阅读说明：本技术 一种高含量大黄素的提取纯化方法 (Extraction and purification method of high-content emodin ) 是由 杨海鹰 王晓龙 杨方超 于 2019-11-15 设计创作，主要内容包括：本发明涉及一种高含量大黄素的提取纯化方法,以解决现有的大黄素提取纯化的方法存在生产成本高、技术难度大的问题。该高含量大黄素的提取纯化方法,包括以下步骤；称取虎杖原料,清洗、晾干、粉碎后得虎杖碎料,并用丙酮浸泡虎杖碎料；使用丙酮提取虎杖碎料并过滤,将滤液合并后浓缩至浸膏,在真空条件下将浸膏干燥,干燥后粉碎；将干燥后的碎料使用乙酸乙酯溶解,加入活性炭除杂并过滤,通过静置的方式滤除沉淀；将沉淀用N-N二甲基甲酰胺溶液回流搅拌热洗涤并趁热抽滤,分别收集不溶物和滤液,将不溶物再次回流搅拌热洗涤,收集沉淀并干燥；将干燥后的沉淀用乙酸乙酯、活性炭结晶,并滤出晶体；用甲醇搅拌洗涤晶体,干燥后即得大黄素。(The invention relates to a method for extracting and purifying high-content emodin, which aims to solve the problems of high production cost and great technical difficulty of the existing method for extracting and purifying emodin. The method for extracting and purifying the high-content emodin comprises the following steps; weighing rhizoma Polygoni Cuspidati raw materials, cleaning, air drying, pulverizing to obtain rhizoma Polygoni Cuspidati crushed material, and soaking the rhizoma Polygoni Cuspidati crushed material with acetone; extracting crushed giant knotweed with acetone, filtering, merging the filtrates, concentrating to obtain an extract, drying the extract under a vacuum condition, drying and crushing; dissolving the dried crushed materials by using ethyl acetate, adding activated carbon to remove impurities, filtering, and filtering out precipitates in a standing mode; refluxing, stirring and hot washing the precipitate with an N-N dimethylformamide solution, carrying out suction filtration while the precipitate is hot, respectively collecting insoluble substances and filtrate, refluxing, stirring and hot washing the insoluble substances again, collecting the precipitate and drying; crystallizing the dried precipitate with ethyl acetate and active carbon, and filtering out crystals; stirring and washing the crystal by using methanol, and drying to obtain the emodin.)

1. A method for extracting and purifying high-content emodin is characterized by comprising the following steps: comprises the following steps;

s1, weighing the giant knotweed raw material, cleaning, airing and crushing to obtain giant knotweed crushed material, and soaking the giant knotweed crushed material in acetone;

s2, extracting polygonum cuspidatum crushed aggregates by using acetone and filtering, merging filtrate and concentrating to obtain extract, drying the extract under a vacuum condition, and crushing the dried extract;

s3, dissolving the dried crushed materials by using ethyl acetate, adding activated carbon to remove impurities and filtering, and filtering out precipitates by standing;

s4, refluxing, stirring and hot washing the precipitate with an N-N dimethylformamide solution, carrying out suction filtration while the precipitate is hot, respectively collecting insoluble substances and filtrate, refluxing, stirring and hot washing the insoluble substances again, collecting the precipitate and drying;

s5, crystallizing the dried precipitate with ethyl acetate and activated carbon, and filtering out crystals;

s6, stirring and washing the crystal by using methanol, and drying to obtain the emodin.

2. The method for extracting and purifying emodin with high content according to claim 1, characterized in that: in step S1, the crushed giant knotweed is soaked in acetone for 8-12 hours.

3. The method for extracting and purifying emodin with high content according to claim 2, characterized in that: in the step S2, acetone with the concentration of 80% and the weight of 12 times of the weight of the crushed materials is used; extracting the crushed material three times at 65 deg.C and pH = 5-6.

4. The method for extracting and purifying emodin with high content according to claim 3, characterized in that: in the step S3, ethyl acetate is used, the weight of the ethyl acetate is 15 times of the weight of the dried crushed aggregates, the dried crushed aggregates are dissolved at the temperature of 80 ℃, and the weight of the added activated carbon is 10% of the weight of the polygonum cuspidatum crushed aggregates.

5. The method for extracting and purifying emodin with high content according to claim 4, wherein the method comprises the following steps: in step S4, the weight of the N-N dimethylformamide solution used was 15 times the weight of the precipitate, and the temperature at the time of the reflux-stirring heat washing was 88 ℃ for 6 hours.

