material powder is directly subjected to step S2.

[0043] Comparative Example 2 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that step S1 is not performed, the Phellodendron chinense raw material powder is directly subjected to the operation in step S2, and the soaking time in dilute sulfuric acid in step S2 is 20 hours.

[0044] Comparative Example 3 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that no entrainer is added in step S1.

[0045] Comparative Example 4 A comprehensive extraction process of berberine is carried out according to the method in Example 1, except that no citric acid is added to the entrainer in step S1.

[0046] Performance Testing The amount of berberine extract, berberine content and berberine yield obtained in the examples and comparative examples of the present application were statistically analyzed, and the statistical results are shown in Table 1 below.

[0047] Table 1: In addition, the content and amount of the acidic polysaccharides obtained in Examples 8-10 of the present application were measured, and the results are shown in Table 2 below.

[0048] Table 2: Test items Example 8 Example 9 Example 10 Acidic polysaccharide content / g 68 56 59 Acidic polysaccharide content / % 83.6 80.5 81.6 According to the test results of Table 1 and Table 2 above, the berberine extract obtained by the method in the embodiment of the present application has a higher content, and the extraction rate of berberine in the present application is higher. At the same time, the polysaccharide extract is processed to obtain an acidic polysaccharide with higher purity, and the extraction of Phellodendron polysaccharide and berberine is achieved. In addition, referring to the test results of Example 1 and Example 4-6, when the two-step stage method of dilute sulfuric acid immersion is used in Example 4-6, not only the dilute sulfuric acid extraction time is shortened compared with Example 1, but also the berberine extraction amount obtained in Example 4 is more, and the berberine extraction rate is higher. Combined with the test results in Example 7, when the two-step acid leaching is used in Example 7 without supplementing dilute sulfuric acid, the berberine extraction effect is reduced. Combined with the test results of Example 8-10, the separation and extraction of acidic polysaccharides can be achieved after the polysaccharide extract is processed in Example 8-10.

[0049] Combining the test results of Example 1 with those of Comparative Examples 1 and 2, it can be seen that when sulfuric acid extraction was directly used without supercritical extraction in Comparative Example 1, the berberine extraction amount was significantly reduced, and its content was significantly reduced. On the one hand, the sulfuric acid extraction time was short, and on the other hand, the presence of tannins and polysaccharides during the extraction process would affect the extraction effect of berberine. Combined with the test results in Comparative Example 2, even if the acid extraction time was longer, although the amount of berberine extract was large, its content was significantly reduced, and the impurity content was large, and the extraction effect of berberine was still poor, which was related to the presence of tannins and polysaccharides. Combined with the test results in Comparative Example 3, no entrainer was added in Comparative Example 3, and no citric acid was added to the entrainer in Comparative Example 4, the amount of berberine extract increased, but its content was significantly reduced, and the extraction effect of berberine was significantly reduced.

[0050] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A comprehensive extraction process of berberine, characterized in that: The following steps are involved: S1. crushing fresh Phellodendron chinense to obtain Phellodendron chinense raw material powder, and then using a mixed solvent of 10-20% methanol solution and citric acid as an entrainer, extracting through supercritical carbon dioxide to obtain an extract, and filtering to obtain Phellodendron chinense filter residue and a polysaccharide extract mainly containing Phellodendron chinense polysaccharide; S2, soaking the Phellodendron chinense residue in step S1 in dilute sulfuric acid to obtain an extract B containing berberine; S3. Add hydrochloric acid to the extract B obtained in step S2 to acidify it to a pH of 2-3, then add sodium chloride for salting out, and then filter and collect the precipitate to obtain a berberine extract.

2. A comprehensive extraction process for berberine according to claim 1, characterized in that: In step S1, the specific operation of supercritical carbon dioxide extraction is: First, the extraction is carried out at an extraction temperature of 30-40° C. and an extraction pressure of 25-30 MPa for 1-2 hours, and then the extraction is carried out at an extraction temperature of 25-30° C. and an extraction pressure of 10-15 MPa for 1-2 hours to obtain an extract.

3. A comprehensive extraction process of berberine according to claim 1, characterized in that: In step S1, the amount of entrainer added is 5-8wt% of the mass of the Cortex Phellodendri raw material powder, and the added mass ratio of the methanol solution to the citric acid is 1:(0.2-0.3).

4. The comprehensive extraction process of berberine according to claim 1, characterized in that: The specific operation of step S2 is: first soak the Phellodendron chinense residue in step S1 in dilute sulfuric acid for 8-10 hours, then add gelatin solution to filter, and obtain a filtrate containing tannins and an extract B containing berberine, wherein the mass concentration of dilute sulfuric acid is 0.1-0.3%, and the amount of dilute sulfuric acid added is 10-15 times the mass of the amount of Phellodendron chinense residue added, the mass concentration of the gelatin solution is 20-30wt%, and the amount of gelatin added is until no precipitation is produced.

5. The comprehensive extraction process of berberine according to claim 1, characterized in that: The specific operation of step S2 is: soak the Phellodendron chinense residue in step S1 in dilute sulfuric acid for 2-3 hours, then add sodium chloride for salting out, the amount of sodium chloride added is 5-10wt% of the Phellodendron chinense residue, and then filter to obtain berberine crude extract A, add dilute sulfuric acid to the filtrate and continue soaking for 2-3 hours, the amount of dilute sulfuric acid added is 0.05-0.1wt% of the mass of the Phellodendron chinense residue, and the mass concentration of the added dilute sulfuric acid is 0.01-0.05%, to obtain berberine crude extract and berberine crude extract B, combine the obtained berberine crude extract A and berberine crude extract B with the berberine crude extract, add gelatin solution and filter, to obtain a filtrate containing tannins and an extract B containing berberine.

6. The comprehensive extraction process of berberine according to claim 1, characterized in that: The polysaccharide extract in step S1 is further subjected to the following separation treatment: Adding a long-chain quaternary ammonium salt to a polysaccharide extract, filtering after standing to obtain an acidic polysaccharide residue and a filtrate, mixing the acidic polysaccharide residue with an inorganic alkali alcohol solution to obtain free acidic polysaccharide, then adding ethanol and filtering, washing the obtained precipitate with water, crystallizing and drying to obtain an acidic polysaccharide; Adding lime milk to the filtrate to adjust the pH to weak alkalinity, leaving it to stand and then filtering it to obtain a polysaccharide precipitate containing neutral polysaccharides and alkaline polysaccharides and an extract A containing berberine; Then in step S3, the obtained extract B is combined with the extract A and then hydrochloric acid is added for acidification.

7. The comprehensive extraction process of berberine according to claim 6, characterized in that: When the polysaccharide extract is separated and treated, the inorganic alkali alcohol solution is an ethanol solution and a sodium hydroxide solution with a mass concentration of 8-15wt%, and the addition of the sodium hydroxide solution makes the pH value 7.5-8.

8. The comprehensive extraction process of berberine according to claim 6, characterized in that: The long-chain quaternary ammonium salt is selected from cetyltrimethylammonium bromide or cetyltrimethylammonium base.

9. The comprehensive extraction process of berberine according to claim 1, characterized in that: The amount of sodium chloride added in step S3 is 3-5wt% of the liquid mixture after adding hydrochloric acid.

10. The comprehensive extraction process of berberine according to claim 1, characterized in that: The precipitate collected after filtration in step S3 is further subjected to heat dissolution, filtration, crystallization and drying treatments in sequence to obtain a berberine extract.

Images