CN112007070A - High-content kudzu root extract, preparation method and application thereof - Google Patents
High-content kudzu root extract, preparation method and application thereof Download PDFInfo
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- CN112007070A CN112007070A CN202010967503.1A CN202010967503A CN112007070A CN 112007070 A CN112007070 A CN 112007070A CN 202010967503 A CN202010967503 A CN 202010967503A CN 112007070 A CN112007070 A CN 112007070A
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- pueraria lobata
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/488—Pueraria (kudzu)
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
- C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
- C07D407/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
The invention discloses a high-content radix puerariae extract, a preparation method and application thereof.
Description
Technical Field
The invention relates to the technical field of biomass extraction, and particularly relates to a high-content radix puerariae extract, a preparation method and application thereof.
Background
Pueraria lobata (Willd.) Ohwi, originally recorded in Shennong Ben Cao Jing, is the dry root of Pueraria lobata (Willd.) Ohwi of Leguminosae, and has the effects of relieving muscles and reducing fever, promoting fluid production to quench thirst, invigorating yang to arrest diarrhea, clearing and activating the channels and collaterals, etc. Research shows that the kudzuvine root contains abundant nutrient components such as flavonoid substances, daidzein, puerarin, a large amount of starch, carbohydrate, protein, various trace elements, amino acid, choline derivatives and the like, and has high nutritional value and medicinal value. The main effective component of the kudzu root is flavonoid, wherein puerarin is the most main active substance. Clinical trial studies showed that: puerarin has effects of reducing oxygen consumption of myocardium, dilating blood vessel, relieving angina pectoris, lowering blood pressure, and treating retinal artery and vein occlusion and sudden deafness. Has high medicinal value in the aspects of biological oxidation resistance, blood vessel expansion, cardiovascular and cerebrovascular disease treatment, cancer cell inhibition and the like. The kudzu root extract has the effects of relieving fever, promoting the production of body fluid, invigorating yang, promoting eruption, reducing blood sugar, protecting heart and cerebral vessels and the like, and is widely applied to the fields of food, health products, medicines and the like.
For convenience of medication, a high-purity puerarin product is concerned in the international market. Since the puerarin extracted solutions obtained by using various extraction methods are solutions containing many components and having low puerarin content, we need to further separate and purify puerarin with high purity. Generally, puerarin can be separated and purified by macroporous adsorption resin method, complexation extraction technology, beta-cyclodextrin bonded stationary phase method, salting out method, ion exchange fiber method, polyphthalamide column chromatography adsorption method and other methods. Different purification methods have respective advantages and disadvantages, but the process has low purity and yield of puerarin, or uses a large amount of acid, alkali and organic solvent in the production process, and has great pollution. Therefore, how to use a simple, feasible and environment-friendly process to separate and purify puerarin with high purity and high yield has important significance for realizing the industrial production of the high-content pueraria extract.
Disclosure of Invention
The invention aims to provide a preparation method of a high-content radix puerariae extract, which is convenient and direct, simple in process, low in equipment requirement, environment-friendly, easy to popularize and suitable for industrial mass production. The kudzu root extract with high puerarin content is obtained by extraction and separation through combined application of extraction, resin purification, membrane separation and recrystallization, and the whole process only uses water and ethanol, so that toxic organic solvents are avoided, and the food safety risk is reduced.
The invention provides the following technical scheme:
a preparation method of a high-content kudzu root extract comprises the following steps:
(1) extraction: crushing a kudzu root medicinal material, adding 5-15 times of 60-80% ethanol solution, and extracting at 70-90 ℃ for 2-3 times, 1-2 hours each time;
(2) and (3) filtering: cooling to 15-25 ℃, passing through a disc centrifuge, and collecting clear liquid;
(3) concentration: concentrating the centrifugate to a liquid-medicine ratio of 1: 4-1: 8;
(4) resin separation: loading the concentrated solution into macroporous resin, washing with water by 1-3 times of column volume to remove impurities, eluting with 10-25% ethanol at 50-70 deg.C, and collecting eluate;
(5) membrane separation: firstly, separating eluent by using a microfiltration membrane, separating the permeate by using a nanofiltration membrane, and collecting the permeate of the nanofiltration membrane;
(6) and (3) recrystallization: concentrating the nanofiltration membrane filtrate until the liquid medicine ratio is 6: 1-3: 1, cooling to 2-10 ℃, standing for crystallization, adding 5-10 times of purified water into crystals, heating for dissolution, cooling to 2-10 ℃, standing for crystallization, and repeating for 2-4 times;
(7) and (3) drying: drying the recrystallized crystal to obtain the radix Puerariae extract.
