CN108362809B - Quality evaluation method of ginkgo leaf and extract and preparation thereof - Google Patents
Quality evaluation method of ginkgo leaf and extract and preparation thereof Download PDFInfo
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- CN108362809B CN108362809B CN201810103484.0A CN201810103484A CN108362809B CN 108362809 B CN108362809 B CN 108362809B CN 201810103484 A CN201810103484 A CN 201810103484A CN 108362809 B CN108362809 B CN 108362809B
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- glucoside
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- kaempferol
- ginkgo
- rhamnoside
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- G—PHYSICS
- G01—MEASURING; TESTING
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N2030/8809—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
- 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
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
A quality evaluation method of ginkgo leaf and an extract and a preparation thereof belongs to the technical field of quality detection of traditional Chinese medicinal materials. The invention realizes the determination of the contents of various flavonoid chemical components in the ginkgo by constructing the high-performance liquid phase fingerprint of the chemical components of the ginkgo and calculating the correction factor by means of a one-test-multiple-evaluation method. The method has simple operation and good stability, can objectively, comprehensively and accurately evaluate the quality of the ginkgo leaves and the extract or preparation thereof, is used for quality control, can solve the problem that the quality of the medicinal materials and the extract or preparation thereof cannot be objectively and reasonably controlled due to the lack of reference substances, and has important significance for controlling the quality and ensuring the curative effect.
Description
Technical Field
The invention relates to the technical field of quality detection of traditional Chinese medicinal materials, in particular to a quality evaluation method of ginkgo leaves and extracts and preparations thereof based on a high performance liquid fingerprint and a one-measurement-multiple-evaluation method.
Technical Field
The folium Ginkgo is Ginkgo of GinkgoaceaeGinkgo bilobaL, the dried leaves have the effects of promoting blood circulation, removing blood stasis, dredging collaterals, relieving pain, astringing lung, relieving asthma, eliminating turbid pathogen, and reducing blood lipid, the ginkgo has rich chemical components including flavone, lactone, phenolic acid, etc., the ginkgo flavone has the most content and various compositions, and the ginkgo extract with the total flavone content of 24 percent can improve various diseases of cardiovascular and cerebrovascular systems, and has no side effect.
At present, in the quality standard of ginkgo leaf of 'Chinese pharmacopoeia' 2015 edition, the content of total flavonoids is evaluated and controlled by measuring the content of quercetin, kaempferol and isorhamnetin after acid hydrolysis, the method is complex in pretreatment, long in time consumption and incapable of fully reflecting the real and accurate content of flavonoid components.
Therefore, the invention provides a quality evaluation method of ginkgo leaves and extracts and preparations thereof based on a high performance liquid fingerprint and a one-measurement-multiple-evaluation method. The method takes one flavone component in ginkgo as an internal reference substance, and realizes the determination of the contents of various flavonoid chemical components in ginkgo by calculating relative correction factors. The method is simple and convenient to operate, has good stability, can objectively, comprehensively and accurately evaluate the quality of the ginkgo leaves and the extract and the preparation thereof, can solve the problem that the quality cannot be objectively and reasonably evaluated due to the lack of reference substances, and has important significance for controlling the quality and ensuring the curative effect.
Disclosure of Invention
The invention aims to provide a content determination method for determining multiple flavone components in ginkgo aiming at the defects of the prior art, which can not only accurately evaluate the quality of ginkgo leaves and extracts thereof and total flavone in preparations, but also solve the problem that the quality of traditional Chinese medicinal materials and preparations thereof cannot be objectively and reasonably controlled due to the lack of reference substances.
In order to achieve the purpose, the invention adopts the following technical scheme:
the method comprises the steps of measuring the content of a plurality of flavonoid chemical components in the ginkgo by utilizing a high performance liquid chromatography technology and a multi-evaluation method, so as to realize objective, comprehensive and accurate quality evaluation of the ginkgo and the extract thereof and the preparation, wherein the content measurement of the flavonoid chemical components refers to the content measurement of the ginkgo and the extract thereof and a plurality of flavonoid reference substances in the preparation, and the number of the flavonoid chemical components is 8-16.
