CN108329369A - A kind of preparation method of high-content rutin - Google Patents

Abstract

The invention discloses a preparation method of high-content rutin, which belongs to the technical field of medicinal chemistry. The key points of the technical scheme of the present invention are: a preparation method of high-content rutin, which comprises acetylating the raw material crude rutin to obtain acetylated rutin; purifying the acetylated rutin to obtain high-purity acetylated rutin; The purified acetylated rutin is hydrolyzed to obtain a high content of rutin. The refining effect of the present invention is better, and the purity of the obtained rutin can reach more than 99.0%, and the yield of the rutin prepared by the method is relatively high, stably maintained at 95%-97%, and the cost of producing rutin is far lower than the existing The production cost of high-content rutin in the reported patents and literature is suitable for industrial application.

Description

A kind of preparation method of high-content rutin

technical field

The invention belongs to the technical field of medicinal chemistry, and in particular relates to a preparation method of high-content rutin.

Background technique

Rutin (Rutin), also known as rutin and purple quercetin, is the 3-O-rutinoside of quercetin, the only flavonoid compound with a wide source, mainly found in the flower buds of the leguminous plant Sophora japonica (Sophora japonica), Fruit (Sophora japonica), Rutaceae Rutaceae whole herb, Hypericaceae red Eclipta whole herb and Polygonaceae buckwheat seedlings. It is yellow or yellow-green powder or very fine needle-like crystal; odorless and tasteless; easy to deteriorate when exposed to light, the color gradually becomes darker in the air, easily soluble in boiling methanol, soluble in hot ethanol, alkaline solution, slightly soluble in hot water Water and cold methanol, insoluble in cold water, ethanol, chloroform, petroleum ether, acetone and ethyl acetate, the maximum ultraviolet absorption of methanol liquid is at 258nm and 361nm. The molecular formula is C ₂₇ H ₃₀ O ₁₆ , CAS NO.153-18-4, the molecular weight is 610.51, and the melting point is 190-192°C. The structural formula is as follows:

As natural traditional Chinese medicine compounds, rutin and its derivatives have a wide range of pharmacological activities, not only anti-myocardial hypoxia, ischemia, anti-arrhythmia, lowering serum cholesterol, inhibiting platelet aggregation, anti-ulcer, anti-tumor, anti-inflammatory, anti- Allergy, anti-aging, anti-radiation, anti-virus and immunity-enhancing functions, clinically used to prevent and treat cerebral hemorrhage, hypertension, retinal hemorrhage, purpura and acute hemorrhagic nephritis, the total effective rate of its treatment of chronic bronchitis is 84.4%; at the same time It also has a significant effect on the treatment or alleviation of modern civilization diseases such as cardiovascular and cerebrovascular diseases, obesity, diabetes and ventilation; it can also be used as a functional food. The pharmaceutical industry at home and abroad uses rutin as a raw material to produce rutin tablets, troxerutin, rutin magnesium chromium salt, multidimensional glucose oxybuterutin and quercetin, which expands the application scope and market of rutin.

At present, my country's rutin production mainly uses Huai Mi as raw material for extraction. Since rutin is easily soluble in lye and precipitates after adding acid, the extraction of rutin in laboratories and enterprises is often carried out by alkali extraction and acid precipitation. Because there are a lot of flavonoids in Sophora japonica, and the structure is similar to rutin, it is difficult to achieve a high rutin content by conventional extraction methods, and the HPLC content is generally 90%-95%, and the impurities in crude rutin The presence of rutin will affect the quality of rutin raw materials and the quality of troxerutin prepared from rutin, so it is particularly important to prepare high-content rutin.

