A method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride with Berberis soulieana Schneid

Abstract

The present invention provides a method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride from Berberis vulgaris. The present invention uses Berberis vulgaris as the raw material, and obtains a membrane-refined liquid through crushing, acid leaching, heat preservation filtration clarification and membrane separation. Thereafter, berberine hydrochloride is produced through crystallization and recrystallization; berbamine is produced through decolorization with an alkaline anion exchange resin and fractional crystallization; jatrorrhizine hydrochloride is produced through macroporous adsorption resin chromatography. The present invention continuously produces and prepares berberine hydrochloride, berbamine and jatrorrhizine hydrochloride, realizing the comprehensive utilization of the raw material of Berberis vulgaris. The present invention can solve the problem of environmental pollution by salt wastewater and realize the step-by-step separation and purification of alkaloids. The present invention combines membrane separation with resin, avoiding repeated crystallization and decolorization with activated carbon, and can significantly improve the yield and quality of berbamine. The present invention prepares high-content jatrorrhizine without using organic solvents other than ethanol, and has broad application prospects in industry.

Description

Technical Field

[0001] The present invention relates to the preparation of heterocyclic compounds, and particularly to a method for producing berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride from Berberis soulieana Schneid. Background Art

[0002] Berberis soulieana Schneid. is the common name of plants in the genus Berberis of the family Berberidaceae. There are about more than 200 species of plants in the genus Berberis. Because the spines on the stems of most varieties are divided into three forks, it is named Berberis soulieana Schneid.. It is widely distributed in North China, Northwest China, and Southwest China. The Berberis soulieana Schneid. included in the Chinese Pharmacopoeia (2020 Edition) is the root of several plants of the same genus such as Berberis soulieana Schneid., Berberis wilsonae Hemsl., Berberis poiretii Schneid., or Berberis vernae Schneid. of the family Berberidaceae. It is stipulated that the content of berberine hydrochloride shall not be less than 0.6%. Although Berberis soulieana Schneid. is the main raw material for the production of berberine hydrochloride (berberine) in China at present, the alkaloid contents of many varieties vary greatly. It mainly contains various alkaloids such as berberine, berbamine, jatrorrhizine, palmatine, and other components. There are many domestic studies on the pharmacology and clinical applications of Berberis soulieana Schneid., especially the alkaloids - berberine hydrochloride and berbamine, while there are few reports on the process technology for extracting and separating berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride from Berberis soulieana Schneid.. The traditional textbooks and literature generally disclose that the separation of alkaloids adopts the salting-out method. For many years, few new purification technologies have been applied to the preparation of Berberis soulieana Schneid. alkaloids, especially the reports on the step-by-step preparation of multiple alkaloid monomer components.

[0003] For example, a Chinese patent application with the publication number CN106674218A discloses a method for separating and extracting berbamine and berberine from Berberis soulieana Schneid. The process is as follows: Take the crude powder of Berberis soulieana Schneid, place it in a glass container, soak it in a 0.2% sulfuric acid solution with a volume 5 - 8 times that of the crude drug for more than 24 hours, then filter the filtrate with a gauze. The filter residue is extracted again with acid water with a volume 3 - 5 times that of the residue in the same way, and the two filtrates are combined in an enamel vat; while stirring, add lime milk to adjust the pH to 12, let it stand for 30 minutes, and then carry out vacuum filtration. Add an 8% sodium chloride solution to the filtrate, let it stand, and wait for the precipitation to be complete; dropwise add concentrated hydrochloric acid to adjust the pH to 8 - 8.5, and keep it warm in a water bath at 80°C for more than 30 minutes. Wait for the precipitation to flocculate, take it out and let it cool, let it stand, and carry out vacuum filtration to obtain the precipitate; dry the obtained precipitate below 60°C, then add distilled water with a volume 30 times that of the precipitate, heat it to boiling, and carry out hot filtration while it is hot. The berberine is in the obtained filtrate, and the precipitate is the crude product of berbamine; Dropwise add concentrated hydrochloric acid to the hot solution containing berberine obtained above to adjust the pH to 2, let it stand for the precipitation of berberine hydrochloride to precipitate, carry out filtration, dry it above 60°C, weigh it, and the yield is generally not less than 0.2%; Dry the obtained precipitate containing berbamine below 60°C, weigh it, place it in a glass container, add ethanol with a volume 20 times that of the precipitate and heat it to dissolve, add 2% activated carbon, then heat it to boil for 5 minutes, carry out hot filtration, evaporate the ethanol in the filtrate until the remaining volume is one-fourth of the original volume, cool it, dropwise add distilled water until no more precipitation occurs, adjust the pH to 8 - 8.5 with 5% sodium hydroxide, heat it in a water bath at 80°C for about 5 minutes, let it stand, carry out filtration, and dry it to obtain white berbamine.

[0004] Although the separation and extraction of berbamine and berberine are carried out in this patent application; however, this patent application uses dilute sulfuric acid for room-temperature leaching, which takes a long time, up to more than 24 hours; this patent application uses lime milk to adjust the pH to 12, and keeps it warm at 80°C for more than 30 minutes when the pH is 8 - 8.5. The alkalization produces browning, which deepens the color of the alkaloids and has a great impact on the color of the purified product; this patent application adds 8% sodium chloride for salting out, with a large amount of salt used, polluting the environment; the purification of berberine in this patent application mainly uses hot water and hydrochloric acid for crystallization, making it difficult to meet the standards of purity and color, and several repeated crystallizations and purifications are required; the purification of berbamine in this patent application only uses hot alcohol and activated carbon for purification and decolorization, making it difficult to meet the standards of purity and color, and several repeated crystallizations and purifications are required; moreover, this patent application only prepares berberine hydrochloride and berbamine, and does not involve jatrorrhizine, resulting in partial waste of resources.

[0005] In view of this, it is necessary to provide a method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride from Berberis soulieana Schneid to solve or at least alleviate the technical problems of how to efficiently produce berberine hydrochloride, berbamine and jatrorrhizine hydrochloride from Berberis soulieana Schneid at the same time, how to ensure the product quality, and how to omit salting out. Summary of the Invention

[0006] The main object of the present invention is to provide a method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride from Berberis soulieana Schneid., aiming to solve the technical problems of how to efficiently produce berberine hydrochloride, berbamine and jatrorrhizine hydrochloride from Berberis soulieana Schneid. simultaneously, how to ensure the product quality, and how to omit salting out.

[0007] To achieve the above object, the present invention provides a method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride from Berberis soulieana Schneid., comprising the steps of:

[0008] S1, obtaining the powder of Berberis soulieana Schneid. containing berberine, berbamine and jatrorrhizine;

[0009] S2, extracting the powder of Berberis soulieana Schneid. to obtain an extract;

[0010] S3, filtering the extract through primary filtration and microfiltration to obtain a clear liquid; passing the clear liquid through a high-temperature ultrafiltration membrane, and collecting the downstream liquid of the membrane; passing the downstream liquid of the membrane through a high-temperature nanofiltration membrane, and collecting the upstream liquid of the membrane as a membrane-refined liquid;

[0011] S4, adjusting the pH of the membrane-refined liquid to 6-7, filtering, and then adjusting the pH to 2-3 with a hydrochloric acid solution; standing and then filtering to collect the first wet precipitate and the first mother liquor; adding an ethanol solution with a concentration of 60-65% to the first wet precipitate, heating and dissolving, standing and then filtering to collect the second wet precipitate and the second mother liquor; concentrating the second mother liquor to make the ethanol concentration in the second mother liquor reach 60-65%, standing and then filtering to collect the third wet precipitate and the third mother liquor; combining the first mother liquor and the third mother liquor to obtain a berberine crystallization mother liquor; combining the second wet precipitate and the third wet precipitate to obtain a wet precipitate containing berberine hydrochloride;

[0012] S5, concentrating the berberine crystallization mother liquor to obtain a first concentrated liquid; passing the first concentrated liquid through an alkaline anion exchange resin column, and collecting an ion exchange resin decolorized liquid containing the feed effluent; adjusting the pH of the ion exchange resin decolorized liquid to 6-7, and concentrating to obtain a second concentrated liquid; standing the second concentrated liquid for at least 4 h, and then adjusting the pH to 8-9 to obtain a third concentrated liquid; standing the third concentrated liquid and then filtering to obtain a berbamine crystallization mother liquor and a wet precipitate containing berbamine;

[0013] S6, passing the berbamine crystallization mother liquor through a macroporous adsorption resin column, and then desorbing to collect a desorbed liquid; concentrating the desorbed liquid, and adjusting the pH to 4-6 with a hydrochloric acid solution to obtain a refined liquid containing jatrorrhizine hydrochloride.

