CN112903887A - Method for establishing folium ginkgo dripping pill HPLC-VWD-ELSD characteristic map and characteristic map thereof - Google Patents
Method for establishing folium ginkgo dripping pill HPLC-VWD-ELSD characteristic map and characteristic map thereof Download PDFInfo
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
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- G01N2030/884—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample organic compounds
Abstract
The invention relates to a method for analyzing a characteristic fingerprint spectrum of a traditional Chinese medicine, in particular to a method for establishing a folium ginkgo dropping pill HPLC-VWD-ELSD characteristic spectrum and an obtained characteristic spectrum. The invention discloses a method for establishing a folium ginkgo dropping pill HPLC-VWD-ELSD characteristic map, which comprises the steps of preparing a test solution, and establishing and determining characteristic map determination conditions, wherein 11 characteristic common peaks are identified in the characteristic map through a reference substance. The method uses the ultrafiltration tube for sample pretreatment, can basically remove the polyethylene glycol 4000(PEG4000) as an auxiliary material, and reduces the influence of the polyethylene glycol 4000 on the response of the lactone components in an ELSD chromatogram.
Description
Technical Field
The invention relates to a method for analyzing a characteristic fingerprint spectrum of a traditional Chinese medicine, in particular to a method for establishing a folium ginkgo dropping pill HPLC-VWD-ELSD characteristic spectrum and an obtained characteristic spectrum.
Background
Folium Ginkgo dripping pill is prepared by heating and melting folium Ginkgo extract and polyethylene glycol 4000, mixing, dripping into methyl silicone oil coolant, removing surface oil stain, or coating with film coat, and is mainly used for treating thoracic obstruction, cardialgia, apoplexy, hemiplegia, and stiff tongue caused by blood stasis obstruction; stable angina pectoris due to coronary heart disease and cerebral infarction.
The traditional Chinese medicine characteristic fingerprint spectrum refers to a chromatogram or a spectrogram which can reflect the chemical characteristics of a traditional Chinese medicine or a preparation thereof and is obtained by a certain analysis method after the traditional Chinese medicine or the preparation thereof is properly processed. The chromatogram fingerprint can well reflect the comprehensiveness and complexity of the traditional Chinese medicine, and is an effective method for evaluating the authenticity and consistency of the traditional Chinese medicine.
The flavone and lactone components are the main chemical components in the ginkgo leaf dripping pill and also are index components for content determination in Chinese pharmacopoeia, and the quality control method of the ginkgo leaf dripping pill mainly comprises the steps of independently determining the flavone and lactone components or establishing a fingerprint of the single component at present. Because the flavonoid components have good ultraviolet absorption, but the lactones have no ultraviolet absorption, the flavonoid and lactone components can be simultaneously detected by using the ultraviolet detector and the evaporative light scattering detector together, the HPLC-VWD-ELSD characteristic spectrum of the ginkgo leaf dropping pill is established, and the chemical components of the ginkgo leaf dropping pill can be comprehensively characterized from the integral angle.
Disclosure of Invention
In order to achieve the purpose, the invention provides a method for establishing a folium ginkgo dropping pill HPLC-VWD-ELSD characteristic map, which comprises the following steps:
(1) preparation of a test solution:
pretreatment of an ultrafiltration membrane: 400 μ L of 50% methanol was placed in Millipore ultrafiltration tube (1.5 mL in outer tube, 3K) and centrifuged at 15000r for 20min, the filtrate was discarded, the inner tube was placed in the outer tube upside down and centrifuged at 1000r for 1min to remove the liquid in the inner tube.
Taking a proper amount of folium ginkgo dripping pills, grinding, taking 0.25g of powder, precisely weighing, placing in a conical flask with a plug, precisely adding 20mL of 50% methanol, carrying out ultrasonic treatment for 60min (40KHz,500W), taking out, cooling to room temperature, shaking up, centrifuging for 20min at 15000r, taking 400 mu L of supernatant liquid, placing in a pretreated ultrafiltration tube, centrifuging for 20min at 15000r, and taking a solution in a filtrate collecting tube to obtain the ginkgo biloba extract dripping pills.
