CN110967426A - Enrichment method and content determination method of free quercetin in safflower injection - Google Patents
Enrichment method and content determination method of free quercetin in safflower injection Download PDFInfo
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Abstract
The invention provides an enrichment method and a content determination method of free quercetin in a safflower injection. The enrichment method of free quercetin in the safflower injection adopts a salting-out adsorption method, excessive salt is added to separate out the free quercetin in the safflower injection and dissolve the free quercetin in acetonitrile, and the enrichment of the free quercetin in the safflower injection can be realized by repeatedly and fully adsorbing the acetonitrile. The acetonitrile-enriched free quercetin in the method has high purity and low impurity content, reduces the interference of impurities in the subsequent determination process of the content of the free quercetin, and improves the detection accuracy. The method for measuring the content of free quercetin in the safflower injection is high in precision, good in repeatability, good in stability and high in accuracy.
Description
Technical Field
The invention relates to the field of detection and analysis, and particularly relates to an enrichment method and a content determination method of free quercetin in a safflower injection.
Background
The safflower injection liquid has complex chemical components and mainly contains 16 compounds. Quercetin is a kind of water insoluble component, and is mainly present in safflower injection as flavonol compound comprising glycoside with quercetin as mother nucleus, and its free component is still present. The free quercetin has a great relationship with the visible foreign matter cause of the safflower injection, and the monitoring of the content of the water-insoluble free quercetin in the safflower injection has great relation with the quality control of the safflower injection.
The related literature indicates that a method for detecting the content of quercetin in a large number of traditional Chinese medicine components is to hydrolyze a flavonol compound consisting of glycoside taking quercetin as a parent body by acid, and no related research on a method for measuring the content of free quercetin in a safflower injection exists at present. In addition, the content of free quercetin in the safflower injection is extremely low, other components also have certain interference on the content determination of the free quercetin, and the difficulty in the content determination of the free quercetin is high.
Disclosure of Invention
The invention aims to provide a method for enriching free quercetin in a safflower injection and a content determination method.
In order to achieve the above object, the present invention provides the following technical solutions.
The invention provides a method for enriching free quercetin in a safflower injection, which comprises the following steps:
step a, adding salt into the safflower injection, and uniformly mixing to obtain a mixed solution A, wherein the salt is in a supersaturated state in the mixed solution A;
and b, adsorbing the free quercetin in the mixed solution A by using acetonitrile.
In a further improvement of the above technical means, the salt is selected from any one of sodium chloride, sodium sulfate and ammonium sulfate.
As a further improvement of the technical proposal, at least 4.5g of sodium chloride is added into 20mL of safflower injection.
As a further improvement of the technical scheme, the mixed solution A needs to be kept still for 30-60 min before acetonitrile is used for adsorption.
As a further improvement of the technical scheme, the volume ratio of the acetonitrile to the safflower injection is (0.5-2): 1.
as a further improvement of the above technical scheme, acetonitrile is used for adsorbing the free quercetin in the mixed solution A for multiple times in the step b;
as a further improvement of the above technical solution, the plurality of times is at least 3 times.
As a further improvement of the above technical solution, the adsorption of the free quercetin in the mixed solution a using acetonitrile includes: and adding acetonitrile into the mixed solution A, uniformly mixing, centrifuging, and centrifuging to obtain an upper layer solution, namely the acetonitrile solution adsorbed with the free quercetin.
The second aspect of the invention provides a method for measuring the content of free quercetin in a safflower injection, which comprises the following steps:
and 2, quantitatively detecting the enriched free quercetin.
As a further improvement of the above technical scheme, the quantitative detection is HPLC detection.
As a further improvement of the above technical solution, step 1 includes: after removing acetonitrile in the acetonitrile solution absorbed with the free quercetin, the acetonitrile solution is dissolved in methanol and then subjected to HPLC detection.
As a further improvement of the above technical scheme, the acetonitrile in the acetonitrile solution is removed by evaporation.
