CN107158048A - A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone - Google Patents

A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone Download PDF

Info

Publication number
CN107158048A
CN107158048A CN201710332354.XA CN201710332354A CN107158048A CN 107158048 A CN107158048 A CN 107158048A CN 201710332354 A CN201710332354 A CN 201710332354A CN 107158048 A CN107158048 A CN 107158048A
Authority
CN
China
Prior art keywords
ginkgo leaf
hours
flavones
reaction
conditions
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201710332354.XA
Other languages
Chinese (zh)
Inventor
何福林
袁志辉
陈小明
蒋琼凤
赵昌会
张瑞
刘伟
唐静
杨芳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hunan University of Science and Engineering
Original Assignee
Hunan University of Science and Engineering
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hunan University of Science and Engineering filed Critical Hunan University of Science and Engineering
Priority to CN201710332354.XA priority Critical patent/CN107158048A/en
Publication of CN107158048A publication Critical patent/CN107158048A/en
Pending legal-status Critical Current

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/16Ginkgophyta, e.g. Ginkgoaceae (Ginkgo family)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/10Preparation or pretreatment of starting material
    • A61K2236/13Preparation or pretreatment of starting material involving cleaning, e.g. washing or peeling
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/10Preparation or pretreatment of starting material
    • A61K2236/15Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/10Preparation or pretreatment of starting material
    • A61K2236/19Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/51Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/53Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

Landscapes

  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

A kind of efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone, takes ginkgo leaf cleaning clean, crushes, and ginkgo leaf, which adds water to mix, loads fermentation tank, in 121 DEG C of sterilizations, then carries out steam blasting;Then add fermentation of Aspergillus niger;Then add mixed enzyme solution, addition is 2% the 5% of reaction substrate, and the reaction time is 36 hours under the conditions of 25 30 DEG C;It is subsequently added β glucuroides, addition is the 1% of reaction substrate, reaction condition is that pH is 4.3 under the conditions of 42 DEG C, and the reaction time is 5 hours;It is subsequently added under the conditions of protease 1%, 25 30 DEG C, the reaction time is 34 hours;Heated up 95 DEG C of 10min, is subsequently added the extraction that extractant carries out flavones, and 12000g centrifugation 20min are taken after supernatant concentration, extracted using alcohol steep, receive to obtain ethanol eluate, concentrated, using freeze-drying, obtain product.