[0014] Further, the Berberis powder is made from Berberis; by mass ratio, the content of berbamine in the Berberis powder is 0.5 - 2.0%; by mass ratio and in the form of hydrochloride, the content of berberine hydrochloride in the Berberis powder is 1.0 - 2.5%, and the content of jatrorrhizine hydrochloride is 0.4 - 0.9%.

[0015] Further, in step S2, the Berberis powder is subjected to the leaching with a sulfuric acid solution containing an antioxidant protector; the concentration of the sulfuric acid solution is 0.2 - 0.7%, the temperature of the leaching is 50 - 65°C, and the total duration of the leaching is not more than 8 h.

[0016] Further, in step S3, the temperature of the extraction solution is controlled at 60 - 65°C, the temperature of the clear liquid is controlled at 60 - 65°C, and the temperature of the membrane downstream liquid is controlled at 60 - 65°C; before the clear liquid passes through the high-temperature ultrafiltration membrane, the pH of the clear liquid is adjusted to 1 - 4; the membrane pore size used for the microfiltration is 200 - 800 nm; the maximum temperature that the high-temperature ultrafiltration membrane can withstand is not lower than 70°C; the cut-off molecular weight of the high-temperature ultrafiltration membrane is 3 - 8 KD; the maximum temperature that the high-temperature nanofiltration membrane can withstand is not lower than 70°C; the cut-off molecular weight of the high-temperature nanofiltration membrane is 150 - 300 D.

[0017] Further, in step S4, an ethanol solution with a concentration of 60 - 65% is added to the first wet precipitate at a ratio of 1 g: 12 - 15 mL; the process of concentrating the second mother liquor includes: concentrating the second mother liquor to 1 / 5 - 1 / 4 of the original volume.

[0018] Further, in step S5, the process of concentrating the berberine crystallization mother liquor includes: concentrating the berberine crystallization mother liquor until there is no alcohol smell, and controlling the Brix of the berberine crystallization mother liquor to be 1 - 3 Brix.

[0019] Further, in step S5, the Brix of the second concentrated liquid is 3 - 8 Brix.

[0020] Further, step S5 further includes: after the first concentrated liquid enters the alkaline anion exchange resin column, the column is washed with purified water, the washing column liquid is collected, and the feed effluent and the washing column liquid are combined as the ion exchange resin decolorizing liquid.

[0021] Further, in step S6, desorption is carried out with an ethanol solution with a concentration of 40 - 45%; the process of concentrating the desorbed liquid includes: concentrating the desorbed liquid to 17 - 25 Baumé degrees;

[0022] Step S6 further includes: after the berbamine crystallization mother liquor enters the macroporous adsorption resin column, before desorption, the column is washed with purified water.

[0023] Further, the wet precipitate containing berberine hydrochloride is dried to obtain a berberine hydrochloride product; the wet precipitate containing berbamine is dried to obtain a berbamine product; the refined solution containing jatrorrhizine hydrochloride is dried to obtain a jatrorrhizine hydrochloride product; the berberine hydrochloride product is bright yellow, the berbamine product is white, and the jatrorrhizine hydrochloride product is red-yellow.

[0024] Compared with the prior art, the present invention has at least the following advantages:

[0025] 1. The present invention can efficiently produce berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride simultaneously from Berberis julianae Schneid., forming an extractive separation and intensive processing production technology system for Berberis julianae Schneid. that can be applied industrially, and solving the technical problems of traditional technologies that do not focus on berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride; the present invention first combines high-temperature resistant membrane separation + alkaline anion exchange resin + macroporous adsorption resin, and combines stepwise separation and stepwise crystallization to simultaneously prepare three alkaloid monomer components from Berberis julianae Schneid. medicinal materials, realizing the comprehensive utilization of resources and creating good economic and social benefits.

[0026] 2. The present invention can improve production efficiency, significantly shorten the extraction time, and reduce equipment occupancy; by extracting with dilute sulfuric acid at 50 - 65 °C, the leaching time of more than 24 hours in the traditional process is shortened to within 8 hours.

[0027] 3. The present invention conducts high-temperature resistant ultrafiltration membrane separation and nanofiltration membrane concentration and refinement at 60 - 65 °C and omits salting out, solving the problem of salt wastewater polluting the environment; at the same time, the present invention combines temperature control with the concentration degree, standing time, resin combination, and stepwise crystallization to produce berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride in sequence, solving the problem that it is difficult to separate and purify monomer alkaloids caused by all alkaloids precipitating out during salting out.

[0028] 4. The present invention can reduce the process steps that deepen the color of alkaloids during production and processing, reduce the browning reaction under alkaline conditions, and simplify the decolorization steps; by adding antioxidant protectants, stepwise crystallization, and resin combination during the extraction stage, the present invention avoids the adsorption loss of alkaloids caused by using activated carbon for decolorization, and while improving the product yield, yellow berberine hydrochloride, white berbamine, and red-yellow jatrorrhizine are prepared, solving the problem of poor product color in the market.

[0029] 5. The present invention does not use high-cost separation technology processes such as silica gel and alumina chromatography, and does not use solvents other than water and ethanol, improving the operability and safety of production. Specific embodiments

[0030] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0031] Moreover, the technical solutions between the various embodiments of the present invention can be combined with each other, but it must be based on the ability of those of ordinary skill in the art to implement. When the combination of technical solutions results in contradictions or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0032] When the embodiments give a numerical range, it should be understood that unless otherwise specified in the present invention, both endpoints of each numerical range and any value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention, based on the understanding of those skilled in the art of the prior art and the description of the present invention, can also use any methods, devices, and materials of the prior art similar to or equivalent to those described in the embodiments of the present invention to implement the present invention.

[0033] The three needles used in the embodiments of the present invention were purchased from Sichuan, and the harvesting time was late December; the purified water used was self-made by a pure water machine; the sulfuric acid, ethanol, ion exchange resin, and macroporous adsorption resin used were obtained through conventional commercial channels unless otherwise specified; in the present invention, the concentration of the sulfuric acid solution is the volume concentration, the concentration of the hydrochloric acid solution is the volume concentration, the concentration of the sodium hydroxide solution is the mass concentration, and the concentration of the ethanol is the volume concentration, and all are prepared with water.

[0034] In the embodiments of the present invention, the content of berberine hydrochloride is determined by high performance liquid chromatography (HPLC) according to the "Berberine Hydrochloride" or "Berberis soulieana Schneid." item in the Chinese Pharmacopoeia 2020 Edition; the content of berbamine is determined by high performance liquid chromatography (HPLC); the simultaneous determination of the content of berberine hydrochloride and jatrorrhizine hydrochloride refers to the high performance liquid chromatography (HPLC) under the item of "Mahonia bealei (Fort.) Carr." in the Chinese Pharmacopoeia 2020 Edition; the simultaneous determination of the content of berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride is carried out by high performance liquid chromatography (HPLC). It should be clear that although the Berberis soulieana Schneid. in the present invention contains berberine, berbamine, and jatrorrhizine, the unified standard for determining the content of berberine and jatrorrhizine in the Chinese Pharmacopoeia 2020 Edition is the hydrochloride salt. Therefore, when expressing the substance content of berberine and jatrorrhizine in Berberis soulieana Schneid. in the present invention, it is calculated in the form of hydrochloride salt; and after the production in the present invention, the obtained substances are berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride.

[0035] The descriptions of berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride are as follows:

[0036] Berberine hydrochloride: Also known as berberine hydrochloride, it is a yellow crystalline powder with an extremely bitter taste. It is easily soluble in boiling water, soluble in hot water, slightly soluble in cold water, and almost insoluble in cold ethanol, chloroform, and ether. It is mainly used as an antibacterial agent and is clinically mainly used for intestinal infections and bacillary dysentery, etc., and has the effect of anti-arrhythmia;

[0037] Chinese name: Berberine hydrochloride;

[0038] English name: Berberine;

[0039] CAS Number: 633-65-8;

[0040] Molecular formula: C 20 H 18 ClNO4;

[0041] Molecular weight: 372.82.

[0042] Berbamine: A bis-benzylisoquinoline alkaloid. The free form is poorly soluble in water, insoluble in alkaline water, and easily soluble in inorganic acid water and ethanol. Its hydrochloride is a white powder, easily soluble in water, and has a slightly bitter taste. It has the effects of promoting hematopoiesis, inhibiting tumors, treating anemia, and muscle relaxation, and is used for preventing and treating leukopenia caused by chemotherapy or radiotherapy in cancer patients, benzene poisoning, leukopenia caused by radioactive substances and drugs, etc.;

[0043] Chinese name: Berbamine;

[0044] English name: Berbamine;

[0045] CAS Number: 6078-17-7;

[0046] Molecular formula: C 37 H 40 N2O6;

[0047] Molecular weight: 681.65.

