CN106166164A - The preparation method of a kind of high-purity ginkgo leaf standard extract and goods thereof - Google Patents
The preparation method of a kind of high-purity ginkgo leaf standard extract and goods thereof Download PDFInfo
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- CN106166164A CN106166164A CN201610621671.9A CN201610621671A CN106166164A CN 106166164 A CN106166164 A CN 106166164A CN 201610621671 A CN201610621671 A CN 201610621671A CN 106166164 A CN106166164 A CN 106166164A
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- 239000000284 extract Substances 0.000 title claims abstract description 55
- 241000218628 Ginkgo Species 0.000 title claims abstract description 54
- 235000011201 Ginkgo Nutrition 0.000 title claims abstract description 54
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 70
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- 238000010992 reflux Methods 0.000 claims abstract description 20
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- 238000001556 precipitation Methods 0.000 claims abstract description 16
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- 238000000703 high-speed centrifugation Methods 0.000 claims abstract description 9
- 238000010828 elution Methods 0.000 claims abstract description 8
- 239000002994 raw material Substances 0.000 claims abstract description 6
- MOLPUWBMSBJXER-YDGSQGCISA-N bilobalide Chemical compound O([C@H]1OC2=O)C(=O)[C@H](O)[C@@]11[C@@](C(C)(C)C)(O)C[C@H]3[C@@]21CC(=O)O3 MOLPUWBMSBJXER-YDGSQGCISA-N 0.000 claims description 26
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 16
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- SVINQHQHARVZFF-UHFFFAOYSA-N Ginkgotoxin Chemical compound COCC1=C(CO)C=NC(C)=C1O SVINQHQHARVZFF-UHFFFAOYSA-N 0.000 claims description 11
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- 239000012141 concentrate Substances 0.000 claims description 8
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- 229930045534 Me ester-Cyclohexaneundecanoic acid Natural products 0.000 description 6
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- 239000000243 solution Substances 0.000 description 6
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- GQODBWLKUWYOFX-UHFFFAOYSA-N Isorhamnetin Natural products C1=C(O)C(C)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 GQODBWLKUWYOFX-UHFFFAOYSA-N 0.000 description 2
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 description 2
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- IYRMWMYZSQPJKC-UHFFFAOYSA-N kaempferol Chemical compound C1=CC(O)=CC=C1C1=C(O)C(=O)C2=C(O)C=C(O)C=C2O1 IYRMWMYZSQPJKC-UHFFFAOYSA-N 0.000 description 2
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- 238000010812 external standard method Methods 0.000 description 1
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- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- -1 has a ginkalide A Chemical class 0.000 description 1
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- 235000008777 kaempferol Nutrition 0.000 description 1
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- UXOUKMQIEVGVLY-UHFFFAOYSA-N morin Natural products OC1=CC(O)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 UXOUKMQIEVGVLY-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- 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
- A61K36/16—Ginkgophyta, e.g. Ginkgoaceae (Ginkgo family)
-
- 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
-
- 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
-
- 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
-
- 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
-
- 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 the preparation method of a kind of high-purity ginkgo leaf standard extract, this preparation method comprises the steps: to weigh raw material, pulverizes and sieves, and puts into and extracts in return tank；Reflux, extract,；Recovered alcohol, is concentrated in vacuo to and obtains the extractum that proportion is 1.0 1.5；Being sufficiently stirred for by extractum pure water, high speed centrifugation removes precipitation, obtains supernatant；Supernatant is crossed macroporous resin, gradient elution, collects 40 60% ethanol eluents, be then acidified, obtain acidifying solution；By acidifying solution precipitates overnight at a temperature of 05 DEG C, collect precipitation；Precipitate pure water is fully washed, then under vacuum condition, is dried to obtain standard Folium Ginkgo extract.The used equipment of the present invention is simple, technique simple possible, it is simple to industrialized production；Solvent for use nontoxic residue-free, does not produce environmental pollution；Folium Ginkgo extract nuisance removal rate of the present invention is high, and institute's production standard extract meets U.S. FDA requirement.
Description
Technical field
The present invention relates to pharmaceutical technology field, be more particularly to the preparation method of a kind of high-purity ginkgo leaf standard extract
And goods.