(2) Preparation of control solutions: taking reference substances of bilobalide, bilobalide J, bilobalide C, bilobalide A, bilobalide B, rutin, quercetin-3-O-glucosyl (1 → 2) rhamnoside, kaempferol-3-O-rutinoside, narcissin, quercetin-3-O-2 '- (6' -p-coumaroyl) -glucosyl-rhamnoside and kaempferol-3-O-2 '- (6' -p-coumaroyl) -glucosyl-rhamnoside, adding 50% methanol to prepare a mother solution with a proper concentration, and properly diluting to prepare a mixed reference substance solution with a final concentration of 10 mu g/mL-100 mu g/mL.
(3) And (3) measuring by using an ultraviolet-evaporative light scattering detector combination method: a chromatographic column: agilent InfinityLab Poroshell 120EC-C18 (3.0X 150mm,2.7 μm); mobile phase: gradient elution was performed with 0.05% aqueous formic acid as mobile phase a and acetonitrile-methanol (8: 2) as mobile phase B according to the following table: 0.4mL/min, column temperature: 35 ℃, detection wavelength: 360 nm; evaporative light scattering detector: n is a radical of2Flow rate: 1.2SLM, evaporating tube temperature: 80 ℃; temperature of the atomizer: 80 ℃.
(4) Establishing a characteristic spectrum: preparing 20 batches of folium ginkgo dropping pills into a test solution according to the step (1), injecting the test solution into a high performance liquid chromatograph, detecting according to the chromatographic conditions of the step (3), respectively recording chromatograms, and analyzing by adopting software of 'traditional Chinese medicine chromatographic fingerprint similarity evaluation system 2012 edition' to obtain a VWD-ELSD standard fingerprint (R) of the folium ginkgo dropping pills, wherein 11 common peaks are determined, the lactones take the peak 1 (bilobalide) as a reference peak, and the flavonoid glycosides take the peak 4 (rutin) as a reference peak.
In addition, the invention also provides a folium ginkgo dropping pill HPLC-VWD-ELSD characteristic map, as shown in figure 8.
The invention discloses a method for establishing a folium ginkgo dropping pill HPLC-VWD-ELSD characteristic map, which comprises the steps of preparing a test solution, and establishing and determining characteristic map determination conditions, wherein 11 characteristic common peaks are identified in the characteristic map through a reference substance.
Compared with the prior art, the invention has the improvement points that:
1. the ultrafiltration tube is used for sample pretreatment, so that the auxiliary material polyethylene glycol 4000(PEG4000) can be basically removed, and the influence of the auxiliary material polyethylene glycol 4000 on the response of the lactone components in an ELSD chromatogram map is reduced.
2. The combination of an ultraviolet detector and an evaporative light scattering detector can simultaneously detect compounds with ultraviolet absorption (flavonoid components) and compounds without ultraviolet absorption (lactone components), and comprehensively analyze chemical components in the ginkgo leaf dropping pills.
3. The method has the advantages of good precision, stability and repeatability, the obtained ginkgo leaf dripping pills have high similarity of characteristic maps, are accurate and reliable, can comprehensively reflect the overall characteristics of the chemical compositions of the ginkgo leaf dripping pills, and can more comprehensively evaluate the quality of the ginkgo leaf dripping pills.
Drawings
FIG. 1 is a comparison chart of different parameter selections of a test solution preparation method;
wherein the extraction conditions are different in FIG. 1A, the extraction solvents are different in FIG. 1B, the extraction time is different in FIG. 1C, and the material-liquid ratio is different in FIG. 1D.
FIG. 2 chromatogram before and after ultrafiltration membrane filtration (before and after removal of PEG4000)
FIG. 3 is a comparison of an embodiment of the invention using three different chromatography columns.
FIG. 4 is a comparison of different mobile phase systems used in the examples of the invention.
FIG. 5 is a comparative graph of different aqueous phases in a mobile phase system for an example of the invention.
FIG. 6 is a comparative graph of an embodiment of the present invention using different flow rate systems.
FIG. 7 is a comparison of different column temperature systems used in examples of the invention.
FIG. 8 is a standard characteristic spectrum of folium Ginkgo dripping pill (1-11 is 11 characteristic common peaks)
FIG. 9 spectrum of ginkgo leaf drop pills
FIG. 10 is an overlay of 20 batches of characteristic spectra of folium Ginkgo dripping pills
Wherein, peak 1: bilobalide; peak 2: bilobalide J; peak 3: bilobalide C; peak 4: rutin; peak 5: quercetin-3-O-glucosyl (1 → 2) rhamnoside; peak 6: kaempferol-3-O-rutinoside; peak 7: narcissus glycosides; peak 8: bilobalide A; peak 9: bilobalide B; peak 10: quercetin-3-O-2 "- (6" -p-coumaroyl) -glucosyl-rhamnoside; peak 11: Kaempferol-3-O-2 '- (6' -p-coumaroyl) -glucosyl-rhamnoside
Detailed Description
The present invention will be further described with reference to the following examples in order to understand the present invention in more detail.