[0048] Jatrorrhizine hydrochloride: Also known as jatrorrhizine, it is a red-yellow needle-like crystal with an extremely bitter taste, soluble in water and ethanol. It has the effects of detoxifying and sterilizing, lowering blood sugar, and positive inotropic effect;

[0049] Chinese name: Jatrorrhizine hydrochloride;

[0050] English name: Jatrorrhizine;

[0051] CAS Number: 3621-38-3;

[0052] Molecular formula: C 20 H20 ClNO4;

[0053] Molecular weight: 465.28.

[0054] As a further illustration of the existing research, the technical content of the existing public disclosure on the extraction of berberine hydrochloride, berbamine, and jatrorrhizine from Berberis vulgaris is mainly as follows:

[0055] The Chinese patent application for invention with the publication number CN111297930A discloses a Berberis vulgaris extract and its preparation method. The preparation method includes the following steps: (1) Raw material treatment: Take the raw material Berberis vulgaris, select and wash it clean; (2) Extraction: Add dilute acid alcohol for reflux extraction; (3) Filtration: Separate the solid and liquid to obtain filtrate a; (4) Concentration: Concentrate filtrate a until it has no alcohol smell, let it stand, and filter to obtain a concentrated solution; (5) Salting out: Add sodium chloride to the concentrated solution, let it stand, and precipitate crystals; (6) Alkalization: Dissolve the crystals in hot water and alkalize them with quicklime, then filter to obtain filtrate b; (7) Acidification: Add dilute hydrochloric acid to filtrate b for acidification, let it stand, precipitate crystals, filter out the crystals, and wash them with water; (8) Drying: Dry the crystals washed with water to obtain the product Berberis vulgaris extract. This patent application extracts Berberis vulgaris alkaloids with dilute acid alcohol, and the use of alcohol makes the production cost relatively high; it prepares the Berberis vulgaris extract by salting out with an acidic solution, alkalizing for impurity removal, and acidifying for crystallization, which is essentially the preparation of berberine hydrochloride. However, during the preparation process, adding a large amount of salt pollutes the environment; alkalization produces browning, which deepens the color of the alkaloids and has a greater impact on the color of the product obtained by crystallization purification; simple single crystallization is difficult to prepare high-purity and good-color berberine hydrochloride, and multiple crystallizations are required; overall, only a single berberine hydrochloride is prepared, and valuable berbamine and jatrorrhizine are discarded, resulting in waste of resources.

[0056] The Chinese invention patent application with publication number CN105440031A discloses a method for purifying berberine hydrochloride from three needles, which comprises the following steps: (1) taking 100-120 parts by weight of root coarse powder, passing it through a 20-mesh sieve, adding 300-400 parts by weight of a 0.5% sulfuric acid aqueous solution to immerse the drug surface, soaking for 6-10 hours, and filtering with absorbent cotton; (2) adding lime milk to the filtrate to neutralize excess sulfuric acid, adjusting the pH to 7, and standing 10-20min; (3) filter out the precipitate, add 200-300 parts by weight of ethanol to the filtrate, stir, heat to 80-90°C and reflux for 20-30min; (4) add ethanol to the filtrate and concentrate under reduced pressure until the filtrate is a brown-red slurry; (5) add 2-4% acetic acid to the brown-red slurry and heat; (6) add hydrochloric acid and acetone until the solution becomes turbid, cool with ice water until berberine hydrochloride precipitates, and dry to obtain fine berberine hydrochloride. This patent application uses dilute sulfuric acid to extract berberine hydrochloride by immersion at room temperature, which is time-consuming; acetic acid, hydrochloric acid and acetone are used for dissolution and purification under acidic conditions, and acetone has certain toxicity; it is miscible with water and a large amount is used until berberine hydrochloride is precipitated, which directly leads to high costs; the low boiling point results in large losses in recovery, making it difficult to use in production; only a single berberine hydrochloride is prepared as a whole, and the valuable berbamine and jatrorrhizine are discarded, resulting in a waste of resources.

[0057] The Chinese invention patent application with the publication number CN103059017A discloses a comprehensive extraction process design of the three needle plant resources. This design route mainly extracts berberine hydrochloride, jatrorrhizine hydrochloride, bamipine hydrochloride, and muscle relaxants simultaneously and efficiently, and also uses berbamine as a raw material to synthesize analgesics. The patent application claims that berberine hydrochloride, berberine, jatrorrhizine and bamipine are comprehensively extracted, but its technical essence lies in repeated salting out, pH adjustment, repeated crystallization of acid water or alcohol water, and activated carbon decolorization; in fact, due to the low content of bamipine in the three needles, there is no extraction value; simple pH adjustment and crystallization are difficult to prepare the three main alkaloids with high purity and good color, and repeated crystallization is required, resulting in unsatisfactory yield and quality, and it is difficult to use in production, and salting out under different pH conditions alone cannot prepare jatrorrhizine with a purity of more than 40%.

[0058] The Chinese patent application with the publication number CN103044417A discloses a method for extracting and purifying berberine from Berberis amurensis Rupr. in the wild in Daxing'an Mountains, especially a new method for purifying berberine by using biological enzymolysis, ultrasonic extraction and column chromatography purification technology. Its advantages are as follows: (1) By applying biological enzymolysis technology, berberine in cells can be fully dissolved; (2) By adopting ultrasonic extraction technology, the heating time of the active ingredients is reduced, the activity loss is small, and the biological activity of the product is high; (3) By using silica gel column chromatography and recrystallization purification technology, the purity of the active ingredient berberine is further improved, which is simple and easy to operate, and the production cost is greatly reduced, and it can be used industrially. However, this patent application only extracts berberine hydrochloride singly and does not involve berbamine and jatrorrhizine, resulting in waste of resources. A large amount of toxic organic solvents are needed for silica gel column chromatography, and there are also safety risks, the preparation cost is high, and it cannot be applied to production.

[0059] The Chinese patent application with the publication number CN104031057A discloses a production method of berbamine, which includes the following steps: S1. Take 250 g of Berberis amurensis Rupr. root powder, add 1750 mL of 2% hydrochloric acid, and place for 22 h; S2. Filter with double-layer gauze, add lime powder to the filtrate to adjust the pH value to 12, and place for 30 min; S3. Siphon and extract, filter out the residue, discard the residue, take the filtrate, add 8% sodium chloride by volume of the filtrate for salting out, after the precipitate precipitates, add hydrochloric acid to adjust the pH value to 8, heat to 80 °C, keep for 30 min, place for 24 h, siphon and filter; S4. Wash and remove impurities, take 1 g and add 20 mL of ether, reflux in a water bath for 30 min; S5. Add 4 mL of petroleum ether, place for 12 h, filter, wash the filter with a small amount of petroleum ether, and dry at 80 °C to obtain berbamine. This patent application only prepares berbamine, ignoring the highly valuable berberine hydrochloride and jatrorrhizine, resulting in serious waste of resources. Adding a large amount of sodium chloride for salting out causes environmental pollution. This patent application uses ether and petroleum ether for decolorization, which is toxic, volatile, flammable and explosive, and the preparation cost is high, and it cannot be applied to production.

[0060] In summary, there is very little existing research on the simultaneous preparation of berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride. The extraction solvents mainly include dilute sulfuric acid and dilute acid alcohol. Comparatively speaking, the production cost of extraction with dilute sulfuric acid is lower, but the time consumption at room temperature is long. Traditional salting-out is generally adopted, with a large amount of sodium chloride used, which is easy to pollute the environment. Alkalinization produces browning, which deepens the color of the alkaloids and has a great impact on the color of the products obtained by crystallization and purification. The use of organic solvents such as acetone, ether, and petroleum ether for decolorization and refinement not only has toxicity, but is also extremely volatile, flammable, and explosive, with a high preparation cost and cannot be applied to production. The use of silica gel chromatography has a high preparation cost and is difficult to implement in production. For simple pH adjustment, crystallization, and decolorization with activated carbon, the purity and color of the alkaloids are difficult to meet the standards, and several repeated crystallization and purification are required, thus greatly reducing the product yield. Existing research generally focuses on experimental techniques, and there is still a certain distance from industrial production. Therefore, with the upgrading and improvement of technology, it is necessary to establish a relatively complete industrial production comprehensive utilization technology system for the extraction, separation, and intensive processing of Berberis soulieana Schneid.