The method specifically comprises the following steps:
1) preparing a reference substance solution: precisely weighing 1-5mg of each flavone reference substance respectively, placing in a volumetric flask, dissolving with 80-100% methanol to constant volume, filtering to obtain stock solutions of each single reference substance, weighing single standard substance solutions with different volumes respectively, mixing and diluting to obtain mixed samples containing multiple flavone components as mixed reference substance solutions for later use;
2) preparing a test solution: precisely weighing folium Ginkgo, semen Ginkgo extract or semen Ginkgo preparation, adding 50-100% methanol, ultrasonic or heating for extraction, filtering, and filtering the filtrate with 0.45um or 0.22um filter membrane to obtain sample solution;
3) calculating a correction factor RCF by taking the mixed reference solution prepared in the step 1) at a concentration of 2-20 μ L, determining chromatogram by high performance liquid chromatography, calculating peak area, selecting one compound as an internal reference, and calculating a relative correction factor f between the internal reference and the other compoundsk/s。
Wherein, the relative correction factor calculation formula is as follows: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesThe peak area of the internal reference substance is shown; csThe concentration of the internal reference substance is shown; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
4) Measuring active ingredients in the sample by taking the sample solution prepared in step 2) 2-20 μ L, measuring chromatogram with high performance liquid chromatography, and calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkPeak area of the measured component, fk/sTo correct the factor, AsThe peak area of the internal reference substance is shown.
The invention accurately evaluates the content of the total flavone of ginkgo by measuring the total amount of 8-16 flavone glycosides and aglycone thereof with high content and good activity in ginkgo leaves and extracts thereof and preparations.
The flavone control product comprises quercetin-3-O- (2' ', 6' ' -di-O- α -L-rhamnosyl) -L-D-glucoside, kaempferol-3-0- (2' ', 6' ' -di-O-L2-L-rhamnosyl) -L-D-glucoside, kaempferol-3-O-L sophoroside, quercetin-3-O-rutinoside-7-O-glucoside, 3' -methyl-myricetin-3-O-rutinoside, quercetin 3-O-glucoside, quercetin 3-O- (2-L-D-glucosyl-L-L-rhamnoside), kaempferol-3-O-rutinoside, isorhamnetin-3-O- β -D-glucoside, kaempferol 3- (2' ' - β -D-glucoside) -L-L-rhamnoside, kaempferol-3-O-6-rhamnoside- (2' ' -p-581-p-rhamnoside), and coumarin-3-O-3-O-glucoside, and coumarin-3-O-3-acyl-5968-rhamnoside.
The invention constructs a characteristic liquid phase fingerprint by a high performance liquid chromatography technology and ensures that the flavonoid contrast component has good separation degree (the separation degree is more than or equal to 1.2).
The detection conditions of the high performance liquid chromatography are as follows: the detector of the high performance liquid chromatograph uses a diode array detector or a universal detector, uses octa-or octadecylsilane chemically bonded silica as a filler, acetonitrile as a mobile phase A, and 0.1% formic acid aqueous solution as a mobile phase B, and performs gradient elution, wherein in the gradient elution process, the proportion of the mobile phase A, B is changed as follows: 0-20min, 15-20% of phase A and 85-80% of phase B; 20-25min, 20-30% of phase A and 80-70% of phase B; 25-30min, 30-35% of phase A and 70-65% of phase B; 30-45min, 35-40% of phase A and 65-60% of phase B. The ultraviolet detector is used for detecting the wavelength of 210nm-360 nm. The flow rate is 0.5ml/min-1.5ml/min, and the column temperature is kept at 20-45 ℃.
The high performance liquid phase fingerprint spectrum is obtained by applying the conditions, the similarity of all spectra is not lower than 0.90, and the RSD of each peak area is not higher than 2%.
The high performance liquid chromatography column model is preferably Gemini C18 (250 mm × 4.6.6 mm, 5.0 μm), the flow rate is preferably 0.8m L/min, the detection wavelength is preferably 360nm, and the column temperature is preferably 35 ℃.
The invention adopts a one-test-multiple-evaluation method, takes one component as an internal reference substance, and performs relative calibrationAnd (4) calculating the content of each compound respectively by using a positive factor. Wherein, the relative correction factor calculation formula is as follows: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesThe peak area of the internal reference substance is shown; csThe concentration of the internal reference substance is shown; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
The invention has the following advantages:
1) compared with the traditional method, the invention solves the problems of complicated operation, lack of reference substances and the like in the multi-component quantification and multi-index quality control of the traditional Chinese medicine, realizes the multi-index synchronous quality control while reducing the detection components, and accurately reflects the quality of the traditional Chinese medicine product.