Description

A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone
Technical field
Flavones is extracted the present invention relates to GINKGO BILOBA EXTRACT extractive technique field, more particularly to one kind high efficiency from ginkgo leaf simultaneously It is converted into the method for glucoside type flavone.
Background technology
GINKGO BILOBA EXTRACT is ginkgo biloba p.e, and it can increase cerebrovascular flow, improves Cerebrovascular disorders function, protects brain Cell, coronary artery dilator prevents angina pectoris and miocardial infarction, prevents thrombosis, improves body immunity.To coronary disease Disease, angina pectoris, cerebral arteriovenous malformation, senile dementia, high blood pressure are highly beneficial per capita.But, passed through in ginkgo biloba p.e Can often adulterate ginkgoic acid.Ginkgoic acid has potential sensitization and mutagenesis and strong cytotoxicity, can cause serious Allergic reaction, gene mutation, neurotrosis, cause nausea and heartburn, anaphylactic shock, anaphylactoid purpura, exfoliative dermatitis, The adverse reactions such as alimentary canal mucous membrane allergy, spasm and neural paralysis.Therefore, it is necessary that these impurity are removed in extracting method 's.The production technology of ginkgo biloba p.e has a lot, external to use organic solvent method more, and representational technique is Germany A series of patent DE2117419, DE1767098, DE3940092 of Schwabe companies;And Indena companies of Italy is special Sharp technique EP0360556, the common feature of these techniques is that substantial amounts of organic solvent is used in production process, such as acetone, ethanol, Carbon tetrachloride, n-hexane, MEK, n-butanol etc., the advantage of technique are good product qualities and stably, have the disadvantage production cost pole Height, production technology security is poor, effect on environment is big.
It is all main at present both at home and abroad that GINKGO BILOBA EXTRACT is extracted by raw material of ginkgo leaf.Isolate and purify the method for GINKGO BILOBA EXTRACT very It is many, including the suction of solvent extraction method, supercritical methanol technology, supercritical ultrasonics technology, microwave radiation technology formulation, microwave method, high-speed counter-current method, macroreticular resin Attached method method, Thin-layer chromatography and enzyme process etc., wherein most ripe and the most frequently used extraction process is solvent extraction method.
Such as A of CN 102805755 are related to a kind of preparation method that high-quality ginkgo flavone is extracted from from ginkgo leaf.With Deionized water extracts crude extract from ginkgo leaf, alcohol precipitation removal of impurities after filtering, fining agent removal of impurities, enriching ginkgetin and lactone, Carry out just separating using the macroreticular resin of polarity, low pole.Resin removes the final ginkgo biloba extract product of ginkgoic acid.The invention It is a technical advantage that:A) macroreticular resin that different performance is used in combination comes enriching ginkgetin and lactone, abjection ginkgoic acid, makes production Product quality is significantly improved, it is ensured that product.B) simplify production technology, shorten the production cycle, production cost is decreased.C) make With nontoxic organic solvent and natural clarifying agent, the residual quantity of poisonous and harmful substance in product is greatly reduced, is improved The quality of product.But need alcohol precipitation removal of impurities, fining agent removal step complicated in separation process.
A kind of preparation technology that GINKGO BILOBA EXTRACT is extracted with calcium carbonate is disclosed in the A of CN 101463051, including:(1) weigh Raw material ginkgo leaf, is crushed;Add water as 6~8 times of ginkgo leaf weight, while adding the carbon of ginkgo leaf weight 0.5~1.5% Sour calcium, boils and is filtered after 1~1.5 hour, obtain an Aqueous extracts, is once to extract;Same method, carries out second extraction, obtains Secondary Aqueous extracts;Merge once, secondary Aqueous extracts, produce amalgamation liquid;(2) amalgamation liquid is concentrated in vacuo at a temperature of 60~70 DEG C To 0.5~0.6 times of amalgamation liquid volume, ethanol is added, is 40~50% to ethanol degree, is sufficiently stirred for, after standing 3~4 hours Filtering;It is diluted with water filtered fluid so that ethanol degree is 15~20%, it is 3.0 that pH value is adjusted to dilute sulfuric acid, must dilute filtrate, will Filtrate is diluted to be adsorbed with DM131 resin columns;(3) it is 3.0 the ethanol water that ethanol degree is 20% to be adjusted into pH value with dilute sulfuric acid, Resin column impurity is eluted, then is rinsed with water to neutrality;(4) finally, with ethanol degree for 70% ethanol water elute effectively into Point, then eluent is concentrated in vacuo at a temperature of 60~70 DEG C, dried, gingko flavone extractive is produced.This method needs big Amount uses acid solution, while yield is nor very high, and complex process is unfavorable for the production of industrially scalable metaplasia.