Preferably, the macroporous resin in the step (4) is selected from AB-8 macroporous resin, D101 macroporous resin, HPD-600 macroporous resin, DA-201 macroporous resin, DM-301 macroporous resin and DS-401 macroporous resin.
Preferably, in the step (5), a microfiltration membrane with the thickness of 20 nm-100 nm is used for separating eluent, a nanofiltration membrane with the thickness of 500D-1000D is used for separating permeate, and the nanofiltration membrane permeate is collected.
As another object of the present invention, the present invention also provides the kudzu root extract obtained by the above preparation method, wherein the puerarin content is greater than 80%.
As another object of the invention, the invention also provides the application of the kudzu root extract as food to be directly eaten, wherein the eating amount is 0.125-0.5 g/day.
As another object of the invention, the invention also provides the application of the kudzu root extract as a common food raw material, wherein the addition amount is 1.0-10.0 g/kg.
As another object of the invention, the invention also provides the application of the kudzu root extract as a functional food raw material, wherein the addition amount is 5.0-50.0 g/kg.
As another object of the invention, the invention also provides the application of the kudzu root extract as a raw material for producing the health wine, wherein the addition amount is 0.5-1.0 g/L.
As another purpose of the invention, the invention also provides application of the kudzu root extract as a raw material to production of healthy white spirit, wherein the addition amount is 0.1-0.5 g/L.
Compared with the prior art, the invention has the following advantages:
the process is simple, the equipment requirement is low, the production period is short, the cost is low, and the method is easy to popularize: compared with the common extraction process of the pueraria extract, the invention utilizes the characteristic that the puerarin is easy to dissolve in ethanol, uses macroporous resin for adsorption and ethanol elution, simultaneously increases the temperature of the ethanol to ensure that the puerarin is completely eluted, further enriches the puerarin by membrane separation, simultaneously carries out decoloration, and finally uses water for recrystallization, so that the prepared pueraria extract has high puerarin content and light color. Production equipment used for production is common and easy to obtain, equipment cost is low, cheap water and ethanol are used as production raw materials, production cost is low, the process is simple, and large-scale production is easy to realize.
(II) mild condition and environmental protection: at present, the pueraria extract is mostly extracted by using toxic organic solvents such as n-butyl alcohol, methanol and the like, and is decolored by using active carbon or hydrogen peroxide, so that the produced waste water and waste have large pollution, and the waste treatment cost is high. The extraction and separation method adopted by the invention only uses water and ethanol, reduces the pollution of the wastewater to the minimum, has mild extraction conditions, does not have drastic temperature difference change among the working procedures, reduces the energy waste, and is beneficial to further reducing the cost and saving the resources.
Drawings
FIG. 1 is a flow chart of the manufacturing process of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
(1) Extraction: extracting 200kg of radix Puerariae with 8 times of 70% ethanol at 80 deg.C for 2 times, each for 2 hr;
(2) and (3) filtering: cooling the extractive solution to 20 deg.C, passing through a disc centrifuge, and collecting clear liquid;
(3) concentration: concentrating the centrifugate to a liquor ratio of 1: 4;
(4) resin separation: loading the concentrated solution on AB-8 macroporous resin, washing with water for 2 times of column volume to remove impurities, eluting with 15% ethanol at 60 deg.C for 4 times of column volume, and collecting eluate;
(5) membrane separation: separating the eluent by using a 50nm microfiltration membrane, separating the permeate by using an 800D nanofiltration membrane, and collecting the permeate of the nanofiltration membrane;
(6) and (3) recrystallization: concentrating the nanofiltration membrane filtrate until the liquid medicine ratio is 6:1, cooling to 2 deg.C, standing for crystallization, adding 8 times of purified water into the crystal, heating for dissolving, cooling to 2 deg.C, standing for crystallization, and repeating for 2 times.
(7) And (3) drying: drying the recrystallized crystal to obtain 4.86kg of radix Puerariae extract with puerarin content of 82.3%.
(8) When the kudzu root extract is directly eaten as food, the recommended eating amount is 0.25 g/day.