6. The method for extracting and purifying emodin with high content according to claim 5, wherein the method comprises the following steps: in step S5, the weight of ethyl acetate used is 40 times of the weight of the dried precipitate, the temperature of ethyl acetate is 80 ℃, and the weight of the added activated carbon is 20% of the weight of the crushed giant knotweed.

7. The method for extracting and purifying emodin with high content according to claim 6, wherein the method comprises the following steps: in step S6, methanol was used in an amount of 3 times the weight of the crystals, and the crystals were washed with stirring at room temperature.

Technical Field

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a method for extracting and purifying high-content emodin.

Background

The emodin is named as cinnabarinin A, and can be used in medical treatment, health care and daily chemicals, for example, hair care and skin care products, and also be coded in natural pigment. Abroad, people use it as a laxative. The emodin has inhibitory effect on Staphylococcus aureus 209P, Streptococcus, diphtheria bacillus, Bacillus subtilis, Bacillus paratyphi, Shigella dysenteriae, Escherichia coli, Bacillus influenzae, pneumococcus, and Cartacoccus; the compound has stronger inhibition effect on common anaerobic bacteria in clinic; the MIC of the compound is slightly higher than that of metronidazole, and 76% -91% of anaerobic bacteria can be inhibited in the concentration of 8 mug/ml. The emodin antibacterial action mechanism is related to inhibition of mitochondrial respiratory chain electron transfer, inhibition of respiration, oxidation and dehydrogenation of amino acid, sugar and protein metabolism intermediate products and the like. The end result of inhibiting nucleic acid and protein synthesis is that bacterial growth is inhibited. Also has spasmolytic, antitussive, and blood pressure lowering effects.

There are many methods for extracting and purifying high-quality emodin, and the common methods include chromatographic column method, and method based on extraction and membrane filtration, but these methods all have the problems of high production cost, large technical difficulty, and difficulty in realizing industrialization. Due to the strict confidentiality of the route of emodin refining process by manufacturers at home and abroad, a few patents are already applied.

The patent with publication number CN 107337594A discloses a method for extracting high-purity emodin, which comprises the following steps: step one, crushing; step two, adjusting the PH value; step three, ultrasonic treatment; step four, membrane treatment; step five, extraction; step six, neutralization treatment; step seven, drying; and step eight, grinding.

The patent with publication number CN 105906493A discloses an extraction process of emodin, which comprises the following steps: firstly, washing rhubarb, drying at 110-130 ℃, crushing to obtain fine powder, distilling the obtained fine powder by a steam distillation method, removing distilled impurities, then adding refined powder into an ethanol aqueous solution, heating, refluxing, extracting, cooling, adjusting the pH of an extracting solution to 3.0-4.0, standing, layering, sending a supernatant into a microfiltration membrane for filtering, then heating and concentrating a microfiltration permeating liquid, finally sending a concentrated solution into a chromatographic column for removing impurities, and spray-drying the obtained permeating liquid to obtain the emodin.

Although the method can obtain the high-content emodin product, the method has the characteristics of overlong column chromatography purification time, difficult realization of industrialization, large solvent consumption, difficult recovery of an extraction solvent, high technical requirement on membrane filtration and the like.

Column chromatography purification time problem: through tests, the target product is obtained by a column chromatography purification method for about 25 days, and the production periods in different seasons are obviously different, so that the supply time in part of seasons is longer.

The membrane filtration extraction problem: the solubilities of chrysophanol, physcion and emodin in the same solvent are very similar, so that the operation difficulty is increased, and the product yield is greatly reduced.

Disclosure of Invention

The invention aims to provide a method for extracting and purifying high-content emodin so as to solve the problems of high production cost and great technical difficulty of the existing method for extracting and purifying emodin.

In order to achieve the purpose, the technical solution of the invention is as follows:

a method for extracting and purifying high-content emodin comprises the following steps;

s1, weighing the giant knotweed raw material, cleaning, airing and crushing to obtain giant knotweed crushed material, and soaking the giant knotweed crushed material in acetone;

s2, extracting polygonum cuspidatum crushed aggregates by using acetone and filtering, merging filtrate and concentrating to obtain extract, drying the extract under a vacuum condition, and crushing the dried extract;

s3, dissolving the dried crushed materials by using ethyl acetate, adding activated carbon to remove impurities and filtering, and filtering out precipitates by standing;

s4, refluxing, stirring and hot washing the precipitate with an N-N dimethylformamide solution, carrying out suction filtration while the precipitate is hot, respectively collecting insoluble substances and filtrate, refluxing, stirring and hot washing the insoluble substances again, collecting the precipitate and drying;

s5, crystallizing the dried precipitate with ethyl acetate and activated carbon, and filtering out crystals;

s6, stirring and washing the crystal by using methanol, and drying to obtain the emodin.

