CN106770865B - A kind of organic acid content testing method in ginkgo biloba p.e - Google Patents
A kind of organic acid content testing method in ginkgo biloba p.e Download PDFInfo
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Abstract
The invention discloses a kind of method for determining organic acid content in ginkgo biloba p.e.This method includes following content：The steps such as the pre-treatment of ginkgo biloba p.e sample, the preparation of need testing solution, efficient liquid phase chromatographic analysis.The present invention carries out ultrasonic extraction with the aqueous solution containing bovine serum albumin(BSA), eliminates the interference of OPC during ginkgo biloba p.e Sample pretreatment；In the preparation process of need testing solution, sample preparation is carried out with the aqueous solution containing 0.1% phosphoric acid, eliminates the interference of part miscellaneous peak, substantially improve the chromatographic behavior and separating effect of chromatographic peak.After the above method is analyzed, ginkgo biloba p.e organic acid collection of illustrative plates can be obtained, and can be with the content of five kinds of Determination of Organic Acids such as simultaneous quantitative shikimic acid, gallic acid, protocatechuic acid, 6 hydroxykynurenic acids, P-hydroxybenzoic acid.
Description
Technical field
The present invention relates to a kind of content assaying method for being used for organic acid in ginkgo biloba p.e.
Background technology
Ginkgo leaf be Ginkgoaceae plant Ginkgo biloba (Ginkgobiloba L.) dried leaf, mildly bitter flavor is mild-natured, have invigorating heart,
Promoting blood circulation and stopping pain, astringe the lung relieving asthma, dampness elimination antidiarrheal the effect of.Ginkgo biloba p.e is clinically used to treat cardiovascular and cerebrovascular disease and nerve
Systemic disease.Ginkgo biloba p.e complicated component, current quality control index only have two constituents：Flavonols glycoside and lactone
Class, both total contents are 30%.Separately there are 7 one-tenth compositions not to be put into quality standard.Existing document report, ginkgo biloba p.e remove
Outside flavonol glycosides and Ginkgolide Component, also containing compositions such as OPC, organic acid, biflavone, isoflavones.Therefore, in order to by
The quality standard surveyed composition, step up and improve ginkgo biloba p.e in step increase ginkgo biloba p.e, to except flavones
Other compositions outside alcohol glycosides and Ginkgolide Component are studied, and are just particularly important.
Document " content of organic acid in high effective liquid chromatography for measuring ginkgo nut " (Journal of Analytical Science, 2011,27 (3):
336-338.) report containing oxalic acid, tartaric acid, malic acid, ascorbic acid, lactic acid, acetic acid, citric acid in gingko, and with one
Kind chromatographic condition determines this 7 kinds of small molecular organic acids simultaneously.Chromatographic condition mobile phase is 3% methanol -0.01mol/
LK2HPO4, that is, buffer salt system is used.
The present inventor team finds under study for action, although containing above-mentioned 7 kinds of small molecular organic acids in gingko, ginkgo leaf carries
Take and these organic acids are but free of in thing, but contain at least five kinds of other organic acid compositions, including shikimic acid, gallic acid, original
Catechuic acid, 6-HKA (6-HKA) and P-hydroxybenzoic acid.These organic acid compositions are respectively provided with preferable biology
Activity：Shikimic acid has the effect such as anti-inflammatory and antalgic, antitumor, antithrombotic, AntiHIV1 RT activity, is synthesis anti-avian influenza virus and A type
The important source material of H1N1 influenza viruses medicine " Tamiflu "；Gallic acid has the effect such as antitumor, liver protection, anti-fungal infection；It is former
Catechuic acid has the multiple efficacies such as anti-inflammatory, protection nerve cell, antiviral, promotion stem cells hyperplasia differentiation；6-HKA is efficient
Glutamate receptor antagonists, there is good protective effect to nerve excitability damage；P-hydroxybenzoic acid has anti-inflammation
Effect.