A kind of technique for producing high-activity ginkgo leaf extract is disclosed in the A of CN 1166675, it is characterised in that described Technique comprises the following steps:(1) extractant sodium sulfite or sodium hydrogensulfite are added in pending ginkgo leaf, addition is pressed W/w ratios are 0.001-0.5, and benefit is filled with water to 100;(2) w/w is pressed, it is 1 that clean ginkgo leaf is pressed into the ratio between raw material and extractant : 5-20, fed intake extraction extraction, and 30-100 DEG C of extraction temperature is handled -20 hours 30 minutes, intermittent stirring raw material in leaching process, 1-10 DEG C/h of cooling rate is controlled, feed liquid is then left and taken, the extractant for adding 5-20 times of ginkgo leaf is carried for the second time Take, merge feed liquid twice;(3) feed liquid for obtaining step (2) is filtered, and removes the impurity below 200 mesh, and feed liquid is cooled to 30 DEG C Below;(4) feed liquid for obtaining step (3) flows through resin absorbing column polishing purification, is washed with water after the removal of impurity, then use 30%-95% lower alcohol rinses resin column, obtains eluent;(5) by the eluent described in step (4) below temperature 50 C It is concentrated in vacuo, recycling design obtains concentrate;(6) concentrate for obtaining step (5) is dried, and (7) obtain step (6) Product use reversed-phased high performace liquid chromatographic quantitative analysis GINKGO BILOBA EXTRACT, terpene lactones, OPC and ginkgolic acid.This method Same yield is not high, also complex in preparation technology, there is huge room for improvement.
In addition, extractant generally has larger toxicity in the prior art, also there is larger harm to the body of people, in life Also there is potential danger during production.
Finally, the metabolic pathway of flavone compound in animal body is different, and only glucoside type flavone can directly be inhaled Take in into animal blood, and the chromocor compound of most of glucoside type can not be entered in blood in human body by small bowel, And only fraction flavones is in the presence of faecal flora, blood could be entered by being hydrolyzed into glucoside member.Therefore, flavone glycoside is dynamic Bioavailability in object is well below glucoside type flavone.Therefore, improve the configuration of flavones, improve its absorption in blood Rate is to improve the important channel of flavonoids product bioavailability.At present, the configuration conversion to flavones glycocide mainly has chemistry Method and bioanalysis, domestic and international many researchers hydrolyze flavone glycoside, the also etherificate of flavones, esterification, acylation using chemical method Deng chemical derivatization reaction, the characteristic of flavones can also be changed.But these methods often shield flavones during the course of the reaction The main functional group of compound --- phenolic hydroxyl group, to anti-oxidant generation harmful effect.Meanwhile, these method byproducts are more, Yi Zao Into environmental pollution.Therefore, in order that flavones in the systemic effect of human body more preferably, a kind of efficient conversion flavones of exploitation is glucoside First type flavones is also the task of top priority.
The content of the invention
Flavones and the method for being converted into glucoside type flavone, main bag are extracted the invention provides a kind of efficiency from ginkgo leaf Crushing is included, is sterilized, steam blasting is digested, and is extracted, Ethanol Treatment, concentration, these steps such as dry.