Preferably, in step S1, the crushed giant knotweed is soaked in acetone for 8-12 hours.

Preferably, the crushed aggregates are extracted three times at a temperature of 65 ℃ and a PH =5-6 using acetone with a concentration of 80% and a weight of 12 times the weight of the crushed aggregates in step S2.

Preferably, in step S3, ethyl acetate is used in an amount of 15 times the weight of the dried crushed material, and the dried crushed material is dissolved at a temperature of 80 ℃, and activated carbon is added in an amount of 10% of the weight of the crushed material.

Preferably, the weight of the N-N dimethylformamide solution used in step S4 is 15 times of the weight of the precipitate, and the temperature at the time of the reflux stirring heat washing is 88 ℃ and the time period is 6 hours.

Preferably, in step S5, the weight of ethyl acetate used is 40 times of the weight of the dried precipitate, the temperature of ethyl acetate is 80 ℃, and the weight of the added activated carbon is 20% of the weight of the crushed giant knotweed.

7. The method for extracting and purifying emodin with high content according to claim 5, wherein the method comprises the following steps: in step S6, methanol was used in an amount of 3 times the weight of the crystals, and the crystals were washed with stirring at room temperature.

The invention has the beneficial effects that:

according to the extraction and purification method of the high-content emodin, provided by the invention, the product is dissolved and separated by adopting the N-N dimethylformamide solvent under the heating condition, and the method is simple to operate, good in effect and environment-friendly; the method has the advantages of less solvent consumption, simple process and easy realization of industrialization, and saves the cost of extraction and purification; the method has the advantages that the product yield is over 95 percent, the solvent recovery rate is over 90 percent, the comprehensive utilization rate is high, and the waste of raw materials can be avoided; the method has the advantages that the production period of the product is short, so that the market demand can be met in time, and the problem of overlong time consumption in the column chromatography purification method is solved.

Drawings

FIG. 1 is a flow chart of the method for extracting and purifying high-content emodin of the present invention.

Detailed Description

The invention is described in further detail below with reference to the following figures and specific examples:

the following is an embodiment of a specific example one of the high emodin extraction and purification methods of the present invention, as shown in fig. 1;

s1, weighing 100g of giant knotweed raw material (emodin content is 2%), cleaning, airing and crushing to obtain giant knotweed crushed material, and soaking the giant knotweed crushed material in acetone for 10 hours;

s2, using 1200g of 80% acetone, extracting polygonum cuspidatum crushed material for three times at the temperature of 65 ℃ and under the condition that the pH is =5.5, filtering, combining the filtrates, concentrating to obtain an extract, drying the extract under the vacuum condition, and crushing the dried extract;

s3, dissolving the dried crushed aggregates by using ethyl acetate at the temperature of 80 ℃, adding 10g of activated carbon to remove impurities and filtering, filtering out precipitates by standing, and measuring the content of the crude emodin product to be 42 percent and 4.5 g of crude emodin product;

wherein the weight of the ethyl acetate is 15 times of the weight of the dried crushed materials;

s4, refluxing, stirring and hot washing the precipitate for 6 hours at the temperature of 88 ℃ by using an N-N dimethylformamide solution, carrying out suction filtration while the precipitate is hot, and respectively collecting insoluble substances and filtrate;

refluxing, stirring and hot washing the insoluble substances again under the same condition, collecting the precipitate and drying; detecting that the content of the emodin is 94 percent, and 1.67 g of crude product;

wherein the weight of the N-N dimethylformamide solution is 15 times of the weight of the precipitate;

s5, crystallizing the dried precipitate by using ethyl acetate with 20g of activated carbon content, and filtering out crystals;

wherein the temperature of the ethyl acetate is 80 ℃, and the weight of the ethyl acetate is 40 times of the weight of the dried precipitate;

s6, stirring and washing the crystal by methanol at normal temperature, and drying to obtain pure emodin HPLC98.3% and 1.56 g;

wherein the weight of methanol used was 3 times the weight of the crystals.

