CN102335209B - Preparation method and product of minimal acid water soluble ginkgo leaf extract - Google Patents
Preparation method and product of minimal acid water soluble ginkgo leaf extract Download PDFInfo
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- CN102335209B CN102335209B CN 201110306111 CN201110306111A CN102335209B CN 102335209 B CN102335209 B CN 102335209B CN 201110306111 CN201110306111 CN 201110306111 CN 201110306111 A CN201110306111 A CN 201110306111A CN 102335209 B CN102335209 B CN 102335209B
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Abstract
The invention discloses a preparation method and product of minimal acid water soluble ginkgo leaf extract. According to the method, the ginkgo leaf is extracted by using water as a solvent, the content of ginkgolic acid is reduced by flocculating settling combined with refrigeration, and gingko flavone and terpene lactone are purified and concentrated by resin column chromatography. The content of ginkgolic acid of the product is less than 0.1ppm, the solubility of the product in cold water is 5g/100ml, and two main problems of high content of ginkgolic acid and poor water solubility of ginkgo leaf extract are solved simultaneously. Compared with the prior art, the extraction and preparation process disclosed by the invention has the advantages of little production investment, simplicity of equipment, flow and operation, low production cost, safety of used reagents, no intervention of toxic organic solvent, stable product quality, strong operability in industrialized production and the like.
Description
Technical field
The present invention relates to a kind of preparation method and product of minimal acid water soluble ginkgo leaf extract, particularly extract from Folium Ginkgo the preparation ginkgoic acid less than 0.1ppm and cold water solubility greater than the method for the Folium Ginkgo extract of 5g/100ml, belong to the Chinese medicine extraction technical field.
Background technology
Semen Ginkgo (Ginkgo biloba L.) is the Ginkgo gymnosperm, and China's Folium Ginkgo resource is extremely abundant, accounts for 70% of whole world Folium Ginkgo resource.Folium Ginkgo extract is a kind of stronger free radical scavenger, and alzheimer disease, aging, neuropathy etc. all are improved effect.Gingko leaf preparation is main person in middle and old age's brain insufficiency, the senile dementia of degenerating and causing for coronary heart diseases and angina pectoris, myocardial infarction, apoplexy, disturbance of cerebral circulation and brain function clinically at present.And these effects of Folium Ginkgo extract are mainly because contain ginkgetin and bilobalide active component.The quality standard of German History watt C Compaq's Folium Ginkgo extract (EGb761) is: Gingko yellow ketoside content 22%-27%; Terpene lactones content 5%-7%, in its terpene lactones, the Ginkgolide A. B. C sum is 2.8%-3.4%, content of bilobalide 2.6%-3.2%, ginkgoic acid content＜5mg/kg (the 5th innings of Kommisson E of German national bureau of drug administration in JIUYUE, 1994).
Ginkgoic acid is 6-alkyl or the salicylic derivant of 6-thiazolinyl in Folium Ginkgo, is a homology mixture, and its side chain is respectively: C
13H
27(13:0), C
15H
31(C15:0), C
15H
29(C15:1), C
17H
33(C17:1), C
17H
31(C17:2) etc.Research finds, ginkgoic acid has immunotoxicity and fetal toxicity effect, can cause dermatitis rhus sample dermatitis, is the allergen in Semen Ginkgo, and the mankind's health is had potential harm.In Germany's Ministry of Public Health regulation Semen Ginkgo extrac (EGB) preparation, the content of ginkgoic acid must be less than 5ppm, and " Chinese pharmacopoeia also requires Ginkgolic Acid in Extract of Ginkgo biloba Leaves less than 10ppm in 2005 editions, requires it less than 5ppm in 2010 editions.Therefore, reduce the content of Ginkgolic Acid in Extract of Ginkgo biloba Leaves, improve the water solublity of Folium Ginkgo extract, to production and the application important in inhibiting of Folium Ginkgo extract.