[0061] The present invention mainly realizes the production of berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride through the following aspects: (1) comprehensively utilizing the medicinal materials resources of Berberis soulieana Schneid, continuously producing and preparing three high-value-added products, namely berberine hydrochloride, berbamine, and jatrorrhizine, to reduce the production cost; (2) improving the production efficiency, reducing the equipment occupation, and changing the traditional long-time impregnation extraction; (3) adding as little salt as possible or not adding salt at all to reduce the pollution of salt wastewater to the environment; (4) improving the product color, reducing the process steps that deepen the color during the production and processing, or simplifying the decolorization steps; (5) increasing the yield of alkaloid products, using innovative technologies such as the combination of membrane separation and resin for separation and purification, and avoiding the process steps that reduce the product yield such as repeated crystallization and decolorization with activated carbon; (6) not using silica gel, alumina chromatography processes and organic solvents other than ethanol, to improve the operability and safety of production.

[0062] The present invention provides a method for producing berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride from Berberis soulieana Schneid, and the steps are as follows:

[0063] S1, Crushing:

[0064] Obtaining the powder of Berberis soulieana Schneid; the powder of Berberis soulieana Schneid uses Berberis soulieana Schneid as the raw material, and the powder of Berberis soulieana Schneid contains berberine, berbamine, and jatrorrhizine; the process of obtaining the powder of Berberis soulieana Schneid includes: crushing the raw material into coarse powder with a size basically the same as that of grains.

[0065] In the present invention, by mass ratio, the content of berbamine in the powder of Berberis soulieana Schneid is 0.5-2.0%; by mass ratio and in the form of hydrochloride, the content of berberine hydrochloride in the powder of Berberis soulieana Schneid is 1.0-2.5%, and the content of jatrorrhizine hydrochloride is 0.4-0.9%.

[0066] For Berberis soulieana Schneid., except for the roots, the stem bark also contains relatively high contents of berberine, berbamine and jatrorrhizine. To expand the medicinal parts for alkaloid extraction from the increasingly scarce medicinal materials and improve the comprehensive utilization of resources, considering the contents of the main alkaloids in Berberis soulieana Schneid. comprehensively, the roots and stem bark of Berberis soulieana Schneid. are used as the extraction raw materials in the present invention.

[0067] S2, acid leaching:

[0068] The Berberis soulieana Schneid. powder is leached with a sulfuric acid solution containing an antioxidant protective agent to obtain an extraction solution; the concentration of the sulfuric acid solution is 0.2 - 0.7%, the temperature of the leaching is 50 - 65°C, and the total duration of the leaching is not more than 8 h, and further can be 5 - 8 h.

[0069] In the present invention, the antioxidant protective agent includes vitamin C and ferrous sulfate, and the mass ratio of vitamin C to ferrous sulfate is 1 - 3:1; the mass percentage of the antioxidant protective agent to the Berberis soulieana Schneid. powder is 1.0 - 4.0%. Stirring is carried out during the leaching process in the present invention, and specifically, intermittent stirring can be carried out, such as: starting stirring every 10 min or carrying out air pressure backwashing for 2 min.

[0070] In the present invention, the number of leaching times is three; during the first leaching, the ratio of the sulfuric acid solution to the Berberis soulieana Schneid. powder is controlled to be 5 - 7 mL:1 g, the Berberis soulieana Schneid. powder is leached in the sulfuric acid solution for 2 - 4 h, and the first filtrate and the first filter residue are collected after filtration; during the second leaching, the ratio of the sulfuric acid solution to the Berberis soulieana Schneid. powder is controlled to be 4 - 6 mL:1 g, the first filter residue is leached in the sulfuric acid solution for 1 - 3 h, and the second filtrate and the second filter residue are collected after filtration; during the third leaching, the ratio of the sulfuric acid solution to the Berberis soulieana Schneid. powder is controlled to be 5 - 7 mL:1 g, the second filter residue is leached in the sulfuric acid solution for 0.5 - 1.5 h, and the third filtrate is collected after filtration; the first filtrate and the second filtrate are combined as the extraction solution, and the third filtrate is used for leaching the next batch of Berberis soulieana Schneid. powder. Of course, without considering the third leaching, the number of leaching times can also be two, and the first leaching and the second leaching are carried out, and then the first filtrate and the second filtrate are combined as the extraction solution.

[0071] In the present invention, during the first leaching, the mass percentage of the antioxidant protective agent to the Berberis soulieana Schneid. powder is 1.0 - 2.0%; during the second leaching, the mass percentage of the antioxidant protective agent to the Berberis soulieana Schneid. powder is 0.3 - 0.8%; during the third leaching, the mass percentage of the antioxidant protective agent to the Berberis soulieana Schneid. powder is 0.1 - 0.3%.

[0072] S3, heat preservation, filtration, clarification and membrane separation:

[0073] S31. Control the temperature of the extraction liquid at 60 - 65°C; subject the extraction liquid to primary filtration and microfiltration in sequence to obtain a clear liquid; the membrane pore size used for the microfiltration is 200 - 800 nm.

[0074] In the present invention, the primary filtration includes three - foot cloth - bag centrifugal filtration or plate - frame filtration, and the microfiltration uses a ceramic membrane with a pore size of 200 - 800 nm; the material of the ceramic membrane is alumina or zirconia.

[0075] In the present invention, when subjecting the liquid after primary filtration to microfiltration, when the volume of the upstream liquid remains only 1 / 30 - 1 / 40 of the extraction liquid, add hot water (the volume of the hot water is 1.5 - 2.5 times the remaining volume of the upstream liquid, and the temperature is 60 - 65°C) to dilute the upstream liquid, and continue to pass through the membrane; when the volume of the upstream liquid remains only 1 / 30 - 1 / 40 of the extraction liquid again, add hot water (the volume of the hot water is 1.5 - 2.5 times the remaining volume of the upstream liquid, and the temperature is 60 - 65°C) to dilute the upstream liquid again, and continue to pass through the membrane.

[0076] S32. Control the temperature of the clear liquid at 60 - 65°C; adjust the pH of the clear liquid to 1 - 4 using an alkaline solution, and then pass the clear liquid through a high - temperature ultrafiltration membrane, and collect the liquid downstream of the membrane.

[0077] The maximum temperature that the high - temperature ultrafiltration membrane can withstand is not lower than 70°C, and the cut - off molecular weight of the high - temperature ultrafiltration membrane is 3 - 8 KD; specifically, the maximum temperature that the high - temperature ultrafiltration membrane can withstand is 70°C, which is prepared by a unique high - temperature - resistant membrane filament and paper - membrane process, and the membrane material is PES (polyethersulfone) or PVDF (polyvinylidene fluoride), and the model is the UE series (such as UE005 - 8040H); that is, the membrane material of the high - temperature ultrafiltration membrane includes one or more of polyethersulfone and polyvinylidene fluoride.

[0078] In the present invention, when passing the clear liquid through the high - temperature ultrafiltration membrane, when the volume of the upstream liquid remains only 1 / 30 - 1 / 40 of the extraction liquid, add hot water (the volume of the hot water is 1.5 - 2.5 times the remaining volume of the upstream liquid, and the temperature is 60 - 65°C) to dilute the upstream liquid, and continue to pass through the membrane; when the volume of the upstream liquid remains only 1 / 30 - 1 / 40 of the extraction liquid again, add hot water (the volume of the hot water is 1.5 - 2.5 times the remaining volume of the upstream liquid, and the temperature is 60 - 65°C) to dilute the upstream liquid again, and continue to pass through the membrane.

[0079] S33. Control the temperature of the liquid upstream of the membrane at 60 - 65°C; pass the liquid upstream of the membrane through a high - temperature nanofiltration membrane, collect the liquid upstream of the membrane, and use the liquid upstream of the membrane as the membrane - refined liquid.

[0080] The maximum temperature that the high-temperature nanofiltration membrane can withstand is not lower than 70 °C, and the molecular weight cut-off of the high-temperature nanofiltration membrane is 150-300 D; specifically, the maximum temperature that the high-temperature nanofiltration membrane can withstand is 70 °C, the membrane material is PES (polyethersulfone), UF series (such as UF8040); that is, the membrane material of the high-temperature nanofiltration membrane includes polyethersulfone.

[0081] S4, Production of berberine hydrochloride:

[0082] S41, Adjust the pH of the membrane refining liquid to 6-7 with an alkali solution, and then filter; adjust the pH of the filtered liquid to 2-3 with a hydrochloric acid solution, and let it stand at 5-25 °C for at least 48 h or 48-72 h to fully precipitate the precipitate; after standing, filter (suction filtration or plate-and-frame filtration) to collect the first wet precipitate and the first mother liquor.