The following is an embodiment of a second specific example of the method for extracting and purifying high-content emodin of the present invention, as shown in fig. 1;

s1, weighing 1kg of giant knotweed raw material (emodin content is 2%), cleaning, airing and crushing to obtain giant knotweed crushed material, and soaking the giant knotweed crushed material in acetone for 10 hours;

s2, extracting polygonum cuspidatum crushed aggregates for three times by using 12kg of 80% acetone at the temperature of 65 ℃ and under the condition that the pH is =5.5, filtering, combining filtrates, concentrating into an extract, drying the extract under the vacuum condition, and crushing the dried extract;

s3, dissolving the dried crushed aggregates by using ethyl acetate at the temperature of 80 ℃, adding 100g of activated carbon to remove impurities and filtering, filtering out precipitates by standing, and measuring the content of crude emodin product to be 41.8 percent and 46 g of crude emodin product;

wherein the weight of the ethyl acetate is 15 times of the weight of the dried crushed materials;

s4, refluxing, stirring and hot washing the precipitate for 6 hours at the temperature of 88 ℃ by using an N-N dimethylformamide solution, carrying out suction filtration while the precipitate is hot, and respectively collecting insoluble substances and filtrate;

refluxing, stirring and hot washing the insoluble substances again under the same condition, collecting the precipitate and drying; detecting 17 g of crude product with the emodin content of 94.1 percent;

wherein the weight of the N-N dimethylformamide solution is 15 times of the weight of the precipitate;

s5, crystallizing the dried precipitate by using ethyl acetate with the content of active carbon of 200g, and filtering out crystals;

wherein the temperature of the ethyl acetate is 80 ℃, and the weight of the ethyl acetate is 40 times of the weight of the dried precipitate;

s6, stirring and washing the crystal by methanol at normal temperature, and drying to obtain pure emodin HPLC98.3% and 15.5 g;

wherein the weight of methanol used was 3 times the weight of the crystals.

The following is an embodiment of a third specific example of the method for extracting and purifying high-content emodin of the present invention, as shown in fig. 1;

s1, weighing 10kg of giant knotweed raw material (emodin content is 2%), cleaning, airing and crushing to obtain giant knotweed crushed material, and soaking the giant knotweed crushed material in acetone for 10 hours;

s2, extracting polygonum cuspidatum crushed aggregates for three times by using 120kg of 80% acetone at the temperature of 65 ℃ and under the condition that the pH is =5.5, filtering, combining filtrates, concentrating into an extract, drying the extract under the vacuum condition, and crushing the dried extract;

s3, dissolving the dried crushed aggregates by using ethyl acetate at the temperature of 80 ℃, adding 1kg of activated carbon to remove impurities and filtering, filtering out precipitates by standing, and measuring the content of crude emodin product to be 42.3 percent and 454 g of crude emodin product;

wherein the weight of the ethyl acetate is 15 times of the weight of the dried crushed materials;

s4, refluxing, stirring and hot washing the precipitate for 6 hours at the temperature of 88 ℃ by using an N-N dimethylformamide solution, carrying out suction filtration while the precipitate is hot, and respectively collecting insoluble substances and filtrate;

refluxing, stirring and hot washing the insoluble substances again under the same condition, collecting the precipitate and drying; detecting that the content of the emodin is 94 percent, and 174 g of crude product;

wherein the weight of the N-N dimethylformamide solution is 15 times of the weight of the precipitate;

s5, crystallizing the dried precipitate by using ethyl acetate with the activated carbon content of 2kg, and filtering out crystals;

wherein the temperature of the ethyl acetate is 80 ℃, and the weight of the ethyl acetate is 40 times of the weight of the dried precipitate;

s6, stirring and washing the crystal by methanol at normal temperature, and drying to obtain pure emodin HPLC98.18% and 155 g pure emodin;

wherein the weight of methanol used was 3 times the weight of the crystals.

The method has the advantages of simple operation, good effect, simple process and easy realization of industrialization; the method has the advantages of high product yield, high comprehensive solvent utilization rate and capability of avoiding waste of raw materials; the method ensures that the production period of the product is short, thereby being capable of meeting the market demand in time.

The above description is only an embodiment of the present invention, and is not intended to limit the scope of the present invention, and all equivalent structural changes made by using the contents of the present specification and the drawings, or applied directly or indirectly to other related technical fields, are included in the scope of the present invention.