CN106880652A - A kind of extracting method of mango leaf polyphenol - Google Patents
A kind of extracting method of mango leaf polyphenol Download PDFInfo
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- CN106880652A CN106880652A CN201510935514.0A CN201510935514A CN106880652A CN 106880652 A CN106880652 A CN 106880652A CN 201510935514 A CN201510935514 A CN 201510935514A CN 106880652 A CN106880652 A CN 106880652A
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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)
- A61K36/22—Anacardiaceae (Sumac family), e.g. smoketree, sumac or poison oak
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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
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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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
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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
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Abstract
The invention discloses a kind of extracting method of mango leaf polyphenol, comprise the following steps：1) it is beaten, digests, obtains mango leaf enzymolysis liquid；2) mango leaf enzymolysis liquid is slightly carried with ethanol；3) by crude extract through membrane filtration, clarified solution is obtained；4) macroporous resin column on clarified solution is purified, elutes, dries, as mango leaf polyphenol.The present invention has the advantages that Extracting temperature is low, applicability is wide, recovery rate is high, easy to operate；Obtained mango leaf polyphenol purity is more than 95%, and recovery rate is more than 6.5%, higher by more than 1.5% than traditional heating restricted-access media rate.
Description
Technical field
The invention belongs to active components of plants extractive technique field, and in particular to a kind of extracting method of mango leaf polyphenol.
Background technology
Plant polyphenol is the focus of the area research such as current functional food and Natural products research exploitation, plant polyphenol New function is constantly found both at home and abroad and the report of plant polyphenol is developed from new plant resource.According to free radical ability, the numerous function of the plant polyphenol reported at present such as prevents cardiovascular and cerebrovascular disease, regulation immunocompetence, antitumor,
It is all relevant with the ability that plant polyphenol removes free radical.The main free radical being detrimental to health in vivo is ultra-oxygen anion free radical and hydroxyl radical free radical.
Plant polyphenol is the important secondary metabolite being widely present in plant; it is distributed widely in the root of plant, leaf, skin, pulp and shell; with different physiological roles such as anti-oxidant, antitumor, protection livers; it is polyphenoils that human body must be absorbed from food daily, is referred to as the 7th nutrient after dietary fiber.
Mango, is aiphyllium, originates in India, and existing torrid zone various regions are planted, cultivated by South China, Guangxi, Guangdong, Hainan various regions village or room residence, somewhat soak the visible plant of being scattered about like the stars of soil.The fruit of mango, leaf and bark can be used as medicine, with economic worth higher.Mango leaf is the leaf of Anacardiaceae Mangifera tropical evergreen megaphanerophyte mango (Mangiferaindica Linn).Rich in polyphenols such as mangiferin, Quercetin, Hyperoside, protocatechuic acid, gallic acids, these polyphenol compounds have the extensive physiologically active such as antiasthmatic-antitussive, analgesia, anti-oxidant, antibacterial, antitumor, treatment cardiovascular and cerebrovascular disease to mango leaf.Mango is abounded with each province and municipality of south China, and byproduct mango leaf resource is quite enriched, China at present to being confined to the exploitation of mango leaf more be the very few several antiasthmatic-antitussives of number preparation.
The content of the invention
The present invention provides a kind of extracting method of mango leaf polyphenol, and mango leaf polyphenol purity is high obtained in the method, high income.
Technical scheme：A kind of extracting method of mango leaf polyphenol, comprises the following steps：
1) it is beaten, digests：Using solid-liquid ratio 1:3-5, will clean the mango leaf that drain and be beaten 3-5min with beater, be subsequently adding after protease is sufficiently stirred for and stand after 1-2 h are digested, and filter to take supernatant enzymolysis liquid with 800-1000 mesh filter screens standby；
2) slightly carry：Ultrasonic extraction is carried out to mango leaf enzymolysis liquid with ethanol, is centrifuged, collect supernatant, ethanol is recovered under reduced pressure, obtain mango leaf crude extract；
3) film is crossed：Supernatant is crossed into film filter to be filtered, clarified solution is obtained；
4) purify：By macroporous resin column on clarified solution, loading flow velocity is 0.4~0.6BV/h, and upper macroporous resin column washes post, then with ethanol elution, collects ethanol eluate, is concentrated under reduced pressure, and is dried, and obtains final product mango leaf polyphenol.
Above-mentioned steps 1) in, described enzymolysis is that every 100 grams of mango leafs add 0.2-0.4g protease, 45-55 DEG C of enzymolysis 1-2h.
