CN109503334B - Preparation process for preparing resveratrol with purity of 99% from polygonum cuspidatum - Google Patents
Preparation process for preparing resveratrol with purity of 99% from polygonum cuspidatum Download PDFInfo
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- CN109503334B CN109503334B CN201811570311.6A CN201811570311A CN109503334B CN 109503334 B CN109503334 B CN 109503334B CN 201811570311 A CN201811570311 A CN 201811570311A CN 109503334 B CN109503334 B CN 109503334B
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- C07C37/004—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring by obtaining phenols from plant material or from animal material
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Abstract
The invention discloses a preparation process for preparing resveratrol with the purity of 99% from giant knotweed, which comprises the steps of adding wine yeast, saccharifying enzyme and aspergillus niger into soaked giant knotweed powder, and fermenting for 6-24 hours at constant temperature in a container at the temperature of 28-50 ℃ to convert analogues of resveratrol into resveratrol; carrying out countercurrent pressurization and low-temperature seepage on the fermented material; and then concentrating and recovering ethanol, dissolving and removing impurities, carrying out alumina column chromatography, crystallizing and recrystallizing to obtain the resveratrol with the purity of more than 99%. The invention improves the purity of the resveratrol to more than 99 percent, improves the product yield from 0.5 to 0.9 percent to more than 1.2 percent, improves the product quality by enabling the whiteness of the resveratrol product to reach more than 80 degrees, is really applied to industrial production, can shorten the production period and improve the production efficiency.
Description
Technical Field
The invention belongs to the field of biochemistry, and particularly relates to a process for extracting resveratrol from giant knotweed rhizome.
Background
Resveratrol is a polyphenol compound with stronger activity, is an important phytoalexin, has obvious effects of inhibiting cancer cells, reducing blood fat, preventing and treating cardiovascular diseases, resisting oxidation, delaying senescence and the like, and is a new green anticancer drug which follows paclitaxel.
Resveratrol was first discovered in 1939, grapes were first discovered to contain this substance in the 20 th century 70 th, and later, plants such as giant knotweed, peanut, mulberry and the like were discovered to contain this substance. Natural resveratrol is a natural active component, can be distributed and biosynthesized in 2 forms of free state (cis form and trans form) and glucoside bound state (cis form and trans form) in plants (such as Chinese medicinal material giant knotweed rhizome), and has antioxidant effect.
At present, most of domestic resveratrol production enterprises adopt giant knotweed rhizome as a raw material for extraction, and the adopted process route is as follows: giant knotweed rhizome material generation → biological conversion → alcohol extraction concentration → active carbon decolorization or extraction → dry column chromatography (silica gel or alumina) → crystallization and recrystallization. The yield of the final product resveratrol in the process route is only 0.5-0.9%, the color is slightly yellowish or grey white, the whiteness is about 70 ℃, the purity of the resveratrol is 97-98%, and the resveratrol is sold as 98% in the market. The price of the product has a larger difference with 99 percent, and how to improve the yield and the product content and simultaneously improve the color and the whiteness of the product to be more than 80 degrees. Is a problem that those skilled in the art and enterprises expect to solve.
Disclosure of Invention
The invention aims to provide a preparation process for preparing resveratrol with the purity of 99% from giant knotweed, which improves the purity of the resveratrol to more than 99%, improves the product yield from 0.5-0.9% to more than 1.2%, enables the whiteness of the resveratrol product to reach more than 80 ℃, improves the product quality, is really applied to industrial production, can shorten the production period and improve the production efficiency.