As a further improvement of the technical scheme, the methanol solution is filtered by a microporous filter membrane and then subjected to HPLC detection, and the pore diameter of the microporous filter membrane is 0.22 μm or 0.45 μm.
As a further improvement of the technical proposal, the aperture of the microporous filter membrane is 0.22 μm.
As a further improvement of the above technical solution, the detection conditions of HPLC include one or more of the following a to f:
a. the chromatographic column is an octadecylsilane chemically bonded silica reverse chromatographic column;
b. the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a phosphoric acid aqueous solution, and the volume ratio of the mobile phase A to the mobile phase B is (45-50) to (55-50); preferably, the concentration of the phosphoric acid aqueous solution is 0.4 wt%;
c. the column temperature is 30 ℃;
d. the column flow rate is 1 mL/min;
e. the sample injection amount is 20-40 mu L;
f. the detector is an ultraviolet detector; preferably, the detection wavelength is 370 nm.
The invention has the beneficial effects that:
the method for enriching the free quercetin in the safflower injection adopts a salting-out adsorption method to enrich the free quercetin in the safflower injection, the purity of the enriched free quercetin is high, the content of impurities is low, the interference of the impurities in the subsequent free quercetin content measurement process is reduced, and the accuracy of the content measurement is improved.
The method for measuring the content of the free quercetin in the safflower injection has the advantages of high precision, good repeatability, good stability and high accuracy.
Drawings
To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, and it should be understood that the following drawings only illustrate some embodiments of the present invention, and therefore should not be considered as limiting the scope of the present invention.
FIG. 1 is a standard operating curve for example 1.
FIG. 2 is a high performance liquid chromatogram obtained for different mobile phases.
FIG. 3 is a high performance liquid chromatography of acetonitrile-enriched free quercetin and a quercetin control.
FIG. 4 is a high performance liquid chromatography of acetone-enriched free quercetin and a quercetin control.
FIG. 5 is a high performance liquid chromatography of ethyl acetate enriched free quercetin and a quercetin control.
Detailed Description
The terms as used herein:
"prepared from … …" is synonymous with "comprising". The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
The conjunction "consisting of … …" excludes any unspecified elements, steps or components. If used in a claim, the phrase is intended to claim as closed, meaning that it does not contain materials other than those described, except for the conventional impurities associated therewith. When the phrase "consisting of … …" appears in a clause of the subject matter of the claims rather than immediately after the subject matter, it defines only the elements described in the clause; other elements are not excluded from the claims as a whole.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when the range "1 ~ 5" is disclosed, the ranges described should be construed to include the ranges "1 ~ 4", "1 ~ 3", "1 ~ 2 and 4 ~ 5", "1 ~ 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.
In these examples, the parts and percentages are by mass unless otherwise indicated.
"part by mass" means a basic unit of measure indicating a mass ratio of a plurality of components, and 1 part may represent any unit mass, for example, 1g or 2.689 g. If we say that the part by mass of the component A is a part by mass and the part by mass of the component B is B part by mass, the ratio of the part by mass of the component A to the part by mass of the component B is a: b. alternatively, the mass of the A component is aK and the mass of the B component is bK (K is an arbitrary number, and represents a multiple factor). It is unmistakable that, unlike the parts by mass, the sum of the parts by mass of all the components is not limited to 100 parts.
"and/or" is used to indicate that one or both of the illustrated conditions may occur, e.g., a and/or B includes (a and B) and (a or B).
The invention provides a method for enriching free quercetin in a safflower injection, which comprises the following steps:
step a, adding salt into the safflower injection, and uniformly mixing to obtain a mixed solution A, wherein the salt is in a supersaturated state in the mixed solution A;
and b, adsorbing the free quercetin in the mixed solution A by using acetonitrile.
The method adopts a salting-out adsorption method, excessive salt is added to separate out free quercetin in the safflower injection and dissolve the free quercetin in acetonitrile, the acetonitrile and the safflower injection are separated into layers, the acetonitrile layer is taken, and the enrichment of the free quercetin in the safflower injection can be realized through repeated and sufficient adsorption of the acetonitrile. The acetonitrile-enriched free quercetin in the method has high purity and low impurity content, reduces the interference of impurities in the subsequent determination process of the content of the free quercetin, and improves the detection accuracy.