In the organic acid content in determining ginkgo biloba p.e, it is found that the most important composition of interference measurement is former cyanine
Element.Therefore, the premise of Accurate Determining organic acid content is that organic acid composition is purified.The conventional way of purification of organic acid
There are two kinds, first, being extracted with buck, extracted after extract solution acid adjustment with organic solvent；Another kind is entered with anion exchange resin
Row purifying.Both have used aggressive solvent or toxic solvents at method, and the former includes sodium hydroxide, ethyl acetate, the latter's bag
Include sodium hydroxide, methanol etc..And both approaches all can not effectively remove the OPC in ginkgo biloba p.e, and
Organic acid also has loss.It is to use polyamide to remove the optimal method of OPC, can adsorb this constituents completely, but together
When, part organic acid can be also lost, such as 6-HKA (6-HKA).A kind of patent " natural 6-HKA
The preparation method and applications of extract " (CN201410007590.0) illustrate that 6-HKA is had to 70% ethanol containing acid
The 6-HKA adsorbed on polyamide could be eluted completely, and now, part OPC composition can be also eluted
Get off, so as to interference measurement.Macroporous resin purification method is a kind of method of conventional purifying OPC, but the pole of OPC
Property span is very big, monomer component, as catechin is slightly soluble in cold water, and high polymer OPC, it is soluble in cold water, and former cyanine
The degree of polymerization of element is higher, and polarity is bigger, therefore macroreticular resin can be used for adsorbing OPC, but the high polymer original that polarity is larger
Anthocyanidin and Determination of Organic Acids are eluted with water or low-concentration ethanol aqueous solution can, still can not remove interference completely.
In summary, the organic acid content in Accurate Determining ginkgo biloba p.e improves silver for the quality of control ginkgo biloba p.e
The standard of apricot leaf extract, there is great meaning；And a kind of removal OPC is found, in Accurate Determining ginkgo biloba p.e
The method of organic acid is current pharmaceuticals industry personnel extremely concern.
The content of the invention
The invention provides a kind of content assaying method of organic acid in ginkgo biloba p.e, it is therefore intended that can quantify and survey
The content of organic acid in ginkgo biloba p.e is determined, to reach the more controllable target of the quality of ginkgo biloba p.e.
The technical solution adopted in the present invention is specific as follows：
The content assaying method of organic acid, comprises the following steps in a kind of ginkgo biloba p.e：
(1) pre-treating method of ginkgo biloba p.e：Ginkgo biloba p.e is taken, is surpassed with the aqueous solution containing bovine serum albumin(BSA)
Sound extracts, filtering；Filtrate is concentrated and dried, and obtains organic acids extract；
(2) preparation of need testing solution：Aqueous dissolution constant volume of the organic acids extract containing 0.05~0.4% phosphoric acid,
Shake up, filter, take subsequent filtrate, produce；
(3) preparation of reference substance solution：Precision weighs shikimic acid, gallic acid, protocatechuic acid, 6- hydroxykynurenes
Acid, the standard items of P-hydroxybenzoic acid are appropriate, with mobile phase (acetonitrile:0.1% phosphate aqueous solution (V/V)：3.5:96.5) dissolve,
Produce；
(4) efficient liquid phase chromatographic analysis：Need testing solution and reference substance solution are drawn respectively, inject high performance liquid chromatography
Instrument, determined by following chromatographic condition, obtain the reference substance chromatogram of five kinds of organic acids and the chromatogram of test sample；
Chromatographic condition is：Stationary phase is using octyl silane group silica gel as filler, and mobile phase A is acetonitrile, Mobile phase B
For 0.1% phosphate aqueous solution；UV-detector, 0~6min of Detection wavelength are 215nm, and 6~25min is 254nm；Column temperature is 30
℃；Sample size is 10ul；Flow velocity is 1.0ml/min；Using isocratic elution, the volume fraction of mobile phase A is 3.5%, Mobile phase B
Volume fraction be 96.5%.
In preparation method of the present invention：
In step 1), the pre-treating method of ginkgo biloba p.e is to take ginkgo biloba p.e 100mg, with containing 0.1~10g/
The aqueous solution 5~100ml, 20~70 DEG C of ultrasonic extraction 5min~3h of L bovine serum albumin(BSA)s, filtering；Filtrate is concentrated and dried, and obtains
Organic acids extract.
The optimal pre-treating method of ginkgo biloba p.e is preferably to take ginkgo biloba p.e 100mg in step 1), with containing 1g/L
The aqueous solution 10ml, the 50 DEG C of ultrasonic extraction 1h of bovine serum albumin(BSA), filtering；Filtrate is concentrated and dried, and obtains organic acids extract.
The preparation method of need testing solution is preferably in step 2), the aqueous solution of the organic acids extract containing 0.1% phosphoric acid
Dissolve and be settled to 10ml, shake up, filter, take subsequent filtrate, produce.