2) The invention combines the high performance liquid chromatography technology with a one-test-and-multiple-evaluation method, double standard-reaching products which meet the requirements of both fingerprint similarity and content are determined as qualified products, and the method has important significance for controlling the quality consistency among different batches of products.
3) The method of the invention takes a flavone component in ginkgo as a reference substance, and simultaneously quantifies the content of a plurality of flavone components such as quercetin and glycosides thereof, kaempferol and glycosides thereof, isorhamnetin and glycosides thereof, ginkgo biflavone and the like in ginkgo by a one-test-multiple-evaluation method, thereby accurately evaluating the content of total flavone in ginkgo leaf and extract thereof and preparations.
4) The one-test-multiple-evaluation method provided by the invention is simple and convenient to operate, and the multi-index quality evaluation method is good in stability, can objectively, comprehensively and accurately evaluate the quality of the ginkgo leaves and the extract and preparation thereof, avoids the evaluation method for measuring the content of flavone component hydrolysate in the traditional method, and can effectively prevent the problems of product counterfeiting, process modification and the like.
Drawings
FIG. 1 is a high performance liquid chromatogram of a Ginkgo biloba extract.
FIG. 2 is a high performance liquid chromatogram of a ginkgo leaf mixed standard consisting of 8 flavonoids.
FIG. 3 is a high performance liquid chromatogram of a ginkgo leaf mixed standard consisting of 10 kinds of flavonoids.
FIG. 4 is a high performance liquid chromatogram of a ginkgo leaf mixed standard consisting of 12 flavonoids.
Detailed Description
The present invention is further illustrated by the following examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure, and which are intended to cover all modifications, equivalents, and equivalents that may be resorted to by those skilled in the art to which the invention pertains.
Chromatographic conditions used in the following examples:
1) the instrument comprises the following steps: high performance liquid chromatography (hua spectrum S6000, dahlianhua spectrum science and technology development limited);
2) analyzing a chromatographic column: gemini C18(250 mm × 4.6.6 mm, 5.0 μm, Philomen, USA);
3) mobile phase: acetonitrile is taken as a mobile phase A, 0.1 percent formic acid aqueous solution is taken as a mobile phase B,
4) gradient conditions: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A
5) Detector conditions: an ultraviolet detector with a detection wavelength of 360nm
6) The flow rate is 0.8 ml/min;
7) the column temperature was 35 ℃.
Example 1
The following detailed description is made with reference to fig. 2 and the embodiments:
1) reference substance
quercetin-3-O- (2'', 6'' -di-O- α -L-rhamnosyl) -L1-D-glucoside (1), kaempferol-3-0- (2'', 6'' -di-O-L0-L-rhamnosyl) -L3-D-glucoside (2), quercetin-3-O-rutinose-7-O-glucoside (3), quercetin 3-O-glucoside (4), isorhamnetin-3-O- β -D-rutinoside (5), kaempferol 3- (2'' - β -D-glucoside) -L2-L-rhamnoside (6), quercetin 3-O- α - (6'' '-p-coumaroyl-glucoside- β -1, 4-rhamnoside) (7), kaempferol 3-O- α - (6' '' -p-coumaroyl-glucoside- β -1, 2-rhamnoside) (8).
2) Preparation of sample solution
Precisely weighing semen Ginkgo extract powder 5.0mg in volumetric flask, adding methanol for dissolving, ultrasonic treating for 5min, diluting to constant volume of 20m L, shaking, filtering with 0.22 μm microporous membrane to obtain test solution, and analyzing and detecting by high performance liquid chromatography.
3) Preparation of control solutions
Precisely weighing 1.0mg of each of the 8 ginkgetin monomer compounds in the step 1), respectively adding methanol into volumetric flasks to dissolve and fix the volume to 2ml, shaking up to prepare stock solutions of all reference substances, wherein the concentration is 0.5mg/m L, precisely measuring the stock solutions of all reference substances to be 2.0m L, placing the stock solutions into the same volumetric flask with the volume of 25m L, adding methanol to dissolve and dilute to a scale, shaking up to obtain a reference substance mixed solution, and feeding the reference substance mixed solution into a high-performance liquid chromatograph.
