CN110772550B - Method for preparing ellagic acid from scalded myrobalan meat and application - Google Patents
Method for preparing ellagic acid from scalded myrobalan meat and application Download PDFInfo
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Abstract
The invention relates to the technical field of traditional Chinese medicine processing and natural product extraction, in particular to a method for preparing ellagic acid from sand-scalded myrobalan meat and application thereof. The method for preparing the ellagic acid comprises the following steps: and (3) placing the myrobalan in river sand at the temperature of 220-260 ℃, scalding for 5-9 min, and peeling off the pulp. Pulverizing into medicinal powder, reflux-extracting with ethanol, gradient-eluting with ethanol of different concentrations by macroporous adsorbent resin method, collecting eluate with ethanol concentration of 90%, recovering ethanol, and drying the residue under reduced pressure to obtain ellagic acid product. The invention greatly improves the content of the ellagic acid in the raw materials by optimizing the processing method and the process parameters, and the ellagic acid is extracted by ethanol reflux and eluted by ethanol solution through a macroporous adsorption resin method, thereby effectively reducing the cost and improving the yield and the purity of the ellagic acid product. The ellagic acid prepared by the invention is used for treating ulcerative colitis, is safe and effective, and is beneficial to improving the life quality of patients.
Description
Technical Field
The invention belongs to the technical field of medicines, relates to the technical field of traditional Chinese medicine processing and natural product extraction, and particularly relates to a method for preparing ellagic acid from fructus chebulae meat processed by sand scalding and an application of the method.
Background
Ellagic acid, also known as gallic acid, is a dimeric derivative of gallic acid, widely found in a variety of plant fruits. Natural ellagic acid was first found in plants such as raspberry, chestnut, pomegranate and the like. Modern pharmacological studies have found that ellagic acid has a variety of biological activities. Firstly, the ellagic acid has obvious inhibiting effect on canceration induced by chemical substances and other multiple canceration, particularly has good inhibiting effect on colon cancer, esophageal cancer, liver cancer, lung cancer, tongue and skin cancer, is called as a weishi in human body, and is called as the only natural variety of anticancer products in nearly ten years by the Brugins cancer research institute at the beginning of this century; in addition, the ellagic acid has strong astringency, has obvious inhibition effect on various bacteria, fungi and yeasts, and can prevent infection and inhibit ulcer; ellagic acid also has antiviral effect, and can inhibit proliferation of HIV; in addition, ellagic acid has the effects of inhibiting tyrosinase activity and blocking melanin generation, has whitening and spot-lightening effects, and is one of the accepted effective whitening components in dermatology. In view of various biological activities of ellagic acid, development and application in the fields of medicines, foods and cosmetics become hot at present, and the ellagic acid has a wide development space.
The myrobalan is dry mature fruit of myrobalan (Terminalia chebula Retz.) or Terminalia tomentosa Retz (Terminalia chebula Retz. var. tomentolla Kurt.) belonging to Combretaceae, has the effects of astringing intestines to check diarrhea, astringing lung to relieve cough, and is clinically used for different purposes of raw and mature, and the ancient books of the herbal such as Ben Cao Tong Xuan and the like are recorded: raw material can clear and promote the circulation of qi, while roasted can warm the stomach and secure the intestines, and the processed myrobalan fruit has the enhanced effect of astringing the intestines to check diarrhea, and is mainly used for treating qi dysentery and intestinal wind to purge blood, which corresponds to modern clinical ulcerative colitis. Modern chemical research shows that the main component of myrobalan is tannin, and the main components are gallnut tannin and ellagitannin, and the content of the main component can reach 23.60% -37.36%. After heating and processing, the tannin component in the myrobalan is respectively converted into gallic acid and ellagic acid, when the processing temperature is higher, the component conversion degree is larger, and the product mainly contains ellagic acid.