Step 2) in, protein and carbohydrate are contained in mango leaf, it is solvent to use ethanol, is unfavorable for the dissolving of protein and carbohydrate, can reduce the dissolution of impurity component, beneficial to subsequent purification.Ethanol volumetric concentration is 40~50% and the solid-liquid ratio of enzymolysis liquid and ethanol is 1ml：2~4ml is preferred, and concentration of alcohol is too high to produce cohesion to protein so that protein is trapped in the internal gutter of particle, and the diffusional resistance of mango leaf polyphenol is increased on the contrary, reduces the yield of finished product.The too low then impurity content of concentration of alcohol is larger, influences the purity of finished product.
Step 3) in, the film filter is 50000 dalton microfiltration membranes.
Step 4) in, washing flow velocity is 2.5~3BV/h, and the elution amount of water is 5~6BV；The elution flow rate of ethanol is 2~2.5BV/h, and the elution amount of ethanol is 4~5BV；The volumetric concentration of ethanol is 65~85%.
Compared with prior art, the present invention has the advantages that Extracting temperature is low, applicability is wide, recovery rate is high, easy to operate；Obtained mango leaf polyphenol purity is more than 95%, and recovery rate is more than 6.5%, higher by more than 1.5% than traditional heating restricted-access media rate.
Specific embodiment
The present invention is further elaborated for specific examples below, but not as a limitation of the invention.
Embodiment 1
1) it is beaten, digests：Take to clean and drain mango leaf 1000g, using solid-liquid ratio 1:3 add pure water, the mango leaf for draining will be cleaned with beater and is beaten 3min, are subsequently adding after 2g protease is sufficiently stirred for and digest 1h at 45 DEG C, then filter to take supernatant enzymolysis liquid with 800 mesh filter screens standby；
2) slightly carry：It is 1ml by the solid-liquid ratio of enzymolysis liquid and ethanol：2ml 40% ethanol of addition carries out ultrasonic extraction to mango leaf enzymolysis liquid, is centrifuged, and collects supernatant, and ethanol is recovered under reduced pressure, and obtains mango leaf crude extract；
3) film is crossed：The dalton micro-filtration film device of supernatant 50000 is filtered, clarified solution is obtained；
4) purify：It is 0.4BV/h by DA201 macroporous resin columns on clarified solution, loading flow velocity, first washes post, washing flow velocity is 2.5 BV/h, and the elution amount of water is 5BV；Then 65% ethanol elution is used, the elution flow rate of ethanol is 2BV/h, and the elution amount of ethanol is 4BV, collects ethanol eluate, is concentrated under reduced pressure, and is dried, and obtains final product 13.78g mango leaf polyphenol.Detected through HPLC, the purity of mango leaf polyphenol is 95.2%, mango leaf polyphenol recovery rate in finished product（Relative to siccative）It is more than 6.5%.
Embodiment 2
1) it is beaten, digests：Take to clean and drain mango leaf 1000g, using solid-liquid ratio 1:5 add pure water, the mango leaf for draining will be cleaned with beater and is beaten 5min, are subsequently adding after 4g protease is sufficiently stirred for and digest 2h at 55 DEG C, then filter to take supernatant enzymolysis liquid with 1000 mesh filter screens standby；
2) slightly carry：It is 1ml by the solid-liquid ratio of enzymolysis liquid and ethanol：4ml 50% ethanol of addition carries out ultrasonic extraction to mango leaf enzymolysis liquid, is centrifuged, and collects supernatant, and ethanol is recovered under reduced pressure, and obtains mango leaf crude extract；
3) film is crossed：The dalton micro-filtration film device of supernatant 50000 is filtered, clarified solution is obtained；
4) purify：It is 0.6BV/h by DA201 macroporous resin columns on clarified solution, loading flow velocity, first washes post, washing flow velocity is 3BV/h, and the elution amount of water is 6BV；Then 85% ethanol elution is used, the elution flow rate of ethanol is 2.5BV/h, and the elution amount of ethanol is 5BV, collects ethanol eluate, is concentrated under reduced pressure, and is dried, and obtains final product 13.88g mango leaf polyphenol.Detected through HPLC, the purity of mango leaf polyphenol is 95.1%, mango leaf polyphenol recovery rate in finished product（Relative to siccative）It is more than 6.5%.