The refining methods of rutin are as follows: (1) Zhao Wenbin et al. reported the method of utilizing molecular size separation, after dissolving rutin with methanol, adding Sephadex column top, eluting with methanol, collecting methanol liquid, Concentrated and crystallized. The principle is that since large molecules cannot enter the interior of the Sephadex gel, they are eluted first, and small molecules are eluted later, so that components with different molecular sizes are separated. (2) Wang Houquan et al. reported that rutin was dissolved in alcohol at room temperature, filtered, the filtrate was left to crystallize, and the mother liquor was concentrated and crystallized again to obtain high-content rutin. The method transforms the two-dimensional planar structure of rutin into a three-dimensional three-dimensional structure, which facilitates the entry of solvent molecules, increases solubility, and can remove some impurities dissolved in alcohol after heating. (3) Shen Jihong et al. reported that rutin was purified by trichloroacetic acid deproteinization and improved Sevage deproteinization. This method is simple and labor-saving, but the reagents used are highly toxic and the purity is only 90%, which is difficult to meet the current requirements. Demand for high levels of rutin. (4) Qian Yuanhua and others reported that high-purity rutin was refined by high-speed countercurrent chromatography, which is characterized by the use of water-containing, multi-component organic solvent system, and the use of high-speed countercurrent chromatography without irreversible adsorption, high recovery and high separation efficiency. Refined to obtain High content of rutin. The purity of the rutin prepared by the method can reach more than 98%, but the use of various organic solvents has a certain impact on the environment, and at the same time, the preparation is only at the gram level, and the cost is high, which is not conducive to industrial production. (5) Liu Zhiqiang uses resin to carry out column chromatography to refine rutin. In this method, the obtained rutin is rinsed with deionized water and dried at 80-90°C for 10-14h, because the long-term drying at a higher temperature is especially air-infected. Or under the condition that light exists, the oxidation or decomposition of part of rutin is easy to occur, so there are certain defects.

In the existing reports, the method of refining rutin with a purity of more than 98% either requires chromatography and is cumbersome to operate; or uses too many toxic organic solvents, which is not conducive to environmental protection; or the cost is high and is not suitable for industrialization.

Contents of the invention

The technical problem solved by the present invention is to provide a method for preparing high-content rutin, which utilizes the performance difference of acylated rutin and acylated impurities to purify rutin, and then hydrolyzes to obtain high-purity rutin. The prepared rutin has the advantages of high yield (above 95%), high purity (above 99.0%), low cost, economy and environmental protection, and suitability for industrialization. It is a refining method with industrial production value, and the cost is only 100 -160 yuan/kg.

The present invention adopts following technical scheme for solving the above-mentioned technical problem, a kind of preparation method of high-content rutin is characterized in that concrete steps are:

(1) Carrying out acetylation treatment of raw material crude rutin to obtain acetylated rutin;

(2) recrystallizing acetylated rutin to obtain purified acetylated rutin;

(3) Hydrolyzing the purified acetylated rutin to obtain high-content rutin.

Further preferably, the specific process of step (1) is as follows: dissolve the raw material crude rutin in an alkaline solution, add the acetylation reagent dropwise under ice-water bath conditions after dissolving, heat to reflux, and add the reaction solution to the ice water after fully reacting. Crystallization in aqueous solution, suction filtration to obtain acetylated rutin.

Further preferably, the crude rutin is commercially available common rutin, its HPLC content is 92%, and the alkaline solution is one of sodium hydroxide solution, pyridine, triethylamine, sodium acetate solution, DMAP or DMSO. one or more species.

Further preferably, the acetylation reagent is acetyl chloride, glacial acetic acid or acetic anhydride.

Further preferably, the feeding ratio of the crude rutin, the alkaline solution and the acetylation reagent is 1mol:3-4L:12mol, and the feeding ratio of the ice-water solution and the crude rutin is 80-150L:1Kg.

Further preferably, the specific process of step (2) is: heating and dissolving the acetylated rutin in a solvent, then adding a mixture of boric acid and activated carbon with a weight ratio of 1:1, filtering, then cooling down and standing for crystallization or adding dropwise to the solvent The purified acetylated rutin was obtained by crystallization at moderate temperature.

Further preferably, the solvent is one or more of isopropanol, methanol, ethanol, water, ethyl acetate or petroleum ether.

Further preferably, the weight of the mixture of boric acid and activated carbon is 5%-10% of the weight of crude rutin.

Further preferably, the specific process of step (3) is; the purified acetylated rutin is dissolved in methanol at room temperature, and then the methanol solution of alkali is added dropwise. The pH value of the mixed system is adjusted to 5-6 by the acidic solution, and the high-content rutin is obtained by suction filtration.

Further preferably, the alkali in the methanol solution of alkali is sodium methoxide, ammonia or imidazole, wherein the molar ratio of alkali to crude rutin is 10-15:1.

Further preferably, the acidic solution is one or more of hydrochloric acid, sulfuric acid, nitric acid or acetic acid.

Compared with the prior art, the present invention has the following advantages: the refining effect of the present invention is better, and the purity of the obtained rutin can reach more than 99.0%. %, the cost is only 5%-10% higher than that of commercially available rutin, the cost of producing rutin is far lower than the production cost of high-content rutin in reported patents and literature, and is suitable for industrial application.