The content assaying method of organic acid in a kind of ginkgo biloba p.e provided by the invention, there is following advantage：
1. it is of the invention compared with the method for conventional purifying organic acid, severe corrosive and the solvent of toxicity are not used, entirely
Pretreatment process only uses second alcohol and water, environment friendly and pollution-free.
2. the present invention removes OPC with bovine serum albumin(BSA), with the aqueous solution of phosphoric acid come sample preparation, fully removal
The impurity of interference organic acid main peak chromatographic behavior, the chromatographic behavior and separating effect of chromatographic peak are substantially improved, so as to reach
The purpose of accurate quantitative analysis organic acid.
3. the mobile phase in the chromatographic condition that assay method of the present invention uses is acetonitrile:0.1% phosphate aqueous solution (V/V)：
3.5:96.5, buffer salt system is not used, is more beneficial for balance, post processing and the preservation of chromatographic column.
Brief description of the drawings：
Fig. 1 is mixing reference substance chromatogram, and No. 1 peak is shikimic acid, No. 2 peaks are gallic acid, No. 3 peaks are protocatechuic acid, 4
Number peak is 6-HKA, No. 5 peaks are P-hydroxybenzoic acid
Fig. 2 is sample chromatogram prepared by purification process 1 (the inventive method), and No. 1 peak is shikimic acid, No. 2 peaks are not eat
Sub- acid, No. 3 peaks are protocatechuic acid, No. 4 peaks are 6-HKA, No. 5 peaks are P-hydroxybenzoic acid
Fig. 3 is sample chromatogram prepared by purification process 2, and No. 1 peak is shikimic acid, No. 2 peaks are gallic acid, No. 3 peaks are
Protocatechuic acid, No. 4 peaks are 6-HKA, No. 5 peaks are P-hydroxybenzoic acid
Fig. 4 is sample chromatogram prepared by purification process 3, and No. 1 peak is shikimic acid, No. 2 peaks are gallic acid, No. 3 peaks are
Protocatechuic acid, No. 4 peaks are 6-HKA, No. 5 peaks are P-hydroxybenzoic acid
Fig. 5 is the sample chromatogram without bovine serum albumin(BSA) pre-treatment, and No. 1 peak is shikimic acid, No. 2 peaks are nutgall
Acid, No. 3 peaks are protocatechuic acid, No. 4 peaks are 6-HKA, No. 5 peaks are P-hydroxybenzoic acid
Fig. 6 is the sample chromatogram with 0.3g/L bovine serum albumin(BSA) pre-treatments, and No. 1 peak is shikimic acid, No. 2 peaks are not eat
Sub- acid, No. 3 peaks are protocatechuic acid, No. 4 peaks are 6-HKA, No. 5 peaks are P-hydroxybenzoic acid
Fig. 7 is the chromatogram with 70% methanol sample preparation sample, and No. 1 peak is shikimic acid, No. 2 peaks are gallic acid, No. 3 peaks are
Protocatechuic acid, No. 4 peaks are 6-HKA, No. 5 peaks are P-hydroxybenzoic acid
Fig. 8 is the chromatogram for using the aqueous solution sample preparation sample containing 0.05% phosphoric acid, and No. 1 peak is shikimic acid, No. 2 peaks are not eat
Sub- acid, No. 3 peaks are protocatechuic acid, No. 4 peaks are 6-HKA, No. 5 peaks are P-hydroxybenzoic acid
Embodiment
Influence of the 1 different purification process of embodiment to organic acidity test
Purification process 1：100mg ginkgo biloba p.es, with the aqueous solution 10ml of the bovine serum albumin(BSA) containing 1g/L, 50 DEG C of ultrasounds
Extract 60min, filtering；Filtrate is with HPD5000 macroporous resin adsorptions, washing, 10% ethanol elution, collect merge absorption raffinate,
Position and 10% alcohol elution are washed, is concentrated and dried, with 0.1% phosphoric acid dissolved residue, and is settled to 10ml.
Purification process 2：100mg ginkgo biloba p.es, with the extraction with aqueous solution of 10ml pH value 13, extract solution concentrated hydrochloric acid
PH value is adjusted to be extracted 2 times to 1, then with 100ml ethyl acetate.Merge ethyl acetate solution twice, be concentrated and dried, with 0.1% phosphoric acid
Dissolved residue, and it is settled to 10ml.
Purification process 3：100mg ginkgo biloba p.es, being extracted with 10ml water, extract solution is adsorbed with anion exchange resin,
After washing, eluted with the methanol solution of 1mol/L sodium hydroxides 70%, collect elution solution, be concentrated and dried, dissolved with 0.1% phosphoric acid
Residue, and it is settled to 10ml.
