CN110824069A - Method for constructing fingerprint of terpene lactones in ginkgo leaf extract or preparation thereof - Google Patents
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Abstract
The invention relates to the technical field of medicine detection, in particular to a method for constructing a terpene lactone fingerprint in a ginkgo leaf extract or a preparation thereof. The construction method adopts a high performance liquid chromatography to construct the fingerprint, wherein the chromatographic conditions of the high performance liquid chromatography are as follows: the stationary phase is a chromatographic column taking octadecyl bonded silica gel as a filler, a mobile phase A is methanol, a mobile phase B is formic acid aqueous solution with the volume concentration of 0.05-0.3%, a detector is an electrospray detector, and the column temperature is 20-40 ℃; the flow rate is 0.5-1mL/min, and gradient elution is carried out. The fingerprint provided by the invention provides scientific basis for comprehensively establishing the quality control standard of the ginkgo leaf extract or the preparation thereof.
Description
Technical Field
The invention relates to the technical field of medicine detection, in particular to a method for constructing a terpene lactone fingerprint in a ginkgo leaf extract or a preparation thereof.
Background
The ginkgo leaves can tonify heart and qi, tonify kidney and moisten lung, strengthen brain and refresh mind, and prolong life, and the modern pharmacological research shows that the ginkgo leaves have the functions of removing oxygen free radicals, reducing blood fat, enhancing the function of a central nervous system, regulating the level of neurotransmitter and hormone, improving the rheological state of blood, resisting inflammation, resisting allergy and the like. At present, the ginkgo leaves are clinically applied to tablets, granules and injections of various brands such as Sitylong, gold dolo, Tianbaoning, Yinchuo, Shuxuening, Luoxin and coronary heart disease, and are mainly used for treating various cardiovascular and cerebrovascular diseases, diabetes, nervous system diseases, respiratory system diseases and the like.
The fingerprint refers to a chromatogram or a spectrogram which can mark the chemical characteristics of certain traditional Chinese medicinal materials or preparations thereof and is obtained by adopting a certain analysis means after the traditional Chinese medicinal materials or the preparations thereof are properly processed, and the method is a comprehensive and quantifiable identification means. At present, the fingerprint spectrum is taken as the quality control method of the traditional Chinese medicine extract and the preparation thereof, and the international consensus is achieved.
High performance liquid chromatography is one of the common means of identifying compounds and determining the structure of substances. In the drug analysis, the characteristic of 'fingerprint' of liquid chromatogram is taken as the basis of drug identification, and is a method commonly adopted by pharmacopoeias of various countries. Although the main active ingredients in the ginkgo leaf extract and the preparation thereof are flavonoid glycoside and terpene lactone active substances, the ginkgo leaf extract and the preparation have far-ranging other ingredients except the active ingredients, so that different instruments and different liquid phase test conditions are adopted for respectively sampling and testing, thereby causing complicated operation steps and insufficient process simplicity.
Therefore, in order to better control the quality of the ginkgo biloba extract and the preparation thereof, ensure the safety of medication, better guide production, ensure that the process control is stricter and more reasonable, and enable consumers to comprehensively know the product quality, a method which is more favorable for product quality detection and defines the types and the contents of specific active ingredients is continuously researched and searched, and a scientific basis is provided for comprehensively establishing the quality control standard of the ginkgo biloba extract and the preparation thereof.
Disclosure of Invention
Aiming at the problems, the invention provides a method for constructing a fingerprint of terpene lactones in a ginkgo leaf extract or a preparation thereof, which provides a scientific basis for comprehensively establishing the quality control standard of the ginkgo leaf extract and the preparation thereof.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
a method for constructing fingerprint of terpene lactones in folium Ginkgo extract or its preparation comprises constructing fingerprint by high performance liquid chromatography, wherein the high performance liquid chromatography has the following chromatographic conditions: the stationary phase is a chromatographic column taking octadecyl bonded silica gel as a filler, a mobile phase A is methanol, a mobile phase B is formic acid aqueous solution with the volume concentration of 0.05-0.3%, a detector is an electrospray detector, and the column temperature is 20-40 ℃; the flow rate is 0.5-1mL/min, and gradient elution is carried out.