Detailed ways

The above-mentioned contents of the present invention are described in further detail below through the embodiments, but this should not be interpreted as the scope of the above-mentioned themes of the present invention being limited to the following embodiments, and all technologies realized based on the above-mentioned contents of the present invention all belong to the scope of the present invention.

Example 1

Add 150mL of pyridine to 30.5g (0.05mol) of crude rutin, stir until dissolved, slowly drop in 61g (0.6mol) of acetic anhydride in an ice-water bath, stir at 70°C for 12h, then drop the reaction solution into 3L of ice-water solution In the process, milky white particles precipitated. After standing at room temperature for crystallization for 4 hours, suction filtration; heated the filter cake with 500mL isopropanol until dissolved, added 3.0g of a mixture of equal mass boric acid and activated carbon, filtered while hot, and then slowly cooled down Crystallization, suction filtration; filter cake is dissolved in 200mL methanol, slowly dropwise thereinto the methanol solution of 32g (0.6mol) sodium methylate again, this moment, solution becomes turbid, adjust the pH value of mixed system with glacial acetic acid to be 5.0, A large amount of yellow precipitates precipitated, left standing at room temperature, and suction filtered to obtain high-content rutin with a yield of 95.3% and a purity of 99.7%.

Example 2

Add 150mL of pyridine to 30.5g (0.05mol) of crude rutin, stir until dissolved, slowly drop in 61g (0.6mol) of acetic anhydride in an ice-water bath, stir at 70°C for 12h, then drop the reaction solution into 3L of ice-water solution In the process, milky white particles were precipitated. After standing at room temperature for crystallization for 4 hours, suction filtration was performed; the filter cake was heated and dissolved with 500 mL of ethyl acetate, and a mixture of 2.5 g of equal mass of boric acid and activated carbon was added, suction filtered while hot, and 500 mL of Slowly cool down and crystallize in petroleum ether, and filter with suction; dissolve the filter cake in 200mL of methanol, and then slowly add 32g (0.6mol) of methanol solution of sodium methoxide dropwise to it, at this time the solution becomes turbid, and the mixed system is adjusted with glacial acetic acid. The pH value was 5.8, and a large amount of yellow precipitates precipitated out. After standing at room temperature, high-content rutin was obtained by suction filtration, with a yield of 95.7% and a purity of 99.4%.

Example 3

Add 150 mL of triethylamine to 30.5 g (0.05 mol) of crude rutin, stir until dissolved, slowly drop 47 g (0.6 mol) of acetyl chloride in an ice-water bath, stir at 70°C for 12 hours, then drop the reaction solution into In 3L of ice-water solution, milky white particles were precipitated. After static crystallization at room temperature for 4 hours, filter with suction; heat the filter cake with 500mL of isopropanol until dissolved, add 1.5g of a mixture of equal mass boric acid and activated carbon, and filter with suction while hot. Then slowly cool down and crystallize, and filter with suction; dissolve the filter cake in 200mL of methanol, and then slowly add 32g (0.6mol) of methanol solution of sodium methoxide dropwise, at this time, the solution becomes turbid, adjust the pH value of the mixed system with dilute hydrochloric acid It was 6.0, and a large amount of yellow precipitates precipitated out. After standing at room temperature, high-content rutin was obtained by suction filtration, with a yield of 97.1% and a purity of 99.0%.

Example 4

Add 150 mL of pyridine to 30.5 g (0.05 mol) of crude rutin, stir until dissolved, slowly add 47 g (0.6 mol) of acetyl chloride dropwise in an ice-water bath, stir at 70°C for 12 hours, then drop the reaction solution into 3 L of ice In the aqueous solution, milky white particles were precipitated. After standing at room temperature for crystallization for 4 hours, suction filtration; the filter cake was heated and dissolved with 500mL ethanol, and a mixture of 3.0g equal mass of boric acid and activated carbon was added, suction filtered while hot, and then dropped into 1L of water Slowly crystallize and filter with suction; dissolve the filter cake in 200mL of methanol, then slowly add 32g (0.6mol) of sodium methoxide in methanol solution dropwise, at this time the solution becomes turbid, adjust the pH value of the mixed system to 5.5 with glacial acetic acid , a large amount of yellow precipitates were precipitated, left standing at room temperature, and suction filtered to obtain high-content rutin with a yield of 96.1% and a purity of 99.8%.