Specifically, extracting flavones the invention provides a kind of efficiency from ginkgo leaf and being converted into the side of glucoside type flavone Method, takes ginkgo leaf cleaning clean, is crushed into 20-100 mesh, and ginkgo leaf adds water mixing, amount of water 40%-60%;Mixture is filled Enter fermentation tank, in 121 DEG C of sterilizations, then carry out steam blasting;Inoculated aspergillus niger, 108Individual/mL seed liquors inoculum concentration is 100mL/ 100kg, fermentation condition is 30 DEG C of 3d;Mixed enzyme solution is then added, addition is the 2%-5% of reaction substrate, in 25-30 DEG C of bar The part lower reaction time is 3-6 hours;Beta-glucosidase is subsequently added, addition is the 1% of reaction substrate, and reaction condition is 42 PH is 4.3 under the conditions of DEG C, and the reaction time is 5 hours;It is subsequently added under the conditions of protease 1%, 25-30 DEG C, the reaction time is 3-4 Hour;Heated up 95 DEG C of 10min, is subsequently added the extraction that extractant carries out flavones, and 12000g centrifugation 20min are taken after supernatant concentration, Extracted using alcohol steep, receive to obtain ethanol eluate, concentrated, using freeze-drying, obtain product.
Still further aspect of the present invention, the steam blasting uses saturated steam for working media, and pressure is 0.73MPa, Dwell time is 70 seconds.
Still further aspect of the present invention, the mixed enzyme solution is cellulase, hemicellulase, pectase these three enzyme liquids Combination;Specific proportion of composing is cellulase:Hemicellulase:Pectase=2:1:5.
Still further aspect of the present invention, described extractant is 4% sodium citrate, and pH is 8.0.Extraction conditions are according to ginkgo The ratio between leaf weight and extractant are 1: 2-4, and the extraction that feeds intake is extracted, and 30-100 DEG C of Extracting temperature is handled 1-5 hours, leaching process Middle interval 10min stirs raw material.
Still further aspect of the present invention, described raw materials for production can be ginkgo fresh leaf or through dry ginkgo leaf, described Leaf needs not move through crushing.
Still further aspect of the present invention, ethanol eluate is concentrated under vacuum.
Still further aspect of the present invention, the aspergillus niger is preserving number CCTCCM2012515.
Still further aspect of the present invention, described production technology will concentration using freeze-drying or spray drying or vacuum drying Liquid is dried.
Still further aspect of the present invention, with GINKGO BILOBA EXTRACT, ginkgolides in high performance liquid chromatography (HPLC) Detection and Extraction thing Method with ginkgolic acid content is respectively:The detection method of GINKGO BILOBA EXTRACT is:Mobile phase:Methanol:Phosphoric acid:Water=57:0.2: 42.8, flow velocity is 1.0ml/min, and Detection wavelength is 370nm, chromatographic column temperature:25 DEG C, sample size:20μl;The detection of ginkgolides Method is:Mobile phase is methanol:Water=25:75, flow velocity 1.0mL/min, detector is ELSD;OPC detection method is inspection Survey wavelength:280nm;30 DEG C of column temperature;Flow rate of mobile phase is 1.0ml/min;Mobile phase:A, 20.0% acetic acid;B:80% acetonitrile;Adopt Use gradient elution:0min-85min, 100%-20%A;6min-85min, 4%-80%B, sample size:10μL;Ginkgolic acid is examined Survey method is methanol:Water=90:10, flow velocity is 1.0mL/min, and Detection wavelength is 310nm.
Compared with prior art, the method for the present invention for extracting flavones has the advantage that:
1st, the present invention has carried out early stage processing using fermentation of Aspergillus niger and composite enzyme solution to ginkgo leaf texture, improves The mass transfer characteristic of material cellular content;Plant cell wall shields, and prevents the random discrepancy of intraor extracellular material, mainly It is made up of cellulose and pectin substance, therefore using cellulase and pectase dissolving cell membrane, plant can be effectively facilitated thin The dissolution of intracellular active ingredient.After fermentation and enzyme liquid mixed processing, with preferable treatment effect.
2nd, the present invention is groped by the substantial amounts of condition of early stage, for the product of above extraction step, using beta-glucosidase Enzyme carries out the conversion of glucoside type flavone, and it is very high that transformation efficiency optimizes, with preferable effect.
3rd, specific processing mode is carried out using steam blasting, the bar of explosion has been carried out especially for ginkgo leaf texture Piece optimization, flavonoid substance that can be in quick release tissue.
4th, sodium citrate have finally chosen by substantial amounts of experiment and is used as extractant, than the such as bisulfite of prior art Sodium has nontoxic effect, use it is more efficient, with more preferable extraction effect.Most importantly can effectively it go Except ginkgoic acid.
