CN110882246B

CN110882246B - Extraction method and application of coptis alkaloid with different biological activities

Info

Publication number: CN110882246B
Application number: CN201811046724.4A
Authority: CN
Inventors: 叶小利; 李学刚; 李滟临; 李亚松
Original assignee: Southwest University
Current assignee: Southwest University
Priority date: 2018-09-08
Filing date: 2018-09-08
Publication date: 2022-12-23
Anticipated expiration: 2038-09-08
Also published as: CN110882246A

Abstract

The invention belongs to the technical field of extraction of active ingredients of traditional Chinese medicines, and particularly relates to an extraction method and application of coptis alkaloids with different biological activities, wherein the extraction method comprises the following steps: (1) coarse extraction: pretreating coptis chinensis, soaking in sulfuric acid, percolating, neutralizing percolate, concentrating and filtering to obtain refined coptis chinensis extract concentrated solution; and (2) extracting active alkaloid I: adding concentrated hydrochloric acid and sodium chloride into the concentrated solution of Coptidis rhizoma, precipitating, and performing solid-liquid separation to obtain solid as active alkaloid I and liquid as mother liquor I; (3) extracting active alkaloid II: extracting the mother liquor I by using an organic solvent, and concentrating the extracted organic phase to obtain active alkaloid II, wherein the extracted water phase is the mother liquor II; (4) extracting active alkaloid III: extracting the mother liquor II by using an organic solvent, and concentrating the extracted organic phase to obtain active alkaloid III; the extraction method can obtain three kinds of Coptidis rhizoma alkaloids with different biological activities, and improve the comprehensive utilization rate of Coptidis rhizoma.

Description

Extraction method and application of coptis alkaloid with different biological activities

Technical Field

The invention belongs to the technical field of extraction of active ingredients of traditional Chinese medicines, and particularly relates to an extraction method and application of coptis alkaloids with different biological activities.

Background

Huang Lian was recorded in Shen nong Ben Cao Jing, listed as the superior. Coptis root, rhizoma Coptidis is bitter in taste and cold in nature, and has the effects of purging pathogenic fire, removing toxic substance, clearing heat, and eliminating dampness. Can be used for treating dysphoria, coma, vexation, insomnia, damp-heat, abdominal distention, emesis, abdominal pain, dysentery, conjunctival congestion, toxic swelling, aphtha, eczema, scald, hematemesis, and epistaxis. With the improvement of modern extraction technology, the development of the extraction technology of the coptis alkaloid is greatly improved.

Patent CN1730480A discloses a method for extracting total alkaloids from coptis, which mainly comprises the following steps: decocting Coptidis rhizoma with water, purifying with macroporous resin, washing with water, eluting with aqueous ethanol, recovering ethanol, concentrating, and drying to obtain the final product. The content of the total alkaloids of the coptis reaches 25 to 100 percent.

Patent CN101269132A discloses a process for extracting total alkaloids of coptis, which mainly comprises: crushing raw materials (coptis chinensis or coptis chinensis fibrous roots or coptis chinensis leaves) (if the coptis chinensis fibrous roots or the coptis chinensis leaves do not need to be crushed), adding a proper amount of 0.01-10% of sulfuric acid for soaking, heating and extracting for four times, pre-concentrating, purifying, precipitating sodium chloride and high-valence metal salt, filtering, drying and obtaining the coptis chinensis total alkaloid product. The main active ingredients of the product are berberine, coptisine, jateorhizine, palmatine, epiberberine and other rhizoma coptidis effective ingredients, the content of the rhizoma coptidis total alkaloids is more than 20%, wherein the content of the berberine is more than 10%. The rhizoma Coptidis total alkaloid produced by this method has low content (25%), and low total alkaloid yield.

The patent CN103393780A discloses a method for extracting high-purity coptis total alkaloid, which increases the content of coptis total alkaloid to 50 percent and comprises the specific processes of raw material (coptis root or coptis fibrous root or coptis ash), crushing (for example, coptis fibrous root or coptis ash does not need to be crushed), adding 0.1 to 5 percent of sulfuric acid for soaking, heating and extracting for four times, filtering, calcium oxide (or calcium hydroxide) for neutralization, filtering and washing for precipitation, preconcentration, acid regulation, sodium chloride precipitation, filtering, mother liquor recovery of alkaloid, precipitation and drying, and coptis total alkaloid product.