Example 1
1 Instrument and reagent
1.1 instruments
One-tenth-of-ten-thousandth analytical balance (Sartorious), one-ten-thousandth analytical balance (Sartorious), Agilent 1260InfinityVWD-ELSD combination, Milli-Q deionized water, high speed refrigerated centrifuge.
1.2 reagent
The ginkgo leaf dripping pill is provided by the pharmaceutical group of Wanbangde, and the sample batch number is shown in Table 1. The methanol, acetonitrile and formic acid are chromatographically pure, the water is ultrapure water, and the other reagents are analytically pure.
TABLE 1 Ginkgo biloba leaf drop pill information sheet
2 methods and results
2.1 examination of preparation method of test solution
Examining the preparation method of the test solution, including the extraction conditions (ultrasound, reflux), the extraction solvents (water, 25% methanol and 50% methanol), the extraction time (20min, 40min and 60min) and the extraction material-liquid ratio (1-40, 1-80 and 1-120), the result is shown in fig. 1, and the final extraction method is determined: 0.25g of sample was weighed and extracted with 20mL of 50% methanol for 60min by sonication (50% methanol solution was observed as the extraction solvent since the ultrafiltration membrane material only passed through < 60% methanol solution).
Preparation of a test solution:
pretreatment of an ultrafiltration membrane: 400 μ L of 50% methanol was placed in Millipore ultrafiltration tube (1.5 mL in outer tube, 3K) and centrifuged at 15000r for 20min, the filtrate was discarded, the inner tube was placed in the outer tube upside down and centrifuged at 1000r for 1min to remove the residual liquid in the inner tube.
Taking a proper amount of folium ginkgo dripping pills, grinding, taking 0.25g of powder, precisely weighing, placing in a conical flask with a plug, precisely adding 20mL of 50% methanol, carrying out ultrasonic treatment for 60min (40KHz,500W), taking out, cooling to room temperature, shaking up, centrifuging for 20min at 15000r, taking 400 mu L of supernatant liquid, placing in a pretreated ultrafiltration tube, centrifuging for 20min at 15000r, and taking a solution in a filtrate collecting tube to obtain the ginkgo biloba extract dripping pills.
Chromatograms before and after ultrafiltration membrane filtration (before and after removal of PEG4000) are shown in fig. 2.
2.2 optimization and selection of chromatographic methods
The mobile phase considers a water-acetonitrile and acid water-acetonitrile-methanol system, the chromatographic columns observe three chromatographic columns of Agilent ZORBAX SB-C18 (2.1X 100mm,1.8 mu m), Agilent Infinity Lab Poroshell 120EC-C18 (2.1X 100mm,2.7 mu m) and Agilent Infinity Lab Poroshell 120EC-C18 (3.0X 150mm,2.7 mu m), Agilent Infinity Lab Poroshell 120EC-C18 (3.0X 150mm,2.7 mu m) are selected as analytical columns, and parameter parts of ELSD including evaporation temperature, atomization temperature and N are considered2Flow rate, etc., to determine the optimum ELSD parameters.
A chromatographic column: agilent InfinityLab Poroshell 120EC-C18 (3.0X 150mm,2.7 μm), mobile phase: gradient elution was performed with 0.05% aqueous formic acid as mobile phase a and acetonitrile-methanol (8: 2) as mobile phase B according to the following table: 0.4mL/min, column temperature: 35 ℃, detection wavelength: 360 nm; evaporative light scattering detector: n is a radical of2Flow rate: 1.2SLM, evaporating tube temperature: 80 ℃, atomizer temperature: 80 ℃.
TABLE 2 elution gradiometer
2.2.1 investigation of different columns
Three different columns (column 1: Agilent ZORBAX SB-C18 (2.1X 100mm,1.8 μm), column 2: Agilent Infinity Lab Poroshell 120EC-C18 (2.1X 100mm,2.7 μm) and column 3: Agilent Infinity Lab Poroshell 120EC-C18 (3.0X 150mm,2.7 μm)) were examined for separation effect and eluted in a gradient according to Table 2. As is clear from FIG. 3, the chromatographic chart obtained in column 3 had a good peak shape and resolution, and therefore, Agilent InfinityLab Poroshell 120EC-C18 (3.0X 150mm,2.7 μm) was selected as the analytical column.