Background technology
Semen Ginkgo standard extract is to be widely used in cardiovascular and cerebrovascular disease medication, is one of widest herbal species, with
The further investigation to Semen Ginkgo extrac, increasingly find that in Semen Ginkgo extrac, active component and some other composition are to human body
Side effect, gingkolic acid makes human body skin generation allergic dermatitis, and 4-O-methyl pyridoxine causes twitching and unconscious effect.Silver
Fructus Pruni extract mainly contain biological activity flavone has a kind more than 30, wherein more than 20 structures planted have been analyzed clear, various researchs
Showing, ginkgetin has a suppression platelet coalescence, regulation blood fat, expands blood vessel, improves cerebral circulation, improves brain function, anti-
The biological activity widely such as atherosclerosis, antitumor；Bilobalide is diterpene-kind compound, mainly has a ginkalide A, B,
These are several for C, J, M and bilobalide, and cerebral nerve is passed by bilobalide is well recognized as PAF receptor antagonist
Pass, restoration and protection cerebral tissue aspect effect huge, cardiovascular and cerebrovascular vessel are had good DEVELOPMENT PROSPECT.China Semen Ginkgo is world's silver
Fructus Pruni main product ground, accounts for the yield in the whole world 70%, but Semen Ginkgo extrac is at China but major part only primary extract, content
Low, lactone content does not reaches requirement, and poisonous and harmful element therein cannot separate, and result produces to external offer primary
Product, are worth the lowest.Semen Ginkgo extrac Study on extraction overview: Semen Ginkgo extrac manufacturer is a lot, and major part is all that water carries
Or low-concentration ethanol extracts, then cross macroporous resin D101, be then spray-dried gained.Other are also in laboratory research rank
Section, does not enter into production application, such as molecular film Filtration, and reverse liquid chromatography, SCF-CO 2 etc. are such
Report is a lot.Now it is badly in need of researching and developing a kind of new method improving flavonoid of ginkgo biloba.
Summary of the invention
(1) to solve the technical problem that
The technical problem to be solved in the present invention is how to improve the content of ginkgetin, effectively removes harmful components, and
Preparation method and the goods thereof of a kind of high-purity ginkgo leaf standard extract are provided.
(2) technical scheme
In order to solve above-mentioned technical problem, the invention provides the preparation side of a kind of high-purity ginkgo leaf standard extract
Method, this preparation method comprises the steps (raw materials used the most commercial obtain):
Step one: prepare: weigh raw material, pulverized 18-22 mesh sieve, and dried, is then put in extraction return tank；
Step 2: reflux, extract: adding the 60-75% ethanol of Folium Ginkgo weight 8-12 times, reflux temperature controls at 65-70
DEG C, return time is 2.5-3.5 hour；
Step 3: secondary back extracts: adding the 60-75% ethanol of Folium Ginkgo weight 4-6 times, temperature controls at 65-70
DEG C, return time is 1.5-2.5 hour；
Step 4: concentrate: merge twice withdrawing fluid, recovered alcohol, be concentrated in vacuo to and obtain the leaching that proportion is 1.0-1.5
Cream；
Step 5: de-bubble: be sufficiently stirred for by the pure water of 8-12 times of weight of extractum, alkalizes to pH 8.0-10 with KOH, adds
Enter 0.2% Tween 80, the automatic scumming of high-speed stirred, and to not producing foam, then high speed centrifugation removes precipitation, obtain on
Clear liquid；
Step 6: macroporous resin filters: supernatant is crossed macroporous resin, gradient elution, collects 40-60% ethanol eluting
Liquid, 5 BV eluents, recovered alcohol, it is sufficiently stirred for 8-12 times of pure water, is then acidified to PH3.5-4.5 with hydrochloric acid, obtains acid
Change liquid；
Step 7: acidifying solution step 6 obtained precipitates overnight at a temperature of 0-5 DEG C, collects precipitation；
Step 8: vacuum drying: fully washing precipitate pure water at a temperature of 0-5 DEG C, pH value is 6.5-7.0, so
Being dried to obtain standard Folium Ginkgo extract, flavonoid of ginkgo biloba 31% under the conditions of final vacuum, bilobalide 11%, gingkolic acid is low
It is less than 1PPM less than 1PPM, bisflavone content in 1PPM, 4-O-methyl pyridoxine；Recovery rate is 2.3%.
Preferably, in step one, described sieve is 20 mesh.
Preferably, in step 2 and step 3, described reflux temperature is 65 DEG C.
Preferably, in step 4, described recovered alcohol is recycled to alcohol content less than 15%.
Preferably, in step 4, described vacuum is-0.08--0.09MPa；Described proportion is 1.15-1.2.