At present about reducing the method for Ginkgolic Acid in Extract of Ginkgo biloba Leaves, existing related invention patent application all discloses macroporous resin column chromatography as 200810022162.X " a kind of low-acid gingko flavones and extracting method thereof " and 02113529.0 " taking off the technique of ginkgoic acid with resin method from Folium Ginkgo extract " and has removed ginkgoic acid; And 02113531.2 " taking off the technique of ginkgoic acid with extraction from Folium Ginkgo extract " and a kind of 03131915.7 " method of removing ginkgolic acid from ginkgo leaf extract by extraction method " disclosed be to adopt petroleum ether and alkane solvents extraction ginkgoic acid.The Ginkgolic Acid in Extract of Ginkgo biloba Leaves content that the disclosed removal ginkgoic acid of aforementioned patent applications technique makes about 1ppm to about 10ppm, but extraction when having production operation safety not high, and easily make the problem that in product, organic solvent residual exceeds standard.Water soluble ginkgo leaf extract relevant patent application is as CN1772009A " a kind of production method of water soluble ginkgo leaf extract ", and the raw material of employing is Folium Ginkgo extract, though and the product water dissolubility greater than 5%, ginkgoic acid is only less than 5ppm; CN101596221A " a kind of preparation technology of water-soluble ginkgo biloba extract used for injection " discloses and has a kind ofly prepared flavone take Folium Ginkgo extract as raw material: the preparation technology of lactone=4: 1 product, adopt ultrafiltration purification in technique, product quality is not controlled the toxic component ginkgoic acid, and the dissolubility of product in water do not offer some clarification on; CN101254220A " a kind of production technology of water soluble ginkgo leaf extract " adopts high concentration ethanol to extract from Folium Ginkgo, after macroporous resin column and prepare the method for water soluble ginkgo leaf extract in conjunction with the polymer nano-scale membrane filtration, the ginkgoic acid of this product is controlled at below 1ppm, water solublity is greater than 5%, but its production cost is higher.
Summary of the invention
The object of the invention is to: preparation method and product that a kind of minimal acid water soluble ginkgo leaf extract is provided.The present invention is directed to that prior art Ginkgolic Acid in Extract of Ginkgo biloba Leaves content is high, the large main bugbear of water solublity bad two, adopt water as effective ingredient in the solvent extraction Folium Ginkgo, and utilize ginkgoic acid not dissolve in water and the method for cold preservation precipitation removal, gained Ginkgolic Acid in Extract of Ginkgo biloba Leaves content is less than 0.1ppm, cold water solubility is greater than 5g/100ml, and residue is less than 0.3%.
Technical scheme of the present invention: the preparation method of minimal acid water soluble ginkgo leaf extract comprises the following steps:
(1) get Folium Ginkgo and make raw material, the water boil that adds 8～12 times of weight extracts 2～3 times, each 1～2 hour, merge aqueous extract, be concentrated into 1/4～1/5 of original volume, standing cooling 12～18 hours, get upper solution and add the flocculating agent flocculation to form precipitation, the filtering precipitation is got supernatant standby;
(2) macroporous resin column on supernatant, wash away polar impurity with deionized water, 40%～50% alcohol solvent eluting, the concentrated ethanol of removing of eluent, low temperature (0 ℃～1 ℃) cold preservation 12～18 hours, to feed temperature be 3 ℃～5 ℃, get supernatant liquid filtering, filtrate vacuum concentration, drying get dry thing;
(3) dry thing with 95% ethanol in 40 ℃ of heating for dissolving, cooled and filtered, the filtrate vacuum concentration namely gets Folium Ginkgo extract to extractum after extract dry.
Preferred preparation method comprises the following steps:
(1) get dry Folium Ginkgo, the deionized water heated and boiled that adds 10 times of weight is extracted 2 times, each 2 hours, merge aqueous extract, be concentrated into 1/5 of original volume, standing cooling 15 hours, get upper solution and add the flocculating agent flocculation to form precipitation, diatomite filtration is got supernatant standby except precipitation;
(2) macroporous resin column on supernatant, deionized water with 2 times of column volumes washes away polar impurity, 40%～50% alcohol solvent eluting of 3～5 times of column volumes, the concentrated ethanol of removing of eluent, low temperature (0 ℃～1 ℃) cold preservation 15 hours, to feed temperature be 3 ℃～5 ℃, get the supernatant diatomite filtration, the filtrate vacuum concentration is to extractum, and the extractum vacuum drying gets dry thing;
(3) by solid-liquid ratio 1: 10 (Kg: L) adding mass fraction in the dry thing is 95% ethanol, in 40 ℃ of heating for dissolving, cooled and filtered, the filtrate vacuum concentration is to extractum, the extractum vacuum drying gets Folium Ginkgo extract.
Flocculating agent described in abovementioned steps (1) is chitin, chitosan or gelatin.
Described in abovementioned steps (2), the model of macroporous resin column is DM130, D101 or AB-8.
Described in abovementioned steps (2) and step (3), the vacuum concentration temperature is 50～65 ℃, and vacuum is 0.07～0.08Mpa.
Described in abovementioned steps (2) and step (3), the vacuum drying temperature is 40～60 ℃, and vacuum is 0.07～0.08Mpa.
The present invention also comprises the Folium Ginkgo extract product that aforementioned preparation method makes.
In aforementioned Folium Ginkgo extract, ginkgoic acid content is less than 0.1ppm, and cold water solubility is greater than 5g/100ml, and residue is less than 0.3%; And flavonoid of ginkgo biloba is 22%～28%, and Ginkgo total lactones content is 6%～10%, and wherein content of bilobalide is greater than 2.8%, and bilobalide content is greater than 3.2%.
