CN113214070B

CN113214070B - Preparation method of aurantio-obtusin and application of aurantio-obtusin in health-care product for relaxing bowels

Info

Publication number: CN113214070B
Application number: CN202110521022.2A
Authority: CN
Inventors: 赵一弘; 雷雨
Original assignee: Beijing Outsell Health Product Development Co ltd
Current assignee: Beijing Outsell Health Product Development Co ltd
Priority date: 2021-05-13
Filing date: 2021-05-13
Publication date: 2022-03-11
Anticipated expiration: 2041-05-13
Also published as: CN113214070A

Abstract

The invention discloses a preparation method of aurantio-obtusin and application of the aurantio-obtusin in a health-care product for relaxing bowels, wherein the preparation method of the aurantio-obtusin comprises the following steps: (1) mixing the coarse powder of the cassia seeds with a solvent, and performing microwave-assisted treatment to obtain a pretreatment solution; (2) heating, refluxing, cooling and filtering the pretreatment liquid obtained in the step (1) to obtain a mixed liquid; (3) carrying out silica gel column chromatography on the mixed solution obtained in the step (2), eluting, and collecting eluent; (4) and (4) concentrating the eluent obtained in the step (3), and recrystallizing to obtain the aurantio-obtusin. The preparation method of the invention obviously improves the yield and purity of the aurantio-obtusin. Compared with the prior art, the laxative health-care product disclosed by the invention has the advantages that the consumption of the senna leaves and the cassia seeds is obviously reduced by compounding the orange cassia tora essence with the senna leaves, the poria cocos, the radix glehniae, the Chinese yam and the like, the effect of relaxing bowel is obvious, and the safety is higher.

Description

Preparation method of aurantio-obtusin and application of aurantio-obtusin in health-care product for relaxing bowels

Technical Field

The invention belongs to the technical field of extraction of active ingredients of traditional Chinese medicines, and particularly relates to a preparation method of aurantio-obtusin and application of the aurantio-obtusin in a health-care product for relaxing bowels.

Background

Cassia seed, originally recorded in Shennong Ben Cao Jing, is a dry mature seed of Cassia obtusifolia Cass ia Obtus ifolia L. or Cassia tora siata a L. of leguminous plants, and has cold, sweet and bitter taste, and enters liver and large intestine meridians. Modern pharmacological research shows that cassia seed has pharmacological activities of reducing blood pressure, reducing blood fat, inhibiting bacteria, relieving asthma, benefiting gallbladder, protecting liver, resisting oxidation, resisting atherosclerosis, enhancing immunity and the like. It contains anthraquinones such as chrysophanol, emodin, rhein, aurantio-obtusin, and obtusin.

The aurantio-obtusin is a yellow acerola anthraquinone compound, is a special component in cassia seed, and has the effects of reducing blood fat, estrogen and antiestrogen activity, reducing blood sugar, and inducing platelet aggregation by adenosine diphosphate (A DP), Arachidonic Acid (AA), collagen (co lla-gen), and the like.

At present, most researches on the extraction process of the cassia seeds take emodin and chrysophanol as indexes, and reports taking the content of orange cassia tora element as an index are hardly seen.

CN111228325A discloses a method for extracting the whole components of cassia seed by fusing supercritical CO₂The extraction technology and the high-pressure-difference low-temperature continuous extraction technology adopt two extraction modes, all effective components in the cassia seeds are respectively extracted and converted into finished products to the greatest extent, the added value of the products is improved, the use amount of organic solvents is reduced, meanwhile, the high extraction rate of various effective components is ensured, the pollution is reduced, and the extraction rates of total flavonoids, polysaccharides, chrysophanol, aurantiamarin, cassia seed glycoside C and cassia seed glycoside B2 are all up to more than 65%. However, the extraction method of the present invention uses expensive equipment and has high preparation cost.

CN101967090A discloses an extraction process of aurantio-obtusin, which comprises the steps of crushing cleaned cassia seeds into coarse powder, leaching in solvent gasoline, discarding the leaching solution, adding residues into an alcohol solvent for ultrasonic extraction, concentrating the extracting solution, performing Soxhlet reflux extraction by using ethyl acetate, concentrating the ethyl acetate extracting solution, passing the ethyl acetate extracting solution through a polyamide chromatographic column, eluting by using petroleum ether-acetone and acetone-methanol, recovering the solvent from the eluent at low temperature under reduced pressure to obtain a crude product, and recrystallizing the crude product by using N-dimethylformamide/acetone to obtain the aurantio-obtusin. The method has simple extraction equipment, the purity of the obtained aurantio-obtusin is between 98.27 and 98.62 percent, but the method has low yield, long extraction time and great harm to the environment and human bodies, and is not suitable for industrial production.