Compared with the prior art, the method for constructing the fingerprint of the terpene lactones in the ginkgo leaf extract or the preparation thereof adopts the high performance liquid chromatography to construct the fingerprint, and has the following advantages:
the fingerprint spectrum construction method provided by the invention can simultaneously determine the content of the 5 terpene lactones in the ginkgo leaf extract and the Shuxuening injection thereof under the same high performance liquid chromatography condition, avoids frequently changing the liquid phase condition during detection, improves the working efficiency, and is suitable for the requirement of industrial mass production.
Preferably, the conditions of the gradient elution are:
0-20min, 20% mobile phase A; 80% mobile phase B;
20-25min, 20% → 30% mobile phase a; 80% → 70%, mobile phase B;
25-39min, 30% → 35% mobile phase a; 70% → 65%, mobile phase B;
39-49min, 35% → 75% mobile phase a; 65% → 25%, mobile phase B;
49-51min, 75% mobile phase A; 25%, mobile phase B;
51-53min, 75% → 40% mobile phase a; 25% → 60%, mobile phase B;
53-60min, 40% mobile phase A; 60% mobile phase B.
Due to the complex components in the ginkgo biloba extract and the Shuxuening injection thereof, the components can not be well separated by using a single mobile phase, and the components with larger difference of the neutral properties of the solution to be detected can be well separated according to the respective properties and the respective proper volume factor k by using the optimal elution time and the optimal mobile phase proportion.
Preferably, the chromatographic conditions of the high performance liquid chromatography are: the stationary phase is a chromatographic column taking octadecyl bonded silica gel as a filler, a mobile phase A is methanol, a mobile phase B is a formic acid aqueous solution with the volume concentration of 0.2 percent, a detector is an electrospray detector, and the column temperature is 30 ℃; the flow rate was 0.5 mL/min.
Preferably, the parameters of the chromatography column are 250mm x 4.6mm, 5 μm.
Preferably, the high performance liquid chromatography is performed by using a sample amount of 2-10. mu.L.
Preferably, the fingerprint of the ginkgo biloba extract has 13 characteristic common peaks, the retention time of the chromatographic peak of the ginkgolide C is 1, and the relative retention times of other characteristic common peaks and the reference peak are respectively as follows: peak 1: 0.165 +/-0.005; peak 2: 0.587 plus or minus 0.003; peak 3: 0.742 +/-0.004; peak 4: 0.888 plus or minus 0.002; peak 5: 1.00; peak 6: 1.672 plus or minus 0.005; peak 7: 1.821 +/-0.006; peak 8: 2.281 plus or minus 0.022; peak 9: 2.320 plus or minus 0.018; peak 10: 2.398 +/-0.021; peak 11: 2.462 +/-0.026; peak 12: 2.567 +/-0.029; peak 13: 2.658 + -0.032.
Preferably, the fingerprint of the ginkgo biloba leaf extract has 13 characteristic common peaks, and the relative peak areas of other characteristic common peaks are 1.490 plus or minus 0.373 (peak 3), 0.394 plus or minus 0.118 (peak 4), 1.649 plus or minus 0.412 (peak 6), 0.957 plus or minus 0.239 (peak 7), 0.912 plus or minus 0.274 (peak 11) and 1.385 plus or minus 0.346 (peak 12) respectively, wherein the peak area of the ginkgolide C (S peak, peak 5) is 1. According to the relevant regulations of 'technical requirements for traditional Chinese medicine injection fingerprint spectrum research', the single peak area accounts for less than 5% of the total peak area of the common peak, and the peak area ratio is not required. Therefore, peak area ratios of peaks 1, 2, 8, 9, 10 and 13 are not required.