Patent CN104644793A discloses a method for efficiently extracting total alkaloids of coptis, which improves the extraction rate of total alkaloids to 98%, and the method comprises the following specific process steps: 1) Crushing the raw materials; 2) Pretreating raw materials; 3) Percolating; 4) Neutralizing percolate; 5) Concentrating; 6) Salting out; 7) Filtering and washing; 8) Treating mother liquor of total alkaloid; 9) And (5) drying. The method has simple operation and low energy consumption, greatly reduces the cost of industrial production, and has the extraction rate of the total alkaloids of the coptis root of more than 98 percent; the main active components of the rhizoma coptidis total alkaloids are berberine, namely berberine, epiberberine, palmatine, jateorhizine and other rhizoma coptidis total alkaloids.

The extraction process disclosed above has improved the extraction rate and content of coptis total alkaloids to a high level, but the above extraction methods are directed to coptis total alkaloids, and if alkaloids with certain biological activity are studied or used, the alkaloids need to be further separated from the alkaloids by using costly chromatographic separation techniques. Modern pharmaceutical research results show that the main active ingredients of the coptis chinensis are coptis chinensis alkaloids which mainly comprise berberine (berberine), palmatine, coptisine, epiberberine, african tetrandrine, jateorhizine, magnoline and the like. Different coptis alkaloids have different biological activities. In addition, the coptis chinensis is a valuable traditional Chinese medicine, if the coptis chinensis alkaloid can be extracted respectively according to the biological activity, the comprehensive medicinal value of the coptis chinensis can be obviously improved, and the cost of a medicinal preparation taking the coptis chinensis as a raw material can be reduced.

Disclosure of Invention

In order to solve the problems, the invention provides an extraction method of coptis alkaloids with different biological activities, which can obtain three coptis alkaloids with different biological activities and improve the comprehensive utilization rate of coptis.

The extraction method of the coptis alkaloid with different biological activities comprises the following steps:

(1) Coarse extraction: pretreating coptis chinensis, soaking in sulfuric acid, percolating, neutralizing percolate, concentrating and filtering to obtain refined coptis chinensis extract concentrated solution;

(2) Extraction of active alkaloid I: adding concentrated hydrochloric acid into the coptis chinensis extraction concentrated solution, wherein the volume ratio of the concentrated hydrochloric acid to the coptis chinensis extraction concentrated solution is 0.5-1%, adding sodium chloride accounting for 5-10% of the weight of the coptis chinensis extraction concentrated solution, precipitating for 0.1-30 days, and performing solid-liquid separation to obtain a solid active alkaloid I and a liquid mother solution I;

(3) Extraction of active alkaloid II: extracting the mother liquor I by using ethyl acetate, dichloromethane, trichloromethane or/and diethyl ether, and concentrating the extracted organic phase to obtain active alkaloid II, wherein the extracted water phase is the mother liquor II;

(4) Extraction of active alkaloid III: extracting the mother liquor II by using ethyl acetate-butanol or butanol, and concentrating the extracted organic phase to obtain the active alkaloid III.

Preferably, the concentration of the coptis chinensis extract concentrated solution is 0.1-5 g crude drug (raw medicinal material)/mL solution.

Preferably, in the steps (3) and (4), the extraction refers to 2 times of extraction with an organic solvent with the same volume as the mother liquor I or II.

Preferably, in the step (2), the volume ratio of the added concentrated hydrochloric acid to the coptis chinensis extract concentrated solution is 0.5%.

Preferably, in the step (2), the weight of the added sodium chloride is 5% of the weight of the coptis concentrated extract.

Preferably, the extraction method further comprises silica gel column chromatography and recrystallization of the active alkaloid II and the active alkaloid III to obtain a refined active alkaloid II and a refined active alkaloid III, wherein the eluent of the silica gel column chromatography is methanol, ethanol or acetone.

Preferably, the solvent for recrystallization is methanol.