4) Calculation of correction factors
Taking 10 mu L of the mixed reference substance solution prepared in the step 3), measuring chromatogram by using high performance liquid chromatography, calculating peak area, selecting No. 4 reference substance as an internal reference substance, and respectively calculating relative correction factor f between the internal reference substance and other compoundsk/s. The relative correction factor calculation formula is: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesPeak area of the internal standard substance; csIs the internal standard concentration; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
4) Determination of other flavone Components
Taking 10 mu L of the test solution prepared in the step 2), measuring a chromatogram by using a high performance liquid chromatography, and respectively calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkPeak area of the measured component, fk/sIs a correction factor，AsThe peak area of the internal reference substance is shown.
The gradient condition is as follows: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A;
example 2
The following detailed description is made with reference to fig. 3 and the embodiments:
1) reference substance
quercetin-3-O- (2' ', 6' ' -di-O- α -L-rhamnosyl) -L1-D-glucoside (1), kaempferol-3-0- (2' ', 6' ' -di-O-L0-L-rhamnosyl) -L3-D-glucoside (2), quercetin-3-O-rutinoside-7-O-glucoside (3), 3' -methyl-myricetin-3-O-rutinoside (4), quercetin 3-O-glucoside (5), kaempferol-3-O-rutinoside (6), isorhamnetin-3-O- β -D-rutinoside (7), kaempferol 3- (2' ' - β -D-glucoside) -L2-L-rhamnoside (8), quercetin 3-O- α - (6' ' ' -p-coumaroyl-glucoside- β -1, 4-rhamnoside) (9), kaempferol 3-O- α - (6' ' ' -p-coumaroyl-glucoside- β -1, 4-rhamnoside) (9 ' ' -rhamnosyl- α -p-O-3-O-rhamnoside) (β).
2) Preparation of sample solution
Precisely weighing semen Ginkgo extract powder 5.0mg in volumetric flask, adding methanol for dissolving, ultrasonic treating for 5min, diluting to constant volume of 20m L, shaking, filtering with 0.22 μm microporous membrane to obtain test solution, and analyzing and detecting by high performance liquid chromatography.
3) Preparation of control solutions
Precisely weighing 1.0mg of each of the 10 ginkgetin monomer compounds in the step 1), respectively adding methanol into volumetric flasks to dissolve and fix the volume to 2.0ml, shaking up to prepare stock solutions of each reference substance, wherein the concentration is 0.5mg/m L, precisely measuring the stock solutions of each reference substance by 2.0m L, placing the stock solutions into the same volumetric flask with the volume of 25m L, adding methanol to dissolve and dilute to a scale, shaking up to obtain a reference substance mixed solution, and feeding a filter membrane into a high performance liquid chromatograph.
4) Calculation of correction factors
Collecting the mixed control solution 10 μ L prepared in step 3), and measuring chromatogram with high performance liquid chromatographyCalculating peak area, selecting No. 5 reference as internal reference, and calculating relative correction factor f between the internal reference and other compoundsk/s. The relative correction factor calculation formula is: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesPeak area of the internal standard substance; csIs the internal standard concentration; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
4) Determination of other flavone Components
Taking 10 mu L of the test solution prepared in the step 2), measuring a chromatogram by using a high performance liquid chromatography, and respectively calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkPeak area of the measured component, fk/sTo correct the factor, AsThe peak area of the internal reference substance is shown.
The gradient condition is as follows: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A;
example 3
The following detailed description is made with reference to fig. 4 and the embodiments:
1) reference substance
quercetin-3-O- (2' ', 6' ' -di-O- α -L-rhamnosyl) -L-D-glucoside (1), kaempferol-3-0- (2' ', 6' ' -di-O-L2-L-3-rhamnosyl) -L-D-glucoside (2), kaempferol-3-O-L5 sophoroside (3), quercetin-3-O-rutinoside-7-O-glucoside (4), 3' -methyl-myricetin-3-O-rutinoside (5), quercetin 3-O-glucoside (6), quercetin 3-O- (2-L-D-glucosyl-L-L-rhamnoside) (7), kaempferol-3 ' ' -O-rutinoside (8), isorhamnetin-3-O- β -D-glucoside (9), kaempferol 3- (2' ' - β -D-6-D-rutinoside) -3985-p-586-rhamnoside (3-O- α), and kaempferol-3-O-636-glucoside (3-O-2 ' ' -6-rhamnoside) (3-2 ' ' -366), and kaempferol-3-O-3-2 ' ' -6-O-glucoside (3-4-L).