Preferably, the fingerprint of the shuxuening injection has 13 characteristic common peaks, the chromatographic peak of the ginkgolide C is taken as a reference peak, and the retention time of the chromatographic peak is 1, and the relative retention times of other characteristic common peaks and the reference peak are respectively as follows: peak 1: 0.164 +/-0.006; peak 2: 0.195 plus or minus 0.01; peak 3: 0.888 plus or minus 0.002; peak 4: 1.00; peak 5: 1.672 +/-0.006; peak 6: 1.822 +/-0.005; peak 7: 2.238 ± 0.018; peak 8: 2.282 +/-0.020; peak 9: 2.321 +/-0.021; peak 10: 2.399 +/-0.013; peak 11: 2.463 +/-0.025; peak 12: 2.568 plus or minus 0.028; peak 13: 2.659 +/-0.030.
Preferably, the fingerprint of the Shuxuening injection has 13 characteristic common peaks, and the relative peak areas of other 6 common peaks are 0.677 + -0.203 (peak 1), 0.401 + -0.120 (peak 3), 1.718 + -0.430 (peak 5), 0.981 + -0.245 (peak 6), 0.862 + -0.259 (peak 11) and 1.310 + -0.328 (peak 12) respectively, wherein the peak area of the ginkgolide C (S peak, peak 4) is 1. According to the relevant regulations of 'technical requirements for traditional Chinese medicine injection fingerprint spectrum research', the single peak area accounts for less than 5% of the total peak area of the common peak, and the peak area ratio is not required. Therefore, peak area ratios of peaks 2, 4, 7-10 and 13 are not required.
Preferably, bilobalide J, bilobalide C, bilobalide A and bilobalide B are used as reference substances, and are dissolved by methanol to obtain reference substance solution.
Preferably, the concentrations of bilobalide, bilobalide J, bilobalide C, bilobalide A and bilobalide B in the control solution are 0.07-0.10mg/mL, 0.015-0.035mg/mL, 0.05-0.08mg/mL, 0.07-0.10mg/mL and 0.05-0.075mg/mL, respectively.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a high performance liquid chromatogram of a control solution provided by an embodiment of the invention;
FIG. 2 is a fingerprint of Ginkgo biloba extract provided in example 1 of the present invention;
fig. 3 is a fingerprint spectrum provided by the shuning injection provided in embodiment 2 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
The embodiment of the invention provides a method for constructing a fingerprint of terpene lactones in a ginkgo leaf extract, which comprises the following steps: the method comprises the following steps:
step a, preparing a ginkgo biloba extract test solution by referring to a preparation method of a ginkgo biloba lactone test solution under the content determination term in a ginkgo biloba extract in the first edition of Chinese pharmacopoeia 2015, which specifically comprises the following steps: precisely weighing 130mg of ginkgo leaf extract, adding 10mL of water, placing in a water bath, warming to dissolve, adding 2 wt% of hydrochloric acid solution for 2 drops, shaking and extracting for 4 times (15mL, 10mL and 10mL) by using ethyl acetate, combining extracting solutions, washing by using 5 wt% of sodium acetate solution for 20mL, separating sodium acetate solution, washing by using ethyl acetate 10mL, combining ethyl acetate 10mL extracting solution and washing solution, washing by using water for 2 times and 20mL each time, separating water solution, washing by using ethyl acetate 10mL, combining ethyl acetate solutions, recovering a solvent till the solution is nearly dry, dissolving a residual liquid by using methanol, transferring the residual liquid into a 25mL measuring flask, adding the methanol to a scale, shaking uniformly, filtering by using a proper amount of 0.45 mu m microporous filter membrane, and taking an obtained subsequent filtrate as a sample solution of the ginkgo leaf extract.
And B, taking bilobalide, bilobalide J, bilobalide C, bilobalide A and bilobalide B as reference substances, and dissolving with methanol to obtain a reference substance solution, wherein the concentrations of the bilobalide, the bilobalide J, the bilobalide C, the bilobalide A and the bilobalide B in the reference substance solution are respectively 0.085mg/mL, 0.025mg/mL, 0.062mg/mL, 0.082mg/mL and 0.063 mg/mL.