The present invention extracts Folium Ginkgo take water as solvent, reduce the content of ginkgoic acid in conjunction with cold preservation with flocculation sediment, with resin column chromatography purification, enriching ginkgetin and terpene lactones, products obtained therefrom is the yellow powder shape, and in product, flavonoid of ginkgo biloba is 22%～28%, Ginkgo total lactones content is 6%～10%, wherein content of bilobalide is greater than 2.8%, and bilobalide content is greater than 3.2%, and ginkgoic acid content is less than 0.1ppm, cold water solubility is greater than 5g/ml, and residue is less than 0.3%.In product, the assay of ginkgoic acid adopts the Europe official method, ginkgoic acid content all meets Europe pharmacopeia and U.S. FDA requirement, and being applicable on the domestic and international market has the medicine such as injection, drop, freeze dried powder of specific (special) requirements and cosmetics, health food etc. to the crude drug water solubility.
The present invention adopts water as extracting solvent, and its cost is lower, and the recycling ginkgoic acid does not dissolve in water and the cold preservation precipitation is removed, and has solved simultaneously that Ginkgolic Acid in Extract of Ginkgo biloba Leaves content is high, the large main bugbear of water solublity bad two.Thereby compared with prior art, extraction and preparation technique of the present invention has the production less investment, equipment, flow process and simple to operate, and production cost is low, uses reagent safety, gets involved constant product quality, the advantages such as suitability for industrialized production strong operability without poisonous organic solvent.
The specific embodiment
Embodiments of the invention 1: get the commercially available oven dry Folium Ginkgo 400kg in Pizhou City, Jiangsu, drop into 6 tons and extract in the pond, add 4 tons of deionized waters to soak 1 hour, heated and boiled was extracted 2 hours, drained extracting solution; The Folium Ginkgo slag such as adds at the water gaging to be repeated to extract once, the aqueous extract merging is concentrated into 1 ton, put into 2 tons of basins standing cooling 15 hours, and got upper solution and add 10% chitin solution flocculation to form precipitation, diatomite filtration is except precipitation, cross AB-8 macroporous resin column on cleaner liquid, wash away polar impurity with 1.2 tons of deionized waters, 2.4 ton of 40%～50% alcohol solvent eluting, eluent is concentrated into without ethanol, low temperature (1 ℃) cold preservation 15 hours, to feed temperature be 5 ℃.Get the supernatant diatomite filtration, (50～65 ℃ of temperature, vacuum 0.07～0.08Mpa) are to extractum, and (40～60 ℃ of temperature, vacuum 0.07～0.08Mpa) gets dry thing 8kg to the extractum vacuum drying for the filtrate vacuum concentration.Adding the 80L mass fraction in the dry thing is 95% ethanol, 40 ℃ of heating for dissolving, solution filters after being cooled to room temperature, filtrate vacuum concentration (50～65 ℃ of temperature, vacuum 0.07～0.08Mpa) is to extractum, (40～60 ℃ of temperature, vacuum 0.07～0.08Mpa) gets golden yellow Folium Ginkgo extract 6.5kg to the extractum vacuum drying.
Embodiments of the invention 2: get the commercially available oven dry Folium Ginkgo 500kg in Pizhou City, Jiangsu, drop into 6 tons and extract in the pond, add 5 tons of deionized waters to soak 1 hour, heated and boiled was extracted 2 hours, drained extracting solution; The Folium Ginkgo slag adds water to be repeated to extract once, the aqueous extract merging is concentrated into 1/5 of original volume, put into 2 tons of basins standing cooling 13 hours, and got upper solution and add 10% gelatin solution flocculation to form precipitation, diatomite filtration is except precipitation, cross DM130 macroporous resin column on cleaner liquid, wash away polar impurity with 1.5 tons of deionized waters, 2.5 ton of 40%～50% alcohol solvent eluting, eluent is concentrated into without ethanol, low temperature (0 ℃～1 ℃) cold preservation 13 hours, to feed temperature be 3 ℃.Get the supernatant diatomite filtration, the filtrate vacuum concentration is dry, gets dry thing 10.2kg.It is 95% ethanol that dry thing adds the 102L mass fraction, 40 ℃ of heating for dissolving, and solution filters after being cooled to room temperature, and the filtrate vacuum concentration is to extractum, and the extractum vacuum drying gets golden yellow Folium Ginkgo extract product 9.6kg.