Zhang Jing reports mixing semen Cassiae powder with 15% sulfuric acid solution, heating in water bath, separating hydrolyzed substances with filter paper and a funnel, distilling with distilled water to obtain anthraquinone aglycone solution, heating and refluxing at 70 ℃ with ethanol, cooling, correcting lost weight with ethanol, separating the mixed solution with a silica gel chromatographic column, collecting the liquid passing through the chromatographic column into a conical flask, eluting with eluent of petroleum ether and acetone at a ratio of 9:1 to obtain aurantio-obtusin, and recrystallizing the separated aurantio-obtusin for many times with an evaporating dish and acetone to obtain aurantio-obtusin with a purity of 99% (research and analysis on separation and extraction of the aurantio-obtusin, China and foreign medicine research, volume 10, and 20 th period). The invention provides a technology for obtaining high-purity aurantio-obtusin to a certain extent, but still has the problem that the yield and the purity of aurantio-obtusin can not be improved simultaneously.

The modern society has fast pace of life and high working pressure, and a lot of people are in sub-health state. The sub-healthy people are most susceptible to gastrointestinal discomfort, constipation, obesity, insomnia, hypertension, hyperlipidemia, cerebrovascular diseases and hypoimmunity. Constipation is a complex symptom common in the clinic and usually manifests as: less defecation and less defecation times; difficult and laborious defecation; the defecation is not smooth; dry stool, hard stool and unclean defecation. The long-term constipation can be accompanied by symptoms of abdominal distension, abdominal pain, bloody stool and the like, and easily induces the attack of cardiovascular and cerebrovascular diseases and serious hemorrhoids, anal fissure, rectocele and other diseases.

In recent years, with the more and more obvious pharmacological action and physiological activity of aurantio-obtusin, the more and more effect of aurantio-obtusin on the aspect of preventing and treating diseases by traditional Chinese medicines is achieved, so that the development of a preparation method of aurantio-obtusin capable of solving the technical problems and the application of aurantio-obtusin in health products for relaxing bowels are very necessary.

Disclosure of Invention

In order to solve the problems, the invention provides a preparation method of the aurantio-obtusin, which has high yield and purity, simple preparation process and low cost, and application of the aurantio-obtusin prepared by the preparation method in health-care products for relaxing bowels.

In order to achieve the purpose, the invention adopts the following technical scheme:

a preparation method of aurantio-obtusin comprises the following steps:

(1) mixing the coarse powder of the cassia seeds with a solvent, and performing microwave-assisted treatment to obtain a pretreatment solution;

(2) heating, refluxing, cooling and filtering the pretreatment liquid obtained in the step (1) to obtain a mixed liquid;

(3) carrying out silica gel column chromatography on the mixed solution obtained in the step (2), eluting, and collecting eluent;

(4) and (4) concentrating the eluent obtained in the step (3), and recrystallizing to obtain the aurantio-obtusin.

Preferably, the particle size of the cassia seed coarse powder in the step (1) is 0.05-0.25 mm.

Preferably, the solvent in the step (1) is ethanol water solution and n-butanol in a volume ratio of 8-10: 1.

Further preferably, the volume concentration of the ethanol water solution is 50-70%.

Preferably, the mass ratio of the cassia seed coarse powder to the solvent in the step (1) is 1: 10-15.

Preferably, the microwave-assisted treatment in step (1) is: the microwave power is 300-400W, and the microwave time is 20-40 s.

Preferably, the heating reflux in step (2) is: reflux extracting at 60-70 deg.C for 15-30 min.

Preferably, the eluent in the step (2) is a mixed solution of petroleum ether and acetone.

Further preferably, the mass ratio of the petroleum ether to the acetone is 6.5:1-8.5: 1.

The invention also provides the orange cassia seed element prepared by the preparation method.

The invention also provides application of the aurantio-obtusin prepared by the preparation method in preparation of health-care products for relaxing bowels.

The invention also provides a health-care product for relaxing bowels, which comprises the orange cassia essence prepared by the preparation method.