5th, extraction process of the present invention need not use macroreticular resin, cost-effective, while being used as extraction, recovery rate with ethanol It is higher, and Extracting temperature is relatively low, organic solvent-free residual, reduces the solvability to impurity, makes chromocor extract once Processing purity reaches more than 98.6%, therefore, and the present invention has the advantages that extract purity is high and recovery rate is good, safe;
6th, using freeze-drying, flavones active ingredient is kept not to be destroyed, bioactivity is high, from unused in technical process Poisonous and hazardous organic solvent, obtained final products organic solvent-free residual;The suitable mankind are safe to use.
Embodiment
With reference to embodiment, the invention will be further described.Embodiment is the optimal embodiment party of displaying Formula, but protection scope of the present invention can not be limited.
The efficiency from ginkgo leaf of embodiment 1 extracts flavones
The ginkgo leaf 100kg for cleaning clean drying is taken, 20 mesh are crushed into, ginkgo leaf adds water mixing, amount of water 50%; Mixture loads fermentation tank, in 121 DEG C of sterilizations, then carries out steam blasting, and steam blasting uses saturated steam to be situated between for work Matter, pressure is 0.73MPa, and the dwell time is 70 seconds;Inoculated aspergillus niger, 108/mL seed liquors inoculum concentration is 100mL/100kg, Fermentation condition is 30 DEG C of 3d;Then add mixed enzyme solution:Cellulase:Hemicellulase:Pectase=2:1:5, addition is The 2% of reaction substrate, the reaction time is 6 hours under the conditions of 30 DEG C, is subsequently added under the conditions of protease 1%, 30 DEG C, during reaction Between be 4 hours;Heat up 95 DEG C of 10min, is subsequently added the sodium citrate that pH is 8.0,4%, and extraction conditions are according to ginkgo leaf weight The ratio between amount and extractant are 1: 3, and the extraction that feeds intake is extracted, and 60 DEG C of Extracting temperature is handled 3 hours, interval 10min in leaching process 50r/min stirs raw material.12000g centrifuges 20min, takes supernatant to use concentration tower to carry out concentration cocnentration factor for 1:10, concentration Liquid is extracted with absolute ethyl alcohol, and the ultimate density of ethanol is 90%, receives to obtain alcohol extract, it is 1 that concentration tower, which carries out concentration cocnentration factor,: 15, it is freeze-dried using atomizing freeze drying machine, obtains obtaining the faint yellow ginkgo biloba p.e products of 4.36kg.
Specific composition employs high performance liquid chromatography and analyzed, and assay is:In total flavone glucoside 38.0%, total terpene Solubility 11.8% in ester 10.6%, OPC 12.3%, ginkgolic acid 0.1ppm, water, wherein ginkgo acid content 0.01ppm It is nearly no detectable.
The reference examples 1 of embodiment 2
The composition and proportioning (being calculated by deal) of extractant:Sodium hydrogensulfite 10kg;Running water 8000kg;pH5.0.
By dry ginkgo leaf 300kg cleanings totally, drainage puts into extractor, adds 94 DEG C of extractant 5000kg, plus Lid is closed, and temperature makes up to 97 DEG C in rise extractor, and keeps 1 atmospheric pressure, is stirred 1 minute per half an hour, controls tank Interior temperature declines 2.5 DEG C per hour, after extracting 16 hours, filters out feed liquid 4400kg.3000kg extractions are added into extractor again Agent solution, carries out second and extracts, feed liquid 3100kg is filtered out after 10 hours, merges feed liquid filter centrifugal coarse filtration twice and goes out 20 The following impurity of mesh, sand filtration machine refined filtration goes out the following impurity of 200 mesh, and feed liquid is cooled into 30 DEG C.
Feed liquid is flowed through into NKA-9 resin absorbing columns with 1500kg/ hours flows, is then washed with water down and is not adsorbed or adsorbs Less impurity, changes the washing of 2 times of column volume deionized waters, then with 70% alcohol flushing resin adsorption column, receive to obtain ethanol elution Liquid 1500kg, 20 times of ethanol eluate is concentrated at a temperature of 43 DEG C using triple effect concentration tower, and concentrate is obtained using freeze-drying Product 9.75kg.Assay:Total flavone glucoside 28.1%, Total terpene lactones 9.2%, OPC 10.3%, ginkgolic acid Solubility 7.7% in 3.8ppm, water.
The reference examples 2 of embodiment 3