2) Preparation of sample solution
Precisely weighing semen Ginkgo extract powder 5.0mg in volumetric flask, adding methanol for dissolving, ultrasonic treating for 5min, diluting to constant volume of 20m L, shaking, filtering with 0.22 μm microporous membrane to obtain test solution, and analyzing and detecting by high performance liquid chromatography.
3) Preparation of control solutions
Precisely weighing 1.0mg of each of the 12 ginkgetin monomer compounds in the step 1), respectively adding methanol into volumetric flasks to dissolve and fix the volume to 2ml, shaking up to prepare stock solutions of all reference substances, wherein the concentration is 0.5mg/m L, precisely measuring 1.5m L of the stock solutions of all reference substances, placing the stock solutions in the same volumetric flask with the volume of 25m L, adding methanol to dissolve and dilute to a scale, shaking up to obtain a reference substance mixed solution, and feeding the reference substance mixed solution into a high-performance liquid chromatograph.
4) Calculation of correction factors
Taking 10 mu L of the mixed reference substance solution prepared in the step 3), measuring chromatogram by using high performance liquid chromatography, calculating peak area, selecting No. 6 reference substance as an internal reference substance, and respectively calculating relative correction factor f between the internal reference substance and other compoundsk/s. The relative correction factor calculation formula is: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesPeak area of the internal standard substance; csIs the internal standard concentration; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
4) Determination of other flavone Components
Taking 10 mu L of the test solution prepared in the step 2), measuring a chromatogram by using a high performance liquid chromatography, and respectively calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkAs a group to be testedPeak area of fraction fk/sTo correct the factor, AsThe peak area of the internal reference substance is shown.
The gradient condition is as follows: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A;
example 4
The invention will be further explained below in connection with fig. 3
Methodological investigation example
1) Linear relationship, minimum detection limit (L OD) and minimum quantitation limit (L OQ) assays
Precisely sucking standard solution 8, 20, 40, 80, 160, 320 and 640ug/ml of 10 compounds serving as reference substances, feeding the standard solution into a liquid chromatograph with the sample amount of 10 u L, recording peak areas, drawing a standard curve by taking the concentration of each reference substance as a horizontal coordinate and the peak area as a vertical coordinate, determining the detection limit of the method by S/N =3, determining the quantification limit of the method by S/N =10, determining parameters such as the quantification limit, the detection limit, the quantification limit and a linear regression equation, and displaying a good linear relation (r) by the result2Greater than or equal to 0.9993), detection limit of 0.13-1.11, quantification limit of 0.43-3.69, RSD of relative correction factor of 0.23-1.34, and relative retention time of 0.05-0.72.
Regression equation, linear range, and minimum detection limit and minimum quantification limit
2) Precision, repeatability, stability and accuracy tests:
precision: taking the reference substance solution, repeatedly injecting samples for 6 times in one day and 6 times in 3 consecutive days under the determined chromatographic conditions, and evaluating the precision in the day and the day by the Relative Standard Deviation (RSD) and Retention Time (RT) of the peak areas of the analyzed components, wherein the precision result shows that the RSD of the A (peak area) and the RT (retention time) in the day is respectively lower than 0.89 and 0.75, and the RSD of the A (peak area) and the RT (retention time) in the day is respectively lower than 1.86 and 0.6, thereby indicating that the precision is good.
And (3) stability test: and (3) taking the same mixed standard solution, injecting the mixed standard solution into a liquid chromatograph at 0, 2, 4, 6, 8, 12, 24 and 48 hours respectively, and observing the stability of the mixed standard solution by using the peak area RSD and the Retention Time (RT) of each component, wherein the RSD of the stability A (peak area) and the RSD of the RT (retention time) are respectively 0.66-1.92 and 0.13-1.18, which indicates that the sample solution is stable within 48 hours.
And (3) repeatability test: taking the mixed standard solution (6 parts in parallel), carrying out sample injection analysis under the chromatographic conditions, and evaluating the stability of the mixed standard solution according to the RSD Retention Time (RT) value of each component content in the sample, wherein the RSD of the repeatability A (peak area) and the RSD of the RT (retention time) are respectively 0.67-1.96 and 0.05-0.51, which shows that the repeatability of the sample solution is good.