The purity of the natural ellagic acid extracted in the prior art is low, generally about 20-30%, although the purity of the ellagic acid can reach 40-50% by only performing one alkali-soluble acid precipitation and one water washing, the purity can be improved by adding the alkali-soluble acid precipitation or water washing step, but the product quality is reduced. At present, tannin is prepared by converting plant tannin through a method of peroxidation or acid hydrolysis, the acid hydrolysis method has the advantages of low conversion rate, a plurality of byproducts, difficult content improvement, low-temperature oxidation, high conversion rate, a few byproducts and easy purity improvement. A preparation method of high-purity ellagic acid (CN101434608A) comprises preparing ellagic acid by acid hydrolysis method, using sulfuric acid and hydrogen peroxide reagent, wherein the hydrolysate contains a large amount of acid agent hydrogen peroxide, which can cause severe environmental pollution, and has low product yield, high requirement on equipment, and is not suitable for popularization and production. A method (CN101759165A) for preparing ellagic acid from tana powder adopts an oxidation method to prepare ellagic acid, although the content can reach 98%, the oxidation process is complex to operate, a large amount of chemical reagents are used, a pyridine toxic solvent is used in the refining process, great harm is caused to human bodies, and environmental pollution can be caused. A method for preparing ellagic acid by enzyme method with pericarpium Granati as raw material (CN101481714A) adopts biological enzyme conversion method to prepare ellagic acid, and has the advantages of complicated operation, strict condition control, long production cycle, and low production efficiency. A method (CN201510156630.2) for preparing high-purity ellagic acid by purifying tannin with macroporous resin method discloses a method for preparing high-purity ellagic acid by purifying with macroporous resin method, and specifically discloses a method for purifying tannin with macroporous adsorption resin technology, which comprises oxidizing tannin in alkaline environment to generate ellagic acid, and refining to obtain ellagic acid product with purity of more than 98%. Although the method can prepare high-purity ellagic acid products, the operation is more complicated, and the technical requirements on resin are higher. A preparation method of ellagic acid from Terminalia chebula Li et Li (CN102020682B) comprises collecting Terminalia chebula leaf coarse powder, performing microwave extraction and multiple ion exchange resin adsorption and elution to obtain ellagic acid, and has the advantages of complicated operation method, low yield, long production period, and unsuitability for industrialized production.
Therefore, the development of a method for preparing ellagic acid, which is simple and convenient to operate, high in yield and suitable for industrial production, is of great significance.
Disclosure of Invention
In view of the problems in the prior art, the invention aims to provide a method for preparing ellagic acid from fructus chebulae meat processed by sand scalding and application thereof. The processing method provided by the invention can convert the tannins in the myrobalan to the ellagic acid to the maximum extent, and then the ellagic acid is enriched and prepared and used for treating the ulcerative colitis. According to the invention, the content of ellagic acid in the raw materials can be obviously improved by carrying out sand-scalding processing on myrobalan, the purity of the ellagic acid product obtained by ethanol extraction and macroporous adsorption resin method enrichment can reach 55-65%, the yield can reach 50-60%, and the ellagic acid product prepared by the preparation method has a good anti-inflammatory effect and can be used for treating ulcerative colitis.
In order to achieve the above object, the present invention provides the following technical solutions.
A processing method of scalded myrobalan meat comprises the following steps: and (3) placing the myrobalan in river sand at the temperature of 220-260 ℃, scalding for 5-9 min, and peeling off the pulp.
Preferably, the processing temperature is 240 deg.C, and the processing time is 6 min.
A method for preparing ellagic acid from scalded myrobalan meat comprises the following steps.
S1 crushing: pulverizing the scalded fructus Chebulae meat into medicinal powder.
S2 extraction: adding the medicine powder of scalded myrobalan meat into an ethanol solution for reflux extraction, and recovering ethanol to obtain a concentrated solution.
S3 enrichment: diluting the obtained concentrated solution with water, loading onto macroporous adsorbent resin, gradient eluting with ethanol of different concentrations, collecting the eluate with ethanol concentration of 90%, and recovering ethanol.
S4, obtaining: drying the residue under reduced pressure to obtain ellagic acid product.
Wherein, the crushing degree of the scalded myrobalan meat in the step S1 is controlled to be 10-24 meshes.
Preferably, the crushing degree of the scalded myrobalan meat in the step S1 is controlled to be 10 meshes.
Further, in the step S2, the concentration of an ethanol solution used for extracting the medicine powder of the scalded myrobalan meat is 40% -70%, the material-liquid ratio is 1 (10-15), the extraction time is 0.5-2 h, and the extraction times is 2-3.
Preferably, in the step S2, the concentration of the ethanol solution used for extracting the medicine powder of the scalded myrobalan meat is 60%, the material-liquid ratio is 1:10, the extraction time is 1h, and the extraction times are 3 times.