Preferably, the laxative health-care product comprises 10-15 parts of senna leaves, 8-15 parts of cassia seeds, 5-15 parts of black tea, 5-15 parts of green tea, 3-12 parts of poria cocos, 3-12 parts of radix glehniae, 20-30 parts of Chinese yam and 5-10 parts of honey.

The preparation method of the laxative health-care product comprises the following steps:

s1, respectively pulverizing folium sennae, black tea, green tea, Poria, radix Glehniae and rhizoma Dioscoreae, sieving with 10-30 mesh sieve, and mixing to obtain mixed powder;

s2, pretreating semen cassiae according to the preparation method to obtain aurantio-obtusin;

s3, mixing the aurantio-obtusin obtained in the step S2 with honey, adding the mixed powder obtained in the step S1, and uniformly mixing to obtain a wet material;

s4, performing irradiation sterilization on the wet material obtained in the step S3, and granulating to obtain the laxative health-care product.

Compared with the prior art, the invention has the beneficial effects that:

(1) the invention carries out auxiliary treatment on the alcohol solution of the cassia seed coarse powder by microwave, then carries out heating reflux, utilizes silica gel column and recrystallization to separate and purify the aurantio-obtusin, and obviously improves the yield and the purity of the aurantio-obtusin by optimizing the parameters of the particle size of the cassia seed coarse powder, the concentration and the dosage of the alcohol solution, the microwave condition, the composition of an eluant and the like.

(2) The preparation method is simple, has low requirements on equipment, short preparation time and low production cost, and simultaneously has less reagent dosage and is beneficial to environmental protection.

(3) Compared with the prior art, the laxative health-care product disclosed by the invention has the advantages that the consumption of the senna leaves and the cassia seeds is obviously reduced by compounding the orange cassia tora essence with the senna leaves, the poria cocos, the radix glehniae, the Chinese yam and the like, the effect of relaxing bowel is obvious, and the safety is higher.

Detailed Description

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.

Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.

When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

Unless otherwise stated, the raw materials used in the present invention are all common commercial products, the diameter-height ratio of the chromatography column in this example is 1:5, and the silica gel is 300 mesh.

Example 1

A preparation method of aurantio-obtusin comprises the following steps:

(1) mixing the components in a mass ratio of 1: 10, mixing the coarse powder of the cassia seeds with a solvent, and performing microwave-assisted treatment to obtain a pretreatment solution;

wherein the particle size of the cassia seed coarse powder is 0.05 mm;

the solvent is ethanol water solution and n-butanol with the volume ratio of 8:1, and the volume concentration of the ethanol water solution is 70%;

the microwave-assisted treatment comprises the following steps: the microwave power is 300W, and the microwave time is 40 s;

(2) carrying out reflux extraction on the pretreatment liquid obtained in the step (1) at 60 ℃ for 30min, cooling and filtering to obtain a mixed liquid;

(3) performing silica gel column chromatography on the mixed solution obtained in the step (2), eluting with petroleum ether and acetone in a mass ratio of 6.5:1, and collecting eluent;

A laxative health product comprises the following components in parts by weight: 10 parts of senna leaves, 15 parts of cassia seeds, 5 parts of black tea, 5 parts of green tea, 3 parts of poria cocos, 3 parts of radix glehniae, 20 parts of Chinese yam and 5 parts of honey;

s1, respectively crushing the senna leaves, the black tea, the green tea, the poria cocos, the radix glehniae and the Chinese yam according to the formula amount, sieving by a 10-mesh sieve, and mixing to obtain mixed powder;

s2, pretreating the cassia seeds according to the formula amount according to the preparation method of the embodiment 1 to obtain the aurantio-obtusin;

Example 2

A preparation method of aurantio-obtusin comprises the following steps:

(1) mixing the components in a mass ratio of 1: 15, mixing the coarse powder of the cassia seeds with a solvent, and performing microwave-assisted treatment to obtain a pretreatment solution;

wherein the particle size of the cassia seed coarse powder is 0.25 mm;

the solvent is ethanol water solution and n-butanol with the volume ratio of 10:1, and the volume concentration of the ethanol water solution is 50%;

the microwave-assisted treatment comprises the following steps: the microwave power is 400W, and the microwave time is 20 s;

(2) carrying out reflux extraction on the pretreatment liquid obtained in the step (1) at 70 ℃ for 15min, cooling and filtering to obtain a mixed liquid;

(3) performing silica gel column chromatography on the mixed solution obtained in the step (2), eluting with petroleum ether and acetone in a mass ratio of 8.5:1, and collecting eluent;