Chromatographic condition is：Stationary phase is using octyl silane group silica gel as filler, and mobile phase A is acetonitrile, Mobile phase B
For 0.1% phosphate aqueous solution；UV-detector, 0~6min of Detection wavelength are 215nm, and 6~25min is 254nm；Column temperature is
30℃；Sample size is 10ul；Flow velocity is 1.0ml/min；Using isocratic elution, the volume fraction of mobile phase A is 3.5%, flowing
Phase B volume fraction is 96.5%.
As a result accompanying drawing 1~4 is seen, accompanying drawing 1 is mixing reference substance chromatogram, and 5 main peak peak types are good, no miscellaneous peak interference；Fig. 2
The sample chromatogram prepared for purification process 1 (the inventive method), 5 main peak peak types are good, no miscellaneous peak interference；Fig. 3 is purifying
Sample chromatogram prepared by method 2, No. 1 peak almost do not have, No. 2 peaks have miscellaneous peak disturb and peak area be significantly less than in Fig. 22
Number peak area, No. 4 peak peak areas are also obviously reduced, and explanation purification process 2 prepares sample, shikimic acid, gallic acid and 6-HKA
There is loss；Fig. 4 is sample chromatogram prepared by purification process 3, and there is miscellaneous peak interference at No. 1 peak, and No. 5 peaks disappear, and No. 4 peak areas subtract
Small, explanation purification process 2 prepares sample, and 6-HKA and P-hydroxybenzoic acid have loss.
Organic acid composition is qualitative and quantitative in the ginkgo biloba p.e of embodiment 2
Ginkgo biloba p.e 100mg is taken, is carried respectively with the aqueous solution 10ml of the bovine serum albumin(BSA) containing 1.0g/L, 50 DEG C of ultrasounds
1h is taken, is filtered；Filtrate is concentrated and dried, and obtains organic acids extract；Aqueous dissolution of the organic acids extract containing 0.1% phosphoric acid
And 10ml is settled to, shake up, filter, take subsequent filtrate；Subsequent filtrate is measured by the chromatographic condition of embodiment 1.
The above method carries out Method validation, including instrument precision, and repeatability, Intermediate precision, linearly, sample-adding reclaim
Rate, durability.
As a result find, shikimic acid, gallic acid, protocatechuic acid, 6-HKA, para hydroxybenzene first are contained in ginkgo biloba p.e
Acid, without forulic acid, chlorogenic acid, 3,5-Dicaffeoylquinic acid, 3,4-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid, caffeic acid, p-Coumaric Acid, vanillic acid, anti-
Bad hematic acid, D-Glucose aldehydic acid, quininic acid.
Illustrate that the inventive method can be with 5 kinds of organic acids in Simultaneous Determination ginkgo biloba p.e by the result of 1~table of table 4
Content, all indexs of Method validation meet States Pharmacopoeia specifications, and this method is accurate, reliable.
Illustrated by the result of table 5, the ginkgo biloba p.e of different batches is containing shikimic acid, gallic acid, protocatechuic acid, 6-
This 5 kinds of organic acids of HKA, P-hydroxybenzoic acid, 5 kinds of organic acid total contents are about 2.2%~3.0%.
15 kinds of organic acid regression equations of table
Table 2 is loaded recovery test result
The Precision Experiment result of table 3
The durability experimental result of table 4
The batch ginkgo biloba p.e organic acid content testing result of table 5 10
The investigation of the bovine serum albumin(BSA) dosage of embodiment 3
Take ginkgo biloba p.e 100mg, respectively with containing 0,0.1,0.3,1.0,3.0,10g/L bovine serum albumin(BSA)s it is water-soluble
Liquid 10ml, 50 DEG C of ultrasonic extraction 1h, filtering；Filtrate is concentrated and dried, and obtains organic acids extract；Organic acids extract, which is used, contains 0.1%
The aqueous dissolution of phosphoric acid is simultaneously settled to 10ml, shakes up, and filtration, takes subsequent filtrate；Subsequent filtrate is surveyed by the chromatographic condition of embodiment 1
It is fixed.