Example 2
(1) Extraction: taking 500kg of kudzuvine root, adding 10 times of 80% ethanol, extracting for 3 times at 75 ℃, each time for 1.5 hours;
(2) and (3) filtering: cooling the extract to 25 ℃, passing through a disc centrifuge, and collecting clear liquid;
(3) concentration: concentrating the centrifugate to obtain a liquid medicine ratio of 1: 5;
(4) resin separation: loading the concentrated solution on AB-8 macroporous resin, washing with water for 1 time of column volume to remove impurities, eluting with 10% ethanol at 65 deg.C for 3 times of column volume, and collecting eluate;
(5) membrane separation: separating the eluent by using a 100nm microfiltration membrane, separating the permeate by using a 600D nanofiltration membrane, and collecting the permeate of the nanofiltration membrane;
(6) and (3) recrystallization: concentrating the nanofiltration membrane filtrate until the ratio of the liquid medicine to the filtrate is 5:1, cooling to 4 deg.C, standing for crystallization, adding 10 times of purified water into the crystal, heating for dissolving, cooling to 4 deg.C, standing for crystallization, and repeating for 3 times.
(7) And (3) drying: drying the recrystallized crystal to obtain 12.98kg radix Puerariae extract with puerarin content of 84.7%.
(8) When the above radix Puerariae extract is used as common food material, its addition amount is 5.0 g/kg.
Example 3
(1) Extraction: taking 1000kg of kudzuvine root, adding 75% ethanol in an amount which is 12 times that of the kudzuvine root, and extracting for 2 times at 85 ℃ for 3 hours each time;
(2) and (3) filtering: cooling the extractive solution to 20 deg.C, passing through a disc centrifuge, and collecting clear liquid;
(3) concentration: concentrating the centrifugate to obtain a liquid medicine ratio of 1: 6;
(4) resin separation: passing the concentrated solution through HPD-600 macroporous resin, washing with water for 3 times of column volume to remove impurities, eluting with 55 deg.C 20% ethanol for 4 times of column volume, and collecting eluate;
(5) membrane separation: separating the eluent by using a 20nm microfiltration membrane, separating the permeate by using a 1000D nanofiltration membrane, and collecting the nanofiltration membrane permeate;
(6) and (3) recrystallization: concentrating the nanofiltration membrane filtrate until the ratio of the liquid medicine to the filtrate is 4:1, cooling to 6 deg.C, standing for crystallization, adding 5 times of purified water into the crystal, heating for dissolving, cooling to 6 deg.C, standing for crystallization, and repeating for 3 times.
(7) And (3) drying: drying the recrystallized crystal to obtain 24.06kg of radix Puerariae extract with puerarin content of 80.8%.
(8) The radix Puerariae extract is added at a ratio of 15g/kg when used as raw material for producing functional food.
Example 4
(1) Extraction: taking 800kg of kudzuvine root, adding 15 times of 60% ethanol, extracting for 3 times at 90 ℃, each time for 2 hours;
(2) and (3) filtering: cooling the extract to 25 ℃, passing through a disc centrifuge, and collecting clear liquid;
(3) concentration: concentrating the centrifugate to obtain a liquid medicine ratio of 1: 8;
(4) resin separation: loading the concentrated solution on DA-201 macroporous resin, washing with water for 2 times of column volume to remove impurities, eluting with 70 deg.C 10% ethanol for 3 times of column volume, and collecting eluate;
(5) membrane separation: separating the eluent by using a 50nm microfiltration membrane, separating the permeate by using a 700D nanofiltration membrane, and collecting the permeate of the nanofiltration membrane;
(6) and (3) recrystallization: concentrating the nanofiltration membrane filtrate until the ratio of the liquid medicine to the filtrate is 3:1, cooling to 8 ℃, standing for crystallization, adding 7 times of purified water into the crystals, heating for dissolution, cooling to 8 ℃, standing for crystallization, and repeating for 3 times.
(7) And (3) drying: drying the recrystallized crystal to obtain 22.24kg of radix Puerariae extract with puerarin content of 82.6%.
(8) The radix Puerariae extract is used as raw material for producing health wine, and the addition amount is 0.7 g/L.
Example 5
(1) Extraction: extracting radix Puerariae 1500kg with 5 times of 80% ethanol at 85 deg.C for 3 times, each for 2 hr;
(2) and (3) filtering: cooling the extractive solution to 20 deg.C, passing through a disc centrifuge, and collecting clear liquid;
(3) concentration: concentrating the centrifugate to a liquor ratio of 1: 7;
(4) resin separation: loading the concentrated solution on DS-401 macroporous resin, washing with water for 1 time of column volume to remove impurities, eluting with 55 deg.C 15% ethanol for 4 times of column volume, and collecting eluate;
(5) membrane separation: separating the eluent by using a 20nm microfiltration membrane, separating the permeate by using a 600D nanofiltration membrane, and collecting the permeate of the nanofiltration membrane;
(6) and (3) recrystallization: concentrating the nanofiltration membrane filtrate until the ratio of the liquid medicine to the filtrate is 3:1, cooling to 10 deg.C, standing for crystallization, adding 6 times of purified water into the crystal, heating for dissolving, cooling to 10 deg.C, standing for crystallization, and repeating for 4 times.
