CN103130818A - Bilobalide B compound and preparation method thereof - Google Patents
Bilobalide B compound and preparation method thereof Download PDFInfo
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- CN103130818A CN103130818A CN2013100984699A CN201310098469A CN103130818A CN 103130818 A CN103130818 A CN 103130818A CN 2013100984699 A CN2013100984699 A CN 2013100984699A CN 201310098469 A CN201310098469 A CN 201310098469A CN 103130818 A CN103130818 A CN 103130818A
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Abstract
The invention relates to a bilobalide B compound which is extracted from ginkgo leaf. The bilobalide B compound contains trace amounts of bilobalide K; and the bilobalide B content is higher than 99%, and the bilobalide K content is lower than 0.6%. The invention also relates to a preparation method of the bilobalide B compound, which comprises the following steps: under stirring conditions, adding kieselguhr and ginkgo leaf extract, heating under reflux at 60-73 DEG C for 2-3.5 hours, filtering and concentrating; adding distilled water, removing impurities, separating out the ethyl acetate layer, passing the ethyl acetate layer through an peracidic aluminum oxide column, eluting with water saturated ethyl acetate, and collecting the eluate; concentrating the eluate under reduced pressure until no ethyl acetate smell is emitted, adding a proper amount of ethanol to dissolve the concentrated eluate, adding water until the ethanol content is 60-70%, standing, precipitating coarse crystal, filtering, taking the coarse crystal, and drying; and recrystallizing the coarse crystal sequentially with ethanol and ethyl acetate to obtain the finished product bilobalide B compound.
Description
Technical field
The invention belongs to medical technical field, be specifically related to a kind of Ginkgolide B compound and preparation method thereof.
Background technology
Ginkgolide B, English name Ginkgolide B; Molecular formula: C
20H
24O
10Molecular weight: 424.40.
Ginkgolide B is the strong platelet activation factor of a kind of activity (PAF) antagonist, can be combined with paf receptor competitively, suppresses the effect of PAF; Therefore, allly relate to the illness relevant to PAF, as: the shocks that thrombosis, brain tissue impairment, organ-graft refection, acute inflammation, heart allergy, intracellular toxin and IgG cause etc., Ginkgolide B all has certain curative effect.
But up to now, the Ginkgolide B single component not being arranged all both at home and abroad is the preparation listing of raw material.Belong to " extracting or pass through new effective monomer and the preparation thereof of fermented extracted in crude substance " under " the not medicine of list marketing at home and abroad " in pharmaceutical chemicals according to this medicine of pharmaceutical chemicals registration classification in China's " medicine registration management way ", belong to pharmaceutical chemicals 1.2 classes.
CN101182325A relates to a kind of intravenous administration bilobalide B and extracting method, Ginkgolide B content 〉=97% wherein, and ginkgoic acid≤2PPM, bilobalide does not detect, can be directly as intravenous injection or intravenous drip bulk drug; Extracting and adopting leaf dry weight 8～25 times of amounts, concentration is 5%-30% aqueous ethanolic solution refluxing extraction several times, filtering separation; Filtrate is concentrated into 0.05～0.2 times of amount of original volume, adds sorbent material fully absorption under whipped state, filter, filter cake is with 3～10 times of amount alcohol reflux several times, and united extraction liquid is concentrated into 0.01～0.2 times of amount of original volume, standing crystallization filters to get bilobalide; Bilobalide carries out the recrystallization several times with the methyl alcohol of 10～50 times of amounts, recrystallization temperature-10 ℃～-25 ℃, and filtering for crystallizing, drying get Ginkgolide B.