In step S3, the dilution degree of the concentrated solution is that the volume of the crude drug and the liquid medicine is 1 (10-20).
Preferably, the dilution degree of the concentrated solution in step S3 is 1:15 of the volume of the crude drug to the liquid medicine.
Further, the model of the macroporous adsorption resin used for purification in the step S3 is NKA-9 or HPD 600.
Further, the ratio of the volume of the sample loading solution to the volume of the resin in the step S3 is 1 (4-8).
Preferably, the ratio of the volume of the sample solution to the volume of the resin in the step S3 is 1: 6.
Further, the concentration and the amount of the eluting alcohol in the step S3 are 0% for eluting 1-3 column volumes, 60% for eluting 1-3 column volumes, and 90% for eluting 10-20 column volumes.
Preferably, the concentration and the amount of the eluting alcohol in step S3 are 0% eluting 3 column volumes, 60% eluting 3 column volumes, and 90% eluting 15 column volumes.
Application of ellagic acid prepared from fructus Chebulae Immaturus in preparing medicine for treating colitis ulcerosa is provided.
Compared with the prior art, the invention has the following beneficial effects.
1) At present, ellagic acid is mostly extracted from pomegranate rind, Chinese chestnut, raspberry and other plants, and the content of ellagic acid in raw materials is not high. The content of ellagic acid in the myrobalan is about 1.31-2.35%, and can be increased to 2.54-3.82% after processing. According to the invention, the processing method and the processing parameters are optimized, and after the medicine is processed by sand scalding, the content of the ellagic acid in the myrobalan meat can reach 8.0%, so that the content of the ellagic acid in the raw materials is greatly improved, and a foundation is laid for improving the yield of the ellagic acid.
2) The method provided by the invention has the advantages that the ethanol is refluxed and extracted, and then the ethanol solution is eluted by a macroporous adsorption resin method, the operation is simple, the obtained ethanol solution can be recycled by recovering the solvent, the macroporous adsorption resin can be repeatedly used, the cost is effectively reduced, and the yield and the purity of the ellagic acid product are improved.
3) The cause of ulcerative colitis is complex, so far, the ulcerative colitis is not completely clear, the disease has many links, the cure difficulty is large, the disease course is long, and the ulcerative colitis is listed as one of modern intractable diseases by the world health organization. The ellagic acid obtained by the invention is used for treating ulcerative colitis, and can overcome the defect of great side effect of the current western medicine treatment. Safe and effective, improves the life quality of patients and assists the healthy Chinese plan.
Drawings
FIG. 1 is a liquid chromatogram of an ellagic acid control.
FIG. 2 is a liquid chromatogram for measuring the content of ellagic acid in raw myrobalan.
FIG. 3 is a liquid chromatogram for measuring the content of ellagic acid in the blanched myrobalan fruit.
FIG. 4 is a liquid chromatogram for determination of ellagic acid content in an aqueous eluate.
FIG. 5 is a liquid chromatogram for determination of ellagic acid content in an eluate of 60% ethanol solution.
FIG. 6 is a liquid chromatogram for determination of ellagic acid content in an eluate of 90% ethanol solution.
FIG. 7 is a liquid chromatogram for determination of ellagic acid content in the product.
Detailed Description
The invention is described in further detail below with reference to the figures and specific embodiments. It should be understood that the scope of the above-described subject matter is not limited to the following examples, and any techniques implemented based on the disclosure of the present invention are within the scope of the present invention. Example 1 Process optimization experiment of a process for the preparation of ellagic acid from blanched myrobalan meat.
1. Preparation method of fructus Chebulae meat by hot stamping with sand, processing temperature screening experiment.
A preparation method of a scalded myrobalan meat comprises the following steps.
(1) Collecting 9 parts of fructus Chebulae (containing ellagic acid 1.572%), placing 20g each in river sand of 180 deg.C, 190 deg.C, 200 deg.C, 210 deg.C, 220 deg.C, 230 deg.C, 240 deg.C, 250 deg.C, 260 deg.C, scalding for 6min, and removing pulp.
(2) Collecting each part of scalded fructus Chebulae, pulverizing, sieving with 60 mesh sieve, precisely weighing 0.1g, adding 25mL 70% methanol water solution, ultrasonic extracting for 20min, and filtering.