[0083] S42, Add an ethanol solution with a concentration of 60-65% to the first wet precipitate at a ratio of 1 g: 12-15 mL, and heat to 80-90 °C to completely dissolve the precipitate; when the first wet precipitate is completely dissolved, let it stand at 5-25 °C for at least 20 h or 22-26 h; after standing, filter (suction filtration or plate-and-frame filtration) to collect the second wet precipitate and the second mother liquor.

[0084] S43, Concentrate the second mother liquor (vacuum decompression concentration) to 1 / 5-1 / 4 of the original volume, and then add high-grade ethanol to the second mother liquor to make the ethanol concentration in the second mother liquor reach 60-65%; let the second mother liquor stand at 5-25 °C for at least 20 h or 22-26 h, and after standing, filter (suction filtration or plate-and-frame filtration) to collect the third wet precipitate and the third mother liquor.

[0085] S44, Combine the first mother liquor and the third mother liquor to obtain the berberine crystallization mother liquor; combine the second wet precipitate and the third wet precipitate to obtain the wet precipitate containing berberine hydrochloride.

[0086] S5, Production of berbamine:

[0087] S51, Decolorization with ion exchange resin: Concentrate the berberine crystallization mother liquor until there is no alcohol smell (vacuum decompression recovery of ethanol until there is no alcohol smell), and continue to concentrate until the Brix of the berberine crystallization mother liquor is 1-3 Brix to obtain the first concentrated solution.

[0088] Feed the first concentrated solution into an alkaline anion exchange resin column at room temperature (e.g., 5 - 25 °C), collect the feed effluent, and use the feed effluent as the decolorized solution of the ion exchange resin. Further, after feeding the first concentrated solution into the alkaline anion exchange resin column, wash the column with 3 - 5 BV of purified water, collect the wash solution of 0 - 3 BV, and combine the feed effluent and the wash solution as the decolorized solution of the ion exchange resin.

[0089] In the present invention, the alkaline anion exchange resin column contains one of gel - type acrylic strong - base anion resin, gel - type acrylic weak - base anion resin, gel - type styrene strong - base anion resin, gel - type styrene weak - base anion resin, macroporous acrylic strong - base anion resin, macroporous acrylic weak - base anion resin, macroporous styrene strong - base anion resin, and macroporous styrene weak - base anion resin. Preferred models: D900, D941, D316, D296, D280.

[0090] S52, fractional crystallization: Adjust the pH of the decolorized solution of the ion exchange resin to 6 - 7, then concentrate it under vacuum to a Brix of 3 - 8 to obtain a second concentrated solution; let the second concentrated solution stand at 5 - 25 °C (preferably 5 - 20 °C) for at least 4 h or 4 - 6 h to precipitate part of it, then adjust the pH of the second concentrated solution to 8 - 9 to obtain a third concentrated solution; let the third concentrated solution stand at 5 - 25 °C (preferably 5 - 20 °C) for at least 10 h or 10 - 15 h to completely precipitate, and then filter (by suction filtration or plate - and - frame filtration) to obtain the berbamine crystallization mother liquor and the wet precipitate containing berbamine. Further, in the present invention, an alkali solution is used to adjust the pH of the decolorized solution of the ion exchange resin to 6 - 7, and an alkali solution is used to adjust the pH of the second concentrated solution to 8 - 9.

[0091] S6, production of jatrorrhizine hydrochloride:

[0092] Feed the berbamine crystallization mother liquor into a macroporous adsorption resin column, then desorb it with an ethanol solution with a concentration of 40 - 45%, and collect the desorbing solution; concentrate the desorbing solution (by vacuum concentration) to 17 - 25 Baume degrees, and adjust the pH to 4 - 6 with a hydrochloric acid solution to obtain a refined solution containing jatrorrhizine hydrochloride. Further, after feeding the berbamine crystallization mother liquor into the macroporous adsorption resin column, before performing the desorption, wash the column with 2 - 4 BV of purified water.

[0093] In the present invention, the resin in the macroporous adsorption resin column is one of non - polar macroporous adsorption resin and weakly polar macroporous adsorption resin; specifically, the resin in the macroporous adsorption resin is one of non - polar D101, LX - 100B, LX - T28, or weakly polar AB - 8.

[0094] In the present invention, the purification of jatrorrhizine hydrochloride is the combined use of macroporous adsorption resin purification after separating berberine hydrochloride and berbamine through the membrane separation in step S3 and the decolorization with ion exchange resin in step S5.

[0095] S7, drying:

[0096] Dry the wet precipitate containing berberine hydrochloride to obtain a berberine hydrochloride product; dry the wet precipitate containing berbamine to obtain a berbamine product; dry the refined solution containing jatrorrhizine hydrochloride to obtain a jatrorrhizine hydrochloride product; the berberine hydrochloride product is bright yellow, the berbamine product is white, and the jatrorrhizine hydrochloride product is red-yellow.

[0097] Specifically, vacuum dry or microwave vacuum dry the wet precipitate containing berberine hydrochloride, the wet precipitate containing berbamine, and the refined solution containing jatrorrhizine at a temperature of 60 - 65°C and a vacuum degree of ≥ -0.085 MPa to obtain bright yellow berberine hydrochloride, white berbamine, and red-yellow jatrorrhizine hydrochloride respectively.

[0098] The present invention continuously produces and prepares three high-value-added products, realizing the comprehensive utilization of raw materials; through special high-temperature-resistant membrane heat preservation separation and crystallization, replacing traditional salting-out, solving the environmental pollution of salt wastewater, and realizing the step-by-step separation and purification of alkaloids; through the combination of membrane separation and resin, avoiding repeated crystallization and decolorization with activated carbon, significantly improving the yield and quality of berbamine, and preparing high-content jatrorrhizine without using organic solvents other than silica gel and ethanol; forming an advanced and practical production technology system for the extraction, separation, and intensive processing of Berberis julianae Schneid.

[0099] As supplementary explanation, in the present invention, the lye in step S3, step S4, and step S5 is a sodium hydroxide solution with a concentration of 1 - 3%; in step S4 and step S6, the concentration of the hydrochloric acid solution is 1 - 3%.

[0100] In step S2 of the present invention, the concentration of the sulfuric acid solution is 0.2 - 0.7%; if it is too low, the dosage is too large, and if it is too high, it will cause waste and increase production costs, and there will be obvious acid corrosion to the pipeline system.

[0101] In step S2 of the present invention, the extraction temperature is 50 - 65°C; the traditional process uses room-temperature dilute acid extraction, which takes more than 24 hours, has a long production cycle, seriously occupies equipment, and has low efficiency; when the temperature exceeds 75°C, the color of the acid extract significantly deepens, which is not conducive to product decolorization and purification, and there is damage to alkaloids, reducing the content of the extract, and the higher the temperature, the more obvious; the extraction temperature of the present invention can extract the three alkaloid components quickly, with stable content, short time, and high extraction efficiency.

[0102] In the step S2 of the present invention, the antioxidant is a substance obtained by compounding vitamin C and ferrous sulfate in a certain proportion; by adding a specific antioxidant, especially a compound substance, not only can the color deepening of the extraction solution be slowed down, but also the alkaloid components can be kept stable and not easily destroyed in a heating environment of 50-65 °C, which is beneficial to decolorization and purification.

[0103] In the step S3 of the present invention, the temperature is maintained at 60-65 °C; at this temperature, the alkaloids from Berberis pruinosa Franch., especially the sulfates of berberine, berbamine and jatrorrhizine, have a relatively large solubility and are not easily precipitated, which is beneficial to membrane separation and purification; when the temperature is at room temperature, the extraction solution will become turbid and post-precipitation will occur, and in severe cases, the membrane pores will be blocked and the equipment will be damaged; when the temperature is too high, it is not conducive to the content stability of the alkaloid components, and it also exceeds the maximum temperature that the special membrane can withstand, so membrane separation cannot be used to purify the alkaloids.

[0104] In the step S3 of the present invention, the liquid obtained after primary filtration is clarified by microfiltration through a ceramic membrane made of alumina or zirconia with a pore size of 200-800 nm; at 60-65 °C, through the microfiltration clarification of the ceramic membrane with this pore size, the precipitate in the extraction solution can be removed to obtain a clear and transparent supernatant without post-precipitation, which is beneficial to membrane separation and purification.