CN101412725A discloses a kind of method of extracting separating bilobalide B from Ginkgo Leaf, and its step is as follows: 1) get Ginkgo Leaf, the use ethanol of different concns is from high to low extracted, and united extraction liquid reclaims ethanol, the extracting solution after being concentrated; 2) add ethyl acetate extraction in the extracting solution after concentrated, be concentrated into medicinal extract after reclaiming organic phase; 3) with step 2) the rear upper selective polarity atresia adsorption resin column of gained medicinal extract dilution, use ethanol elution, after the gained elutriant reclaims ethanol, then obtain the bilobalide based composition with alcohol crystal; 4) with silicagel column in the crystallization of bilobalide based composition, with the mixed solution wash-out of normal hexane and ethyl acetate, collect elutriant, obtain the elutriant of enrichment Ginkgolide B; 5) recycling step 4) in solvent in the elutriant collected, then use alcohol crystal, obtain the Ginkgolide B monomer.
CN1680392A relates to employing high-speed countercurrent chromatography (HSCCC) and separate a kind of preparation method of high purity ginkgo lactone from Ginkgo Leaf.With three solvent systems, after separating through twice HSCCC respectively, can obtain Ginkgolide A, Ginkgolide B, ginkalide C, bilobalide J and bilobalide, purity all reaches more than 99%.
At present, medicinal Ginkgolide B mainly extracts from Ginkgo Leaf, but the content due to Ginkgolide B in Ginkgo Leaf is extremely low, and Ginkgolide B and bilobalide K and ginkgoic acid, bilobalide exist jointly, each monomer structure is very similar again, polarity difference is very little, extracts comparatively difficulty of highly purified Ginkgolide B monomer from Ginkgo Leaf, particularly with separate the comparatively difficulty of Ginkgolide B with bilobalide K.the Ginkgolide B that extracts in above-mentioned patent and prior art pure not high enough, and Ginkgolide B is poorly soluble, therefore can only be as the oral medication raw material and can not be as the bulk drug of injection, and the bioavailability of oral administration is lower, can not give full play to the drug effect of Ginkgolide B, adopt injection can greatly improve its bioavailability, and clinical applicable crowd---ischemic encephalopathy patient, the patient time window that comprises the acute ischemic stroke cerebral infarction is narrow, and need quick acting, general oral preparations is slow with respect to the injection onset, and after making injection, the administration onset is rapid, in addition, may there be the oral pharmaceutical dysphagia in the ischemic encephalopathy patient, make injection and do not have the problem of dysphagia.But injection is very high to the purity requirement of Ginkgolide B, therefore, provides a kind of highly purified Ginkgolide B compound to have very important significance.
In view of this, special proposition the present invention.
Summary of the invention
The first purpose of the present invention is to overcome the deficiencies in the prior art, and a kind of Ginkgolide B compound is provided, and in described Ginkgolide B compound, the content of Ginkgolide B is higher.
The second purpose of the present invention is to provide a kind of preparation method of above-mentioned Ginkgolide B compound.
In order to realize purpose of the present invention, the following technical scheme of special employing:
A kind of Ginkgolide B compound, described Ginkgolide B compound extracts acquisition from Ginkgo Leaf, the bilobalide K that contains trace in described Ginkgolide B compound, the content of described Ginkgolide B is greater than 99%, the content of described bilobalide K is lower than 0.6%, and the structural formula of described Ginkgolide B compound is:
Bilobalide K is very similar to the structure of Ginkgolide B, and both polarity difference is very little, and in the Ginkgolide B compound that extracts from Ginkgo Leaf, the content of bilobalide K is higher, and the Ginkgolide B that obtain high-content is comparatively difficult.Ginkgolide B compound provided by the invention is to extract from the natural drug Ginkgo Leaf, separate, refining forming, wherein the purity of Ginkgolide B is greater than 99%, compared with prior art, purity is very high, and the content of bilobalide K is very low, can be used as the bulk drug of injection formulations, greatly improved the bioavailability of Ginkgolide B, and, in the Ginkgolide B compound, the kind of impurity and content also have considerable influence to the stability of Ginkgolide B compound, dopant species in Ginkgolide B compound provided by the invention is few, content is low, has better stability with respect to prior art, and the quality approach of this product and stability test result show that this product is quality controllable, stable, can be used as the bulk drug of injection formulations, the basis of good validity and security and quality controllability is provided for this product clinical application, greatly improved patient's drug safety.