Preferably, in step 6, the macroporous resin used is D201 macroporous resin.
Preferably, described preparation method comprises the steps:
Step one: prepare: weigh raw material, pulverized 20 mesh sieves, and dried, is then put in extraction return tank；
Step 2: reflux, extract: adding 65% ethanol of Folium Ginkgo weight 10 times, reflux temperature controls at 65 DEG C, backflow
Time is 3 hours；
Step 3: secondary back extracts: adding 65% ethanol of Folium Ginkgo weight 5 times, temperature controls at 65 DEG C, during backflow
Between be 2 hours；
Step 4: concentrate: merging twice withdrawing fluid, recovered alcohol is recycled to alcohol content less than 15%, is concentrated in vacuo
To obtaining the extractum that proportion is 1.15-1.2；
Step 5: de-bubble: be sufficiently stirred for by the extractum pure water of 10 times of weight, with KOH alkalization to pH 9.0, adds
0.2% Tween 80, the automatic scumming of high-speed stirred, and to not producing foam, then high speed centrifugation removes precipitation, obtains supernatant
Liquid；
Step 6: macroporous resin filters: supernatant is crossed D201 macroporous resin, gradient elution, collects 40-60% ethanol and washes
De-liquid, 5 BV eluents, recovered alcohol, then it is sufficiently stirred for 10 times of pure water, is then acidified to PH4.0 with hydrochloric acid, obtains acid
Change liquid；
Step 7: acidifying solution step 6 obtained precipitates overnight at a temperature of 0-5 DEG C, collects precipitation；
Step 8: vacuum drying: fully washing precipitate pure water at a temperature of 0-5 DEG C, pH value is 7.0, the most very
Being dried to obtain standard Folium Ginkgo extract, flavonoid of ginkgo biloba 31% under empty condition, bilobalide 11%, gingkolic acid is less than
1PPM, 4-O-methyl pyridoxine is less than 1PPM, and bisflavone content is less than 1PPM；Recovery rate is 2.3%.
Present invention also offers the Folium Ginkgo mark obtained by preparation method of described high-purity ginkgo leaf standard extract
Quasi-extract.
(3) beneficial effect
The used equipment of the present invention is simple, technique simple possible, it is simple to industrialized production；Solvent for use nontoxic residue-free,
Do not produce environmental pollution；Semen Ginkgo nuisance removal rate of the present invention is high, and institute's production standard extract meets U.S. FDA requirement.
Detailed description of the invention
Below in conjunction with embodiment, embodiments of the present invention are described in further detail.Following example are used for illustrating this
Invention, but can not be used for limiting the scope of the present invention.
Solvent, KOH, HCL of being used below are food grade or analytical pure, must not use technical grade.
Embodiment 1
1, take Folium Ginkgo (doing) 500KG, (HPLC detection is containing ginkgetin 0.72%, terpene dilactone 0.29%), pulverize
Cross 20 mesh sieves, put into reflux, extract, tank；
2, reflux, extract, for the first time: 65% ethanol 5000 kilograms, temperature controls 65 DEG C of return times 3 hours；
3, second time backflow: 65% ethanol 2500 kilograms, temperature controls at 65 DEG C, refluxes 2 hours；
4, concentrate: merge twice withdrawing fluid, recovered alcohol, be concentrated in vacuo to the extractum 173 kilograms of proportion 1.2.
5, de-bubble: 173 kilograms of extractum add 1730 kilograms of pure water, KOH alkalizes to PH9.0, adds 0.2% Tween 80, at a high speed
Stirring automatic scumming, to not producing foam, then high speed centrifugation removes precipitation, obtains supernatant 1860 kilograms.
6, macroporous resin filters: supernatant crosses D201 macroporous resin, gradient elution, collects 40%-60% ethanol eluent,
5 BV eluents, are sufficiently stirred for 10 times of water after recovered alcohol, and then hydrochloric acid is acidified to PH4.0；
7, acidifying solution is precipitates overnight at 0-5 DEG C, collects precipitation 39 kilograms；
8, washing vacuum drying: precipitate pure water is fully washed at 0-5 DEG C, PH to 6.5-7.0, the most very
It is dried to obtain standard Folium Ginkgo extract under empty condition, analyzes to obtain flavonoid of ginkgo biloba 30.69%, bilobalide through HPLC
11.72%, gingkolic acid, 4-O-methyl pyridoxine, bisflavone content is below 1PPM, receives to obtain Semen Ginkgo standard extract 11.36
Kilogram.
