CN103242279A - Method for extracting and purifying flavonoid substances in hawthorn - Google Patents
Method for extracting and purifying flavonoid substances in hawthorn Download PDFInfo
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Abstract
The invention relates to a method for extracting and purifying flavonoid substances in hawthorn. The common method for extracting flavone has the overall defects that the obtained total flavone is more in impurity, low in color value, poor in selectivity and low in target product concentration. The method provided by the invention comprises the following steps of: soaking dry hawthorn powder in an excessive dilute alkali solution, heating, preserving heat, extracting and filtering; carrying out vacuum concentration on filtrate, and adding ethanol; refrigerating at 4 DEG C, and removing precipitates; carrying out vacuum concentration until the weight is constant, and then drying to form a flavone extract; dissolving the extract in ethanol, adding a metal salt solution, and adjusting the pH value until a flocculent precipitate appears; and dissolving the precipitate in ethanol, adding EDTA (ethylene diamine tetraacetic acid) to dissociate the complexation of metal ions, carrying out rotary evaporation to dry the solution, then dissolving with methanol, filtering, carrying out vacuum concentration until the weight is constant, and drying to finish the purification. According to the method, a metal ion complexation method is adopted to extract and purify the flavonoid substances in the hawthorn, the flavone concentration in the extract can be improved by 9.34%, and the process is simple and has a significant purification effect. Compared with the existing purification method, the method is greatly improved in cost, operation and effect.
Description
Technical field
The present invention relates to a kind of extracting and purifying method, be specifically related to the extracting and purifying method of Flavonoid substances in a kind of hawthorn.
Background technology
Flavones is a class polyphenolic substance, extensively is present in the various plants, has stronger physiologically active, receives publicity as a kind of healthcare products in recent years.Hawthorn (Hawthorn) is " integration of drinking and medicinal herbs " plant, and Flavonoid substances (Flavonoids) content is higher, mainly comprises Vitexin, Quercetin and Quercetin 3-galactoside.From hawthorn, extract flavonoid compound and can improve the added value of hawthorn.At present, the overall fault of common flavones extracting method is that the total flavones impurity of gained is many, the look valency is low, and is the most general with the hot water extraction method in the solvent-extraction process, but its poor selectivity, and target product concentration is on the low side.Alkali liquor extraction method can increase the extraction choice of Solvent, thereby improves target product concentration, but selects suitable extracting solution to become the key problem of extraction process.
Summary of the invention
The extracting and purifying method that the purpose of this invention is to provide Flavonoid substances in the hawthorn of flavones concentration in a kind of effective raising extract.
The technical solution adopted in the present invention is:
The extracting and purifying method of Flavonoid substances in the hawthorn is characterized in that:
Realized by following steps:
Step 1: extract:
(1) dried hawthorn fruit is immersed in the dilute alkaline soln, is heated to 70-90 ℃ of insulation, Flavonoid substances fully is dissolved in the water, so extract three times, preceding twice 1h, 30min filters the back merging filtrate for the third time;
(2) filtrate is carried out concentrating under reduced pressure in Rotary Evaporators, makes filter liquor concentration reach 40%-50%, adds ethanolic soln in concentrated solution, makes the alcoholic degree of mixed solution reach 70%;
(3) mixed solution is refrigerated 12h under 4 ℃ of temperature, remove precipitation;
(4) will remove the mixed solution that precipitates and place Rotary Evaporators to be evaporated to constant weight, reclaim the ethanol in the solution simultaneously;
(5) concentrated solution that step 4 is obtained places loft drier to carry out drying, treats to take out after it forms flavones medicinal extract shape;
Step 2: purifying:
(1) flavones medicinal extract is dissolved in the ethanolic soln, makes the quality volumetric concentration of flavones medicinal extract in the mixed solution reach 10mg/mL;
(2) add the metal salt solution of 20-70 mg/mL in the mixed solution, regulate the pH value to flocks occurring;
(3) collecting precipitation, and be dissolved in the ethanolic soln, adding EDTA ultra-sonic oscillation reaction in 60 ℃ of water-baths dissolves it fully, the addition of EDTA is 40% of the middle flavones medicinal extract amount of step (1), places Rotary Evaporators to carry out concentrating under reduced pressure, to the solution rotating evaporate to dryness, use dissolve with methanol then, filter, filtrate is rotated evaporate to dryness again, and reclaim methyl alcohol.Use dissolve with methanol again, filter, filtrate is concentrated and is drying to obtain product behind the purifying.