Preferably, the content of described Ginkgolide B is greater than 99.5%, and the content of described bilobalide K is lower than 0.4%.
Preferably, do not contain bilobalide in described Ginkgolide B compound.In Ginkgolide B compound provided by the invention, the kind of impurity is few, content is low, and is quality controllable, be a kind of bulk drug of injection formulations of excellent property, and impurity is less on the impact of the stability of Ginkgolide B.
Preferably, in described Ginkgolide B compound the content of total ginkgolic acids less than 0.12ppm.
The present invention is with reference to " total ginkgolic acids Limit Test method under Folium Ginkgo extract item of Chinese pharmacopoeia version in 2010, result to determination of raw material shows, this product contains the amount of total ginkgolic acids less than 0.12ppm, must not cross 10/1000000ths limit regulation far below the Folium Ginkgo extract total ginkgolic acids, Ginkgolide B provided by the invention is safe and effective.
In the present invention, the specific optical rotation of the methanol solution of described Ginkgolide B compound in the time of 20 ℃ is-57 °～-61 °.
The solubleness of Ginkgolide B under dehydrated alcohol is low than methyl alcohol, and the concentration that is mixed with 10mg/ml is more difficult, and the specific rotation numerical value of 5mg/ml is lower, error increases, thereby to select methyl alcohol be solvent, is mixed with the solution of 10mg/ml, and the specific optical rotation of measuring in the time of 20 ℃ is-57 °～-61 °.Bilobalide K is dextro-rotatory substance, therefore the foreign matter content of bilobalide K is larger to the specific optical rotation value effect of Ginkgolide B compound, the specific optical rotation of Ginkgolide B compound of the present invention is than " specific optical rotation of the Ginkgolide B compound of stipulating in Chinese pharmacopoeia narrows down a lot for the scope of-48 ° to-63 °, the content that bilobalide K in Ginkgolide B compound provided by the invention is described is lower, and the purity of Ginkgolide B is higher.
The present invention also provides a kind of preparation method of described Ginkgolide B compound, comprising:
(1) first add diatomite in ethyl acetate under agitation condition, then add Folium Ginkgo extract, in 60～73 ℃ of reflux 2-3.5 hours, filter, concentrated; Add diatomaceous amount be Folium Ginkgo extract weight 0.8-1.2 doubly, the liquid-solid ratio of ethyl acetate and Folium Ginkgo extract is 15-20ml/g;
(2) add the distilled water removal of impurities again, tell ethyl acetate layer, ethyl acetate layer peracidity alumina column is used the water saturation eluent ethyl acetate, collects elutriant;
(3) elutriant is evaporated to without ethyl acetate flavor, adds appropriate ethanol and makes it dissolving, then add water to make and contain the alcohol amount and be 60～70%, and is standing, makes and separates out coarse crystallization, filters, and gets coarse crystallization, oven dry;
(4) coarse crystallization is used ethanol and re-crystallizing in ethyl acetate successively, gets finished product Ginkgolide B compound.
The present invention adopts ethyl acetate to extract Ginkgolide B from Folium Ginkgo extract, Folium Ginkgo extract is added in ethyl acetate, can form the large material of a large amount of viscosity, therefore the technique of not utilizing subsequent technique and clearing out a gathering place add diatomite to help the dispersion of Folium Ginkgo extract in ethyl acetate; When extracting in a large number, directly diatomite is mixed with Folium Ginkgo extract, the dispersion and the mixing that are unfavorable for Folium Ginkgo extract, therefore after adding ethyl acetate in extractor, first add diatomite under agitation condition, then add Folium Ginkgo extract, Folium Ginkgo extract can be uniformly dispersed in ethyl acetate quickly, be conducive to the stripping of Ginkgolide B in ethyl acetate, improve the content of Ginkgolide B in crude product.