In order to achieve the technical purpose, the technical scheme adopted by the invention is as follows:
a preparation process for preparing resveratrol with purity of 99% from rhizoma Polygoni Cuspidati comprises the following steps:
(1) enzyme conversion: soaking rhizoma Polygoni Cuspidati powder in water; placing the soaked giant knotweed rhizome powder in a container with the temperature controlled at 28-50 ℃ by using special enzyme, fermenting at constant temperature for 6-24 hours to convert the resveratrol analogue into resveratrol, wherein the special enzyme is wine yeast, saccharifying enzyme and aspergillus niger;
(2) and (3) countercurrent pressurization low-temperature leakage: uniformly loading the fermented polygonum cuspidatum material into a leakage column with the diameter-height ratio of 1:4-5, adding a mixed reagent of ethyl acetate and petroleum ether which is 3-5 times of the weight of the polygonum cuspidatum in total, and slowly passing through a polygonum cuspidatum layer to obtain leakage liquid;
(3) concentrating and recovering ethanol; concentrating the leakage solution of rhizoma Polygoni Cuspidati, and recovering reagent until no reagent exists to obtain extract;
(4) dissolving and removing impurities; dissolving the extract with 60-80% ethanol at room temperature, and filtering to obtain filtrate;
(5) chromatography: introducing the filtrate into an alumina chromatographic column, and eluting with 80-95% ethanol after the whole liquid is introduced into the alumina chromatographic column until no resveratrol is detected;
(6) crystallizing and recrystallizing;
(7) and (4) drying the crystals at low temperature in vacuum to obtain a finished product, wherein the resveratrol is more than or equal to 99%.
Preferably, the ratio of the wine yeast, the saccharifying enzyme and the aspergillus niger is 2:2: 1.
Preferably, the weight ratio of the ethyl acetate to the petroleum ether is 6-4: 1.
As a preferred preparation process for preparing the resveratrol with the purity of 99% from the giant knotweed rhizome, the giant knotweed rhizome root powder is dried before the step (1) and is crushed until more than 90% of the powder can pass through a 20-mesh sieve.
As another preferable preparation process for preparing resveratrol with the purity of 99% from giant knotweed rhizome, the feeding ratio of the special enzyme to the giant knotweed rhizome powder is 1: 99-101.
As another preferable preparation process for preparing resveratrol with the purity of 99% from giant knotweed rhizome, the feeding ratio of the giant knotweed rhizome powder to the water is 10: 2-3.
As another preferable preparation process for preparing the resveratrol with the purity of 99% from the giant knotweed rhizome, the fermentation temperature in the step (1) is 45 degrees, and the fermentation is carried out for 15 hours at constant temperature.
As another preferable selection of the preparation process for preparing the resveratrol with the purity of 99% from the giant knotweed rhizome, in the step (5), the filtrate is fed into an alumina chromatographic column, sectional collection is started when liquid flows out, and the alumina column is replaced when red to black liquid flows out; after all the liquid enters the alumina column, eluting with 80-95% ethanol until no resveratrol is detected.
Has the advantages that: (1) the yield of 98% resveratrol of finished products is improved, the yield is improved from 0.5-0.9% to more than 1.2%, and is improved by 0.3-0.7% compared with the yield, namely 3-7 kg of 99% resveratrol can be obtained from the same raw material per ton, and a new value of 4800-10200 yuan is created. (2) The color of the 99 percent resveratrol product is improved, and the beige to offwhite color of common varieties is removed, so that the whiteness of the final product reaches over 80 percent of high-quality resveratrol. (3) The extraction step is reduced, the production operation and production links are simplified, the production period is shortened by 23 percent, the large-scale production is easy to control and realize, and the labor safety and protection are improved.
Detailed Description
In order that those skilled in the art can better understand the present invention, the following embodiments are provided to further illustrate the present invention.