In (1) of step 1, dilute alkaline soln is chosen the NaOH solution that massfraction is 0.4%-0.6%.
In (2) of step 1, the massfraction of ethanolic soln is 95%.
In (1) of step 2, the massfraction of ethanolic soln is 60%.
In (2) of step 2, metal salt solution is selected from copper-bath, solution of zinc sulfate, liquor alumini chloridi, ferric chloride Solution.
In (3) of step 2, the massfraction of ethanolic soln is 30%.
The present invention has the following advantages:
The present invention adopts the Flavonoid substances in the metal ion complexation extraction hawthorn and carries out purifying, and flavones concentration can improve 9.34% in the extract, and technology is simple and purification effect is remarkable.Than existing purification process, bigger improvement is arranged on cost, operation and effect.
Embodiment
The present invention will be described in detail below in conjunction with embodiment.
The extracting and purifying method of Flavonoid substances in the hawthorn involved in the present invention, realized by following steps:
Step 1: extract:
(1) dried hawthorn fruit is immersed in the dilute alkaline soln, is heated to 70-90 ℃ of insulation, Flavonoid substances fully is dissolved in the water, so extract three times, preceding twice 1h, 30min filters the back merging filtrate for the third time;
Dilute alkaline soln is chosen the NaOH solution that massfraction is 0.4%-0.6%.
(2) filtrate is carried out concentrating under reduced pressure in Rotary Evaporators, makes filter liquor concentration reach 40%-50%, adds ethanolic soln in concentrated solution, makes the alcoholic degree of mixed solution reach 70%;
The massfraction of ethanolic soln is 95%.
(3) mixed solution is refrigerated 12h under 4 ℃ of temperature, remove precipitation.
(4) will remove the mixed solution that precipitates and place Rotary Evaporators to be evaporated to constant weight, reclaim the ethanol in the solution simultaneously.
(5) concentrated solution that step 4 is obtained places loft drier to carry out drying, treats to take out after it forms flavones medicinal extract shape.
Step 2: purifying:
(1) flavones medicinal extract is dissolved in the ethanolic soln, makes the quality volumetric concentration of flavones medicinal extract in the mixed solution reach 10mg/mL;
The massfraction of ethanolic soln is 60%.
(2) add the metal salt solution of 20-70 mg/mL in the mixed solution, regulate the pH value to flocks occurring;
Metal salt solution is selected from copper-bath, solution of zinc sulfate, liquor alumini chloridi, ferric chloride Solution.
(3) collecting precipitation, and be dissolved in the ethanolic soln, adding EDTA ultra-sonic oscillation reaction in 60 ℃ of water-baths dissolves it fully, the addition of EDTA is 40% of the middle flavones medicinal extract amount of step (1), places Rotary Evaporators to carry out concentrating under reduced pressure, to the solution rotating evaporate to dryness, use dissolve with methanol then, filter, filtrate is rotated evaporate to dryness again, and reclaim methyl alcohol.Use dissolve with methanol again, filter, filtrate is concentrated and is drying to obtain product behind the purifying.
The massfraction of ethanolic soln is 30%.
Embodiment 1:
Step 1: extract:
(1) dried hawthorn fruit is immersed in the dilute alkaline soln, is heated to 70 ℃ of insulations, Flavonoid substances fully is dissolved in the water, so extract three times, preceding twice 1h, 30min filters the back merging filtrate for the third time;
It is 0.4% NaOH solution that dilute alkaline soln is chosen massfraction.
(2) filtrate is carried out concentrating under reduced pressure in Rotary Evaporators, makes filter liquor concentration reach 40%, adds ethanolic soln in concentrated solution, makes the alcoholic degree of mixed solution reach 70%;
The massfraction of ethanolic soln is 95%.
(3) mixed solution is refrigerated 12h under 4 ℃ of temperature, remove precipitation.
(4) will remove the mixed solution that precipitates and place Rotary Evaporators to be evaporated to constant weight, reclaim the ethanol in the solution simultaneously.
(5) concentrated solution that step 4 is obtained places loft drier to carry out drying, treats to take out after it forms flavones medicinal extract shape.
Step 2: purifying:
(1) flavones medicinal extract is dissolved in the ethanolic soln, makes the quality volumetric concentration of flavones medicinal extract in the mixed solution reach 10mg/mL;
The massfraction of ethanolic soln is 60%.