In above-mentioned preparation method's step (1), described Folium Ginkgo extract is to extract to obtain from Ginkgo Leaf, and extracting method can be with reference to any method of the prior art, and those skilled in the art do not need to pay performing creative labour.
The concentrated solution that step (1) obtains has precipitate, precipitate affects column chromatography, therefore added the water extraction in prior to concentrated solution in step (2) before carrying out column chromatography, can make precipitate water-soluble, branch vibration layer, be conducive to remove a large amount of impurity in concentrated solution, and the viscosity that reduces concentrated solution when removing precipitate, more be conducive to follow-up column chromatography technique.
extract the preparation Ginkgolide B from Ginkgo Leaf, the preparation method is different, the content of Ginkgolide B and wherein kind and the content difference of impurity is huge in the compound that obtains, the present invention is by a large amount of experiments, extraction preparation method to Ginkgolide B constantly improves and optimizes, particularly including selecting solvent and chromatographic column, and to the design of whole extraction route, finally obtained the preparation method of the compound of a preparation high-content ginkgo lactone B, and in the Ginkgolide B compound that obtains, the kind of impurity is few, content is low, measure through HPLC-ELSD, the purity of Ginkgolide B compound monomer reaches more than 99%, the content of bilobalide K is lower than 0.6%, can be used as the bulk drug of injection formulations, greatly improved patient's drug safety.In addition, the method is simple to operation, preparation cycle is short, has greatly reduced the cost of preparation Ginkgolide B compound from Ginkgo Leaf, has further reduced the cost that contains this Ginkgolide B compound formulation.
In above-mentioned preparation method's step (4), coarse crystallization uses ethyl alcohol recrystallization more than 3 times, re-crystallizing in ethyl acetate more than 1 time successively, and in the finished product that obtains, the content of Ginkgolide B is greater than 99.5%, and the content of described bilobalide K is lower than 0.4%.
In described step (1), preferred, add diatomaceous amount be Folium Ginkgo extract weight 0.9-1.1 doubly; The liquid-solid ratio of ethyl acetate and Folium Ginkgo extract is 16-18ml/g.
In described step (1), preferred, it is 3-5ml/g that filtrate is concentrated into liquid-solid ratio.
In described step (2), preferred, the blade diameter length ratio of acidic alumina column is 1:4～1:10, and aluminum oxide is 200～300 orders, and the weight ratio of applied sample amount and aluminum oxide is 1:7～10, and elution speed is 1/4～1/2BV/h; Preferred, the blade diameter length ratio of described acidic alumina column is 1:4～1:7, and the weight ratio of applied sample amount and aluminum oxide is 1:8.
By above-mentioned preferred technical scheme, the present invention obtains not contain bilobalide in the Ginkgolide B finished product, and the content of total ginkgolic acids is less than 0.12ppm.
Compared with prior art, beneficial effect of the present invention is:
In Ginkgolide B compound provided by the invention, the content of Ginkgolide B is very high, and the kind of impurity is few, content is low, and particularly the content of bilobalide K is very low, can be used as the bulk drug of injection, improved the bioavailability of bulk drug and patient's drug safety.
In addition, the preparation method of Ginkgolide B compound provided by the invention is simple to operation, and particularly diatomaceous employing is conducive to the carrying out of follow-up column chromatography technique, has improved the efficient of preparation, has reduced the cost of preparation.
Embodiment
The method for chromatographic determination of bilobalide and bilobalide adopts the HPLC-ELSD chromatographic process: be weighting agent with octadecylsilane chemically bonded silica; Take methyl alcohol/tetrahydrofuran (THF) (2:1): water=32:68 as moving phase, and the light scattering detector detection (reference conditions: Alltech3300: 60 ℃ of drift tube temperatures, air velocity 1.5L/min, gain is 8; Agilent380-LC: 30 ℃ of atomization temperatures, 50 ℃ of drift tube temperatures, carrier gas flux 1.5SLM, the PMT gain is 8).Integrative approach: area external standard method.