Example 5

Add 150mL of pyridine to 30.5g (0.05mol) of crude rutin, stir until dissolved, slowly drop in 61g (0.6mol) of acetic anhydride in an ice-water bath, stir at 70°C for 12h, then drop the reaction solution into 3L of ice-water solution In the process, milky white particles were precipitated. After standing at room temperature for crystallization for 4 hours, suction filtration; the filter cake was heated to dissolve with 500mL isopropanol, and a mixture of 1.5g equal mass of boric acid and activated carbon was added, suction filtered while hot, and then slowly cooled Crystallize and filter with suction; dissolve the filter cake in 200mL of methanol, then slowly add dropwise a methanol solution of 15g (0.22mol) of imidazole and 6g (0.35mol) of ammonia, at this time the solution becomes cloudy, adjust the mixed system with dilute hydrochloric acid The pH value of the product was 5.0, and a large amount of yellow precipitates were precipitated. After standing at room temperature, high-content rutin was obtained by suction filtration, with a yield of 97.2% and a purity of 99.3%.

Example 6

Add 150 mL of pyridine to 30.5 g (0.05 mol) of crude rutin, stir until dissolved, slowly add 47 g (0.6 mol) of acetyl chloride dropwise in an ice-water bath, stir at 70°C for 12 hours, then drop the reaction solution into 3 L of ice In the aqueous solution, milky white particles were precipitated. After standing at room temperature for crystallization for 4 hours, suction filtration; the filter cake was heated and dissolved with 500mL ethanol, and a mixture of 3.0g equal mass of boric acid and activated carbon was added, suction filtered while hot, and then dropped into 1L of water Crystallization, suction filtration; The filter cake is dissolved in 200mL methanol, then slowly drips the methanol solution of 41g (0.6mol) imidazole therein, this moment, the solution becomes cloudy, adjust the pH value of the mixed system with glacial acetic acid to be 5.6, there is A large amount of yellow precipitates were precipitated, left standing at room temperature, and then filtered with suction to obtain high-content rutin with a yield of 95.8% and a purity of 99.5%.

Example 7

Add 150 mL of pyridine to 30.5 g (0.05 mol) of crude rutin, stir until dissolved, slowly drop into 35 mL (0.6 mol) of glacial acetic acid in an ice-water bath, stir at 70°C for 12 hours, then drop the reaction solution into 3 L of ice In the aqueous solution, milky white particles were precipitated. After standing at room temperature for crystallization for 4 hours, filter with suction; heat the filter cake with 500mL of isopropanol until dissolved, add 1.5g of a mixture of equal mass boric acid and activated carbon, filter while hot, and then slowly Cool down and crystallize, filter with suction; dissolve the filter cake in 200mL of methanol, and then slowly add 32g (0.6mol) of sodium methoxide in methanol solution dropwise, at this time the solution becomes cloudy, adjust the pH value of the mixed system to 5.0 with glacial acetic acid , a large amount of yellow precipitates were precipitated, left standing at room temperature, and suction-filtered to obtain high-content rutin with a yield of 96.1% and a purity of 99.7%.

Example 8

Add 150 mL of pyridine to 30.5 g (0.05 mol) of crude rutin, stir until dissolved, slowly drop into 35 mL (0.6 mol) of glacial acetic acid in an ice-water bath, stir at 70°C for 12 hours, then drop the reaction solution into 3 L of ice In the aqueous solution, milky white particles were precipitated. After standing at room temperature for crystallization for 4 hours, filter with suction; heat the filter cake with 500mL of isopropanol until dissolved, add 1.5g of a mixture of equal mass boric acid and activated carbon, filter while hot, and then slowly Cool down and crystallize, filter with suction; dissolve the filter cake in 200mL of methanol, then slowly add dropwise a methanol solution of 15g (0.22mol) imidazole and 6g (0.35mol) of ammonia, at this time the solution becomes cloudy, adjust the mixture with dilute hydrochloric acid The pH value of the system was 5.5, and a large amount of yellow precipitates precipitated out. After standing at room temperature, high-content rutin was obtained by suction filtration, with a yield of 97.2% and a purity of 99.6%.

Decaacetylated rutin ¹ H NMR (400MHz, CDCl ₃ )δ7.96(dd, J=8.6,2.1Hz,1H),7.91(d,J=2.1Hz,1H),7.34(d,J=8.6 Hz, 1H), 7.31(d, J=2.2Hz, 1H), 6.83(d, J=2.2Hz, 1H), 5.43(d, J=7.8Hz, 1H), 5.28(t, J=9.5Hz, 1H), 5.17(dd, J=9.7, 7.9Hz, 1H), 5.09(s, 1H), 5.07(d, J=3.5Hz, 1H), 4.95(t, J=9.6Hz, 2H), 4.52( s,1H),3.65(dq,J=12.6,6.3Hz,1H),3.55(ddd,J=20.5,10.0,3.1Hz,2H),3.27(dd,J=11.0,5.6Hz,1H),2.44 (s,3H),2.34(d,J=3.1Hz,6H),2.30(s,3H),2.14(s,3H),2.09(s,3H),2.02(s,6H),1.95(d, J＝6.4Hz, 6H), 1.06(d, J＝6.3Hz, 3H).