Embodiment 2
1, take Folium Ginkgo (doing) 500KG, (HPLC detection is containing ginkgetin 0.76%, terpene dilactone 0.31%), pulverize
Cross 20 mesh sieves, put into reflux, extract, tank；
2, reflux, extract, for the first time: 65% ethanol 5000 kilograms, temperature controls 65 DEG C of return times 3 hours；
3, second time backflow: 65% ethanol 2500 kilograms, temperature controls at 65 DEG C, refluxes 2 hours；
4, concentrate: merge twice withdrawing fluid, recovered alcohol, be concentrated in vacuo to the extractum 179 kilograms of proportion 1.2.
5, de-bubble: 179 kilograms of extractum add 1790 kilograms of pure water, KOH alkalizes to PH9.0, adds 0.2% Tween 80, at a high speed
Stirring automatic scumming, to not producing foam, then high speed centrifugation removes precipitation, obtains supernatant 1910 kilograms.
6, macroporous resin filters: supernatant crosses D201 macroporous resin, gradient elution, collects 40%-60% ethanol eluent,
5 BV eluents, are sufficiently stirred for 10 times of water after recovered alcohol, and then hydrochloric acid is acidified to PH4.0；
7, acidifying solution is precipitates overnight at 0-5 DEG C, collects precipitation 46 kilograms；
8, washing vacuum drying: precipitate pure water is fully washed at 0-5 DEG C, PH to 6.5-7.0, the most very
It is dried to obtain standard Folium Ginkgo extract under empty condition, analyzes to obtain flavonoid of ginkgo biloba 30.27%, bilobalide through HPLC
10.97%, gingkolic acid, 4-O-methyl pyridoxine, bisflavone content is below 1PPM, receives to obtain Semen Ginkgo standard extract 12.77
Kilogram.
Embodiment 3
1, take Folium Ginkgo (doing) 500KG, (HPLC detection is containing ginkgetin 0.69%, terpene dilactone 0.25%), pulverize
Cross 20 mesh sieves, put into reflux, extract, tank；
2, reflux, extract, for the first time: 65% ethanol 5000 kilograms, temperature controls 65 DEG C of return times 3 hours；
3, second time backflow: 65% ethanol 2500 kilograms, temperature controls at 65 DEG C, refluxes 2 hours；
4, concentrate: merge twice withdrawing fluid, recovered alcohol, be concentrated in vacuo to the extractum 165 kilograms of proportion 1.2.
5, de-bubble: 165 kilograms of extractum add 1650 kilograms of pure water, KOH alkalizes to PH9.0, adds 0.2% Tween 80, at a high speed
Stirring automatic scumming, to not producing foam, then high speed centrifugation removes precipitation, obtains supernatant 1790 kilograms.
6, macroporous resin filters: supernatant crosses D201 macroporous resin, gradient elution, collects 40%-60% ethanol eluent,
5 BV eluents, are sufficiently stirred for 10 times of water after recovered alcohol, and then hydrochloric acid is acidified to PH4.0；
7, acidifying solution is precipitates overnight at 0-5 DEG C, collects precipitation 33 kilograms；
8, washing vacuum drying: precipitate pure water is fully washed at 0-5 DEG C, PH to 6.5-7.0, the most very
It is dried to obtain standard Folium Ginkgo extract under empty condition, analyzes to obtain flavonoid of ginkgo biloba 30.19%, bilobalide through HPLC
11.05%, gingkolic acid, 4-O-methyl pyridoxine, bisflavone content is below 1PPM, receives to obtain Semen Ginkgo standard extract 10.15
Kilogram.
Comparative example 1
Using the Semen Ginkgo extraction and purification process of Shandong Agricultural University, alcohol steep-resin adsorption method, technique is:
Folium Ginkgo 12 times of foliage weight of 70% ethanol, 55-60 DEG C extracts 3 hours, extracts 2 times, merging filtrate, be concentrated into
Proportion 1.2,10 times of water add extractum dilution and are sufficiently stirred for, and high speed centrifugation precipitates, and removes supernatant, crosses D101 macroporous resin column, water
Washing resin is to colourless, then with 70% ethanol eluting, obtains eluent, concentrates eluent, is spray-dried to obtain Folium Ginkgo extract.
Specifically extract to obtain Semen Ginkgo by 500 kg silver Folium Pruni (analyzing flavones content 0.81%, lactone 0.32%) process above
Leaf extract 16.7 kilograms.