2.2.2 investigation of different mobile phase systems
2.2.2.1 investigation of the mobile phase System-organic phase-0.05% formic acid Water
The influence of the change of the organic phase in the mobile phase system on the chromatographic peak was examined, and the gradient elution was performed as shown in Table 2. Considering VWD and ELSD chromatogram (figure 4) comprehensively, when pure acetonitrile is used as an organic phase, the separation effect of flavonoid glycosides in the first 10min is poor; after methanol is mixed, retention is enhanced, and the separation degree is increased; the ratio of acetonitrile to methanol was further investigated when acetonitrile: methanol-8: 2, the separation degree and the peak shape of each identification peak are better, and the elution time is proper, so that the ratio of acetonitrile: methanol-8: 2 as organic phase.
2.2.2.2 investigation of the mobile phase system-acetonitrile: methanol-8: 2-aqueous phase
The influence of the change of the water phase in the mobile phase system on the chromatographic peak is examined, and the gradient elution is carried out according to the table 2. The peak shape was improved by adding formic acid to the aqueous phase. As can be seen from FIG. 5, the separation degree of peak 7 (narcissus) can be improved by adding 0.05% formic acid into the aqueous phase; when the concentration of formic acid is increased to 0.1%, the difference is not too large with 0.05% formic acid, so under the condition of ensuring the acidity bearing capacity of a chromatograph and a chromatographic column, acetonitrile is selected to be used: methanol-8: 2-0.05% formic acid aqueous solution as mobile phase.
2.2.3 investigating different flow Rate regimes
An Agilent 1260 high performance liquid chromatograph is adopted to examine the influence of different flow rates (0.3mL/min, 0.4mL/min and 0.5mL/min) on the separation degree of each chromatographic peak, and gradient elution is carried out according to the table 2. The experimental results show (as shown in fig. 6), in the VWD chromatogram, peaks 4 and 7 both include small peaks, and in the ELSD chromatogram, a flow rate of 0.4mL/min achieves a better resolution (peaks 2 and 3), and a flow rate of 0.4mL/min is selected under comprehensive consideration.
2.2.4 investigation of different column temperature systems
The influence of different column temperatures (30 ℃, 35 ℃ and 40 ℃) on the separation degree of each chromatographic peak is examined by adopting an Agilent 1260 high performance liquid chromatograph, and gradient elution is carried out according to the table 2. As is clear from fig. 7, when the column temperature was 30 ℃ and 40 ℃, the separation effect of peak 4 and peak 11 was not good, and the separation of the peaks at 35 ℃ was good, and 35 ℃ was selected as the detection temperature in consideration of the temperature resistance of the column.
2.2.5 investigating ELSD parameters
The evaporation temperature (60 ℃, 70 ℃ and 80 ℃) and the atomization temperature (60 ℃, 70 ℃ and 80 ℃) of ELSD and N were examined2Flow rates (1.2SLM, 1.4SLM and 1.6SLM) are shown in the table below, where the evaporation temperature is 80 deg.C, the atomization temperature is 80 deg.C, and N is2When the flow rate is 1.2SLM, the peak area of each lactone component is high.
3 methodology examination
3.1 precision test
Taking 0.25g of folium ginkgo dripping pills (S12), precisely weighing, preparing a sample solution according to the sample solution preparation method, precisely sucking 2 mu L of subsequent filtrate, injecting into a liquid chromatograph, continuously injecting for 6 times, recording the retention time and the peak area of 11 common peaks, and calculating the relative retention time, the relative peak area and the RSD value thereof, wherein the results are shown in tables 3 and 4. Experimental results show that the method is good in precision.
TABLE 3 results of precision examination (relative retention time)
TABLE 4 results of precision examination (relative peak area)
3.2 repeatability test
Taking 6 parts of folium Ginkgo dripping pills (S12) of the same batch, each part being 0.25g, precisely weighing, preparing a sample solution according to the above "sample solution preparation method", precisely sucking 2 μ L of subsequent filtrate, injecting into a liquid chromatograph, recording the retention time and peak area of 11 common peaks, calculating the relative retention time, relative peak area and RSD value thereof, and finding the results in tables 5 and 6. Experimental results show that the method has good repeatability.