Embodiment 3: get dry Folium Ginkgo 300kg, drop into and extract in the pond, add 3 tons of deionized water heated and boiled to extract 2 hours, drain extracting solution; The Folium Ginkgo slag adds water to be repeated to extract once, the aqueous extract merging is concentrated into 1/4 of original volume, put into basin standing cooling 18 hours, and got upper solution and add 10% chitosan solution to carry out flocculation sediment, the filtering precipitation, D101 macroporous resin column on supernatant, deionized water with 2 times of column volumes washes away polar impurity, 40%～50% alcohol solvent eluting of 5 times of column volumes, the concentrated ethanol of removing of eluent, low temperature (0 ℃～1 ℃) cold preservation 18 hours, to feed temperature be 4 ℃.Get the supernatant diatomite filtration, the filtrate vacuum concentration is to extractum, and the extractum vacuum drying gets dry thing 6.1kg.It is 95% ethanol that dry thing adds the 61L mass fraction, in 40 ℃ of heating for dissolving, and cooled and filtered, the filtrate vacuum concentration is to extractum, and the extractum vacuum drying gets yellow powder shape Folium Ginkgo extract 5.3kg.
Adopt the assay method under Chinese Pharmacopoeia version Folium Ginkgo extract item in 2010, measure ginkgoic acid, ginkgetin and bilobalide content and the cold water solubility of embodiment 1-3 gained Folium Ginkgo extract product, measurement result is as follows:
Claims (6)
1. the preparation method of minimal acid water soluble ginkgo leaf extract is characterized in that: comprise the following steps:
(1) get Folium Ginkgo and make raw material, the water boil that adds 8～12 times of weight extracts 2～3 times, each 1～2 hour, merge aqueous extract, be concentrated into 1/4～1/5 of original volume, standing cooling 12～18 hours, get upper solution and add the flocculating agent flocculation to form precipitation, the filtering precipitation is got supernatant standby;
(2) macroporous resin column on supernatant, wash away polar impurity with deionized water, 40%～50% alcohol solvent eluting, the concentrated ethanol of removing of eluent, 0 ℃～1 ℃ deepfreeze 12～18 hours, to feed temperature be 3 ℃～5 ℃, get supernatant liquid filtering, filtrate vacuum concentration, drying get dry thing;
(3) dry thing with 95% ethanol in 40 ℃ of heating for dissolving, cooled and filtered, the filtrate vacuum concentration namely gets Folium Ginkgo extract to extractum after extract dry.
2. the preparation method of minimal acid water soluble ginkgo leaf extract according to claim 1 is characterized in that: comprise the following steps:
(1) get dry Folium Ginkgo, the deionized water heated and boiled that adds 10 times of weight is extracted 2 times, each 2 hours, merge aqueous extract, be concentrated into 1/5 of original volume, standing cooling 15 hours, get upper solution and add the flocculating agent flocculation to form precipitation, diatomite filtration is got supernatant standby except precipitation;
(2) macroporous resin column on supernatant, deionized water with 2 times of column volumes washes away polar impurity, 40%～50% alcohol solvent eluting of 3～5 times of column volumes, the concentrated ethanol of removing of eluent, 0 ℃～1 ℃ deepfreeze 15 hours, to feed temperature be 3 ℃～5 ℃, get the supernatant diatomite filtration, the filtrate vacuum concentration is to extractum, and the extractum vacuum drying gets dry thing;
(3) adding mass fraction in the dry thing by solid-liquid ratio 1Kg: 10L is 95% ethanol, in 40 ℃ of heating for dissolving, and cooled and filtered, the filtrate vacuum concentration is to extractum, and the extractum vacuum drying gets Folium Ginkgo extract.
3. the preparation method of described minimal acid water soluble ginkgo leaf extract according to claim 1 and 2, it is characterized in that: described flocculating agent is chitin, chitosan or gelatin.
4. the preparation method of described minimal acid water soluble ginkgo leaf extract according to claim 1 and 2, it is characterized in that: described in step (2), the model of macroporous resin column is DM130, D101 or AB-8.
5. the preparation method of described minimal acid water soluble ginkgo leaf extract according to claim 1 and 2, it is characterized in that: described in step (2) and step (3), the vacuum concentration temperature is 50～65 ℃, and vacuum is 0.07～0.08Mpa.
6. the preparation method of minimal acid water soluble ginkgo leaf extract according to claim 2, it is characterized in that: described in step (2) and step (3), the vacuum drying temperature is 40～60 ℃, and vacuum is 0.07～0.08Mpa.
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| CN101194918B (en) * | 2007-05-09 | 2011-04-27 | 广西师范大学 | Technique for removing ginkgolic acid in folium ginkgo extract |
| CN101254220A (en) * | 2008-04-08 | 2008-09-03 | 浙江康恩贝植物提取物有限公司 | Production process of water soluble ginkgo leaf extractive |
| CN102145028A (en) * | 2011-03-21 | 2011-08-10 | 贵州大学 | Method for reducing ignition residues in ginkgo leaf extract |
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