(3) And (3) filtering the subsequent filtrate with a 0.45-micron microporous filter membrane, respectively sucking 10 μ L of the filtrate, injecting into a high performance liquid chromatograph, measuring the peak area of the ellagic acid chromatogram, and calculating the content of the ellagic acid by an external standard one-point method, wherein the results are shown in Table 1.
TABLE 1 comparison of ellagic acid content in different temperature processed products
As can be seen from Table 1, the content of ellagic acid in the product is increased by 3.8-5.0 times compared with that in the raw product when the myrobalan is processed at 220-260 ℃. The conversion rate of other components to ellagic acid in the processing process is related to the temperature, and the conversion rate is obviously higher than that at the low temperature above 220 ℃.
2. A preparation method of fructus Chebulae meat processed by hot pressing, TANGSHENZHENG, is disclosed.
A preparation method of a scalded myrobalan meat comprises the following steps.
(1) Collecting 200g fructus Chebulae (containing ellagic acid 1.572%), placing in river sand at 240 deg.C, scalding for 4min, 5min, 6min, 7min, 8min, 9min, and 10min respectively to obtain about 20g, and removing pulp.
(2) Collecting each part of scalded fructus Chebulae, pulverizing, sieving with 60 mesh sieve, precisely weighing 0.1g, adding 25mL 70% methanol water solution, ultrasonic extracting for 20min, and filtering.
(3) Filtering the filtrate with 0.45 μm microporous membrane, respectively sucking 10 μ L of filtrate, injecting into high performance liquid chromatograph, measuring ellagic acid chromatographic peak area, and calculating ellagic acid content by external standard one-point method. The results are shown in Table 2.
TABLE 2 comparison of ellagic acid content in the preparations processed at different times
As can be seen from Table 2, the myrobalan is processed at 240 ℃ for 5-9 min, and the ellagic acid content of the product is increased by 4.2-5.0 times compared with that of the raw product. Wherein the processing time is short, and the conversion rate of other components to ellagic acid is low; the processing time is long, organic matters are carbonized at high temperature, and ellagic acid is damaged to a certain extent, so that the time needs to be reasonably controlled.
As can be seen from the results obtained in experiments 1 and 2, the optimal condition for sand-scalding fructus Chebulae should be processing at 240 deg.C for 6 min.
3. Method for preparing ellagic acid from scalded fructus Chebulae meat-ethanol concentration screening experiment.
A method for extracting ellagic acid from fructus Chebulae meat by blanching comprises the following steps.
(1) Scalding fructus Chebulae in sand at 240 deg.C for 6min, pulverizing, and sieving with 60 mesh sieve.
(2) Weighing 10 parts of crushed myrobalan meat, adding 50mL of ethanol solutions with the concentrations of 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% and 90% respectively into 2g of each part, refluxing and extracting for 1h, and filtering.
(3) Each of the subsequent filtrates was diluted appropriately, and the diluted solution was passed through a 0.45 μm microporous membrane, 10. mu.L of the diluted solution was sucked and injected into a high performance liquid chromatograph to measure the ellagic acid content, and the results are shown in Table 3.
TABLE 3 comparison of ellagic acid content extracted with solvents of different concentrations
As can be seen from Table 3, the ellagic acid is extracted by 40-70% ethanol solution, the extraction efficiency is high, and the optimal extraction solvent can be determined by the highest extraction efficiency of 60% ethanol solution.
4. Method for preparing ellagic acid from scalded myrobalan meat-optimal condition screening experiment for extracting ellagic acid with 60% ethanol solution.
A method for extracting ellagic acid from fructus Chebulae meat by blanching comprises the following steps.
(1) Processing fructus Chebulae in river sand at 240 deg.C for 6min, and smashing to obtain pulp.
(2) Selecting 60% ethanol solution to extract ellagic acid by orthogonal design method, and extracting with L shown in Table 39(34) The design scheme is used for carrying out experiments.
(3) The content of ellagic acid was measured by HPLC method from each extract, and the results are shown in table 4.