As a result accompanying drawing 2, accompanying drawing 5 and accompanying drawing 6 are seen.Accompanying drawing 2 is the sample chromatogram with 1.0g/L bovine serum albumin(BSA) pre-treatments
Figure, 5 main peak peak types are good, no miscellaneous peak interference, with the sample chromatogram of 3.0 or 10g/L bovine serum albumin(BSA) pre-treatments with attached
Fig. 2.Accompanying drawing 5 is the sample chromatogram without bovine serum albumin(BSA) pre-treatment, and 1,2, No. 4 peak has obvious miscellaneous peak to disturb；Accompanying drawing 6
To there is obvious miscellaneous peak to disturb with the sample chromatogram 1 of 0.3g/L bovine serum albumin(BSA) pre-treatments and No. 2 peaks, with 0.1g/L ox bloods
The sample chromatogram of pure albumen pre-treatment is substantially the same as accompanying drawing 6.Illustrate that bovine serum albumin(BSA) dosage in more than 1.0g/L, can just be gone
Except the miscellaneous peak interference at 1~No. 4 peak.
The investigation of the Extraction solvent amount of embodiment 4
Take ginkgo biloba p.e 100mg, respectively with 5,10,20,50,100ml bovine serum albumin(BSA)s containing 1.0g/L it is water-soluble
Liquid, 50 DEG C of ultrasonic extraction 1h, filtering；Filtrate is concentrated and dried, and obtains organic acids extract；Organic acids extract, which is used, contains 0.1% phosphoric acid
Aqueous dissolution and be settled to 10ml, shake up, filter, take subsequent filtrate；Subsequent filtrate is measured by the chromatographic condition of embodiment 1.
The result of table 6 illustrates that Extraction solvent amount is 5ml, still has part organic acid to fail to extract, therefore, selective extraction
Quantity of solvent is 10ml.
The investigation of the Extraction solvent amount of table 6
| Extraction solvent amount (ml) | 5 kinds of organic acid total contents (%) |
| 5 | 2.029 |
| 10 | 2.285 |
| 20 | 2.288 |
| 50 | 2.286 |
| 100 | 2.290 |
The investigation of the Extracting temperature of embodiment 5
Take ginkgo biloba p.e 100mg, with the aqueous solution 10ml of the bovine serum albumin(BSA) containing 1.0g/L, respectively 20,30,
40th, 50,60,70 DEG C of ultrasonic extraction 1h, filtering；Filtrate is concentrated and dried, and obtains organic acids extract；Organic acids extract is with containing
The aqueous dissolution of 0.1% phosphoric acid is simultaneously settled to 10ml, shakes up, and filtration, takes subsequent filtrate；Subsequent filtrate presses the chromatographic condition of embodiment 1
It is measured.
The result of table 7 illustrates that when Extracting temperature is less than 50 DEG C, part organic acid fails to extract, therefore, selective extraction temperature
50 DEG C of degree.
The investigation of the Extracting temperature of table 7
| Extracting temperature (DEG C) | 5 kinds of organic acid total contents (%) |
| 20 | 1.863 |
| 30 | 1.906 |
| 40 | 2.080 |
| 50 | 2.285 |
| 60 | 2.281 |
| 70 | 2.283 |
The investigation of the extraction time of embodiment 6
Take ginkgo biloba p.e 100mg, with the aqueous solution 10ml of the bovine serum albumin(BSA) containing 1.0g/L, 50 DEG C respectively ultrasound carry
Take 0.5,1,2h, 3h, filtering；Filtrate is concentrated and dried, and obtains organic acids extract；Water of the organic acids extract containing 0.1% phosphoric acid
Solution dissolves and is settled to 10ml, shakes up, and filtration, takes subsequent filtrate；Subsequent filtrate is measured by the chromatographic condition of embodiment 1.
The result of table 8 illustrates that when extraction time is 0.5h, part organic acid fails to extract, therefore, the selective extraction time
1h。
The investigation of the extraction time of table 8
| Extraction time (h) | 5 kinds of organic acid total contents (%) |
| 0.5 | 1.642 |
| 1 | 2.286 |
| 2 | 2.290 |
| 3 | 2.285 |
The phosphoric acid concentration that embodiment 7 prepares need testing solution is investigated
Ginkgo biloba p.e 100mg is taken, with the aqueous solution 10ml of the bovine serum albumin(BSA) containing 1.0g/L, 50 DEG C of ultrasonic extractions
1h, filtering；Filtrate merges absorption raffinate, washing position with HPD5000 macroporous resin adsorptions, washing, 10% ethanol elution, collection
With 10% alcohol elution, it is concentrated and dried, obtains organic acids extract；Organic acids extract respectively with 70% methanol, water, contain
0.05th, the aqueous dissolution of 0.1,0.2,0.4% phosphoric acid and 10ml is settled to, shaken up, filtered, take subsequent filtrate；Subsequent filtrate is by real
The chromatographic condition of example 1 is applied to be measured.