(1) 100 kilograms of ginkgo leaves are crushed with pulverizer, poured into extractor, add 700 kg of water, while adding 1.5 Kilogram calcium carbonate, heating, water boil 1.2 hours after filter, obtain an Aqueous extracts;Same method is extracted once again, merges one Secondary, secondary Aqueous extracts, produce amalgamation liquid;
(2) by amalgamation liquid in 65 DEG C of Zhen Kong Nong Shrink tank Zhong Nong Shrink to 50L, ethanol is added, is 45% to ethanol degree, fully Stirring, filters after standing 3.5 hours, abandons alcohol sediment;It is diluted with water filtered fluid so that ethanol degree is 18%, is adjusted with dilute sulfuric acid It is 3.0 to pH value, filtrate must be diluted;Dilution filtrate is adsorbed with DM131 resin columns, resin column blade diameter length ratio is l:5, flow velocity lBv/ H, resin demand is that resin demand is 50 kilograms (volume is 50.1L);
(3) it is 3.0 that the ethanol water for being 20% by 100L ethanol degree, which is adjusted to pH value with dilute sulfuric acid, elution impurity 2 hours, Elution flow rate is 1BV/h, then is rinsed with water to neutrality;
(4) it is 70% ethanol water elution active ingredient 3 hours with 400L ethanol degree, flow velocity 2BV/h then will elution 6, (Zhen Kong Nong Shrink, drying at a temperature of TC obtain 2.5 kilograms of gingko flavone extractive, wherein total flavone glucoside is 25.3%, total terpene to liquid Lactone is 7.2%, and OPC 9.0%, ginkgolic acid is 3.4ppm.
A kind of efficiency from ginkgo leaf of embodiment 4 extracts flavones and the method for being converted into glucoside type flavone
The ginkgo leaf 100kg for cleaning clean drying is taken, 20 mesh are crushed into, ginkgo leaf adds water mixing, amount of water 50%; Mixture loads fermentation tank, in 121 DEG C of sterilizations, then carries out steam blasting, and steam blasting uses saturated steam to be situated between for work Matter, pressure is 0.73MPa, and the dwell time is 70 seconds;Inoculated aspergillus niger, 108Individual/mL seed liquors inoculum concentration is 100mL/100kg, Fermentation condition is 30 DEG C of 3d;Then add mixed enzyme solution:Cellulase:Hemicellulase:Pectase=2:1:5, addition is The 2% of reaction substrate, the reaction time is 6 hours under the conditions of 30 DEG C;Beta-glucosidase is subsequently added, addition is reaction bottom The 1% of thing, reaction condition is that pH is 4.3 under the conditions of 42 DEG C, and the reaction time is 5 hours;It is subsequently added protease 1%, 30 DEG C of bars Under part, the reaction time is 4 hours;95 DEG C of heating, 10min is subsequently added the sodium citrate that pH is 8.0,4%, and extraction conditions are It is 1: 3 according to the ratio between ginkgo leaf weight and extractant, the extraction that feeds intake is extracted, 60 DEG C of Extracting temperature is handled 3 hours, leaching process Middle interval 10min 50r/min stir raw material.12000g centrifuges 20min, takes supernatant to carry out concentration concentration using concentration tower Than for 1:10, concentrate is extracted with absolute ethyl alcohol, and the ultimate density of ethanol is 90%, receives to obtain alcohol extract, and concentration tower is carried out It is 1 to concentrate cocnentration factor:15, it is freeze-dried using atomizing freeze drying machine, obtains obtaining the faint yellow ginkgo leaf extractions of 4.37kg Produce product.
Specific composition employs high performance liquid chromatography and analyzed, and assay is:In total flavone glucoside 38.2%, total terpene Solubility 11.7% in ester 10.5%, OPC 12.4%, ginkgolic acid 0.1ppm, water, wherein ginkgo acid content 0.01ppm It is nearly no detectable.Glucoside type flavone accounting is 92.3%. wherein in total flavone glucoside
By above example as can be seen that the extract of the present invention has preferable extraction effect, particularly ginkgolic acid Tool is significantly reduced trend, while glucoside type flavone accounting is higher in total flavone glucoside, this also illustrates that the glucoside type of the present invention is yellow Ketone transformation efficiency is also higher, has preferably synergy between this explanation each enzyme of the invention, while in extraction process, glucoside member The loss of type flavones is relatively low, with preferable extraction effect.And the overall extraction step of the present invention is also simple, is suitable for work Industry metaplasia is produced.The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention Any modifications, equivalent substitutions and improvements made within refreshing and principle etc., should be included in the scope of the protection.