Precision, repeatability and stability test
Accuracy experiment: through the accuracy of the sample adding recovery rate experimental evaluation method, 1g of a proper amount of ginkgo medicinal material powder with known content is precisely weighed, a certain amount of reference solution is added, samples are prepared according to the sample solution, the samples are prepared into low, medium and high concentrations (50%, 80% and 100%), the sample adding recovery rate is calculated, the recovery rate = (measured value-original value)/the added standard value is 98.8-102.1, the RSD is 0.32-1.76, and the result shows that the method has good accuracy.
Applicability examination of the method:
in order to detect the new method, the laboratory has examined the different liquid chromatographs, the different chromatographic columns and other influencing factors (wavelength, flow rate, column temperature, gradient of mobile phase, gradient time program), the obtained relative correction factor (RPF) and the relative retention time (RSD).
Accurate positioning of chromatographic peaks is a precondition for ensuring application of a one-test-multiple-evaluation method, and positioning is carried out by combining relative retention time value (RRT) parameters with integral features of a chromatogram. RRT = RTx/RTs，RTxRetention time of other chromatographic peaks RTs: retention time of chromatographic peak of internal standard substance, RSD range of RCF of different liquid chromatographs0.12-1.18, and the RSD of RRT is in the range of 0.2-1.36, so that different liquid chromatographs have little influence on the method. The RSD of RCF of different wavelength is in the range of 0.26-2.03, and the RSD of RRT is in the range of 0.53-1.83. The RSD of RCF of different flow rate is in the range of 0.53-1.72, the RSD of RRT is in the range of 0.33-1.65, the RSD of RCF of different column temperature is in the range of 0.65-2.02, the RSD of RRT is in the range of 0.50-1.67, the RCF of mobile phase acid is in the range of 0.65-1.77, the RRT is in the range of 0.46-2.30, and the method is slightly influenced.
Comparison of one-test-multiple evaluation method with external standard method (mg/g)
Note: peak 5 is quercetin 3-O-glucoside
The correlation between the method and the result obtained by the external standard method is represented by the Pearson correlation coefficient, so that the method and the result obtained by the external standard method have no obvious difference.
Claims (1)
1. A quality evaluation method of a ginkgo leaf extract is characterized in that the method is used for measuring the content of a plurality of flavonoid chemical components in the ginkgo leaf extract by utilizing a high performance liquid chromatography technology and a one-measurement-multiple-evaluation method, and comprises the following steps:
1) reference substance
quercetin-3-O- (2' ', 6' ' -di-O- α -L-rhamnosyl) -L-D-glucoside (1), kaempferol-3-O- (2' ', 6' ' -di-O-L2-L-rhamnosyl) -L-D-glucoside (2), kaempferol-3-O-L-sophoroside (3), quercetin-3-O-rutinoside-7-O-glucoside (4), 3' -methyl-myricetin-3-O-rutinoside (5), quercetin-3-O-glucoside (6), quercetin-3-O- (2-L-D-glucosyl-L-L-rhamnoside) (7), kaempferol-3-O-rutinoside (8), isorhamnetin-3-O- β -D-glucoside (9), kaempferol-3- (2' ' -34-D-rhamnoside) -736-rhamnosyl) -L-D-glucoside (1), kaempferol-3-O-glucoside (3-O-636), kaempferol-3-O-glucoside (3-O- α), kaempferol-3-O-glucoside (3- α), kaempferol-3-4-rhamnoside (3-p-366- α, 3-rhamnoside) (3611-rhamnoside-p-rhamnoside) (3-3611- α);
2) preparation of test solution
Precisely weighing folium Ginkgo extract powder 5.0mg in volumetric flask, adding methanol for dissolving, performing ultrasonic treatment for 5min, diluting to constant volume of 20m L, shaking, and filtering with 0.22 μm microporous membrane to obtain sample solution;
3) preparation of Mixed control solutions