Optionally, the salt is selected from any one of sodium chloride, sodium sulfate and ammonium sulfate.
When sodium chloride is used as the salt, sodium chloride is supersaturated in the mixed solution A when at least 4.5g of sodium chloride is added to 20mL of the safflower injection.
Optionally, the mixed solution A is required to be still standing before the acetonitrile is used for adsorption, and the still standing time is preferably 30-60 min. Free quercetin in the safflower injection can be fully separated out by standing.
Optionally, the volume ratio of the acetonitrile to the safflower injection is (0.5-2): 1.
optionally, adsorbing the free quercetin in the mixed solution A for multiple times by using acetonitrile in the step b; preferably, the plurality of times is at least 3 times.
The free quercetin in the safflower injection can be more fully adsorbed by multiple times of adsorption, and when the adsorption times are more than 4 times, the free quercetin in the safflower injection can be completely adsorbed.
Alternatively, adsorbing the free quercetin in the mixed solution a using acetonitrile includes: and adding acetonitrile into the mixed solution A, uniformly mixing, centrifuging, and centrifuging to obtain an upper layer solution, namely the acetonitrile solution adsorbed with the free quercetin. The centrifugal rotating speed is 4000-6000 r/min, and the centrifugal time is 10-20 min; preferably, the centrifugal speed is 5000r/min, and the centrifugal time is 15 min.
The second aspect of the invention provides a method for measuring the content of free quercetin in a safflower injection, which comprises the following steps:
and 2, quantitatively detecting the enriched free quercetin.
The quantitative method may be a quantitative method commonly used in the art, and may be a quantitative detection capable of realizing the quantitative detection of quercetin, optionally, the quantitative detection is HPLC detection.
Optionally, step 1 comprises: after removing acetonitrile in the acetonitrile solution absorbed with the free quercetin, the acetonitrile solution is dissolved in methanol and then subjected to HPLC detection.
Optionally, the acetonitrile in the acetonitrile solution is removed by evaporation.
Optionally, the methanol solution is filtered by using a microfiltration membrane and then subjected to HPLC detection, wherein the pore diameter of the microfiltration membrane is 0.22 μm or 0.45 μm; preferably, the pore size of the microfiltration membrane is 0.22 μm.
Optionally, the detection conditions of HPLC comprise one or more of the following a to f:
a. the chromatographic column is an octadecylsilane chemically bonded silica reverse chromatographic column;
b. the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a phosphoric acid aqueous solution, and the volume ratio of the mobile phase A to the mobile phase B is (45-50) to (55-50); preferably, the concentration of the phosphoric acid aqueous solution is 0.4 wt%;
c. the column temperature is 30 ℃;
d. the column flow rate is 1 mL/min;
e. the sample injection amount is 20-40 mu L;
f. the detector is an ultraviolet detector; preferably, the detection wavelength is 370 nm.
The HPLC detection method has the advantages of high precision, good repeatability, good stability and high accuracy.
Embodiments of the present invention will be described in detail below with reference to specific examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Acetonitrile: and (4) carrying out chromatographic purification.
Methanol: and (5) analyzing and purifying.
Acetone: and (5) analyzing and purifying.
Ethyl acetate: and (5) analyzing and purifying.
The quercetin standard substance comprises the following components: purity is more than or equal to 99.1 percent, and the China institute for food and drug testing.
Safflower injection: huarun Sanjiu Yaan pharmaceutical Co., Ltd.
Example 1
The method for measuring the content of free quercetin in the safflower injection comprises the following steps:
(1) enriching free quercetin in the safflower injection: placing 20mL of the safflower injection into a centrifuge tube, adding 6g of sodium chloride, shaking up, placing for 30-60 min, adding 40mL of chromatographic acetonitrile, shaking for 5min, centrifuging (the centrifugal rotation speed is 5000r/min, the centrifugal time is 15min), layering, and absorbing the upper acetonitrile solution; then, 30mL and 20mL of acetonitrile were added and adsorbed for 1 time, and the acetonitrile solutions were combined.