Claims (7)

1. a kind of efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone, main to include crushing, sterilize, steam Steam explosion is broken, and is fermented, and is digested, and is extracted, Ethanol Treatment, concentration, drying and other steps.
2. a kind of efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone, take ginkgo leaf cleaning clean, powder It is broken into as 20-100 mesh, ginkgo leaf adds water mixing, amount of water 40%-60%;Mixture loads fermentation tank, is sterilized at 121 DEG C, with After carry out steam blasting;Inoculated aspergillus niger, 108Individual/mL seed liquors inoculum concentration is 100mL/100kg, and fermentation condition is 30 DEG C of 3d; Mixed enzyme solution is then added, addition is the 2%-5% of reaction substrate, the reaction time is 3-6 hours under the conditions of 25-30 DEG C, It is subsequently added beta-glucosidase, addition is the 1% of reaction substrate, reaction condition is that pH is 4.3 under the conditions of 42 DEG C, during reaction Between be 5 hours;It is subsequently added under the conditions of protease 1%, 25-30 DEG C, the reaction time is 3-4 hours;95 DEG C of heating, 10min, with The extraction that extractant carries out flavones is added afterwards, and 12000g centrifugation 20min are taken after supernatant concentration, extracted, received using alcohol steep Alcohol extract, concentration, using freeze-drying, obtains product;
Wherein, the steam blasting uses saturated steam for working media, and pressure is 0.73MPa, and the dwell time is 70 seconds;
The enzyme liquid is cellulase, hemicellulase, the combination of pectase these three enzyme liquids;Specific proportion of composing is fiber Plain enzyme:Hemicellulase:Pectase=2:1:5;
Described extractant is 4% sodium citrate, and pH is 8.0;It according to the ratio between ginkgo leaf weight and extractant is 1 that extraction conditions, which are, : 2-4, the extraction that feeds intake is extracted, and 30-100 DEG C of Extracting temperature is handled 1-5 hours, and interval 10min stirrings raw material is in leaching process Can.
3. according to the method for claim 1 or 2, described raw materials for production are ginkgo fresh leafs or through dry ginkgo leaf.
4. method according to claim 3, alcohol extract is concentrated under vacuum.
5. according to the method for claim 1 or 2 or 4, wherein the freeze-drying is that under vacuum will carry out concentrate Dry.
6. the gingko flavone extractive that the method according to claim 2 or 4 is prepared.
7. purposes of the obtained chromocor extract prepared described in claim 6 in health products or medicine are prepared.
CN201710332354.XA 2017-05-12 2017-05-12 A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone Pending CN107158048A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710332354.XA CN107158048A (en) 2017-05-12 2017-05-12 A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710332354.XA CN107158048A (en) 2017-05-12 2017-05-12 A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone

Publications (1)

Publication Number Publication Date
CN107158048A true CN107158048A (en) 2017-09-15

Family

ID=59814998

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710332354.XA Pending CN107158048A (en) 2017-05-12 2017-05-12 A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone

Country Status (1)

Country Link
CN (1) CN107158048A (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108840727A (en) * 2018-07-19 2018-11-20 长沙学院 A kind of plant nutrient agent and preparation method thereof producing high-quality balsam pear
CN111213512A (en) * 2020-01-13 2020-06-02 广东省农业科学院果树研究所 Planting method for improving total flavone content and antioxidant activity of wampee fruits
CN113295798A (en) * 2021-05-28 2021-08-24 浙江树人学院(浙江树人大学) Sample chromatographic analysis method based on measurement of sample component retention time

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102351826A (en) * 2011-09-26 2012-02-15 重庆大学 Extraction method of ginkgo leaf flavonoids based on steam explosion
CN103263448A (en) * 2013-03-01 2013-08-28 南京林业大学 Fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and application
CN104906153A (en) * 2015-06-09 2015-09-16 邳州鑫源生物制品有限公司 Technological method for efficiently extracting ginkgo flavone

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102351826A (en) * 2011-09-26 2012-02-15 重庆大学 Extraction method of ginkgo leaf flavonoids based on steam explosion
CN103263448A (en) * 2013-03-01 2013-08-28 南京林业大学 Fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and application
CN104906153A (en) * 2015-06-09 2015-09-16 邳州鑫源生物制品有限公司 Technological method for efficiently extracting ginkgo flavone

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108840727A (en) * 2018-07-19 2018-11-20 长沙学院 A kind of plant nutrient agent and preparation method thereof producing high-quality balsam pear
CN111213512A (en) * 2020-01-13 2020-06-02 广东省农业科学院果树研究所 Planting method for improving total flavone content and antioxidant activity of wampee fruits
CN111213512B (en) * 2020-01-13 2021-10-12 广东省农业科学院果树研究所 Planting method for improving total flavone content and antioxidant activity of wampee fruits
CN113295798A (en) * 2021-05-28 2021-08-24 浙江树人学院(浙江树人大学) Sample chromatographic analysis method based on measurement of sample component retention time

Similar Documents

Publication Publication Date Title
CN110304994B (en) Method for extracting high-purity cannabidiol from industrial cannabis sativa
CN104710391B (en) Method for extracting luteolin and beta-sitosterol from peanut shells
CN107115367A (en) A kind of high efficiency from ginkgo leaf extracts the fermentation method for producing of flavones
CN101402657A (en) Process for producing chestnut shell polyphenol
CN104906153A (en) Technological method for efficiently extracting ginkgo flavone
CN104258193B (en) A kind of pharmaceutical composition for treating neurasthenia and prevention senile dementia
CN107158048A (en) A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone
CN102675398B (en) A kind of method extracting momordica grosvenori glycoside V and farnesol from Grosvenor Momordica
CN103432562A (en) Method for extracting fresh ginger polyphenol from fresh ginger
CN102002083B (en) Method for extracting high-purity rutin with flash-extraction technology
CN106349405A (en) Method for extracting pectin from shaddock peel through enzymolysis and ultrasonic waves
CN103356740A (en) Preparation method of baicalein and scutellaria baicalensis flavone total-aglycone extractives
CN107903293A (en) A kind of extraction process of Camellia nitidissima saponin extract
CN104000935A (en) Method for extracting anti-oxidative phenolic acids from potato peel slag
CN106749487A (en) A kind of method that separating ursolic acid is extracted from seabuckthorn fruit peel
CN107375356B (en) Method for simultaneously preparing high-purity total flavonol glycosides and ginkgolides
CN103396461B (en) A kind of separating and purifying method of secoisolariciresinol diglycoside
CN106810619A (en) A kind of gingko episperm pectin and polypentenol extracting method
CN111018929B (en) Process for extracting and separating isocoryzanol
CN107383152A (en) The preparation method of golden flower Tea Saponins
CN102952002A (en) Method for refining magnolol through supersonic extraction separating technology
CN107344930A (en) A kind of method that young fustic is extracted from Rhus succedanea
CN110903168B (en) Method for subcritical extraction of solanesol in waste tobacco leaves
CN102772452A (en) Method for producing ultra-low-acid ginkgo leaf extract
CN108178775A (en) The method that cape jasmine extracts gardenoside and ursolic acid

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20170915