TABLE 4 ellagic acid extraction condition optimization
As can be seen from Table 4, the myrobalan meat is pulverized into 10-24-mesh medicinal powder, the extraction is carried out for 2-3 times at a material-liquid ratio of more than 1:10, the ellagic acid content obtained by extraction can exceed 7% each time of more than 0.5h, the ellagic acid content in the myrobalan meat obtained by an optimal processing technology is 8%, and the extraction rate can reach more than 85%. Wherein the extraction is carried out for 3 times with the crushing granularity of 10 meshes and the material-liquid ratio of 1:10, and the extraction efficiency is the highest each time for 1 h.
5. Method for preparing ellagic acid from scalded myrobalan meat-eluent volume screening experiment.
A method for enriching and preparing ellagic acid from fructus Chebulae meat by blanching with sand comprises the following steps.
(1) Processing fructus Chebulae in river sand at 240 deg.C for 6min, and smashing to obtain pulp.
(2) Pulverizing fructus Chebulae, sieving with 10 mesh sieve, weighing 20g medicinal powder, adding 60% ethanol solution at a material-to-liquid ratio of 1:10, reflux extracting for 3 times (1 hr each time), mixing extractive solutions, recovering ethanol, and adding water to concentrate to a volume of 200ml volumetric flask (containing ellagic acid with concentration of 7.14 mg. ml-1)。
(3) Diluting 2ml of concentrated solution according to the crude drug volume of 1:15 to obtain 3ml of diluted solution, washing the top end of the treated NKA-9 macroporous adsorption resin with water for 5 column volumes (5BV) according to the crude drug volume of 1:6 to obtain a fraction of each column volume; washing 5 column volumes (5BV) with 60% ethanol solution, and collecting fractions of each column volume; finally, the column was washed with 90% ethanol solution until no ellagic acid flowed out, and the fractions of each column volume were taken.
(4) Each column volume fraction eluted from each of the collected solutions was appropriately diluted and the amount of ellagic acid was measured by HPLC, and the results are shown in Table 5.
TABLE 5 ellagic acid content in each fraction
As can be seen from Table 5, after loading, the sample is washed with water, the large polar components are eluted first, the ellagic acid is slightly lost, when the elution reaches 3 times the resin volume (3BV), the large polar components are basically eluted, and the washing volume is not more than 3BV to reduce the loss of the ellagic acid; removing the medium polar components by using a 60% ethanol solution, gradually eluting the medium polar components along with the increase of the elution volume, but accumulating and increasing the loss of the ellagic acid, wherein the elution volume of the 60% ethanol is not more than 3BV for considering the purity and the yield of the ellagic acid preparation; and finally, eluting the ellagic acid adsorbed on the macroporous adsorption resin by using a 90% ethanol solution, wherein when the elution volume reaches more than 10BV, the detection amount of the ellagic acid is gradually reduced, the ellagic acid is basically and completely eluted to 20BV, the volume of the eluent does not need to be increased, the production cost is also taken into consideration, and the elution of 15BV is more appropriate.
6. A method for preparing ellagic acid from scalded myrobalan meat comprises the following steps.
(1) Taking 20ml of the concentrated solution in 5 in example 1, washing the top of the treated NKA-9 macroporous adsorption resin with water for 3BV firstly, wherein the raw medicine amount is 1:10 and the liquid medicine volume is 1: 6; washing with 60% ethanol solution for 3 BV; finally, washing 15BV with 90% ethanol solution, collecting 90% ethanol eluent, and mixing.
(2) Recovering solvent, and drying the residue under reduced pressure to obtain ellagic acid product.
7. A method for preparing ellagic acid from scalded myrobalan meat comprises the following steps.
(1) Taking 20ml of the concentrated solution in the example 5, diluting according to the crude drug amount, namely the volume of the liquid medicine is 1:15 to obtain 30ml of liquid medicine, and firstly washing the top end of the treated NKA-9 macroporous adsorption resin with water for 3BV according to the volume of the liquid medicine, namely the volume of the resin, being 1: 6; washing with 60% ethanol solution for 3 BV; finally, washing 15BV with 90% ethanol solution, collecting 90% ethanol eluent, and mixing.
(2) Recovering solvent, and drying the residue under reduced pressure to obtain ellagic acid product.