[0105] In the step S3 of the present invention, the high-temperature ultrafiltration membrane can withstand a maximum temperature of up to 70 °C, with a cut-off molecular weight of 3-8 KD. It is prepared by using a unique high-temperature resistant membrane filament and paper membrane process, and the membrane material is PES or PVDF, with the model of UE series (such as UE005-8040H). This special specification of ultrafiltration membrane can preliminarily purify the alkaloids from Berberis pruinosa Franch. Because the maximum temperature that a common ultrafiltration membrane can withstand is 45 °C, and under the condition of 45 °C, due to the solubility difference of different alkaloid sulfates, especially the sulfate of berberine, when the ultrafiltration membrane runs to a smaller volume, it will precipitate and block the membrane pores with other impurities, which not only damages the equipment and makes the process impossible to proceed, but also reduces the product yield; a cut-off molecular weight of 3-8 KD can remove most of the impurities and pigments, while the sulfate of the alkaloids from Berberis pruinosa Franch. can completely pass through, thus obtaining a relatively high-purity total alkaloid from Berberis pruinosa Franch.

[0106] In the step S3 of the present invention, the high-temperature nanofiltration membrane can withstand a maximum temperature of up to 70 °C, with a molecular weight cut-off of about 200 D. The membrane material is PES, and it belongs to the UF series (such as UF8040). This specification of nanofiltration membrane can concentrate the ultrafiltration separation liquid of Berberis alkaloids. Since the maximum temperature that a common nanofiltration membrane can withstand is 45 °C, and at 45 °C, due to the solubility differences of different alkaloid sulfates, especially berberine sulfate, during the concentration process of the nanofiltration membrane, it will precipitate and block the membrane pores with other impurities, which not only damages the equipment and makes the process impossible to proceed, but also reduces the product yield; a molecular weight cut-off of about 200 D can completely retain the Berberis alkaloids. At 60 - 65 °C, concentration with the high-temperature resistant nanofiltration membrane can replace the traditional vacuum decompression concentration to obtain the membrane refined liquid.

[0107] In the step S4 of the present invention, before the crystallization of berberine hydrochloride, the pH of the membrane refined liquid needs to be adjusted to 6 - 7 with dilute alkali, filtered, and then the pH is adjusted to 2 - 3 with dilute hydrochloric acid. The solubility of Berberis alkaloid sulfate is greater than that of the hydrochloride, and the solubility differences of berberine, berbamine, and jatrorrhizine sulfate are not significant, which is not conducive to the crystallization and precipitation of monomer alkaloids; however, the solubility of berberine hydrochloride in cold and hot water varies greatly, and the hydrochloride is beneficial to separate berberine from the total alkaloids first, while berbamine and jatrorrhizine remain in the solution. Therefore, it is necessary to convert the sulfate into the hydrochloride.

[0108] In the step S4 of the present invention, the pH for the crystallization condition of berberine hydrochloride is 2 - 3, and it is naturally cooled to room temperature and left standing for at least 48 h or 48 - 72 h; the recrystallization condition is 60 - 65% ethanol, heated at 80 - 90 °C to completely dissolve the precipitate, and left standing after natural cooling to room temperature; the stepwise crystallization condition is that the recrystallization mother liquor is concentrated to 1 / 5 - 1 / 4 of the original volume, and high-grade ethanol is added to adjust the ethanol concentration to 60 - 65%, and left standing after natural cooling to room temperature.

[0109] After separation and concentration by the high-temperature resistant ultrafiltration membrane and nanofiltration membrane in the above-mentioned process, when it is cooled naturally to room temperature at a certain volume and allowed to stand for a sufficient time, berberine hydrochloride can crystallize out completely without adding sodium chloride for salting out, while only a small amount of berbamine hydrochloride and jatrorrhizine are co-precipitated with berberine hydrochloride. This is significantly different from the traditional salting out phenomenon in which a large amount of berbamine and jatrorrhizine are co-precipitated with berberine hydrochloride. In the present invention, the high-temperature resistant membrane separation method, due to improving the purity of the alkaloids in Berberis vulgaris and adjusting the crystallization volume to a more suitable concentration, can not only replace the traditional salting out to completely precipitate berberine hydrochloride, but also obtain berberine hydrochloride crystals with a higher purity, avoiding the defects of salting out most other alkaloids, such as berbamine and jatrorrhizine, which increases the difficulty of later purification and the environmental pollution caused by the traditional salting out with a large amount of salt. Recrystallization with 60-65% ethanol and fractional crystallization can obtain berberine hydrochloride with a purity of more than 97%, and at the same time solve the practical problem that it is not easy to recycle high-concentration ethanol in industrial production due to the use of ethanol with a concentration of more than 80% in the traditional process, and is completely separated from the small amount of berbamine hydrochloride and jatrorrhizine precipitated in the crystallization stage.

[0110] In step S5 of the present invention, the crystallization conditions of berbamine are first decolorized by ion exchange resin, and then subjected to fractional crystallization with a pH of 6-7, a concentration of 3-8 Brix, standing for more than 4 h and a pH of 8-9, standing for more than 10 h.

[0111] In the present invention, fractional separation is the key to continuously preparing the alkaloids in Berberis vulgaris. The mother liquor remaining after the separation of berberine hydrochloride contains a large amount of pigments. Although these pigments have basically the same molecular weight and / or molecular structure as the alkaloids, and still coexist with the alkaloids and are not easy to separate after the above-mentioned membrane separation, a certain specification and model of basic anion exchange resin column can exchange and adsorb these pigments without adsorbing the alkaloid components. Therefore, the alkaloids mainly composed of berbamine and jatrorrhizine can be separated from most of the pigments by the basic anion exchange resin column, so as to obtain alkaloids with good color and high purity. When the pH is 6-7, part of berbamine will crystallize out. Although the crystallization rate is slow, the color and crystal form are good. When the pH is further adjusted to 8-9, it can be completely precipitated after a certain time. The fractional crystallization by adjusting the pH has better color and crystal form of the crystals and higher purity up to more than 98% compared with the crystallization precipitation by adjusting the pH to 8-9 at one time. Therefore, after high-temperature resistant membrane separation and decolorization by basic anion exchange resin column, combined with fractional crystallization technology, it is possible to solve the problems of repeatedly adjusting the pH, adding activated carbon for decolorization, and repeatedly crystallizing with alcohol and water in the traditional technology for refining berbamine, which affect the yield and production cost, by simply adjusting the pH step by step once.

[0112] In step S5 of the present invention, the resin in the alkaline anion exchange resin column can be gel-type and macroporous acrylic weak base anion resins, acrylic strong and weak base anion resins, gel-type and macroporous styrene-based strong base anion resins, macroporous acrylic strong base anion resins, macroporous styrene-based weak base anion resins, etc. Preferred models: D900, D941, D316, D296, D280. The crystallization mother liquor after separating berberine hydrochloride contains more pigments, and it is necessary to further separate the pigments from alkaloids represented by berbamine and jatrorrhizine. There are literature reports on purifying alkaloids with cation exchange resins. Although cation resins do not adsorb pigments, the alkaloids in Berberis sargentiana Schneid. exchange and adsorb with cation resins and are difficult to resolve, and the overall desorption rate is less than 20%, resulting in a large amount of loss and being unable to be applied in production. Through a large number of comparative experiments, it is found that by changing the method, first remove the pigments, that is, use anion exchange resin to exchange and adsorb pigments, and this resin does not adsorb the alkaloids in Berberis sargentiana Schneid., so that the alkaloids represented by berbamine and jatrorrhizine flow out from the anion resin column, thus realizing the separation of alkaloids and pigments.

[0113] In step S6 of the present invention, the resin in the macroporous adsorption resin column is non-polar D101, LX-100B, LX-T28, etc., and weakly polar AB-8, etc.; the desorbing ethanol concentration is 40-45%. In the present invention, a macroporous adsorption resin of a certain specification and model has a strong adsorption for jatrorrhizine, and has a small adsorption for other impurities in the mother liquor after separating berbamine. After the previous high-temperature membrane separation, ion exchange resin and other refining and decolorization process steps, and stepwise separation of berberine hydrochloride and berbamine, the impurities can be separated by a macroporous adsorption resin of a specific specification and model. When the ethanol concentration is relatively high, the jatrorrhizine and impurities adsorbed on the resin will be desorbed together, while controlling an appropriate ethanol concentration can desorb only jatrorrhizine without desorbing most of the impurities, thereby preparing jatrorrhizine with a purity of more than 80%.

[0114] The following are specific examples of the present invention:

[0115] Example 1

[0116] A method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride from Berberis sargentiana Schneid., the steps are as follows:

[0117] S1, Crushing:

[0118] Take a total of 800 kg of dried Berberis sargentiana Schneid. medicinal materials and crush them into coarse powder basically the size of grains with a hammer mill.