(2) Preparation of a test solution: placing the acetonitrile solution adsorbed with free quercetin in an evaporation dish, blowing with nitrogen at normal temperature, evaporating to remove acetonitrile, dissolving the rest solid with a small amount of methanol, transferring to a 2mL volumetric flask, adding methanol to constant volume to scale, shaking, and filtering with microporous (0.22 μm) filter membrane.
(3) Preparation of standard working solution: quercetin control 1.527mg was placed in a 50mL volumetric flask and diluted to the mark with methanol to give a standard stock solution (30.27. mu.g/mL). Then, an appropriate amount of standard stock solution was taken, and the standard stock solution was diluted to 3.027. mu.g/mL, 6.055. mu.g/mL, 9.082. mu.g/mL, 12.108. mu.g/mL, and 15.135. mu.g/mL for linear studies.
(4) And (4) HPLC detection:
the test solution and the standard working solution were subjected to HPLC analysis.
The HPLC operating conditions were as follows:
a chromatographic column: phenomenex Gemini-C18(4.6 mm. times.250 mm, 5.0 μm);
mobile phase: 48% methanol and 52% 0.4 wt% phosphoric acid;
column temperature: 30 ℃;
column flow rate: 1 mL/min;
a detector: an ultraviolet detector;
detection wavelength: 370 nm.
And (3) detection: precisely sucking 20 μ L of each of the test solution and the standard working solution, injecting into a gas chromatograph, measuring, and recording the peak area of quercetin.
According to the measurement result of the standard working solution, the sample volume of the quercetin (i.e. the mass of the quercetin in the 20 μ L standard working solution) is taken as an abscissa (X), the total peak area is taken as an ordinate (Y), a standard working curve (fig. 1) is drawn as Y2190X +2.471, and the correlation coefficient r is 0.9995, so that the correlation requirement required by detection can be met.
Substituting the test result of the test sample into the standard working curve shown in fig. 1 to obtain the mass of the quercetin in the test sample, and calculating to obtain the content of the quercetin in the test sample, wherein the content of the quercetin is the mass of the quercetin/the volume of the test sample.
The content of free quercetin in 10 batches of safflower injection is detected by using the method. The results are shown in Table 1.
TABLE 1 measurement of free Quercetin content in safflower injection
Example 2
Precision test
Precisely absorbing 20 mu L of quercetin standard stock solution with the concentration of 6.055 mu g/mL, continuously feeding samples for 6 times, detecting the content of quercetin, calculating the RSD of the peak area, and calculating the result by using the HPLC conditions as in example 1, wherein the result is shown in Table 2.
TABLE 2 results of precision measurements
| |
1 | 2 | 3 | 4 | 5 | 6 | RSD(％) |
| Peak area | 266.231 | 267.68 | 262.243 | 265.397 | 264.726 | 270.854 | 1.1 |
Example 3
Repeatability test
Taking safflower injection with batch KY180801, enriching free quercetin according to the method in the example 1, and preparing 6 parts of test sample according to the preparation method of the test sample. 20 mu L of sample injection is respectively sucked, the content of the quercetin component of the test sample is measured, the HPLC condition is the same as that in example 1, the peak area RSD is calculated, and the result is shown in Table 3, which indicates that the method has good repeatability.
TABLE 3 results of repeated measurements
| |
1 | 2 | 3 | 4 | 5 | 6 | RSD(％) |
| Peak area | 177.233 | 174.806 | 179.483 | 175.539 | 182.105 | 172.011 | 2.0 |
Example 4
Stability test
According to the method in the embodiment 1, safflower injection with KY180801 batch is taken to prepare a test solution, samples are injected for 0 hour, 2 hours, 4 hours, 8 hours, 16 hours and 24 hours (20 mu L), the peak area of quercetin of the test solution is recorded, the RSD is calculated, the HPLC conditions are the same as the embodiment 1, and the results are shown in the table 4, which indicates that the method has good stability.