8. A method for preparing ellagic acid from scalded myrobalan meat comprises the following steps.
(1) Taking 20ml of the concentrated solution in the example 5, diluting according to the crude drug volume of 1:20 to obtain 40ml of liquid medicine, and firstly washing the top end of the treated NKA-9 macroporous adsorption resin with water for 3BV according to the liquid medicine volume of 1:6 to obtain a solution; washing with 60% ethanol solution for 3 BV; finally, washing 15BV with 90% ethanol solution, collecting 90% ethanol eluent, and mixing.
(2) Recovering solvent, and drying the residue under reduced pressure to obtain ellagic acid product.
9. A method for preparing ellagic acid from scalded myrobalan meat comprises the following steps.
(1) Taking 20ml of the concentrated solution in the example 5, and mixing the raw materials according to the weight ratio: diluting the liquid medicine with the volume of 1:15 to obtain 30ml of liquid medicine, and washing the top end of the treated NKA-9 macroporous adsorption resin with water for 3BV with the liquid medicine volume of 1:4 and the resin volume of 1: 4; washing with 60% ethanol solution for 3 BV; finally, washing 15BV with 90% ethanol solution, collecting 90% ethanol eluent, and mixing.
(2) Recovering solvent, and drying the residue under reduced pressure to obtain ellagic acid product.
10. A method for preparing ellagic acid from scalded myrobalan meat comprises the following steps.
(1) Taking 20ml of the concentrated solution in the example 5, diluting according to the crude drug amount, namely the volume of the liquid medicine is 1:15 to obtain 30ml of liquid medicine, and firstly washing the top end of the treated NKA-9 macroporous adsorption resin with water for 3BV according to the volume of the liquid medicine, namely the volume of the resin, being 1: 8; washing with 60% ethanol solution for 3 BV; finally, washing 15BV with 90% ethanol solution, collecting 90% ethanol eluent, and mixing.
(2) Recovering solvent, and drying the residue under reduced pressure to obtain ellagic acid product.
11. A method for preparing ellagic acid from scalded myrobalan meat comprises the following steps.
(1) Taking 20ml of the concentrated solution in the example 5, diluting according to the crude drug volume of 1:15 to obtain 30ml of liquid medicine, and firstly washing the top end of the processed HPD600 type macroporous adsorption resin with water for 3BV according to the liquid medicine volume of 1:6 to obtain a solution; washing with 60% ethanol solution for 3 BV; finally, washing 15BV with 90% ethanol solution, collecting 90% ethanol eluent, and mixing.
(2) Recovering solvent, and drying the residue under reduced pressure to obtain ellagic acid product.
And (3) measuring the content of ellagic acid in the product obtained by decompression drying after the 90% ethanol eluent and the recovered eluent obtained in the methods 6 to 11 in the example 1 are measured by an HPLC method. The specific experimental conditions were as follows: a chromatographic column: ecosil C18 column (150 mm. times.4.6 mm, 5 μm); mobile phase: 0.3% phosphoric acid aqueous solution (A) -methanol (B) (0-8 min: B% 5.0% → 10.0; 8-15 min: B% 10.0% → 25.0%; 15-25 min: B% 25.0%; 25-30 min: B% 25.0% → 30.0%; 30-50 min: B% 30.0% → 45.0%; 50-55 min: B% 45.0%); the detection wavelength is 270 nm; the flow rate is 1 ml/min-1(ii) a The column temperature is 30 ℃; the amount of the sample was 10. mu.l, and the results are shown in Table 6.
TABLE 6 comparison of the effects of yield and purity
According to the methods 6, 7 and 8, the extracting solution is diluted to different degrees before purification by macroporous adsorption resin, the product purity is higher when the dilution ratio is larger, but the corresponding yield is reduced, so the dilution ratio is not too large, namely the dilution ratio is not required to be too high in the range of 1 (10-20), and the yield and the purity are both higher when the dilution ratio is 1:15, and the optimal dilution ratio is obtained.
According to the methods 7, 9 and 10, the sample loading amount is constant, the purity of the product is higher when the dosage of the macroporous adsorption resin is larger, the corresponding yield is reduced, the dosage of the eluent is increased due to the excessive dosage of the resin, and the material cost and the working hour are consumed, so that the dosage of the resin is not excessive, the volume of the liquid medicine is 1 (4-8) of the volume of the resin, and the yield and the purity are higher when the volume is 1:6, and the optimal dosage of the resin is obtained.