(2) add the metal salt solution of 20mg/mL in the mixed solution, regulate the pH value to flocks occurring;
Metal salt solution is chosen copper-bath.
(3) collecting precipitation, and be dissolved in the ethanolic soln, adding EDTA ultra-sonic oscillation reaction in 60 ℃ of water-baths dissolves it fully, the addition of EDTA is 40% of the middle flavones medicinal extract amount of step (1), places Rotary Evaporators to carry out concentrating under reduced pressure, to the solution rotating evaporate to dryness, use dissolve with methanol then, filter, filtrate is rotated evaporate to dryness again, and reclaim methyl alcohol.Use dissolve with methanol again, filter, filtrate is concentrated and is drying to obtain product behind the purifying.
The massfraction of ethanolic soln is 30%.
Embodiment 2:
Step 1: extract:
(1) dried hawthorn fruit is immersed in the dilute alkaline soln, is heated to 80 ℃ of insulations, Flavonoid substances fully is dissolved in the water, so extract three times, preceding twice 1h, 30min filters the back merging filtrate for the third time;
It is 0.5% NaOH solution that dilute alkaline soln is chosen massfraction.
(2) filtrate is carried out concentrating under reduced pressure in Rotary Evaporators, makes filter liquor concentration reach 45%, adds ethanolic soln in concentrated solution, makes the alcoholic degree of mixed solution reach 70%;
The massfraction of ethanolic soln is 95%.
(3) mixed solution is refrigerated 12h under 4 ℃ of temperature, remove precipitation.
(4) will remove the mixed solution that precipitates and place Rotary Evaporators to be evaporated to constant weight, reclaim the ethanol in the solution simultaneously.
(5) concentrated solution that step 4 is obtained places loft drier to carry out drying, treats to take out after it forms flavones medicinal extract shape.
Step 2: purifying:
(1) flavones medicinal extract is dissolved in the ethanolic soln, makes the quality volumetric concentration of flavones medicinal extract in the mixed solution reach 10mg/mL;
The massfraction of ethanolic soln is 60%.
(2) add the metal salt solution of 45 mg/mL in the mixed solution, regulate the pH value to flocks occurring;
Metal salt solution is chosen solution of zinc sulfate.
(3) collecting precipitation, and be dissolved in the ethanolic soln, adding EDTA ultra-sonic oscillation reaction in 60 ℃ of water-baths dissolves it fully, the addition of EDTA is 40% of the middle flavones medicinal extract amount of step (1), places Rotary Evaporators to carry out concentrating under reduced pressure, to the solution rotating evaporate to dryness, use dissolve with methanol then, filter, filtrate is rotated evaporate to dryness again, and reclaim methyl alcohol.Use dissolve with methanol again, filter, filtrate is concentrated and is drying to obtain product behind the purifying.
The massfraction of ethanolic soln is 30%.
Embodiment 3:
Step 1: extract:
(1) dried hawthorn fruit is immersed in the dilute alkaline soln, is heated to 90 ℃ of insulations, Flavonoid substances fully is dissolved in the water, so extract three times, preceding twice 1h, 30min filters the back merging filtrate for the third time;
It is 0.6% NaOH solution that dilute alkaline soln is chosen massfraction.
(2) filtrate is carried out concentrating under reduced pressure in Rotary Evaporators, makes filter liquor concentration reach 50%, adds ethanolic soln in concentrated solution, makes the alcoholic degree of mixed solution reach 70%;
The massfraction of ethanolic soln is 95%.
(3) mixed solution is refrigerated 12h under 4 ℃ of temperature, remove precipitation.
(4) will remove the mixed solution that precipitates and place Rotary Evaporators to be evaporated to constant weight, reclaim the ethanol in the solution simultaneously.
(5) concentrated solution that step 4 is obtained places loft drier to carry out drying, treats to take out after it forms flavones medicinal extract shape.
Step 2: purifying:
(1) flavones medicinal extract is dissolved in the ethanolic soln, makes the quality volumetric concentration of flavones medicinal extract in the mixed solution reach 10mg/mL;
The massfraction of ethanolic soln is 60%.
(2) add the metal salt solution of 70 mg/mL in the mixed solution, regulate the pH value to flocks occurring;
Metal salt solution is chosen liquor alumini chloridi.