Rutin ¹ H NMR (400MHz, DMSO-d6) δ12.60(s,1H,C5-H),10.84(s,1H,C7-H),9.69(s,1H,C4'-H),9.20 (s,1H,C3'-H),7.55(d,J=2.1Hz,1H,C6'-H),7.53(s,1H,C2'-H),6.84(d,J=8.3Hz,1H ,C5'-H),6.38(d,J=2.0Hz,1H,C8-H),6.19(d,J=2.0Hz,1H,C6-H),5.34(d,J=7.3Hz,1H) ,5.30(d,J=4.1Hz,1H),5.13(d,J=4.2Hz,1H),5.09(d,J=5.8Hz,1H),4.55(d,J=5.1Hz,1H),4.42 (d,J=5.8Hz,1H),4.37(d,J=3.9Hz,2H),3.70(d,J=10.2Hz,1H),3.10–3.00(m,2H),0.99(d,J= 6.2Hz,3H,CH3 _- R).

The above embodiments have described the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. What are described in the above embodiments and description are only to illustrate the principles of the present invention. Without departing from the scope of the principle of the present invention, there will be various changes and improvements in the present invention, and these changes and improvements all fall within the protection scope of the present invention.

Claims (10)

1. a kind of preparation method of high-content rutin, it is characterised in that the specific steps are：

（1）Crude material rutin progress acetylation process is obtained into acetylation rutin；

（2）Acetylation rutin is recrystallized to give acetylation rutin after purification；

（3）It hydrolyzes acetylation rutin after purification to obtain high-content rutin.

2. the preparation method of high-content rutin according to claim 1, it is characterised in that step（1）Detailed process be： Crude material rutin is dissolved in alkaline solution, acetylation reagent is added dropwise under the conditions of ice-water bath after dissolving, is heated to flowing back, fill Divide after reaction and reaction solution is added to crystallization in ice water solution, suction filtration obtains acetylation rutin.

3. the preparation method of high-content rutin according to claim 2, it is characterised in that：The crude product rutin is commercially available Common rutin, HPLC contents be 92%, the alkaline solution be sodium hydroxide solution, pyridine, triethylamine, sodium acetate solution, It is one or more in DMAP or DMSO.

4. the preparation method of high-content rutin according to claim 2, it is characterised in that：The acetylation reagent is second Acyl chlorides, glacial acetic acid or aceticanhydride.

5. the preparation method of high-content rutin according to claim 2, it is characterised in that：The crude product rutin, alkalinity The charge ratio of solution and acetylation reagent is 1mol:3-4L:12mol, the ice water solution are matched with feeding intake for crude product rutin Than for 80-150L:1Kg.

6. the preparation method of high-content rutin according to claim 1, it is characterised in that step（2）Detailed process be： The heating of acetylation rutin is dissolved in solvent, it is 1 to add weight ratio:Then 1 boric acid and the mixture of activated carbon, filtering cool down It stands crystallization or is added drop-wise in solvent the crystallization that cools down and obtain acetylation rutin after purification.

7. the preparation method of high-content rutin according to claim 6, it is characterised in that：The boric acid and activated carbon The weight of mixture is the 5%-10% of crude product rutin weight.

8. the preparation method of high-content rutin according to claim 6, it is characterised in that：The solvent be isopropanol, It is one or more in methanol, ethyl alcohol, water, ethyl acetate or petroleum ether.

9. the preparation method of high-content rutin according to claim 1, it is characterised in that step（3）Detailed process be； Acetylation rutin after purification is dissolved in methanol at room temperature, then the methanol solution of alkali is added dropwise, solution becomes cloudy at this time, has big It measures yellow mercury oxide to be precipitated, then it is 5-6 to adjust the pH value of mixed system with acid solution, suction filtration obtains high-content rutin.

10. the preparation method of high-content rutin according to claim 9, it is characterised in that：The methanol solution of the alkali Middle alkali is that the molar ratio of sodium methoxide, ammonia or imidazoles, wherein alkali and crude product rutin is 10-15:1, the acid solution be hydrochloric acid, It is one or more in sulfuric acid, nitric acid or acetic acid.