The inventor carries out systematic investigation on the adsorption performance of the tannin in the myrobalan by using different types of macroporous adsorption resins in earlier work, and the result shows that the adsorption and desorption performances of the two resins, namely NKA-9 and HPD600, on the tannin in the myrobalan are good, so that the enrichment capacities of the two resins on the ellagic acid are compared, and the result is shown by a method 7 and a method 11, and the two resins can better enrich the ellagic acid.
Example 2 ellagic acid pharmacological study experiment-replication of the mouse model of ulcerative colitis using Dextran Sodium Sulfate (DSS) modelling.
Taking 90 mice, randomly dividing the mice into a blank group, a model group, a positive drug group and 6-11 groups of the method in example 1, wherein 10 mice in each group are taken, after 4 days of free drinking of 4% DSS aqueous solution (blank group drinking water), the blank group and the model group are respectively gavaged to be given with physiological saline, the positive drug group is given with sulfasalazine aqueous solution, the example groups are respectively gavaged to be given with the obtained ellagic acid product, and the ellagic acid product prepared by the method 6-11 in example 1 is prepared into the ellagic acid with the concentration of about 1.7 mg/ml-1The administration amount of the solution (2) was calculated as 0.1ml per 10g of body weight by gavage, and the administration was carried out for 7 days. Mice were sacrificed 4h after the last dose and the colon portion was excised. The contents of TNF-. alpha.and IL-1. beta. in colon tissue were measured by ELISA method, and the results are shown in Table 7.
TABLE 7 comparison of the effects of the pharmacological actions
Note: using the LSD test, compared to the blank group,#P<0.05，##P<0.01; in comparison with the set of models,*P<0.05，**P<0.01。
as can be seen from the results in Table 7, the content of inflammatory factors TNF-alpha and IL-1 beta in the colon of the ulcerative colitis model animal was reduced after the administration of the ellagic acid product, indicating that the ellagic acid product acts to reduce the inflammatory response.
Claims (9)
1. A preparation method of a hot-stamped myrobalan flesh is characterized by placing myrobalan in river sand at 220-260 ℃, stamping for 5-9 min, and stripping flesh.
2. A method for preparing an ellagic acid-containing extract from the meat of myrobalan blanched by the method of claim 1, comprising the steps of:
s1 crushing: pulverizing the scalded myrobalan meat into medicinal powder;
s2 extraction: adding the medicine powder of scalded myrobalan meat into an ethanol solution for reflux extraction, and recovering ethanol to obtain a concentrated solution;
s3 enrichment: diluting the obtained concentrated solution with water, loading onto macroporous adsorbent resin, gradient eluting with ethanol of different concentrations, collecting eluate with ethanol concentration of 90%, and recovering ethanol;
s4, obtaining: drying the residue under reduced pressure to obtain product containing ellagic acid extract.
3. The method for preparing the ellagic acid-containing extract from the blanched myrobalan meat as claimed in claim 2, wherein the degree of pulverization of the blanched myrobalan meat in S1 is controlled to 10-24 mesh.
4. The method for preparing the ellagic acid-containing extract from the blanched myrobalan meat as claimed in claim 2, wherein the concentration of an ethanol solution used for extracting the medicine powder of the blanched myrobalan meat in S2 is 40% -70%, the material-liquid ratio is 1 (10-15), the extraction time is 0.5-2 h, and the extraction times are 2-3 times.
5. The method for preparing the ellagic acid-containing extract from the blanched myrobalan flesh as claimed in claim 2, wherein the dilution degree of the concentrated solution in S3 is the crude drug amount, and the volume of the liquid medicine is 1 (10-20).
6. The method of claim 2, wherein the purification in S3 is performed using a macroporous adsorbent resin of NKA-9 or HPD 600.
7. The method for preparing the ellagic acid-containing extract from the blanched myrobalan meat according to claim 2, wherein the ratio of the volume of the sample solution to the volume of the resin in S3 is 1 (4-8).
8. The method of claim 2, wherein the concentration and amount of the eluting alcohol in S3 is 0% elution for 1-3 column volumes, 60% elution for 1-3 column volumes, and 90% elution for 10-20 column volumes.
9. Use of an ellagic acid-containing extract prepared from the preparation method of any one of claims 2-8 in the preparation of a medicament for treating ulcerative colitis.
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