A laxative health product comprises the following components in parts by weight: 15 parts of senna leaves, 8 parts of cassia seeds, 15 parts of black tea, 15 parts of green tea, 12 parts of poria cocos, 12 parts of radix glehniae, 20 parts of Chinese yam and 10 parts of honey;

s1, respectively crushing the senna leaves, the black tea, the green tea, the poria cocos, the radix glehniae and the Chinese yam according to the formula amount, sieving by a 30-mesh sieve, and mixing to obtain mixed powder;

Example 3

A preparation method of aurantio-obtusin comprises the following steps:

(1) mixing the components in a mass ratio of 1: 12, mixing the coarse powder of the cassia seeds with a solvent, and performing microwave-assisted treatment to obtain a pretreatment solution;

wherein the particle size of the cassia seed coarse powder is 0.15 mm;

the solvent is ethanol water solution and n-butanol with the volume ratio of 9:1, and the volume concentration of the ethanol water solution is 60%;

the microwave-assisted treatment comprises the following steps: the microwave power is 338W, and the microwave time is 30 s;

(2) carrying out reflux extraction on the pretreatment liquid obtained in the step (1) at 65 ℃ for 20min, cooling and filtering to obtain a mixed liquid;

(3) performing silica gel column chromatography on the mixed solution obtained in the step (2), eluting with petroleum ether and acetone in a mass ratio of 7:1, and collecting eluent;

A laxative health product comprises the following components in parts by weight: 12 parts of senna leaves, 12 parts of cassia seeds, 10 parts of black tea, 10 parts of green tea, 8 parts of poria cocos, 5 parts of radix glehniae, 25 parts of Chinese yam and 8 parts of honey;

s1, respectively crushing the senna leaves, the black tea, the green tea, the poria cocos, the radix glehniae and the Chinese yam according to the formula amount, sieving by a 20-mesh sieve, and mixing to obtain mixed powder;

Comparative example 1

The difference between the comparative example and the example 3 is that in the preparation method of the aurantio-obtusin, the particle size of the semen cassiae coarse powder is 0.3 mm;

the solvent is ethanol water solution and n-butanol with the volume ratio of 15: 1;

the power of the microwave-assisted treatment is 500W, and the microwave time is 15 s.

Comparative example 2

The difference between the comparative example and the example 3 is that in the preparation method of the aurantio-obtusin, the particle size of the semen cassiae coarse powder is 0.01 mm;

the solvent is ethanol water solution and n-butanol with the volume ratio of 5: 1;

the power of the microwave auxiliary treatment is 250W, and the microwave time is 50 s.

Comparative example 3

CN107361173A a laxative health product prepared in example 3.

Experimental example I evaluation of physical and chemical indexes of orange-yellow cassia seed extract

Examples 1-3 and comparative examples 1-2 were prepared to obtain aurantio-obtusin with yield and purity measurements under chromatographic conditions: column size (4.6 mm. times.250 mm, 5 μm), column temperature of column: the mobile phase of the chromatograph is methanol and 0.12 percent phosphoric acid solution at 36 ℃, the flow rate is controlled to be 1.1ml/min, and the detection wavelength is 255 nm. The results are shown in Table 1.

TABLE 1

The invention carries out auxiliary treatment on the alcohol solution of the cassia seed coarse powder by microwave, then carries out heating reflux, utilizes silica gel column and recrystallization to separate and purify the aurantio-obtusin, and obviously improves the yield and the purity of the aurantio-obtusin by optimizing the parameters of the particle size of the cassia seed coarse powder, the concentration and the dosage of the alcohol solution, the microwave condition, the composition of an eluant and the like.

Meanwhile, the preparation method is simple, has low requirements on equipment, short preparation time and low production cost, and simultaneously has small reagent dosage and is beneficial to environmental protection.

Experimental example evaluation of Bianduo stool function

See the defecation function test method specified in the "test and evaluation of health food" (2003 edition).

1. Preparation of test article

15g of each of the laxative health products prepared in examples 1-3 and comparative examples 1-3 was taken, 10 times of water was added, the mixture was soaked at 85 ℃ for 30min, filtered by gauze, repeatedly soaked for 2 times, 3 times of filtrates were combined, and concentrated in a water bath to 30ml, which is equivalent to 0.5g crude drug/ml.

2. Laboratory animal

Clean grade ICR mice, male, weighing 18-22 g.