(7) And (3) drying: drying the recrystallized crystal to obtain 37.78kg of radix Puerariae extract with puerarin content of 83.4%.
(8) When the radix Puerariae extract is used as raw material for producing health Chinese liquor, the addition amount is 0.25 g/L.
The above-mentioned embodiments only express the embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (9)
1. A preparation method of a high-content kudzu root extract is characterized by comprising the following steps:
(1) extraction: crushing a kudzu root medicinal material, adding 5-15 times of 60-80% ethanol solution, and extracting at 70-90 ℃ for 2-3 times, 1-2 hours each time;
(2) and (3) filtering: cooling to 15-25 ℃, passing through a disc centrifuge, and collecting clear liquid;
(3) concentration: and concentrating the centrifugate until the ratio of the medicine liquid is 1: 4-1: 8.
(4) Resin separation: and (3) passing the concentrated solution through macroporous resin, washing with water by 1-3 times of the column volume to remove impurities, eluting with 10-25% ethanol at 50-70 ℃, and collecting the eluent.
(5) Membrane separation: firstly, separating eluent by using a microfiltration membrane, separating the permeate by using a nanofiltration membrane, and collecting the permeate of the nanofiltration membrane;
(6) and (3) recrystallization: concentrating the nanofiltration membrane filtrate until the liquid medicine ratio is 6: 1-3: 1, cooling to 2-10 ℃, standing for crystallization, adding 5-10 times of purified water into crystals, heating for dissolution, cooling to 2-10 ℃, standing for crystallization, and repeating for 2-4 times;
(7) and (3) drying: drying the recrystallized crystal to obtain the radix Puerariae extract.
2. The method according to claim 1, wherein the macroporous resin in the step (4) is selected from AB-8 macroporous resin and D101 macroporous resin、HPD-600 macroporous resin, DA-201 macroporous resin, DM-301 macroporous resin or DS-401 macroporous resin.
3. The preparation method according to claim 1, wherein in the step (5), the eluent is separated by using a microfiltration membrane with the diameter of 20nm to 100nm, the permeate is separated by using a nanofiltration membrane with the diameter of 500D to 1000D, and the permeate of the nanofiltration membrane is collected.
4. The pueraria lobata extract prepared by the method according to any one of claims 1 to 3, wherein the puerarin content is greater than 80%.
5. The use of the pueraria lobata extract of claim 4 as a food for direct consumption, wherein the amount of the pueraria lobata extract consumed is 0.125 to 0.5 g/day.
6. The use of the pueraria lobata extract as a common food material according to claim 4, wherein the amount of the pueraria lobata extract added is 1.0 to 10.0 g/kg.
7. The use of the pueraria lobata extract as a functional food material according to claim 4, wherein the amount of the pueraria lobata extract added is 5.0 to 50.0 g/kg.
8. The use of the pueraria lobata extract as a raw material for producing a health wine according to claim 4, wherein the amount of the pueraria lobata extract added is 0.5 to 1.0 g/L.
9. The use of the pueraria lobata extract as a raw material for producing healthy white spirit according to claim 4, wherein the addition amount is 0.1-0.5 g/L.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112569271A (en) * | 2020-12-15 | 2021-03-30 | 吉首大学 | High-content pueraria flavonid and preparation method thereof |
| CN112586738A (en) * | 2020-12-15 | 2021-04-02 | 吉首大学 | Pueraria flavone buccal tablet and preparation method thereof |
| CN113813301A (en) * | 2021-08-30 | 2021-12-21 | 云南中医药大学 | Application of radix Puerariae extract in preparing medicine or health product for regulating intestinal flora and reducing blood lipid |
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| CN112569271A (en) * | 2020-12-15 | 2021-03-30 | 吉首大学 | High-content pueraria flavonid and preparation method thereof |
| CN112586738A (en) * | 2020-12-15 | 2021-04-02 | 吉首大学 | Pueraria flavone buccal tablet and preparation method thereof |
| CN113813301A (en) * | 2021-08-30 | 2021-12-21 | 云南中医药大学 | Application of radix Puerariae extract in preparing medicine or health product for regulating intestinal flora and reducing blood lipid |
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