TABLE 5 repeatability test results (relative retention time)
TABLE 6 repeatability test results (relative peak area)
3.3 stability test
Taking a next sample solution of the repeatability item, precisely absorbing 2 mu L of the sample solution at 0h, 4h, 6h, 10h, 12h and 24h respectively, injecting the sample solution into a liquid chromatograph, recording the retention time and the peak area of 11 common peaks, and calculating the relative retention time, the relative peak area and the RSD value thereof, wherein the results are shown in tables 7 and 8. Experimental results show that the method is good in stability.
TABLE 7 stability test results (relative Retention time)
TABLE 8 stability test results (relative peak area)
4. Sample assay
Preparing a sample solution from 20 batches of folium ginkgo dropping pills according to the sample solution preparation method, carrying out sample injection analysis according to a chromatographic method, respectively recording chromatograms, analyzing by adopting software of a traditional Chinese medicine chromatogram fingerprint similarity evaluation system 2012 version, generating a reference spectrum (figure 10) by a median method, obtaining an HPLC-VWD-ELSD standard characteristic spectrum (R) of the folium ginkgo dropping pills (figure 8), determining 11 common peaks, analyzing the similarity between 20 batches of folium ginkgo dropping pills and the reference characteristic spectrum, wherein the lactones use peak 1 as a reference peak, the flavonoid glycosides use peak 4 as a reference peak, the relative retention time of each peak is shown in a table 9, analyzing the similarity between 20 batches of folium ginkgo dropping pills and the reference characteristic spectrum, and the similarity data are shown in a table 10 and a table 11, wherein the result shows that the folium ginkgo dropping pills in different batches have higher similarity (0.9) with the reference spectrum.
TABLE 920 relative retention time of 11 common fingerprint peaks in Ginkgo biloba leaf drop pills
TABLE 1020 batches of folium Ginkgo dripping pills similarity results (VWD)
TABLE 1120 batch Ginkgo leaf drop pill similarity results (ELSD)
Claims (4)
1. A method for establishing a folium ginkgo dropping pill HPLC-VWD-ELSD characteristic map comprises the following steps:
(1) preparation of a test solution:
pretreatment of an ultrafiltration membrane: taking 400 mu L of 50% methanol in a Millipore ultrafiltration tube, centrifuging for 20min at 15000r, discarding filtrate, placing the inner tube in the outer tube in a reverse manner, centrifuging for 1min at 1000r, and removing liquid in the inner tube;
taking a ginkgo leaf dropping pill, grinding, taking 0.25g of powder, precisely weighing, placing in a conical flask with a plug, precisely adding 20mL of 50% methanol, carrying out ultrasonic treatment for 60min, taking out, cooling to room temperature, shaking up, carrying out 15000r, centrifuging for 20min, taking 400 mu L of supernatant liquid, placing in a pretreated ultrafiltration tube, carrying out 15000r and centrifuging for 20min, and taking a solution in a filtrate collecting tube to obtain the ginkgo leaf dropping pill;
(2) preparation of control solutions:
taking reference substances of bilobalide, bilobalide J, bilobalide C, bilobalide A, bilobalide B, rutin, quercetin-3-O-glucosyl (1 → 2) rhamnoside, kaempferol-3-O-rutinoside, narcissin, quercetin-3-O-2 '- (6' -p-coumaroyl) -glucosyl-rhamnoside and kaempferol-3-O-2 '- (6' -p-coumaroyl) -glucosyl-rhamnoside, adding 50% methanol to prepare a mother solution with a proper concentration, and properly diluting to prepare a mixed reference substance solution with a final concentration of 10 mu g/mL-100 mu g/mL;
(3) the high performance liquid chromatography-evaporative light scattering detector combined method comprises the following steps:
a chromatographic column: agilent InfinityLab Poroshell 120 EC-C18; mobile phase: gradient elution was performed with 0.05% aqueous formic acid as mobile phase a and acetonitrile-methanol as mobile phase B according to the following table: 0.4mL/min, column temperature: 35 ℃, detection wavelength: 360 nm; evaporative light scattering detector: n is a radical of2Flow rate: 1.2SLM, evaporating tube temperature: 80 ℃; temperature of the atomizer: 80 ℃;
(4) establishing a characteristic spectrum:
preparing 20 batches of folium ginkgo dropping pills into a test solution according to the step (1), injecting the test solution into a high performance liquid chromatograph, detecting according to the chromatographic conditions of the step (3), respectively recording chromatograms, and analyzing by adopting software of a traditional Chinese medicine chromatographic fingerprint similarity evaluation system 2012 version to obtain a VWD-ELSD standard fingerprint (R) of the folium ginkgo dropping pills, wherein 11 common peaks are determined, the lactones take the peak 1 as a reference peak, and the flavonoid glycosides take the peak 4 as a reference peak.