Preferably, in the step (1), the coptis chinensis pretreatment refers to slicing or crushing coptis chinensis, coptis chinensis fibrous roots, coptis chinensis ash or coptis chinensis stems and leaves.

Preferably, in the step (1), the sulfuric acid soaking refers to submerging and soaking for 24 hours by using 0.5% sulfuric acid (v/v).

Preferably, the percolate neutralization means that the collected percolate is neutralized by lime or calcium hydroxide until the pH value is 2-10.

The active alkaloid I, the active alkaloid II refined product, the active alkaloid III or the active alkaloid III refined product extracted by the extraction method also belongs to the protection scope of the invention.

Preferably, the active alkaloid I mainly contains berberine and coptisine, the active alkaloid II mainly contains magnoline, and the active alkaloid III mainly contains epiberberine and palmatine.

The application of the extraction method and/or the active alkaloid I extracted by the extraction method in preparing a medicament for treating diabetes, hyperlipidemia, obesity or cardiovascular diseases also belongs to the protection scope of the invention.

The application of the extraction method and the active alkaloid II or the active alkaloid II refined product extracted by the extraction method in preparing the medicine for treating eczema also belongs to the protection scope of the invention.

The application of the extraction method and the active alkaloid III or the active alkaloid III refined product extracted by the extraction method in the preparation of the medicine for treating acne also belongs to the protection scope of the invention.

The invention has the beneficial effects that:

1. the extraction method provided by the invention directly divides the coptis alkaloid into three active alkaloids I, II and III with different biological activities at the extraction stage, and the total alkaloids of the coptis obtained by combining salting-out products and mother liquor do not need to be separated again, so that the separation and extraction difficulty is reduced, meanwhile, the extraction method not only can obtain berberine and coptisine with higher purity, but also can obtain magnoline and palmatine with lower content of coptis, and is beneficial to further preparation of medicaments;

2. the three active alkaloids obtained by the extraction method have excellent biological activity, wherein the active alkaloid I mainly contains berberine and coptisine, has more ideal effects of reducing blood sugar and blood fat and losing weight compared with single berberine, can be used as substitutes of products such as coptis chinensis, coptis chinensis concentrated products, berberine and the like, and is a high-quality raw material for treating diabetes, hyperlipidemia, obesity, cardiovascular diseases and resisting bacteria; the refined active alkaloid II mainly contains magnoline, the purity is as high as 95%, and compared with coptis chinensis and berberine, the refined active alkaloid II can be used for treating eczema more effectively; the active alkaloid III mainly contains the palmatine and epiberberine with the content of 90 percent, the further refined palmatine and epiberberine have the purities of more than 90 percent, and compared with berberine and coptisine, the refined palmatine can more effectively treat acne.

3. The extraction method provided by the invention realizes the comprehensive utilization of the coptis alkaloid, all the alkaloids in the coptis are respectively utilized, no waste is generated, and the full utilization of the coptis resource is realized.

Drawings

FIG. 1 is an HPLC chromatogram of the concentrated solution of Coptidis rhizoma prepared in example 1;

FIG. 2 is an HPLC chromatogram of active alkaloid I extracted in example 1;

FIG. 3 is an HPLC profile of the active alkaloid II extracted in example 1;

FIG. 4 is an HPLC chromatogram of a refined active alkaloid II extracted in example 1;

FIG. 5 is an HPLC profile of the active alkaloid III extracted in example 1;

FIG. 6 is an HPLC profile of the active alkaloid III (palmatine site) extracted in example 1;

FIG. 7 is an HPLC chromatogram of a refined product of active alkaloid III (palmatine site) extracted in example 1;

FIG. 8 is an HPLC chromatogram of active alkaloid III (epiberberine site) extracted in example 1;

FIG. 9 is an HPLC chromatogram of a refined product of active alkaloid III (epiberberine fraction) extracted in example 1.

Detailed Description

In the following, the technical solutions in the embodiments of the present invention will be clearly and completely described, and it is obvious that the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.

In addition, unless otherwise specifically indicated, various starting materials, reagents, instruments and equipment used in the present invention may be commercially available or prepared by existing methods.