The assay of total ginkgolic acids is with reference to " total ginkgolic acids Limit Test method under Folium Ginkgo extract item of Chinese pharmacopoeia version in 2010: take octadecylsilane chemically bonded silica as weighting agent, take methyl alcohol-tetrahydrofuran (THF)-water (22:10:68) as moving phase; With light scattering detector detect (reference conditions: Alltech3300: 60 ℃ of drift tube temperatures, air velocity 1.5L/min, gain is 8; Agilent380-LC: 30 ℃ of atomization temperatures, 50 ℃ of drift tube temperatures, carrier gas flux 1.5SLM, the PMT gain is 8).Integrative approach: area external standard method.
The measuring method of specific optical rotation is with reference to " Chinese pharmacopoeia version appendix VI E in 2010, reference conditions: WZZ-3 automatic polarimeter solvent: methanol solution, concentration: 10mg/ml, temperature: 20 ℃.
Embodiment 1
The preparation of Ginkgolide B compound:
First add diatomite 40g in the 750ml ethyl acetate under agitation condition, then add Folium Ginkgo extract 50g, in 60 ℃ of reflux 3.5 hours, filter, concentrated, it is 5ml:1g that filtrate is concentrated into liquid-solid ratio; Add the approximately distilled water removal of impurities of 1/10 volume in concentrated solution, tell ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:4, aluminum oxide is 200 orders, and the weight ratio of applied sample amount and aluminum oxide is 1:7, uses the water saturation eluent ethyl acetate, elution speed is 1/4BV/h, collects elutriant; Elutriant is evaporated to without ethyl acetate flavor, adds appropriate ethanol and makes it dissolving, then add water to make and contain the alcohol amount and be 60%, and is standing, makes and separates out coarse crystallization, filters, and gets coarse crystallization, oven dry; Coarse crystallization is used ethanol and re-crystallizing in ethyl acetate successively, gets finished product Ginkgolide B compound.Sample detection Ginkgolide B content is 99.0%, and the content of bilobalide K is 0.6%.The specific optical rotation of finished product is-61 °.
Embodiment 2
The preparation of Ginkgolide B compound:
First add diatomite 40g in the 1000ml ethyl acetate under agitation condition, then add Folium Ginkgo extract 50g, in 73 ℃ of reflux 2 hours, filter, concentrated, it is 4ml:1g that filtrate is concentrated into liquid-solid ratio; Add the approximately distilled water removal of impurities of 1/9 volume in concentrated solution, tell ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:4, aluminum oxide is 200 orders, and the weight ratio of applied sample amount and aluminum oxide is 1:7, uses the water saturation eluent ethyl acetate, elution speed is 1/4BV/h, collects elutriant; Elutriant is evaporated to without ethyl acetate flavor, adds appropriate ethanol and makes it dissolving, then add water to make and contain the alcohol amount and be 60%, and is standing, makes and separates out coarse crystallization, filters, and gets coarse crystallization, oven dry; Coarse crystallization is used ethyl alcohol recrystallization 3 times and re-crystallizing in ethyl acetate 1 time successively, gets finished product Ginkgolide B compound.Sample detection Ginkgolide B content is 99.5%, and the content of bilobalide K is 0.4%.The specific optical rotation of finished product is-59.3 °.