Example 1
(1) Taking 500kg of dried giant knotweed rhizome, and crushing until the giant knotweed rhizome passes through a 20-mesh sieve to be more than or equal to 90 percent;
(2) enzyme conversion: adding 150kg of water to soak giant knotweed rhizome powder, adding 2kg of wine yeast, 2kg of saccharifying enzyme and 1kg of aspergillus niger to the soaked giant knotweed rhizome powder, uniformly stirring, placing in a container with the temperature controlled at 45 ℃, and fermenting at constant temperature for 15 hours to convert the resveratrol analogue into resveratrol;
(3) and (3) countercurrent pressurization low-temperature leakage: uniformly loading the fermented giant knotweed material into a leakage column with the diameter-height ratio of 1:4, adding a mixed reagent of ethyl acetate and petroleum ether which is 4 times of the weight of the giant knotweed in total, and slowly passing through a giant knotweed layer to obtain leakage liquid; wherein the ratio of ethyl acetate to petroleum ether is 6: 1;
(4) concentrating and recovering ethanol; concentrating the leakage solution of rhizoma Polygoni Cuspidati, and recovering reagent until no reagent exists to obtain extract;
(5) dissolving and removing impurities; dissolving the extract with 70% ethanol at room temperature, stirring for 30 min, and filtering to obtain filtrate;
(6) chromatography: feeding the filtrate into an alumina chromatographic column, starting sectional collection when liquid flows out, and replacing the alumina column when red liquid flows out to black liquid; eluting with 85% ethanol after all liquid enters alumina column until no resveratrol is detected;
(7) and (3) crystallization and recrystallization: concentrating the eluent on the alumina column to 45% of alcohol concentration, crystallizing and recrystallizing, standing for 10 hours at 20 ℃, then filtering, and recrystallizing;
(8) and (3) drying the crystals at low temperature in vacuum to obtain 6.15kg of the product, wherein the detected content of the resveratrol is 99.3 percent, and the whiteness is 82 degrees.
Example 2
(1) Taking 500kg of dried giant knotweed rhizome, and crushing until the giant knotweed rhizome passes through a 20-mesh sieve to be more than or equal to 90 percent;
(2) enzyme conversion: adding 145kg of water to soak the giant knotweed rhizome powder, adding 2kg of wine yeast, 2kg of saccharifying enzyme and 1kg of aspergillus niger to the soaked giant knotweed rhizome powder, uniformly stirring, placing in a container with the temperature controlled at 42 ℃, and fermenting at constant temperature for 17 hours to convert the resveratrol analogue into resveratrol;
(3) and (3) countercurrent pressurization low-temperature leakage: uniformly loading the fermented giant knotweed material into a leakage column with the diameter-height ratio of 1:5, adding a mixed reagent of ethyl acetate and petroleum ether which is 5 times of the weight of the giant knotweed in total, and slowly passing through a giant knotweed layer to obtain leakage liquid; wherein the ratio of ethyl acetate to petroleum ether is 4: 1;
(4) concentrating and recovering ethanol; concentrating the leakage solution of rhizoma Polygoni Cuspidati, and recovering reagent until no reagent exists to obtain extract;
(5) dissolving and removing impurities; dissolving the extract with 70% ethanol at room temperature, stirring for 30 min, and filtering to obtain filtrate;
(6) chromatography: feeding the filtrate into an alumina chromatographic column, starting sectional collection when liquid flows out, and replacing the alumina column when red liquid flows out to black liquid; eluting with 85% ethanol after all liquid enters alumina column until no resveratrol is detected;
(7) and (3) crystallization and recrystallization: concentrating the eluent on the alumina column to alcohol concentration of 38%, crystallizing and recrystallizing, standing at 20 deg.C for 15 hr, filtering, and recrystallizing;
(8) and (3) drying the crystals at low temperature in vacuum to obtain 6.15kg of the product, wherein the detected content of the resveratrol is 99.3 percent, and the whiteness is 82 degrees.
The preparation process for preparing the resveratrol with the purity of 99 percent from the giant knotweed rhizome provided by the invention is described in detail above. The description of the specific embodiments is only intended to facilitate an understanding of the method of the invention and its core ideas. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.