Example 1 extraction of active alkaloids I, II, and III, respectively

Slicing 10kg of coptis, immersing for 24 hours in 0.5% sulfuric acid, percolating, collecting 10 liters of percolate every day, and continuously collecting for 5 days; neutralizing percolate with lime to pH 2, and filtering; washing the residue with appropriate amount of water for 1 time, and mixing the filtrate and washing solution; concentrating the combined neutralized solution under reduced pressure to obtain Coptidis rhizoma extract concentrated solution with concentration of 0.1g crude drug (raw material)/mL, cooling, and filtering; adding concentrated hydrochloric acid 0.5 vol% into concentrated extractive solution of Coptidis rhizoma, adding sodium chloride 5 wt% of the concentrated solution, precipitating total alkaloids, and standing for 0.1 day; naturally filtering the precipitate, washing with water, and oven drying to obtain active alkaloid I. Referring to fig. 2, the hplc analysis content shows that berberine content is 55%, coptisine content is 20%, palmatine content is 5%, epiberberine content is 3%, jateorhizine content is 2%, african tetrandrine content is 1%, and magnoline content is 0.5%.

Extracting the mother liquor after the active alkaloid I is extracted for 3 times by using ethyl acetate with the same volume, recovering the ethyl acetate, and drying to obtain the active alkaloid II. Referring to fig. 3, the content of magnoline is 12%, berberine is 1%, coptisine is 0.5%, palmatine is 1%, epiberberine is 1%, jateorhizine is 0.2%, and tetrandrine is 0.3% by hplc analysis. 10g of active alkaloid II, and stirring the active alkaloid II with 20g of silica gel; putting 30g of silica gel on the bottom of the silica gel column, then putting the raw materials and the silica gel on the upper part, and eluting with methanol; collecting magnoline part, and recovering methanol to obtain magnoline. Referring to fig. 4, the content of magnoline was 65% by hplc. Recrystallizing in methanol once to obtain pure magnoline (refined Coptidis rhizoma active alkaloid II). Referring to fig. 5, the content of magnoline was 95% by hplc analysis.

Extracting the mother liquor after the active alkaloid II is extracted, extracting for 3 times by using butanol with the same volume, recovering the butanol, and drying to obtain the active alkaloid III. Referring to fig. 6, the content of palmatine 15%, epiberberine 12%, berberine 1%, coptisine 1%, jateorhizine 2%, and african tetrandrine 0.8% by hplc analysis. 10g of active alkaloid III, and stirring the materials by using 20g of silica gel; putting 30g of silica gel on the bottom of the silica gel column, then putting the raw materials and the silica gel on the upper part, and eluting with methanol; collecting palmatine part and epiberberine part, respectively, and recovering methanol to obtain 2 components of palmatine and epiberberine. Referring to fig. 6 and 8, the content of palmatine was 55% and epiberberine was 50% by hplc analysis. Recrystallizing in methanol once to obtain pure palmatine and epiberberine. Referring to fig. 7 and 9, the content of palmatine was 90% and epiberberine was 92% by hplc analysis.

Example 2 extraction of active alkaloid I, II, and III, respectively

Soaking herba Scutellariae Barbatae 10kg in 0.5% sulfuric acid for 24 hr, percolating, collecting 10L percolate every day, and continuously collecting for 5 days; neutralizing percolate with lime to pH 10, and filtering; washing the residue with appropriate amount of water for 3 times, and mixing the filtrate and washing liquid; concentrating the combined neutralized solution under reduced pressure to concentration of 5g crude drug (raw material medicine)/mL to obtain Coptidis rhizoma extract concentrated solution, cooling, and filtering; adding concentrated hydrochloric acid 0.5 vol% into the concentrated solution, adding sodium chloride 5 wt% of the concentrated solution, precipitating total alkaloids, and standing for 30 days; naturally filtering the precipitate, washing with water, and oven drying to obtain active alkaloid I. HPLC analysis content, berberine content 30%, coptisine content 40%, palmatine 0.1%, epiberberine 5%, jateorhizine 1%, african tetrandrine 1.5%, and magnoline 0.2%.