Experimental example 1
Ginkgo total flavones HPLC measures Beijing Food Research Inst. of nutrition unit such as (food science and technology 2006) 03 Wang Jing
1, detection method:
1) material and instrument
Phosphoric acid, hydrochloric acid, methanol, standard substance (kaempferol, Quercetin, isorhamnetin), pure water
WATER 2487 type high pressure liquid chromatography, ultrasonic wave concussion device, DHT magnetic stirring apparatus
2) method
Chromatographic condition: chromatographic column diamond C18 (250*4.6 5 microns) column temperature 40 DEG C, detects wavelength 370nm
Flowing phase: methanol: 0.4% phosphoric acid=60:40 (volume ratio), flow velocity 1.0ML/MIN, sample size 20UL
3) quantitative approach: external standard method
4) calculate:
Total flavones=2.51 Quercetin amount+2.63 kaempferide amount+2.39 isorhamnetin amount
Experimental example 2
Bilobalide detects
RP-HPLC measures method (Cui great Ming, Changchun traditional Chinese medical science pharmaceutical university, Jilin Agriculture University of bilobalide content
Report 2013,35 (4))
Experimental example 3
4-O-methyl pyridoxine measures
GC-MS detects 4-O-methyl pyridoxine
Experimental example 4
Gingkolic acid measures
RP-HPLC measures the method (Yan Hongfeng, Huaiyin, Jiangsu Province engineering college journal 2,008 10) of gingkolic acid content
Table: testing result
| Sample | Ginkgetin | Bilobalide | 4-O-methyl pyridoxine | Gingkolic acid |
| Embodiment 1 | 30.69% | 11.72% | Less than 1PPM | Less than 1PPM |
| Embodiment 2 | 30.27% | 10.97% | Less than 1PPM | Less than 1PPM |
| Embodiment 3 | 30.19% | 11.05% | Less than 1PPM | Less than 1PPM |
| Comparative example | 24.9% | 6.82% | 15 mgs/kg | 27 mgs/kg |
In table, result shows: contrast traditional extraction technique, and present invention harmful substance in removing Semen Ginkgo extrac has substantially
Improvement effect has good popularization and application meaning.
Embodiment of above is merely to illustrate the present invention, rather than limitation of the present invention.Although with reference to embodiment to this
Bright be described in detail, it will be understood by those within the art that, technical scheme is carried out various combination,
Amendment or equivalent, without departure from the spirit and scope of technical solution of the present invention, all should contain the right in the present invention and want
Ask in the middle of scope.
Claims (8)
1. the preparation method of a high-purity ginkgo leaf standard extract, it is characterised in that this preparation method comprises the steps:
Step one: prepare: weigh raw material, pulverized 18-22 mesh sieve, and dried, is then put in extraction return tank；
Step 2: reflux, extract: adding the 60-75% ethanol of Folium Ginkgo weight 8-12 times, reflux temperature controls at 65-70 DEG C,
Return time is 2.5-3.5 hour；
Step 3: secondary back extracts: adding the 60-75% ethanol of Folium Ginkgo weight 4-6 times, temperature controls at 65-70 DEG C, returns
The stream time is 1.5-2.5 hour；
Step 4: concentrate: merge twice withdrawing fluid, recovered alcohol, be concentrated in vacuo to and obtain the extractum that proportion is 1.0-1.5；
Step 5: de-bubble: be sufficiently stirred for by the pure water of 8-12 times of weight of extractum, with KOH alkalization to pH 8.0-10, adds
0.2% Tween 80, the automatic scumming of high-speed stirred, and to not producing foam, then high speed centrifugation removes precipitation, obtains supernatant
Liquid；
Step 6: macroporous resin filters: supernatant crosses macroporous resin, gradient elution, collects 40-60% ethanol eluent, 5
BV eluent, recovered alcohol, it is sufficiently stirred for 8-12 times of pure water, is then acidified to PH3.5-4.5 with hydrochloric acid, obtains acidifying solution；
Step 7: acidifying solution step 6 obtained precipitates overnight at a temperature of 0-5 DEG C, collects precipitation；
Step 8: vacuum drying: fully washing precipitate pure water at a temperature of 0-5 DEG C, pH value is 6.5-7.0, the most very
Being dried to obtain standard Folium Ginkgo extract, flavonoid of ginkgo biloba 31% under empty condition, bilobalide 11%, gingkolic acid is less than
1PPM, 4-O-methyl pyridoxine is less than 1PPM, and bisflavone content is less than 1PPM；Recovery rate is 2.3%.