(3) collecting precipitation, and be dissolved in the ethanolic soln, adding EDTA ultra-sonic oscillation reaction in 60 ℃ of water-baths dissolves it fully, the addition of EDTA is 40% of the middle flavones medicinal extract amount of step (1), places Rotary Evaporators to carry out concentrating under reduced pressure, to the solution rotating evaporate to dryness, use dissolve with methanol then, filter, filtrate is rotated evaporate to dryness again, and reclaim methyl alcohol.Use dissolve with methanol again, filter, filtrate is concentrated and is drying to obtain product behind the purifying.
The massfraction of ethanolic soln is 30%.
Embodiment 4:
Step 1: extract:
(1) dried hawthorn fruit is immersed in the dilute alkaline soln, is heated to 90 ℃ of insulations, Flavonoid substances fully is dissolved in the water, so extract three times, preceding twice 1h, 30min filters the back merging filtrate for the third time;
It is 0.6% NaOH solution that dilute alkaline soln is chosen massfraction.
(2) filtrate is carried out concentrating under reduced pressure in Rotary Evaporators, makes filter liquor concentration reach 50%, adds ethanolic soln in concentrated solution, makes the alcoholic degree of mixed solution reach 70%;
The massfraction of ethanolic soln is 95%.
(3) mixed solution is refrigerated 12h under 4 ℃ of temperature, remove precipitation.
(4) will remove the mixed solution that precipitates and place Rotary Evaporators to be evaporated to constant weight, reclaim the ethanol in the solution simultaneously.
(5) concentrated solution that step 4 is obtained places loft drier to carry out drying, treats to take out after it forms flavones medicinal extract shape.
Step 2: purifying:
(1) flavones medicinal extract is dissolved in the ethanolic soln, makes the quality volumetric concentration of flavones medicinal extract in the mixed solution reach 10mg/mL;
The massfraction of ethanolic soln is 60%.
(2) add the metal salt solution of 70 mg/mL in the mixed solution, regulate the pH value to flocks occurring;
Metal salt solution is chosen ferric chloride Solution.
(3) collecting precipitation, and be dissolved in the ethanolic soln, adding EDTA ultra-sonic oscillation reaction in 60 ℃ of water-baths dissolves it fully, the addition of EDTA is 40% of the middle flavones medicinal extract amount of step (1), places Rotary Evaporators to carry out concentrating under reduced pressure, to the solution rotating evaporate to dryness, use dissolve with methanol then, filter, filtrate is rotated evaporate to dryness again, and reclaim methyl alcohol.Use dissolve with methanol again, filter, filtrate is concentrated and is drying to obtain product behind the purifying.
The massfraction of ethanolic soln is 30%.
It is cited that content of the present invention is not limited to embodiment, and the conversion of any equivalence that those of ordinary skills take technical solution of the present invention by reading specification sheets of the present invention is claim of the present invention and contains.
Claims (6)
1. the extracting and purifying method of Flavonoid substances in the hawthorn is characterized in that:
Realized by following steps:
Step 1: extract:
(1) dried hawthorn fruit is immersed in the dilute alkaline soln, is heated to 70-90 ℃ of insulation, Flavonoid substances fully is dissolved in the water, so extract three times, preceding twice 1h, 30min filters the back merging filtrate for the third time;
(2) filtrate is carried out concentrating under reduced pressure in Rotary Evaporators, makes filter liquor concentration reach 40%-50%, adds ethanolic soln in concentrated solution, makes the alcoholic degree of mixed solution reach 70%;
(3) mixed solution is refrigerated 12h under 4 ℃ of temperature, remove precipitation;
(4) will remove the mixed solution that precipitates and place Rotary Evaporators to be evaporated to constant weight, reclaim the ethanol in the solution simultaneously;
(5) concentrated solution that step 4 is obtained places loft drier to carry out drying, treats to take out after it forms flavones medicinal extract shape;
Step 2: purifying:
(1) flavones medicinal extract is dissolved in the ethanolic soln, makes the quality volumetric concentration of flavones medicinal extract in the mixed solution reach 10mg/mL;
(2) add the metal salt solution of 20-70 mg/mL in the mixed solution, regulate the pH value to flocks occurring;
(3) collecting precipitation, and be dissolved in the ethanolic soln, adding EDTA ultra-sonic oscillation reaction in 60 ℃ of water-baths dissolves it fully, the addition of EDTA is 40% of the middle flavones medicinal extract amount of step (1), places Rotary Evaporators to carry out concentrating under reduced pressure, to the solution rotating evaporate to dryness, use dissolve with methanol then, filter, filtrate is rotated evaporate to dryness again, and reclaim methyl alcohol;
Use dissolve with methanol again, filter, filtrate is concentrated and is drying to obtain product behind the purifying.