Extracting the mother liquor after the active alkaloid I is extracted for 1 time by using dichloromethane with the same volume, recovering the dichloromethane, and drying to obtain the active alkaloid II. HPLC analysis content shows that magnoline accounts for 10%, berberine accounts for 0.8%, coptisine accounts for 1.7%, palmatine 0.1%, epiberberine 0.9%, jateorhizine 0.5%, and tetrandrine 0.4%.10g of active alkaloid II, and stirring the active alkaloid II with 20g of silica gel; putting 30g of silica gel on the bottom of the silica gel column, then putting the raw materials and the silica gel mixed on the upper part, and eluting with ethanol; collecting magnoline part, and recovering ethanol to obtain magnoline. The content of magnoline was 60% by HPLC analysis. Recrystallizing in methanol once to obtain pure magnoline. The content of magnoline was 90% by HPLC analysis.

Extracting the mother liquor after the active alkaloid II is extracted for 1 time by using ethyl acetate-butanol (1). HPLC analysis content, palmatine 1%, epiberberine 25%, berberine 1%, coptisine 1%, jateorhizine 2%, and African tetrandrine 0.8%.10g of active alkaloid III, and stirring the active alkaloid III with 20g of silica gel; putting 30g of silica gel on the bottom of the silica gel column, then putting the raw materials and the silica gel on the upper part, and eluting with ethanol; collecting 2 parts of the palmatine part and the epiberberine part respectively, and recovering ethanol to obtain palmatine and epiberberine. HPLC analysis shows that palmatine accounts for 50% and epiberberine accounts for 65%. Recrystallizing in methanol once to obtain pure palmatine and epiberberine. The content of palmatine is 90% by HPLC analysis, and the content of epiberberine is 95%.

EXAMPLE 3 extraction of active alkaloids I, II, and III, respectively

Crushing 10kg of coptis leaves, immersing for 24 hours in 0.5% sulfuric acid, percolating, and collecting 10 liters of percolate every day for 5 days continuously; neutralizing percolate with lime to pH 5, and filtering; washing the residue with appropriate amount of water for 2 times, and mixing the filtrate and washing solution; concentrating the combined neutralized solution under reduced pressure to concentration of 1g crude drug (raw material)/mL to obtain Coptidis rhizoma extract concentrated solution, cooling, and filtering; adding concentrated hydrochloric acid 0.5 vol%, adding sodium chloride 5 wt%, precipitating total alkaloids, and standing for 10 days; naturally filtering the precipitate, washing with water, and oven drying to obtain active alkaloid I.

HPLC analysis content, berberine content 50%, coptisine content 5%, palmatine 3%, epiberberine 5%, jateorhizine 1%, tetrandrine 0.5%, and magnoline 0.2%.

Extracting the mother liquor after the active alkaloid I is extracted for 2 times by using chloroform with the same volume, recovering the chloroform, and drying to obtain the active alkaloid II. HPLC analysis content shows that magnoline content is 15%, berberine content is 1.2%, coptisine content is 1%, palmatine 1%, epiberberine 2%, jateorhizine 0.8%, and African tetrandrine 0.4%.10g of active alkaloid II, and stirring the active alkaloid II with 20g of silica gel; putting 30g of silica gel on the bottom of the silica gel column, then putting the raw materials and the silica gel on the upper part, and eluting with isopropanol; collecting magnoline part, and recovering isopropanol to obtain magnoline. The content of magnoline was 67% by HPLC analysis. Recrystallizing in methanol once to obtain pure magnoline. The content of magnoline was 97% by HPLC analysis.

Extracting the mother liquor after the extraction of the active alkaloid II for 2 times by using ethyl acetate-butanol (1. HPLC analysis content, palmatine 10%, epiberberine 5%, berberine 1%, coptisine 1%, jateorhizine 1%, and African tetrandrine 0.8%.10g of active alkaloid III, and stirring the materials by using 20g of silica gel; putting 30g of silica gel on the bottom of the silica gel column, then putting the raw materials and the silica gel on the upper part, and eluting with isopropanol; collecting 2 parts of palmatine part and epiberberine part, respectively, and recovering isopropanol to obtain palmatine and epiberberine. The content of palmatine is 55% and the content of epiberberine is 60% by HPLC analysis. Recrystallizing in methanol once to obtain pure palmatine and epiberberine. Content analysis by HPLC showed that palmatine was 93% and epiberberine was 95%.