[0119] Detect the content of the coarse powder of Berberis sargentiana Schneid. medicinal materials. By mass ratio, the content of berbamine is 0.87%; by mass ratio and in the form of hydrochloride, the content of berberine hydrochloride is 1.58%, and the content of jatrorrhizine hydrochloride is 0.65%.

[0120] S2, Acid leaching extraction:

[0121] Put the coarse powder into a 6m 3 enamel extraction tank and extract with dilute sulfuric acid.

[0122] In this example, the extraction is carried out 3 times, and the third extraction is a double extraction; the extraction temperature is 56°C; the stirring method during extraction is: start stirring for 2 minutes every 10 minutes; during the 3 extractions, the addition amounts of the antioxidant protectant are 10.0 kg, 4.5 kg, and 1.5 kg respectively, and the antioxidant protectant is composed of vitamin C and ferrous sulfate in a mass ratio of 2:1; during the 3 extractions, the extraction times are 3.0 h, 2.0 h, and 1.0 h respectively; the concentration of the dilute sulfuric acid used in the 3 extractions is 0.3%; during the 3 extractions, the addition amounts of the dilute sulfuric acid are 4800 L, 4000 L, and 4800 L respectively; after each extraction, filter, and the filter residue enters the next extraction. Combine the filtrates of the first 2 extractions to obtain 7100 L of extraction solution.

[0123] S3, Heat preservation, filtration, clarification and membrane separation:

[0124] S31, Keep the temperature of the extraction solution at 62°C; first carry out plate and frame filtration on the extraction solution, and then micro-filter and clarify the filtrate through a ceramic membrane with a zirconia material and a pore size of 500 nm; when there is only 200 L of the upstream liquid left, add 400 L of hot water at 65°C to dilute the upstream liquid and continue to pass through the membrane; when there is again only 200 L of the upstream liquid left, add another 400 L of hot water at 65°C to dilute the upstream liquid and continue to pass through the membrane; collect the downstream liquid of the membrane to obtain the clear liquid;

[0125] S32, Keep the temperature of the clear liquid at 62°C; adjust the pH of the clear liquid to 3 with 2% sodium hydroxide solution, and pass it through a high-temperature ultrafiltration membrane that can withstand a maximum temperature of 70°C and has a cut-off molecular weight of 5 KD (membrane material PVDF, model UE005 - 8040H); when there is only 200 L of the upstream liquid left, add 400 L of hot water at 62°C to dilute the upstream liquid and continue to pass through the membrane; when there is again only 200 L of the upstream liquid left, add another 400 L of hot water at 62°C to dilute the upstream liquid and continue to pass through the membrane; collect the downstream liquid of the membrane, about 8200 L;

[0126] S33, Keep the temperature of the downstream liquid of the membrane (from step S32) at 62°C; pass the downstream liquid of the membrane through a high-temperature nanofiltration membrane that can withstand a maximum temperature of 70°C and has a cut-off molecular weight of 200 D (material PES, model UF8040), and collect the upstream liquid of the membrane to obtain the refined membrane liquid, about 450 L.

[0127] S4, Production of berberine hydrochloride:

[0128] S41. Adjust the pH of the membrane-refined liquid to 6 with 2% sodium hydroxide solution, perform plate-and-frame filtration, then adjust the pH to 2 with 2% hydrochloric acid solution, naturally cool to room temperature (in this example, the liquid temperature at room temperature is about 15 °C), and let it stand at room temperature for 65 h to fully precipitate the sediment. Then perform plate-and-frame filtration, separately collect the wet sediment and the mother liquor, weigh them and measure the volume, obtaining 34.96 kg of wet sediment and 420 L of mother liquor;

[0129] S42. Add 450 L of ethanol with a concentration of 64%, heat and stir at 85 °C to completely dissolve the wet sediment in step S41, naturally cool to room temperature, and let it stand at room temperature for 24 h. Then perform plate-and-frame filtration, separately collect the wet sediment and the mother liquor, weigh them and measure the volume, obtaining 17.20 kg of wet sediment and 440 L of mother liquor;

[0130] S43. Vacuum-concentrate the 440 L of mother liquor in step S42 to 100 L, add high-proof ethanol to adjust the ethanol concentration to 60%, naturally cool to room temperature, and let it stand at room temperature for 24 h. Then perform plate-and-frame filtration, separately collect the wet sediment and the mother liquor, obtaining 6.40 kg of wet sediment and 110 L of mother liquor;

[0131] S44. Combine the mother liquors in steps S41 and S43 to obtain about 530 L of berberine crystallization mother liquor; combine the wet sediments in steps S42 and S43 to obtain 23.6 kg of bright yellow berberine hydrochloride (wet sediment containing berberine hydrochloride).

[0132] S5. Production of berbamine:

[0133] S51. Decolorization with ion-exchange resin: Vacuum-decompress and recover ethanol from the berberine crystallization mother liquor at 62 °C until there is no alcohol smell, add water to remove alcohol, concentrate to 450 L (2.9 Brix), and cool to room temperature; then, pass it through a column filled with 75 kg of alkaline anion-exchange resin of model D900. After feeding, wash the column with 300 L of purified water (4 BV), collect the feed effluent and 200 L of column wash liquor (2.7 BV), and combine them to obtain the ion-exchange resin decolorized liquid;

[0134] S52. Fractional crystallization: Adjust the pH of the ion-exchange resin decolorized liquid to 7 with 2% sodium hydroxide solution, vacuum-decompress and concentrate to 200 L (6 Brix), let it stand at room temperature for 5 h to precipitate some white sediment; then, continue to adjust the pH to 9 with 2% sodium hydroxide solution, let it stand at room temperature for 12 h to completely precipitate the sediment; finally, perform plate-and-frame filtration, separately collect the wet sediment and the mother liquor, obtaining 13.30 kg of white berbamine (wet sediment containing berbamine), and about 190 L of berbamine crystallization mother liquor.

[0135] S6. Production of jatrorrhizine hydrochloride:

[0136] Feed the berbamine crystallization mother liquor into a non-polar macroporous adsorption resin column of model LX-T28 with a capacity of 200 kg. After feeding, wash the column with 600 L of purified water (3 BV), and then desorb with 800 L of ethanol with a concentration of 40%. Collect the desorbing solution, concentrate it under vacuum to 18 Baumé, and adjust the pH to 4 with a 2% hydrochloric acid solution to obtain a refined solution containing jatrorrhizine.

[0137] S7, Drying:

[0138] Dry the above-mentioned wet precipitate containing berberine hydrochloride to obtain berberine hydrochloride product; dry the above-mentioned wet precipitate containing berbamine to obtain berbamine product; dry the above-mentioned refined solution containing jatrorrhizine hydrochloride to obtain jatrorrhizine hydrochloride product; the drying method is: microwave vacuum drying, temperature 62 °C, vacuum degree ≥ -0.085 MPa.

[0139] In this example, the mass of the berberine hydrochloride product is 10.62 kg, the mass of the berbamine product is 5.95 kg, and the mass of the jatrorrhizine hydrochloride product is 4.32 kg; in this example, the berberine hydrochloride product is bright yellow, the berbamine product is white, and the jatrorrhizine hydrochloride product is red-yellow.

[0140] In the berberine hydrochloride product of this example, the mass proportion of berberine hydrochloride is 97.86%; in the berbamine product of this example, the mass proportion of berbamine is 98.25%; in the jatrorrhizine hydrochloride product of this example, the mass proportion of jatrorrhizine hydrochloride is 82.20%.

[0141] Comparative Example 1

[0142] Compared with Example 1, in this comparative example, the fractional crystallization in step S52 is adjusted to direct crystallization, and other conditions remain unchanged.

[0143] In this comparative example, the process of direct crystallization is as follows: directly adjust the pH of the decolorized solution of ion exchange resin to 9 with a 2% sodium hydroxide solution, concentrate it under vacuum to 200 L, and let it stand at room temperature for 17 h; then, perform plate and frame filtration, and collect the wet precipitate and mother liquor separately to obtain 13.46 kg of white berbamine (wet precipitate containing berbamine) and approximately 190 L of berbamine crystallization mother liquor.

[0144] In this comparative example, the mass of the berbamine product is 6.02 kg, and the mass proportion of berbamine in the berbamine product is 97.10%.

[0145] Comparative Example 2

[0146] Compared with Example 1, in this comparative example, the basic anion exchange resin column in step S51 is adjusted to a granular activated carbon column, and other conditions remain unchanged.

[0147] In this comparative example, the mass of the berbamine product is 4.62 kg, the mass fraction of berbamine in the berbamine product is 96.30%, and the color is off-white.