Claims (4)
1. A preparation process for preparing resveratrol from giant knotweed rhizome is characterized by comprising the following steps:
(1) soaking rhizoma Polygoni Cuspidati powder in water; placing the special enzyme and the soaked giant knotweed rhizome powder in a container with the temperature controlled at 28-50 ℃, fermenting at constant temperature for 6-24 hours, and converting the resveratrol analogue into resveratrol, wherein the special enzyme is wine yeast, saccharifying enzyme and aspergillus niger; wherein the ratio of the wine yeast, the saccharifying enzyme and the aspergillus niger is 2:2: 1;
(2) uniformly loading the fermented polygonum cuspidatum material into a leakage column with the diameter-height ratio of 1:4-5, adding a mixed reagent of ethyl acetate and petroleum ether which is 3-5 times of the weight of the polygonum cuspidatum in total, and slowly passing through a polygonum cuspidatum layer to obtain leakage liquid;
(3) concentrating the leakage solution of rhizoma Polygoni Cuspidati, and recovering reagent until no reagent exists to obtain extract;
(4) dissolving the extract with 60-80% ethanol at room temperature, and filtering to obtain filtrate;
(5) feeding the filtrate into an alumina chromatographic column, starting sectional collection when liquid flows out, and replacing the alumina column when red liquid flows out to black liquid; eluting with 80-95% ethanol after all liquid is fed into alumina column until no resveratrol is detected;
(6) crystallizing and recrystallizing;
(7) and (4) drying the crystals at low temperature in vacuum to obtain a finished product, wherein the resveratrol is more than or equal to 99%.
2. The process for preparing resveratrol from giant knotweed rhizome according to claim 1, wherein the giant knotweed rhizome root powder is dried and pulverized until more than 90% of the powder can pass through a 20-mesh sieve before step (1).
3. The process for preparing resveratrol from giant knotweed rhizome according to claim 1, wherein the feeding ratio of the special enzyme to the giant knotweed rhizome powder is 1: 99-101.
4. The process for preparing resveratrol from giant knotweed rhizome according to claim 1, wherein the feeding ratio of giant knotweed rhizome powder to water is 10: 2-3.
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| CN201811570311.6A CN109503334B (en) | 2018-12-21 | 2018-12-21 | Preparation process for preparing resveratrol with purity of 99% from polygonum cuspidatum |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102242159A (en) * | 2011-05-16 | 2011-11-16 | 张家界湘汇生物有限责任公司 | Method for improving resveratrol content in giant knotweed by two-step microbial conversion |
| CN102250969A (en) * | 2011-05-16 | 2011-11-23 | 张家界湘汇生物有限责任公司 | Process for preparing high-purity resveratrol from giant knotweed |
| CN104388478A (en) * | 2014-11-24 | 2015-03-04 | 苏州嘉禧萝生物科技有限公司 | Method for improving yield of resveratrol through fermentation of aspergillus niger |
| CN107497134A (en) * | 2017-07-24 | 2017-12-22 | 张家界湘汇生物有限责任公司 | The new method for extracting of effective ingredients in plant |
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- 2018-12-21 CN CN201811570311.6A patent/CN109503334B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102242159A (en) * | 2011-05-16 | 2011-11-16 | 张家界湘汇生物有限责任公司 | Method for improving resveratrol content in giant knotweed by two-step microbial conversion |
| CN102250969A (en) * | 2011-05-16 | 2011-11-23 | 张家界湘汇生物有限责任公司 | Process for preparing high-purity resveratrol from giant knotweed |
| CN104388478A (en) * | 2014-11-24 | 2015-03-04 | 苏州嘉禧萝生物科技有限公司 | Method for improving yield of resveratrol through fermentation of aspergillus niger |
| CN107497134A (en) * | 2017-07-24 | 2017-12-22 | 张家界湘汇生物有限责任公司 | The new method for extracting of effective ingredients in plant |
Non-Patent Citations (1)
| Title |
|---|
| Biotransformation of polydatin to resveratrol in Polygonum cuspidatum roots by highly immobilized edible Aspergillus niger and Yeast;Shuang Jin 等;《Bioresource Technology》;20130315;第767页Table 1 和2 * |
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