EXAMPLE 4 extraction of active alkaloid I, II, and III, respectively

Immersing 10kg of ash residue in 0.5% sulfuric acid for 24 hours, percolating, collecting 10 liters of percolating solution every day, and continuously collecting for 5 days; neutralizing percolate with lime to pH3, and filtering; washing the residue with appropriate amount of water for 2 times, and mixing the filtrate and washing solution; concentrating the combined neutralized solution under reduced pressure to obtain concentrated Coptidis rhizoma solution with concentration of 0.5g crude drug (raw material)/mL, cooling, and filtering; adding concentrated hydrochloric acid 0.5 vol%, adding sodium chloride 5 wt% of the concentrated solution, precipitating total alkaloids, and standing for 2 days; naturally filtering the precipitate, washing with water, and oven drying to obtain active alkaloid I. HPLC analysis content, berberine content 60%, coptisine content 25%, palmatine 5%, epiberberine 4%, jateorhizine 1%, african tetrandrine 0.5%, and magnoline 0.6%.

Extracting the mother liquor after extracting the active alkaloid I with equal volume of diethyl ether for 2 times, recovering diethyl ether, and drying to obtain active alkaloid II. HPLC analysis content shows that magnoline accounts for 11%, berberine accounts for 1.2%, coptisine accounts for 1%, palmatine 1%, epiberberine 1%, jateorhizine 0.8%, and african tetrandrine 0.4%.10g of active alkaloid II, and stirring the active alkaloid II with 20g of silica gel; putting 30g of silica gel on the bottom of the silica gel column, then putting the raw materials and the silica gel on the upper part, and eluting with acetone; collecting magnoline part, and recovering acetone to obtain magnoline. Content by HPLC analysis, magnoline was 56%. Recrystallizing in methanol once to obtain pure magnoline. The content was analyzed by HPLC and magnoline was 92%.

Extracting the mother liquor after extracting the active alkaloid II with butanol of the same volume for 2 times, recovering the solvent, and drying to obtain the active alkaloid III. HPLC analysis content, palmatine 8%, epiberberine 7%, berberine 1%, coptisine 1%, jateorhizine 0.9%, and African tetrandrine 0.8%.10g of active alkaloid III, and stirring the materials by using 20g of silica gel; putting 30g of silica gel on the bottom of the silica gel column, then putting the raw materials and the silica gel on the upper part, and eluting with acetone; collecting 2 parts of the palmatine part and the epiberberine part respectively, and recovering acetone to obtain palmatine and epiberberine. Content by HPLC analysis, palmatine was 57%, and epiberberine was 62%. Recrystallizing in methanol once to obtain pure palmatine and epiberberine. Content analysis by HPLC showed that palmatine was 93% and epiberberine was 94%.

EXAMPLE 5 comparative experiments on the treatment of related diseases with active alkaloid I obtained by the present invention

(1) And hypoglycemic Activity test

The test materials are active alkaloid I (the sum of alkaloids such as berberine, coptisine, palmatine and epiberberine is 95%), berberine (content 95%) and coptis (total alkaloid content 10%) prepared in the embodiment 4 of the invention.

The blood sugar reducing experimental method is carried out according to the research guiding principle of natural medicines (traditional Chinese medicines) and new medicines: KK-ay mice were divided into 4 groups: control group, active alkaloid group I, berberine group, and rhizoma Coptidis group. Active alkaloid group I (500 mg/kg), berberine group (500 mg/kg), and Coptidis rhizoma group medicinal group (5000 mg/kg); after the mice are raised for 4 weeks, the mice are continuously fed for 4 weeks until the blood sugar is higher than 10, and then the fasting blood is collected to detect the blood fat index, and the result is shown in table 1.

TABLE 1 comparison of the hypoglycemic Effect

Group of	Before administration (mmol/L)	After administration(mmol/L)
			Control group	11.51±2.20	11.83±2.40
Active alkaloid I (500 mg/kg)	11.61±2.51	6.61±1.70
			Berberine (500 mg/kg)	11.70±2.40	7.45±2.10
Coptis (5000 mg/kg)	11.59±2.30	7.05±2.11 ^×

As can be seen from Table 1, the active alkaloid I has the best hypoglycemic activity, and is superior to the berberine group due to the same dosage of rhizoma Coptidis. The active alkaloid I obtained by the invention has better hypoglycemic activity.