Embodiment 3
1) it is beaten, digests：Take to clean and drain mango leaf 1000g, using solid-liquid ratio 1:4 add pure water, the mango leaf for draining will be cleaned with beater and is beaten 4min, be subsequently adding after 3g protease is sufficiently stirred for and enzymolysis 1.5h is stood at 50 DEG C, then filter to take supernatant enzymolysis liquid with 900 mesh filter screens standby；
2) slightly carry：It is 1ml by the solid-liquid ratio of enzymolysis liquid and ethanol：3ml 45% ethanol of addition carries out ultrasonic extraction to mango leaf enzymolysis liquid, is centrifuged, and collects supernatant, and ethanol is recovered under reduced pressure, and obtains mango leaf crude extract；
3) film is crossed：The dalton micro-filtration film device of supernatant 50000 is filtered, clarified solution is obtained；
4) purify：It is 0.5BV/h by DA201 macroporous resin columns on clarified solution, loading flow velocity, first washes post, washing flow velocity is 2.8BV/h, and the elution amount of water is 5.5BV；Then 75% ethanol elution is used, the elution flow rate of ethanol is 2.3BV/h, and the elution amount of ethanol is 4.5BV, collects ethanol eluate, is concentrated under reduced pressure, and is dried, and obtains final product 14.26g mango leaf polyphenol.Detected through HPLC, the purity of mango leaf polyphenol is 95.8%, mango leaf polyphenol recovery rate in finished product（Relative to siccative）It is more than 6.5%.
Embodiment 4
The mango leaf polyphenol that above-described embodiment 1-3 is obtained is to superoxide anion O2 -The measure of elimination effect：
In the range of experimental concentration, mango leaf polyphenol is to O2 -Scavenging activity it is stronger, in 0.5mg/mL~2.5mg/mL concentration ranges clearance rate with concentration rise and improve trend it is obvious.When mango leaf polyphenol concentration reaches
During 2.5mg/mL, to O2 -Clearance rate more than 90%.
Embodiment 5
Measure of the mango leaf polyphenol that above-described embodiment 1-3 is obtained to hydroxyl radical free radical OH elimination effects：
Mango leaf polyphenol concentration rising with concentration to OH clearance rates in the range of 0.5mg/mL~2.0mg/mL and significantly improving.When mango leaf polyphenol concentration reaches 2.0mg/mL, clearance rate is more than 90%.
Claims (5)
1. a kind of extracting method of mango leaf polyphenol, it is characterised in that：Comprise the following steps：
1) it is beaten, digests：Using solid-liquid ratio 1:3~5, the mango leaf for draining will be cleaned with beater and is beaten 3~5min, it is subsequently adding after protease is sufficiently stirred for and is digested, it is standby to filter to take supernatant enzymolysis liquid with 800-1000 mesh filter screens；
2) slightly carry：Ultrasonic extraction is carried out to mango leaf enzymolysis liquid with ethanol, is centrifuged, collect supernatant, ethanol is recovered under reduced pressure, obtain mango leaf crude extract；
3) film is crossed：Supernatant is crossed into film filter to be filtered, clarified solution is obtained；
4) purify：By macroporous resin column on clarified solution, loading flow velocity is 0.4~0.6BV/h, and upper macroporous resin column washes post, then with ethanol elution, collects ethanol eluate, is concentrated under reduced pressure, and is dried, and obtains final product mango leaf polyphenol.
2. the extracting method of a kind of mango leaf polyphenol according to claim 1, it is characterised in that：Step 1) in, described enzymolysis is that every 100 grams of mango leafs add 0.2~0.4g protease, 45~55 DEG C of 1~2h of enzymolysis.
3. the extracting method of a kind of mango leaf polyphenol according to claim 1, it is characterised in that：Step 2) in, the volumetric concentration of the ethanol is 40~50%, and enzymolysis liquid is 1ml with the solid-liquid ratio of ethanol：2~4ml.
4. the extracting method of a kind of mango leaf polyphenol according to claim 1, it is characterised in that：Step 3) in, the film filter is 50000 dalton microfiltration membranes.
5. the extracting method of a kind of mango leaf polyphenol according to claim 1, it is characterised in that：Step 4) in, washing flow velocity is 2.5~3BV/h, and the elution amount of water is 5~6BV；The elution flow rate of ethanol is 2~2.5BV/h, and the elution amount of ethanol is 4~5BV；The volumetric concentration of ethanol is 65~85%.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107788531A (en) * | 2017-11-10 | 2018-03-13 | 西华大学 | Method for extracting polyphenol from mango kernels |
| CN111991432A (en) * | 2020-09-03 | 2020-11-27 | 昆明理工大学 | Method for extracting mango leaf polyphenol |
-
2015
- 2015-12-15 CN CN201510935514.0A patent/CN106880652A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107788531A (en) * | 2017-11-10 | 2018-03-13 | 西华大学 | Method for extracting polyphenol from mango kernels |
| CN111991432A (en) * | 2020-09-03 | 2020-11-27 | 昆明理工大学 | Method for extracting mango leaf polyphenol |
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Application publication date: 20170623 |