2. The method of establishing according to claim 1, wherein: the volume ratio of the mobile phase B, acetonitrile-methanol is 8: 2.
3. Folium Ginkgo dripping pill HPLC-VWD-ELSD characteristic map is shown in FIG. 8.
4. The folium ginkgo dripping pill HPLC-VWD-ELSD characteristic map obtained by the establishing method of claim 1.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113358786A (en) * | 2021-06-07 | 2021-09-07 | 中国药科大学 | Method for measuring flavonol glycoside component content in ginkgo leaf dripping pills |
| CN115112783A (en) * | 2022-03-01 | 2022-09-27 | 江苏康缘药业股份有限公司 | Ginkgo leaf detection method and construction and application of fingerprint spectrum with simultaneous reflection of multiple components |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1853674A (en) * | 2005-02-07 | 2006-11-01 | 贵阳云岩西创药物科技开发有限公司 | Quality controlling method of Xingdan injection |
| CN104569170A (en) * | 2013-10-29 | 2015-04-29 | 贵州益佰制药股份有限公司 | Method for detecting ginkgo leaf extract and diphyridamole injection preparation |
| CN105891356A (en) * | 2016-03-31 | 2016-08-24 | 河北神威药业有限公司 | Detecting method for measuring content of flavonoid compounds and terpene lactone compounds in ginkgo bioba leaf extract at same time |
| CN105891355A (en) * | 2016-03-31 | 2016-08-24 | 河北神威药业有限公司 | Detecting method for measuring content of flavonoid compounds and terpene lactone compounds in ginkgo leaf extract or preparations of ginkgo leaf extract at same time |
| CN107884483A (en) * | 2017-09-27 | 2018-04-06 | 黑龙江珍宝岛药业股份有限公司 | The content assaying method and purposes of flavones ingredient in a kind of ginkgo leaf and its preparation |
| CN111983077A (en) * | 2020-08-18 | 2020-11-24 | 贵州益佰制药股份有限公司 | Detection method of ginkgo dipyridamole injection preparation |
-
2020
- 2020-12-28 CN CN202011597316.5A patent/CN112903887B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1853674A (en) * | 2005-02-07 | 2006-11-01 | 贵阳云岩西创药物科技开发有限公司 | Quality controlling method of Xingdan injection |
| CN104569170A (en) * | 2013-10-29 | 2015-04-29 | 贵州益佰制药股份有限公司 | Method for detecting ginkgo leaf extract and diphyridamole injection preparation |
| CN105891356A (en) * | 2016-03-31 | 2016-08-24 | 河北神威药业有限公司 | Detecting method for measuring content of flavonoid compounds and terpene lactone compounds in ginkgo bioba leaf extract at same time |
| CN105891355A (en) * | 2016-03-31 | 2016-08-24 | 河北神威药业有限公司 | Detecting method for measuring content of flavonoid compounds and terpene lactone compounds in ginkgo leaf extract or preparations of ginkgo leaf extract at same time |
| CN107884483A (en) * | 2017-09-27 | 2018-04-06 | 黑龙江珍宝岛药业股份有限公司 | The content assaying method and purposes of flavones ingredient in a kind of ginkgo leaf and its preparation |
| CN111983077A (en) * | 2020-08-18 | 2020-11-24 | 贵州益佰制药股份有限公司 | Detection method of ginkgo dipyridamole injection preparation |
Non-Patent Citations (1)
| Title |
|---|
| 陈凡: "银杏叶提取物的HPLC-DAD-ELSD指纹图谱研究", 《药物分析杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113358786A (en) * | 2021-06-07 | 2021-09-07 | 中国药科大学 | Method for measuring flavonol glycoside component content in ginkgo leaf dripping pills |
| CN115112783A (en) * | 2022-03-01 | 2022-09-27 | 江苏康缘药业股份有限公司 | Ginkgo leaf detection method and construction and application of fingerprint spectrum with simultaneous reflection of multiple components |
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