TABLE 4 stability test results
| |
1 | 2 | 3 | 4 | 5 | 6 | RSD(％) |
| Content (wt.) | 177.153 | 177.055 | 176.249 | 173.503 | 170.862 | 165.973 | 2.5 |
Example 5
Recovery test
Precisely sucking 20mL 6 parts of lot number KY180801 flos Carthami injection (free quercetin content of 0.39 μ g/mL), respectively adding 1mL reference substance solution (7.8 μ g/mL), and preparing into test sample according to test sample preparation method. The recovery rate was calculated by performing the test in the same manner as in example 1, and it can be seen from table 5 that the average recovery rate was 99.4% and RSD was 2.5%, and the method of the present invention was excellent in accuracy.
TABLE 5 recovery test results
Example 6
Durability test
The chromatographic column adopts Phenomenex Gemini-C18(4.6mm × 250mm, 5.0 μm), Waters symmerrysheldTMRP18(4.6 mm. times.250 mm, 5.0 μm) and AcclaimTM120C18(4.6mm × 250mm, 5.0 μm) three kinds of chromatographic columns, the content of the safflower injection sample with batch KY180801 is measured, the detection method is the same as that in example 1, the result is shown in Table 6, and the content obtained by the detection of the three kinds of chromatographic columns is stable, and the RSD is 1.93%.
TABLE 6 investigation result of free quercetin component HPLC detection chromatographic column durability
Example 7
Influence of acetonitrile adsorption frequency
Taking 10 finished safflower injection products with the batch of KY180801, uniformly mixing, respectively taking 4 parts, each part being 20mL, and measuring the content of free quercetin in the products.
A, B, C acetonitrile are used for adsorption times different from each other in the preparation process of the test article, D times are the same, but the amounts of the acetonitrile used for the second time and the third time are different from each other and are the same as those in example 1.
Test article A: adsorbing for 1 time by 40 mL;
and (3) test article B: adsorbing for 2 times by 40 mL;
and a test article C: adsorbing for 3 times by 40 mL;
and (3) a test article D: 40mL, 30mL, and 20mL were each adsorbed once.
The results of measuring the content of free quercetin in the four test samples are shown in table 7, and it can be seen from table 7 that the content of quercetin in the test samples C and D is higher than that in the former test sample, which indicates that the free quercetin can be completely adsorbed by the safflower injection after salting out and adsorption for more than three times. In order to ensure complete adsorption, the salting-out adsorption frequency of the content detection sample is set to be 3 times.
TABLE 7 examination of the adsorption frequency of safflower injection in acetonitrile salting-out
Example 8
Influence of the Mobile phase
And (4) carrying out HPLC detection by using different flow relative standard substances, and inspecting the influence of different flow relative detection results. Other conditions of HPLC were the same as in example 1.
The mobile phases are respectively:
(1) 45% methanol and 55% 0.4 wt% phosphoric acid;
(2) 48% methanol and 52% 0.4 wt% phosphoric acid;
(3) 50% methanol and 50% 0.4 wt% phosphoric acid.
The results are shown in fig. 2, in which the curves are the results of detection of the mobile phases (2), (1) and (3) from left to right, and it can be seen that when methanol is present at a ratio of 45 to 50%, the tailing is small and the retention time RSD is within 2%. The effect is best when the methanol proportion is 48%, and the tailing factor is minimum.
Comparative example 1 Effect of different adsorbents
The procedure of example 1 was repeated except that acetone and ethyl acetate were used as the adsorbents in place of acetonitrile to adsorb free quercetin. The safflower injection used in the comparative example has a lot number of KY 180801.
The high performance liquid chromatography of acetonitrile-enriched free quercetin and quercetin control product is shown in FIG. 3. The high performance liquid chromatography of acetone-enriched free quercetin and quercetin control is shown in FIG. 4. The high performance liquid chromatography of the ethyl acetate-enriched free quercetin and the quercetin control product is shown in FIG. 5.