Embodiment 3
The preparation of Ginkgolide B compound:
First add diatomite 60g in the 850ml ethyl acetate under agitation condition, then add Folium Ginkgo extract 50g, in 70 ℃ of reflux 2.5 hours, filter, concentrated, it is 3ml:1g that filtrate is concentrated into liquid-solid ratio; Add the approximately distilled water removal of impurities of 1/11 volume in concentrated solution, tell ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:10, aluminum oxide is 300 orders, and the weight ratio of applied sample amount and aluminum oxide is 1:10, uses the water saturation eluent ethyl acetate, elution speed is 1/2BV/h, collects elutriant; Elutriant is evaporated to without ethyl acetate flavor, adds appropriate ethanol and makes it dissolving, then add water to make and contain the alcohol amount and be 70%, and is standing, makes and separates out coarse crystallization, filters, and gets coarse crystallization, oven dry; Coarse crystallization is used ethyl alcohol recrystallization 3 times and re-crystallizing in ethyl acetate 1 time successively, gets finished product Ginkgolide B compound.Sample detection Ginkgolide B content is 99.6%, and the content of bilobalide K is 0.3%.Bilobalide does not detect, and the content of total ginkgolic acids is 0.12ppm, and the content of bilobalide K is 0.3%.The specific optical rotation of finished product is-57.8 °.
Embodiment 4
The preparation of Ginkgolide B compound:
First add diatomite 45g in the 900ml ethyl acetate under agitation condition, then add Folium Ginkgo extract 50g, in 73 ℃ of reflux 2 hours, filter, concentrated, it is 3.5ml:1g that filtrate is concentrated into liquid-solid ratio; Add the approximately distilled water removal of impurities of 1/10 volume in concentrated solution, tell ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:7, aluminum oxide is 300 orders, and the weight ratio of applied sample amount and aluminum oxide is 1:8, uses the water saturation eluent ethyl acetate, elution speed is 1/2BV/h, collects elutriant; Elutriant is evaporated to without ethyl acetate flavor, adds appropriate ethanol and makes it dissolving, then add water to make and contain the alcohol amount and be 70%, and is standing, makes and separates out coarse crystallization, filters, and gets coarse crystallization, oven dry; Coarse crystallization is used ethyl alcohol recrystallization 3 times and re-crystallizing in ethyl acetate 1 time successively, gets finished product Ginkgolide B compound.Sample detection Ginkgolide B content is 99.6%, and the content of bilobalide K is 0.3%.Bilobalide does not detect, and the content of total ginkgolic acids is 0.12ppm.The specific optical rotation of finished product is-58.4 °.
Embodiment 5
The preparation of Ginkgolide B compound:
First add diatomite 55g in the 900ml ethyl acetate under agitation condition, then add Folium Ginkgo extract 50g, in 72 ℃ of reflux 2 hours, filter, concentrated, it is 3.5ml:1g that filtrate is concentrated into liquid-solid ratio; Add the approximately distilled water removal of impurities of 1/10 volume in concentrated solution, tell ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:4, aluminum oxide is 300 orders, and the weight ratio of applied sample amount and aluminum oxide is 1:8, uses the water saturation eluent ethyl acetate, elution speed is 1/2BV/h, collects elutriant; Elutriant is evaporated to without ethyl acetate flavor, adds appropriate ethanol and makes it dissolving, then add water to make and contain the alcohol amount and be 65%, and is standing, makes and separates out coarse crystallization, filters, and gets coarse crystallization, oven dry; Coarse crystallization is used ethyl alcohol recrystallization 3 times and re-crystallizing in ethyl acetate 1 time successively, gets finished product Ginkgolide B compound.Sample detection Ginkgolide B content is 99.6%, and bilobalide does not detect, and the content of bilobalide K is 0.3%, and the content of total ginkgolic acids is 0.10ppm.The specific optical rotation of finished product is-57 °～-61 °.The specific optical rotation of finished product is-57 °.
Experimental example 1
This experimental example has prepared Ginkgolide B with reference to the method for following documents, and adopts the testing method in the present invention, has measured the content of Ginkgolide B, bilobalide, bilobalide K, total ginkgolic acids in the Ginkgolide B finished product, the results are shown in Table 1.