[0148] Comparative Example 3

[0149] Compared with Example 1, in this comparative example, the use of the alkaline anion exchange resin column in step S51 is omitted (the production process of berbamine is correspondingly adjusted), and other conditions remain unchanged.

[0150] In this comparative example, the production process of berbamine is as follows: The berberine crystallization mother liquor is vacuum-decompressed to recover ethanol at 62°C until there is no alcohol smell, water is added to remove alcohol, concentrated to 450 L, and cooled to room temperature. Then, the pH is adjusted to 7 with 2% sodium hydroxide solution, and vacuum-decompressed and concentrated to 200 L (6.5 Brix), and left to stand at room temperature for 5 h to precipitate some yellowish-white precipitates; afterwards, the pH is continuously adjusted to 9 with 2% sodium hydroxide solution and left to stand at room temperature for 12 h; finally, plate and frame filtration is carried out, and the wet precipitate and the mother liquor are collected separately to obtain 13.56 kg of white berbamine (wet precipitate containing berbamine) and about 190 L of berbamine crystallization mother liquor.

[0151] In this comparative example, the mass of the berbamine product is 6.06 kg, the mass fraction of berbamine in the berbamine product is 96.50%, and the color is yellowish-white.

[0152] Comparative Example 4

[0153] Compared with Example 1, in this comparative example, the extraction temperature in step S2 is adjusted to 15°C, and other conditions remain unchanged.

[0154] In this comparative example, the mass of the berberine hydrochloride product is 7.65 kg, the mass of the berbamine product is 4.65 kg, and the mass of the jatrorrhizine hydrochloride product is 3.28 kg; the results show that: when extracting for the same time as in Example 1 at room temperature, the yields of the three alkaloids are all less than 70%, and sufficient extraction cannot be carried out.

[0155] Comparative Example 5

[0156] Compared with Example 1, in this comparative example, the temperature of the clear liquid in step S32 is adjusted to 42°C, and other conditions remain unchanged.

[0157] In this comparative example, after the ultrafiltration membrane operates for 1 h, the flux drops from 1200 L / h to 350 L / h, which drops significantly and cannot operate normally, and needs to be shut down for cleaning, and the upstream liquid volume is greater than 1200 L.

[0158] Comparative Example 6

[0159] Compared with Example 1, in this comparative example, the temperature of the membrane downstream liquid (from step S32) in step S33 is adjusted to 42°C, and other conditions remain unchanged.

[0160] In this comparative example, after the nanofiltration membrane operates for 30 minutes, the flux drops from 1000 L / h to 250 L / h, showing a significant decrease and being unable to operate normally. It is necessary to stop the machine for cleaning, and the upstream liquid volume is greater than 3000 L.

[0161] In the above technical solution of the present invention, the above are only the preferred embodiments of the present invention, and do not limit the patent scope of the present invention accordingly. Any equivalent structural transformation made using the description of the present invention under the technical concept of the present invention, or direct / indirect application in other related technical fields, is included in the patent protection scope of the present invention.

Claims

1. A method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride using Berberis soulieana Schneid., characterized in that, Including the steps: S1, obtaining Berberis powder containing berberine, berbamine and jatrorrhizine; S2, extracting the Berberis powder to obtain an extraction solution; S3, obtaining a clear solution after primary filtration and microfiltration of the extraction solution; Passing the clear solution through a high-temperature ultrafiltration membrane and collecting the downstream solution of the membrane; Passing the downstream solution of the membrane through a high-temperature nanofiltration membrane and collecting the upstream solution of the membrane as a membrane-refined solution; S4, adjusting the pH of the membrane-refined solution to 6-7, filtering, and then adjusting the pH to 2-3 with a hydrochloric acid solution; Filtering after standing, and collecting the first wet precipitate and the first mother liquor; Adding an ethanol solution with a concentration of 60-65% to the first wet precipitate for heating and dissolution, filtering after standing, and collecting the second wet precipitate and the second mother liquor; concentrating the second mother liquor so that the ethanol concentration in the second mother liquor reaches 60-65%, filtering after standing, and collecting the third wet precipitate and the third mother liquor; combining the first mother liquor and the third mother liquor to obtain a berberine crystallization mother liquor; Combining the second wet precipitate and the third wet precipitate to obtain a wet precipitate containing berberine hydrochloride; S5, concentrating the berberine crystallization mother liquor to obtain a first concentrated solution; passing the first concentrated solution through an alkaline anion exchange resin column and collecting an ion exchange resin decolorized solution containing the feed effluent; Adjusting the pH of the ion exchange resin decolorized solution to 6-7, concentrating to obtain a second concentrated solution; after standing the second concentrated solution for at least 4 h, adjusting the pH to 8-9 to obtain a third concentrated solution; Filtering after standing the third concentrated solution to obtain a berbamine crystallization mother liquor and a wet precipitate containing berbamine; The step S5 further includes: after passing the first concentrated solution through the alkaline anion exchange resin column, washing the column with purified water, collecting the washing solution of the column, and combining the feed effluent and the washing solution of the column as the ion exchange resin decolorized solution; S6, passing the berbamine crystallization mother liquor through a macroporous adsorption resin column, then desorbing, and collecting the desorbed solution; concentrating the desorbed solution and adjusting the pH to 4-6 with a hydrochloric acid solution to obtain a refined solution containing jatrorrhizine hydrochloride; In the step S6, desorbing with an ethanol solution with a concentration of 40-45%; the process of concentrating the desorbed solution includes: concentrating the desorbed solution to 17-25 Baumé; The step S6 further includes: after passing the berbamine crystallization mother liquor through the macroporous adsorption resin column, washing the column with purified water before performing the desorption.

2. The method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride using Berberis soulieana Schneid., as claimed in claim 1, wherein, The Berberis powder uses Berberis as the raw material; by mass ratio, the content of berbamine in the Berberis powder is 0.5-2.0%; by mass ratio and in the form of hydrochloride, the content of berberine hydrochloride in the Berberis powder is 1.0-2.5%, and the content of jatrorrhizine hydrochloride is 0.4-0.9%.

3. The method for producing berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride using Berberis soulieana Schneid., as claimed in claim 1, wherein In the step S2, the Berberis powder is extracted with a sulfuric acid solution containing an antioxidant protector; the concentration of the sulfuric acid solution is 0.2-0.7%, the temperature of the extraction is 50-65°C, and the total duration of the extraction is not more than 8 h.

4. The method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride using Berberis julianae Schneid., as claimed in claim 1, wherein In the step S3, control the temperature of the extraction liquid to be 60 - 65°C, control the temperature of the clear liquid to be 60 - 65°C, and control the temperature of the liquid downstream of the membrane to be 60 - 65°C; before the clear liquid passes through the high-temperature ultrafiltration membrane, adjust the pH of the clear liquid to 1 - 4; the membrane pore size used for the microfiltration is 200 - 800 nm; the maximum temperature that the high-temperature ultrafiltration membrane can withstand is not lower than 70°C; the molecular weight cut-off of the high-temperature ultrafiltration membrane is 3 - 8 KD; the maximum temperature that the high-temperature nanofiltration membrane can withstand is not lower than 70°C; the molecular weight cut-off of the high-temperature nanofiltration membrane is 150 - 300 D.

5. The method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride using Berberis soulieana Schneid., as claimed in claim 1, wherein In the step S4, add an ethanol solution with a concentration of 60 - 65% to the first wet precipitate at a ratio of 1 g:12 - 15 mL. The process of concentrating the second mother liquor includes: concentrating the second mother liquor to 1 / 5 - 1 / 4 of its original volume.

6. The method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride using Berberis soulieana Schneid., as claimed in claim 1, wherein In the step S5, the process of concentrating the berberine crystallization mother liquor includes: concentrating the berberine crystallization mother liquor until there is no alcohol smell, and controlling the Brix of the berberine crystallization mother liquor to be 1 - 3 Brix.

7. The method for producing berberine hydrochloride, berbamine and jatrorrhizine hydrochloride using Berberis soulieana Schneid. as claimed in claim 1, wherein In the step S5, the Brix of the second concentrated liquid is 3 - 8 Brix.

8. The method for producing berberine hydrochloride, berbamine, and jatrorrhizine hydrochloride using Berberis soulieana Schneid. as claimed in any one of claims 1-7, characterized in that, Dry the wet precipitate containing berberine hydrochloride to obtain a berberine hydrochloride product; dry the wet precipitate containing berbamine to obtain a berbamine product; dry the refined liquid containing jatrorrhizine hydrochloride to obtain a jatrorrhizine hydrochloride product; the berberine hydrochloride product is bright yellow, the berbamine product is white, and the jatrorrhizine hydrochloride product is red-yellow.