(2) Blood lipid lowering activity test

The test materials are active alkaloid I (the sum of alkaloids such as berberine, coptisine, palmatine and epiberberine is 95%), berberine (content 95%) and coptis (total alkaloid content 10%) prepared in example 4.

The blood fat reducing experimental method is carried out according to the research guiding principle of natural medicines (traditional Chinese medicines) and new medicines: the golden hamsters were divided into 5 groups, one group (10) given normal diet, and one group (40) given high fat diet; after 4 weeks, the blood lipid index was measured, and then the golden hamster that was successfully molded was divided into 4 groups (8 to 10 mice per group): high fat control group, rhizoma coptidis total alkaloid group, berberine group and rhizoma coptidis total alkaloid crude product group. The drug group is administered with 500mg/Kg of drug; the feeding is continuously carried out for 4 weeks, and then the fasting blood is collected to detect the blood fat index, and the result is shown in the table 2.

TABLE 2 comparison of hypolipidemic effects

As can be seen from Table 2, compared with the normal control group, the blood lipid of each test group is increased, which indicates that the molding is successful; compared with the high-fat group, the Cholesterol (TC) of each drug group is reduced, which shows that the drug group has the activity of reducing blood fat; wherein the blood lipid reducing activity of the active alkaloid I is superior to that of berberine and is also superior to that of coptis chinensis.

(3) And antibacterial activity test

The antibacterial experimental method is carried out according to the research guiding principle of natural medicines (traditional Chinese medicines) and new medicines: coli as a bacterial model, and the activity of Qikangershire was studied by detecting MIC: after the bacteria were cultured, the concentration of the bacteria was adjusted to 1X 10 ⁸ Per mL; 50 microliters of bacteria were not added to the 96-well plate, and then drugs with different concentrations were added, and the bacteria were cultured for 24 hours, and the growth of the bacteria was observed, and the MIC was determined, and the results are shown in Table 3.

TABLE 3 comparison of antibacterial (E.coli) Effect (μ g/mL)

Active alkaloid I	250
		Berberine	500
Coptis chinensis	≥2000

As can be seen from Table 3, the antibacterial activity of active alkaloid I is the highest, followed by berberine, and the antibacterial activity of Coptidis rhizoma is the lowest.

EXAMPLE 6 comparison of Effect of active alkaloid II on eczema treatment

The test materials were refined active alkaloid II (magnoline 90%), berberine (content 95%), and Coptidis rhizoma (total alkaloid content 10%) obtained in example 2.

The antibacterial experimental method is carried out according to the research guiding principle of natural medicines (traditional Chinese medicines) and new medicines: 40 mice, 10 controls, active alkaloid II (10), berberine (10), coptis (10). Mice had hair removed, and the control group had only hair removed, and did not cause sensitization and excitation; the drug groups were sensitized at the shaved area with Dinitrochlorobenzene (DNCB) acetone solution (acetone: glycerol = 3), mice were stimulated on the back with 2%,200ul continuously for 3 days, after which the back was induced every other day with 0.5% DNCB, 40ul;

0.5% DNCB,20ul excited the left ear. After the molding is successful, the molding part is coated with a medicament (the concentration of the medicament is 5%), and the times of the mouse licking the wound are observed. The results are shown in Table 4 below.

TABLE 4 comparison of the effects on eczema treatment (5 min/second)

Model set	10.5±3.4
		Refined active alkaloid II	2.2±1.4
Berberine	6.5±2.2
		Coptis chinensis	5.4±3.1

As can be seen from Table 4, all three drugs have the effect of treating eczema, but the activity of the refined active alkaloid II for treating eczema is the highest and is obviously higher than that of berberine and coptis chinensis; the second is coptis root, and the activity of berberine is the lowest.