Sample 1 is the Ginkgolide B finished product with reference to the method preparation of the embodiment 1 of patent application CN101182325A;
Sample 2 is the Ginkgolide B finished product with reference to the method preparation of the embodiment 1 of patent application CN101412725A;
Sample 3 is the Ginkgolide B finished product with reference to the method preparation of the embodiment 1 of patent application CN1680392A;
Sample 4 is the Ginkgolide B finished product with reference to the method preparation of the embodiment 1 of patent application CN101302222A;
Sample 5 is the Ginkgolide B finished product with reference to the method preparation of the embodiment 1 of patent application CN102627656A;
Table 1
| ? | Ginkgolide B | Bilobalide | Bilobalide K | Total ginkgolic acids |
| Sample 1 | 97.8% | Do not detect | 1.1% | 2ppm |
| Sample 2 | 98.3% | 0.5% | 0.6% | 8ppm |
| Sample 3 | 98.7% | 0.3% | 0.5% | 10ppm |
| Sample 4 | 91.5% | 1.7% | 2.4% | 15ppm |
| Sample 5 | 98.6% | 0.5% | 0.5% | 7ppm |
The measurement result of table 1 shows, compare with the prior art of above-mentioned patent application, the Ginkgolide B compound purity of the present invention's preparation is high, content impurity is few, particularly the content of bilobalide, bilobalide K, total ginkgolic acids is very low, can be used as the bulk drug of injection, greatly improved patient's drug safety.
Experimental example 2
This experimental example has been tested the stability of the Ginkgolide B compound of the present invention's preparation.
Sample 1 is embodiment 1 product, and sample 2 is embodiment 2 products; Sample 3 is embodiment 5 products;
The HPLC purity that sample 4 obtains for the method that adopts patent CN101182325A embodiment 1 is 97.8% Ginkgolide B;
The HPLC purity that sample 5 obtains for the method that adopts patent CN1680392A embodiment 1 is 99.0% Ginkgolide B;
Sample is respectively got 1g, and this experiment is carried out according to 2005 editions second appendix XIX C medicine stability test governing principle of Chinese Pharmacopoeia, and result is as follows:
Table 2, accelerated test result
Table 3, long-term test results
In the Ginkgolide B compound, as bilobalide K, bilobalide and the total ginkgolic acids of impurity, the stability of Ginkgolide B there is certain impact, in apricot lactone B compound provided by the invention, kind and the content of impurity are less, the purity of Ginkgolide B is higher, impurity is less to the stability influence of Ginkgolide B, investigate by accelerated test and experiment for long-term stability, result shows that Ginkgolide B Compound Phase of the present invention has better stability for prior art, accelerates, test of long duration purity content is little.
Claims (10)
1. Ginkgolide B compound, it is characterized in that, described Ginkgolide B compound extracts acquisition from Ginkgo Leaf, the bilobalide K that contains trace in described Ginkgolide B compound, the content of described Ginkgolide B is greater than 99%, the content of described bilobalide K is lower than 0.6%, and the structural formula of described Ginkgolide B compound is:
2. Ginkgolide B compound according to claim 1, is characterized in that, the content of described Ginkgolide B is greater than 99.5%, and the content of described bilobalide K is lower than 0.4%.
3. Ginkgolide B compound according to claim 1, is characterized in that, do not contain bilobalide in described Ginkgolide B compound.
4. Ginkgolide B compound according to claim 1, is characterized in that, in described Ginkgolide B compound, the content of total ginkgolic acids is less than 0.12ppm.
5. according to claim 1-4 described Ginkgolide B compounds of any one, is characterized in that, the specific optical rotation of the methanol solution of described Ginkgolide B compound in the time of 20 ℃ is-57 °～-61 °.