As a result accompanying drawing 2, accompanying drawing 7 and accompanying drawing 8 are seen.Accompanying drawing 2 is the chromatogram with the aqueous solution sample preparation sample containing 0.1% phosphoric acid
Figure, 5 main peak peak types are good, the interference of no miscellaneous peak, with 0.2 or 0.4% phosphoric acid aqueous solution sample preparation sample the same accompanying drawing of chromatogram
2.Accompanying drawing 7 is the chromatogram with 70% methanol sample preparation sample, and No. 4 peaks have obvious miscellaneous peak to disturb；Accompanying drawing 8 is to use to contain 0.05% phosphorus
The chromatogram of the aqueous solution sample preparation sample of acid, there is miscellaneous peak interference at No. 1 peak, with the chromatogram of water sample preparation sample with accompanying drawing 8.
Claims (4)
1. the content assaying method of organic acid, comprises the following steps in a kind of ginkgo biloba p.e：
(1) pre-treating method of ginkgo biloba p.e：Ginkgo biloba p.e is taken, is carried with the aqueous solution ultrasound containing bovine serum albumin(BSA)
Take, filter；Filtrate is concentrated and dried, and obtains organic acids extract；
(2) preparation of need testing solution：Aqueous dissolution constant volume of the organic acids extract containing 0.05~0.4% phosphoric acid, shakes up,
Filtration, takes subsequent filtrate, produces；
(3) preparation of reference substance solution：Precision weighs shikimic acid, gallic acid, protocatechuic acid, 6-HKA, right
The standard items of hydroxybenzoic acid are appropriate, with flowing phased soln, produce；
(4) efficient liquid phase chromatographic analysis：Need testing solution and reference substance solution are drawn respectively, are injected high performance liquid chromatograph, are pressed
Following chromatographic condition measure, obtain the reference substance chromatogram of five kinds of organic acids and the chromatogram of test sample；
Chromatographic condition is：Stationary phase is using octyl silane group silica gel as filler, and mobile phase A is acetonitrile, and Mobile phase B is
0.1% phosphate aqueous solution；UV-detector, 0~6min of Detection wavelength are 215nm, and 6~25min is 254nm；Column temperature is 30 DEG C；
Sample size is 10 μ l；Flow velocity is 1.0ml/min；Using isocratic elution, the volume fraction of mobile phase A is 3.5%, Mobile phase B
Volume fraction is 96.5%.
2. the content assaying method of organic acid in ginkgo biloba p.e according to claim 1, it is characterised in that described
In step (1), the pre-treating method of ginkgo biloba p.e is to take ginkgo biloba p.e 100mg, with containing 0.1~10g/L cow's serums
The aqueous solution 5~100ml, 20~70 DEG C of ultrasonic extraction 5min~3h of albumin, filtering；Filtrate is concentrated and dried, and obtains organic acid and carries
Take thing.
3. the content assaying method of organic acid in ginkgo biloba p.e according to claim 1 or 2, it is characterised in that institute
In the step of stating (1), the pre-treating method of ginkgo biloba p.e is to take ginkgo biloba p.e 100mg, pure with ox blood containing 1g/L
The aqueous solution 10ml, the 50 DEG C of ultrasonic extraction 1h of albumen, filtering；Filtrate is concentrated and dried, and obtains organic acids extract.
4. the content assaying method of organic acid in ginkgo biloba p.e according to claim 1, it is characterised in that described
The preparation method of need testing solution is in step (2), and organic acids extract is used the aqueous dissolution containing 0.1% phosphoric acid and is settled to
10ml, shake up, filter, take subsequent filtrate, produce.
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| CN109369373B (en) * | 2018-12-18 | 2021-01-08 | 长江师范学院 | Method for preparing shikimic acid extract from ginkgo leaf extract chromatography waste liquid |
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| CN113582835A (en) * | 2021-08-05 | 2021-11-02 | 湖北金日生态能源股份有限公司 | Method for extracting shikimic acid from ginkgo leaf waste residue and application |
| WO2023081198A1 (en) * | 2021-11-03 | 2023-05-11 | Lanny Leo Johnson | Methods and compositions including protocatechuic acid crystals for the treatment of conditions caused by an enveloped virus |
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