EXAMPLE 7 comparison of the Effect of active alkaloid III on the treatment of acne (against Propionibacterium acnes)

The test materials were the refined active alkaloid III (palmatine 90%), berberine (95%), and Coptidis rhizoma (total alkaloid 10%) obtained in example 2.

Propionibacterium acnes is the primary pathogen that causes acne. The antibacterial experimental method is carried out according to the research guiding principle of natural medicines (traditional Chinese medicines) and new medicines: taking propionibacterium acnes as a bacterial model, and researching the activity of a fine active alkaloid III by detecting MIC: after the bacteria were cultured, the concentration of the bacteria was adjusted to 1X 10 ⁸ Per mL; 50 microliters of bacteria were added to a 96-well plate without being empty, then drugs of different concentrations were added, the culture was carried out for 24 hours, the growth conditions of the bacteria were observed, and the MIC thereof was judged, with the results shown in Table 5.

TABLE 5 comparison of antibacterial (Propionibacterium acnes) effectiveness (μ g/mL)

Refined product of active alkaloid III	250
		Berberine	500
Coptis chinensis	≥2000

As can be seen from Table 5, the antibacterial activity of active alkaloid I is highest, followed by berberine and the antibacterial activity of Coptidis rhizoma is lowest.

Claims

1. The extraction method of the coptis alkaloid with different biological activities is characterized by comprising the following steps:

(1) Coarse extraction: pretreating coptis chinensis, soaking in sulfuric acid, percolating, neutralizing percolate, concentrating, and filtering to obtain refined coptis chinensis extract concentrated solution;

(2) Extraction of active alkaloid I: adding concentrated hydrochloric acid into the coptis chinensis extraction concentrated solution, wherein the volume ratio of the concentrated hydrochloric acid to the coptis chinensis extraction concentrated solution is 0.5-1%, adding sodium chloride accounting for 5-10% of the weight of the coptis chinensis extraction concentrated solution, precipitating for 0.1-30 days, and carrying out solid-liquid separation to obtain a solid, namely an active alkaloid I, and a liquid, namely a mother solution I, wherein the active alkaloid I mainly contains berberine and coptisine;

(3) Extraction of active alkaloid II: extracting the mother liquor I by using ethyl acetate, dichloromethane, trichloromethane or/and diethyl ether, and concentrating the extracted organic phase to obtain active alkaloid II, wherein the extracted water phase is the mother liquor II, and the active alkaloid II mainly contains magnoline;

(4) Extraction of active alkaloid III: extracting the mother liquor II by adopting ethyl acetate-butanol or butanol, and concentrating the extracted organic phase to obtain active alkaloid III, wherein the active alkaloid III mainly contains epiberberine and palmatine.

2. The extraction method according to claim 1, wherein the concentration of the concentrated solution of Coptidis rhizoma is 0.1-5 g crude drug/mL.

3. The extraction method according to claim 2, wherein in the steps (3) and (4), the extraction is 2 times of extraction with an organic solvent having the same volume as the mother liquor I or II.

4. The extraction method according to claim 2, wherein the volume ratio of the concentrated hydrochloric acid to the coptis chinensis extract concentrate in the step (2) is 0.5%, and the weight of the sodium chloride added in the step (2) is 5% of the weight of the coptis chinensis extract concentrate.

5. The extraction method as claimed in claim 1, further comprising silica gel column chromatography and recrystallization of the active alkaloid II and the active alkaloid III to obtain a refined product of the active alkaloid II and a refined product of the active alkaloid III, wherein the eluent of the silica gel column chromatography is methanol, ethanol or acetone, and the solvent for recrystallization is methanol.

6. The extraction method according to claim 2, wherein in the step (1), the coptis chinensis pretreatment is to slice or crush coptis chinensis, coptis chinensis fibrous roots, coptis chinensis ash or coptis chinensis stems and leaves, in the step (1), the sulfuric acid soaking is to submerge and soak the coptis chinensis, i.e. 0.5% sulfuric acid (v/v) is used for 24 hours, and the percolate neutralization is to neutralize the collected percolate with lime or calcium hydroxide until the pH value is 2-10.

7. Use of the active alkaloid II extracted by the extraction method of any one of claims 1 to 6 in the preparation of a medicament for treating eczema.