6. the preparation method of the described Ginkgolide B compound of claim 1-5 any one, is characterized in that, described preparation method comprises:
(1) first add diatomite in ethyl acetate under agitation condition, then add Folium Ginkgo extract, in 60～73 ℃ of reflux 2-3.5 hours, filter, concentrated; Add diatomaceous amount be Folium Ginkgo extract weight 0.8-1.2 doubly, the liquid-solid ratio of ethyl acetate and Folium Ginkgo extract is 15-20ml/g;
(2) add the distilled water removal of impurities again, tell ethyl acetate layer, ethyl acetate layer peracidity alumina column is used the water saturation eluent ethyl acetate, collects elutriant;
(3) elutriant is evaporated to without ethyl acetate flavor, adds appropriate ethanol and makes it dissolving, then add water to make and contain the alcohol amount and be 60～70%, and is standing, makes and separates out coarse crystallization, filters, and gets coarse crystallization, oven dry;
(4) coarse crystallization is used ethanol and re-crystallizing in ethyl acetate successively, gets finished product Ginkgolide B compound.
7. preparation method according to claim 6, is characterized in that, in described step (1), add diatomaceous amount be Folium Ginkgo extract weight 0.9-1.1 doubly; The liquid-solid ratio of ethyl acetate and Folium Ginkgo extract is 16-18ml/g.
8. preparation method according to claim 6, is characterized in that, in described step (1), it is 3-5ml/g that filtrate is concentrated into liquid-solid ratio.
9. preparation method according to claim 6, is characterized in that, in described step (2), the blade diameter length ratio of acidic alumina column is 1:4～1:10, aluminum oxide is 200～300 orders, and the weight ratio of applied sample amount and aluminum oxide is 1:7～10, and elution speed is 1/4～1/2BV/h; Preferably, the blade diameter length ratio of described acidic alumina column is 1:4～1:7, and the weight ratio of applied sample amount and aluminum oxide is 1:8.
10. preparation method according to claim 6, is characterized in that, in described step (4), coarse crystallization uses ethyl alcohol recrystallization more than 3 times, re-crystallizing in ethyl acetate more than 1 time successively.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110627806A (en) * | 2019-09-29 | 2019-12-31 | 上海信谊百路达药业有限公司 | Bilobalide B compound and preparation method thereof |
| CN110627807A (en) * | 2019-09-29 | 2019-12-31 | 上海信谊百路达药业有限公司 | Bilobalide B raw material and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005046829A2 (en) * | 2003-11-12 | 2005-05-26 | The Trustees Of Columbia University In The City Of New York | Separation of ginkgolides and bilobalide from g. biloba |
| CN1651436A (en) * | 2004-12-07 | 2005-08-10 | 王敬勉 | Extraction method of ginkgolactone B and ginkgo lactone in ginkgo leaf |
| CN101412725A (en) * | 2007-10-15 | 2009-04-22 | 桂林市振达生物科技有限责任公司 | Method for extracting and separating bilobalide B from ginkgo leaf |
-
2013
- 2013-03-26 CN CN201310098469.9A patent/CN103130818B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005046829A2 (en) * | 2003-11-12 | 2005-05-26 | The Trustees Of Columbia University In The City Of New York | Separation of ginkgolides and bilobalide from g. biloba |
| WO2005046829A3 (en) * | 2003-11-12 | 2005-11-10 | Univ Columbia | Separation of ginkgolides and bilobalide from g. biloba |
| CN1651436A (en) * | 2004-12-07 | 2005-08-10 | 王敬勉 | Extraction method of ginkgolactone B and ginkgo lactone in ginkgo leaf |
| CN101412725A (en) * | 2007-10-15 | 2009-04-22 | 桂林市振达生物科技有限责任公司 | Method for extracting and separating bilobalide B from ginkgo leaf |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110627806A (en) * | 2019-09-29 | 2019-12-31 | 上海信谊百路达药业有限公司 | Bilobalide B compound and preparation method thereof |
| CN110627807A (en) * | 2019-09-29 | 2019-12-31 | 上海信谊百路达药业有限公司 | Bilobalide B raw material and preparation method thereof |
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| CN103130818B (en) | 2014-03-05 |
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