Precisely weighing 1.0mg of each of the 12 reference substances in the step 1), respectively placing the reference substances in volumetric flasks, adding methanol for dissolving, fixing the volume to 2ml, shaking up to prepare stock solutions of the reference substances, wherein the concentration of the stock solutions is 0.5mg/m L, precisely weighing 1.5m L of the stock solutions of the reference substances, placing the stock solutions in the same volumetric flask with the volume of 25m L, adding methanol for dissolving, diluting to the scale, and shaking up to obtain mixed reference substance solutions;
4) calculation of correction factors
Taking 10 mu L of the mixed reference substance solution prepared in the step 3), measuring chromatogram by using high performance liquid chromatography, calculating peak area, selecting the No. 6 reference substance quercetin-3-O-glucoside as an internal reference substance, and calculating relative correction factor f between the internal reference substance and other compounds respectivelyk/s；
The relative correction factor calculation formula is: f. ofk/s＝fk/fs＝AkCs/(AsCk) (ii) a In the formula: a. thesPeak area of the internal standard substance; csIs the internal standard concentration; a. thekThe peak area of a certain component k to be measured; ckThe concentration of a certain component k to be measured;
5) determination of other flavone Components
Taking 10 mu L of the test solution prepared in the step 2), measuring a chromatogram by using a high performance liquid chromatography, and respectively calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkPeak area of the measured component, fk/sTo correct the factor, AsIs the peak area of the internal reference substance;
the high performance liquid chromatography adopts an analytical chromatographic column: gemini C18；
Mobile phase: acetonitrile is taken as a mobile phase A, and 0.1% formic acid aqueous solution is taken as a mobile phase B;
mobile phase gradient conditions: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103110670A (en) * | 2012-10-23 | 2013-05-22 | 北京华润高科天然药物有限公司 | Preparation method for efficiently extracting separating high-purity flavone components from ginkgo leaf |
| CN104523771A (en) * | 2015-01-13 | 2015-04-22 | 中国药科大学 | Ginkgo leaf total flavones and preparing method thereof |
| CN106706809A (en) * | 2016-12-26 | 2017-05-24 | 河北神威药业有限公司 | Method for simultaneously determining contents of multiple components in Shuxuening injection |
| CN106770828A (en) * | 2016-12-26 | 2017-05-31 | 河北神威药业有限公司 | It is a kind of while the method for determining multicomponent content in ginkgo biloba p.e and its preparation |
| CN107064397A (en) * | 2017-06-16 | 2017-08-18 | 黑龙江珍宝岛药业股份有限公司 | A kind of Shu Xuening injection method of quality control that many evaluation amount methods and finger-print are surveyed based on one |
-
2018
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103110670A (en) * | 2012-10-23 | 2013-05-22 | 北京华润高科天然药物有限公司 | Preparation method for efficiently extracting separating high-purity flavone components from ginkgo leaf |
| CN104523771A (en) * | 2015-01-13 | 2015-04-22 | 中国药科大学 | Ginkgo leaf total flavones and preparing method thereof |
| CN106706809A (en) * | 2016-12-26 | 2017-05-24 | 河北神威药业有限公司 | Method for simultaneously determining contents of multiple components in Shuxuening injection |
| CN106770828A (en) * | 2016-12-26 | 2017-05-31 | 河北神威药业有限公司 | It is a kind of while the method for determining multicomponent content in ginkgo biloba p.e and its preparation |
| CN107064397A (en) * | 2017-06-16 | 2017-08-18 | 黑龙江珍宝岛药业股份有限公司 | A kind of Shu Xuening injection method of quality control that many evaluation amount methods and finger-print are surveyed based on one |
Non-Patent Citations (4)
| Title |
|---|
| Rapid and Sensitive Determination of Major Active Ingredients and Toxic Components in Leaves Extract (EGb 761) by a Validated UPLC–MS-MS Method;Yao Wang et al;《Journal of Chromatographic Science》;20170109;第55卷(第4期);第459-464页 * |
| 一测多评法同时测定舒血宁注射液中6种黄酮类成分;张方 等;《中国药师》;20151231;第18卷(第10期);第1652-1656页 * |
| 一测多评法测定银杏叶提取物中7种黄酮类成分含量;杨艳模 等;《中国实验方剂学杂志》;20140430;第20卷(第8期);第82-85页 * |
| 指纹图谱结合一测多评模式在中药鱼腥草质量评价中的应用研究;何兵 等;《中国中药杂志》;20130831;第38卷(第16期);第2682-2689页 * |
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