It is known in the art that the solubility of quercetin in acetone is 16.91 mg/mL at maximum－1As can be seen from fig. 1, 4 and 5, the peak area difference in the hplc chromatogram of acetonitrile, acetone and ethyl acetate adsorbed free quercetin is within 5%, but the number of peaks is the least when acetonitrile is used, which indicates that acetonitrile can not only adsorb free quercetin in the safflower injection well, but also eliminate the influence of other components in the safflower injection well.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Furthermore, those skilled in the art will appreciate that while some embodiments herein include some features included in other embodiments, rather than other features, combinations of features of different embodiments are meant to be within the scope of the invention and form different embodiments. For example, in the claims above, any of the claimed embodiments may be used in any combination. The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.
Claims (10)
1. A method for enriching free quercetin in a safflower injection comprises the following steps:
step a, adding salt into the safflower injection, and uniformly mixing to obtain a mixed solution A, wherein the salt is in a supersaturated state in the mixed solution A;
and b, adsorbing the free quercetin in the mixed solution A by using acetonitrile.
2. The enrichment method according to claim 1, wherein the salt is selected from any one of sodium chloride, sodium sulfate, ammonium sulfate;
preferably, at least 4.5g of sodium chloride is added to 20mL of the safflower injection.
3. The enrichment method according to claim 1, wherein the mixed solution A is allowed to stand for 30-60 min before being adsorbed by acetonitrile.
4. The enrichment method according to claim 1, wherein the volume ratio of the acetonitrile to the safflower injection is (0.5-2): 1.
5. the enrichment method according to claim 1, wherein acetonitrile is used in step b to adsorb free quercetin in the mixed solution A for a plurality of times;
preferably, the plurality of times is at least 3 times.
6. The enrichment method according to claim 1, wherein adsorbing free quercetin in the mixed solution a using acetonitrile comprises: and adding acetonitrile into the mixed solution A, uniformly mixing, centrifuging, and centrifuging to obtain an upper layer solution, namely the acetonitrile solution adsorbed with the free quercetin.
7. A method for measuring the content of free quercetin in a safflower injection comprises the following steps:
step 1, enriching free quercetin in a safflower injection to be tested by adopting the enrichment method of any one of claims 1 to 6;
and 2, quantitatively detecting the enriched free quercetin.
8. The assay of claim 7, wherein the quantitative assay is an HPLC assay.
9. The content measurement method according to claim 8, wherein step 1 comprises: removing acetonitrile in the acetonitrile solution adsorbed with the free quercetin, dissolving the acetonitrile in methanol, and then carrying out HPLC detection;
preferably, the methanol solution is filtered by using a microfiltration membrane and then subjected to HPLC detection, wherein the pore diameter of the microfiltration membrane is 0.22 μm or 0.45 μm;
preferably, the pore size of the microfiltration membrane is 0.22 μm.
10. The content measurement method according to claim 8 or 9, wherein the detection conditions of HPLC include one or more of the following a to f:
a. the chromatographic column is an octadecylsilane chemically bonded silica reverse chromatographic column;
b. the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a phosphoric acid aqueous solution, and the volume ratio of the mobile phase A to the mobile phase B is (45-50) to (55-50); preferably, the concentration of the phosphoric acid aqueous solution is 0.4 wt%;
c. the column temperature is 30 ℃;
d. the column flow rate is 1 mL/min;
e. the sample injection amount is 20-40 mu L;
f. the detector is an ultraviolet detector; preferably, the detection wavelength is 370 nm.
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| CN114236028A (en) * | 2021-12-23 | 2022-03-25 | 华润三九（雅安）药业有限公司 | Preparation method of test sample and determination method of content of free flavone in ginkgo leaf preparation |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114236028A (en) * | 2021-12-23 | 2022-03-25 | 华润三九（雅安）药业有限公司 | Preparation method of test sample and determination method of content of free flavone in ginkgo leaf preparation |
| CN114236028B (en) * | 2021-12-23 | 2023-09-08 | 华润三九（雅安）药业有限公司 | Preparation method of sample and determination method of free flavone content in ginkgo leaf preparation |
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