The preparation method of high-purity ginkgo leaf standard extract the most according to claim 1, it is characterised in that in step one
In, described sieve is 20 mesh.
The preparation method of high-purity ginkgo leaf standard extract the most according to claim 1, it is characterised in that in step 2
With in step 3, described reflux temperature is 65 DEG C.
The preparation method of high-purity ginkgo leaf standard extract the most according to claim 1, it is characterised in that in step 4
In, described recovered alcohol is recycled to alcohol content less than 15%.
The preparation method of high-purity ginkgo leaf standard extract the most according to claim 1, it is characterised in that in step 4
In, described vacuum is-0.08--0.09MPa；Described proportion is 1.15-1.2.
The preparation method of high-purity ginkgo leaf standard extract the most according to claim 1, it is characterised in that in step 6
In, the macroporous resin used is D201 macroporous resin.
7. according to the preparation method of the high-purity ginkgo leaf standard extract described in any one of claim 1-6, it is characterised in that
This preparation method comprises the steps:
Step one: prepare: weigh raw material, pulverized 20 mesh sieves, and dried, is then put in extraction return tank；
Step 2: reflux, extract: adding 65% ethanol of Folium Ginkgo weight 10 times, reflux temperature controls at 65 DEG C, return time
It it is 3 hours；
Step 3: secondary back extracts: adding 65% ethanol of Folium Ginkgo weight 5 times, temperature controls at 65 DEG C, and return time is
2 hours；
Step 4: concentrate: merging twice withdrawing fluid, recovered alcohol is recycled to alcohol content less than 15%, is concentrated in vacuo to
It is the extractum of 1.15-1.2 to proportion；
Step 5: de-bubble: be sufficiently stirred for by the extractum pure water of 10 times of weight, with KOH alkalization to pH 9.0, adds 0.2% and tells
Temperature 80, the automatic scumming of high-speed stirred, and to not producing foam, then high speed centrifugation removes precipitation, obtains supernatant；
Step 6: macroporous resin filters: supernatant is crossed D201 macroporous resin, gradient elution, collects 40-60% ethanol eluting
Liquid, 5 BV eluents, recovered alcohol, then it is sufficiently stirred for 10 times of pure water, is then acidified to PH4.0 with hydrochloric acid, is acidified
Liquid；
Step 7: acidifying solution step 6 obtained precipitates overnight at a temperature of 0-5 DEG C, collects precipitation；
Step 8: vacuum drying: fully washing precipitate pure water at a temperature of 0-5 DEG C, pH value is 7.0, then vacuum bar
Be dried to obtain standard Folium Ginkgo extract under part, flavonoid of ginkgo biloba 31%, bilobalide 11%, gingkolic acid less than 1PPM,
4-O-methyl pyridoxine is less than 1PPM, and bisflavone content is less than 1PPM；Recovery rate is 2.3%.
8. the Folium Ginkgo mark obtained by preparation method of the high-purity ginkgo leaf standard extract described in any one of claim 1-7
Quasi-extract.
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| CN106727746A (en) * | 2017-04-07 | 2017-05-31 | 浙江新昌天然保健品有限公司 | The preparation method of low sour water soluble ginkgo leaf extract |
| CN109718141A (en) * | 2019-03-14 | 2019-05-07 | 上海琴馨文化传播有限公司 | A kind of facial mask containing ginkgo biloba p.e |
| CN111004335A (en) * | 2019-11-29 | 2020-04-14 | 湖南绿蔓生物科技股份有限公司 | Bletilla striata oligosaccharide extracted from rhizome of bletilla striata and preparation method thereof |
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| CN105267257A (en) * | 2015-12-01 | 2016-01-27 | 黑龙江珍宝岛药业股份有限公司 | Preparation method of ginkgo leaf extract |
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| CN106727746A (en) * | 2017-04-07 | 2017-05-31 | 浙江新昌天然保健品有限公司 | The preparation method of low sour water soluble ginkgo leaf extract |
| CN109718141A (en) * | 2019-03-14 | 2019-05-07 | 上海琴馨文化传播有限公司 | A kind of facial mask containing ginkgo biloba p.e |
| CN111004335A (en) * | 2019-11-29 | 2020-04-14 | 湖南绿蔓生物科技股份有限公司 | Bletilla striata oligosaccharide extracted from rhizome of bletilla striata and preparation method thereof |
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