3. Experimental methods

ICR mice are randomly divided into 8 groups, 10 mice are per group, and defecation experiments are carried out, wherein test groups 1-3 are respectively subjected to intragastric administration of the test substances of the defecation health-care products prepared in examples 1-3, comparison groups 1-3 are respectively subjected to intragastric administration of the test substances of the defecation health-care products prepared in comparative examples 1-3, the administration amount is 10 times (18.5g/kgd) of the recommended dose of a human body, a negative control group and a model control group are respectively subjected to intragastric administration of distilled water with the same volume, and continuous administration is carried out for 7 days. After 7 days of administration, mice in each group are fasted for 16 hours without water prohibition, and a model control group, experimental groups 1-3 and comparison groups 1-3 are respectively gavaged with compound diphenoxylate (10mg/Kg bw), and a negative control group is gavaged with distilled water with the same volume as the stomach.

After 0.5h of administration of the compound diphenoxylate, the mice of the negative control group and the model control group are subjected to intragastric gavage by ink, the test objects containing the ink are respectively intragastric-gavage of the experimental group 1-3 and the comparative group 1-3, timing is started after intragastric-gavage, the mice are placed in a metabolism cage, normal diet is recovered, the black defecation time of the first grain of each mouse, the number of the defecation grains within 6h and the total weight of the excrement are observed and recorded. The results are shown in Table 2.

TABLE 2

Group of	First granule defecation time/min	Number of black-discharged excrement particles/particle	Total weight of black excrement/g
				Model control group	261.3±52.6**	11±2.6**	0.3745±0.1246*
Negative control group	176.4±19.2	20±3.2	0.5123±0.1210
				Example 1	193.4±18.4#	18±3.1#	0.4779±0.1227#
Example 2	197.1±17.1#	17±2.9#	0.4712±0.1231#
				Example 3	182.2±16.3#	19±2.7#	0.4926±0.1188#
Comparative example 1	243.6±43.6	14±3.1	0.3935±0.1144
				Comparative example 2	248.7±44.7	13±3.0	0.4015±0.1186
Comparative example 3	251.8±46.2	12±3.4	0.3884±0.1120

Note: compared with a negative control group, P is less than 0.01, and the difference is very significant; compared with the model control group, # P < 0.05, it is significantly different.

As can be seen from the table above, compared with the model control group, the first grain black stool discharging time, the number of grains discharging the stools within 6 hours and the total weight of the stools of the mice of the implementing groups 1 to 3 are significantly different (P is less than 0.05), which indicates that the health-care product for relaxing bowels prepared by the invention has good defecation effect.

Meanwhile, compared with a model control group, the first grain defecation time, the number of grains in defecation within 6h and the total weight of excrement of the mice in the control group 1-2 have no significant difference, and the preparation method of the aurantio-obtusin has an important influence on the defecation effect of the defecation health-care product claimed by the invention.

Compared with the prior art, the laxative health-care product disclosed by the invention has the advantages that the consumption of the senna leaves and the cassia seeds is obviously reduced by compounding the orange cassia tora essence with the senna leaves, the poria cocos, the radix glehniae, the Chinese yam and the like, the effect of relaxing bowel is obvious, and the safety is higher.

The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.

Claims

1. A health product for relaxing bowels is characterized by comprising the following components in parts by weight: 10-15 parts of senna leaves, 8-15 parts of semen cassiae, 5-15 parts of black tea, 5-15 parts of green tea, 3-12 parts of poria cocos, 3-12 parts of radix glehniae, 20-30 parts of Chinese yam and 5-10 parts of honey, wherein the semen cassiae is prepared by adopting the following preparation method to obtain orange-yellow cassia seed essence;

the preparation method of the aurantio-obtusin comprises the following steps:

(4) concentrating the eluent obtained in the step (3), and recrystallizing to obtain the aurantio-obtusin;

the particle size of the cassia seed coarse powder in the step (1) is 0.05-0.25 mm;

in the step (1), the solvent is ethanol water solution and n-butanol with the volume ratio of 8-10: 1;

the microwave auxiliary treatment in the step (1) comprises the following steps: the microwave power is 300-400W, and the microwave time is 20-40 s.

2. The laxative health product of claim 1, wherein the mass ratio of the cassia seed coarse powder to the solvent in the step (1) is 1: 10-15.

3. The laxative health product of claim 1, wherein the heating reflux in step (2) is: reflux extracting at 60-70 deg.C for 15-30 min.

4. The laxative health product of claim 1, wherein the eluent is a mixture of petroleum ether and acetone.