2. according to the extracting and purifying method of Flavonoid substances in the hawthorn described in the claim 1, it is characterized in that:
In (1) of step 1, dilute alkaline soln is chosen the NaOH solution that massfraction is 0.4%-0.6%.
3. according to the extracting and purifying method of Flavonoid substances in the hawthorn described in claim 1 or 2, it is characterized in that:
In (2) of step 1, the massfraction of ethanolic soln is 95%.
4. according to the extracting and purifying method of Flavonoid substances in the hawthorn described in the claim 3, it is characterized in that:
In (1) of step 2, the massfraction of ethanolic soln is 60%.
5. according to the extracting and purifying method of Flavonoid substances in the hawthorn described in the claim 4, it is characterized in that:
In (2) of step 2, metal salt solution is selected from copper-bath, solution of zinc sulfate, liquor alumini chloridi, ferric chloride Solution.
6. according to the extracting and purifying method of Flavonoid substances in the hawthorn described in the claim 5, it is characterized in that:
In (3) of step 2, the massfraction of ethanolic soln is 30%.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104643065A (en) * | 2015-01-28 | 2015-05-27 | 郑州大学 | Hawthorn omega-3 soft capsule and preparation method thereof |
| CN105687297A (en) * | 2016-02-17 | 2016-06-22 | 蕴彤本草（武汉）生物科技有限公司 | Mung bean extract and preparation method thereof |
| CN109042765A (en) * | 2018-09-29 | 2018-12-21 | 河北工程大学 | A kind of pharmaceutical composition and preparation method thereof of environmentally protective environment disinfectant |
| CN109528843A (en) * | 2018-11-28 | 2019-03-29 | 河北科技大学 | A kind of extracting method of flavone of hawthorn fruit |
| CN112353724A (en) * | 2020-11-20 | 2021-02-12 | 太和康美(北京)中医研究院有限公司 | Plant extract and preparation method and application thereof |
| CN116211986A (en) * | 2023-04-03 | 2023-06-06 | 北京安德普泰医疗科技有限公司 | A Curcumae rhizoma extract, and its preparation method and application |
| CN116407583A (en) * | 2023-04-26 | 2023-07-11 | 华北理工大学 | Method for extracting total flavonoids from hawthorns |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104643065A (en) * | 2015-01-28 | 2015-05-27 | 郑州大学 | Hawthorn omega-3 soft capsule and preparation method thereof |
| CN105687297A (en) * | 2016-02-17 | 2016-06-22 | 蕴彤本草（武汉）生物科技有限公司 | Mung bean extract and preparation method thereof |
| CN105687297B (en) * | 2016-02-17 | 2020-07-14 | 蕴彤本草（武汉）生物科技有限公司 | Mung bean extract and preparation method thereof |
| CN109042765A (en) * | 2018-09-29 | 2018-12-21 | 河北工程大学 | A kind of pharmaceutical composition and preparation method thereof of environmentally protective environment disinfectant |
| CN109528843A (en) * | 2018-11-28 | 2019-03-29 | 河北科技大学 | A kind of extracting method of flavone of hawthorn fruit |
| CN112353724A (en) * | 2020-11-20 | 2021-02-12 | 太和康美(北京)中医研究院有限公司 | Plant extract and preparation method and application thereof |
| CN116211986A (en) * | 2023-04-03 | 2023-06-06 | 北京安德普泰医疗科技有限公司 | A Curcumae rhizoma extract, and its preparation method and application |
| CN116211986B (en) * | 2023-04-03 | 2024-04-30 | 北京安德普泰医疗科技有限公司 | A Curcumae rhizoma extract, and its preparation method and application |
| CN116407583A (en) * | 2023-04-26 | 2023-07-11 | 华北理工大学 | Method for extracting total flavonoids from hawthorns |
| CN116407583B (en) * | 2023-04-26 | 2024-03-26 | 华北理工大学 | Method for extracting total flavonoids from hawthorns |
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Effective date of registration: 20210527 Address after: No.157, Xiguan new village, Jincheng City, Shanxi Province 048000 Patentee after: Shanxi Kanglisheng Pharmaceutical Co.,Ltd. Address before: 710021 Shaanxi city of Xi'an province Weiyang University City Patentee before: SHAANXI University OF SCIENCE & TECHNOLOGY |