And c, respectively carrying out high performance liquid chromatography analysis on the test solution and the reference solution by adopting high performance liquid chromatography to construct a fingerprint, wherein the chromatographic conditions of the high performance liquid chromatography are as follows: the chromatographic column is a YMC-Pack ODS column (250mm multiplied by 4.6mm, 5 mu m), the mobile phase A is methanol, the mobile phase B is formic acid aqueous solution with the volume concentration of 0.2%, the detector is an electrospray detector, and the column temperature is 30 ℃; the flow rate is 0.5mL/min, the sample injection amount is 5 mu L, and the gradient elution conditions are as follows:
0-20min, 20% mobile phase A; 80% mobile phase B;
20-25min, 20% → 30% mobile phase a; 80% → 70%, mobile phase B;
25-39min, 30% → 35% mobile phase a; 70% → 65%, mobile phase B;
39-49min, 35% → 75% mobile phase a; 65% → 25%, mobile phase B;
49-51min, 75% mobile phase A; 25%, mobile phase B;
51-53min, 75% → 40% mobile phase a; 25% → 60%, mobile phase B;
53-60min, 40% mobile phase A; 60% mobile phase B.
The high performance liquid chromatography chromatogram of the control solution is shown in FIG. 1, wherein peak BB is bilobalide; peak GJ is bilobalide J; peak GC is bilobalide C; peak GA is bilobalide A; peak GB is ginkgolide B.
The method for constructing the fingerprint of the ginkgo biloba extract is adopted to establish the high-efficiency liquid phase spectrum of 10 batches of ginkgo biloba extracts, and the fingerprint of 10 batches of samples is synthesized by adopting a Chinese medicine chromatography fingerprint similarity evaluation system of the national pharmacopoeia committee, as shown in figure 2, the HPLC fingerprint of the ginkgo biloba extract consisting of 13 common peaks is generated, wherein the chromatographic peak of No. 5 is ginkgolide C. And calculating the similarity by taking the reference spectra generated by 10 batches of samples as a reference, wherein the similarity is more than 0.90.
As can be seen from fig. 2, the fingerprint of the ginkgo biloba extract has 13 characteristic common peaks, wherein the No. 5 chromatographic peak is ginkgolide C, the chromatographic peak of ginkgolide C is taken as a reference peak, and the relative retention times of the other characteristic common peaks and the reference peak are respectively as follows: peak 1: 0.165; peak 2: 0.587; peak 3: 0.742; peak 4: 0.888; peak 5: 1.00; peak 6: 1.672 a; peak 7: 1.821; peak 8: 2.281; peak 9: 2.320 of; peak 10: 2.398; peak 11: 2.462; peak 12: 2.567, respectively; peak 13: 2.658.
Test example 1 reproducibility test of ginkgo biloba leaf extract
Taking the same batch of samples, preparing 6 parts of test solution according to the step a in the construction method of the terpene lactones fingerprint in the ginkgo biloba leaf extract in the example 1, and performing repeatability inspection under the same chromatographic conditions as the example 1, wherein the results are shown in tables 1 and 2, and the RSD of the relative retention time and the relative peak area of the common peak is less than 5%, which indicates that the method has good repeatability.
TABLE 1 characteristic common peak relative retention time
Note: the retention time of ginkgolide C (S peak, peak 5) was 1.
TABLE 2 characteristic common peaks relative peak area
Note: the peak area of ginkgolide C (S peak, peak 5) is 1 percent, and the area of other common peak accounts for less than 5 percent of the total peak area, so the calculation is not carried out.
Test example 2 precision test of ginkgo biloba leaf extract
The sample solution prepared in example 1 was taken, and sample injection was repeated 6 times under the same chromatographic conditions as in example 1, precision investigation was performed, and the consistency of the relative retention time and the relative peak area of the characteristic common peak was examined, and the results are shown in tables 3 and 4, respectively, and the RSD of the relative retention time and the relative peak area of the characteristic common peak was less than 5%, indicating that the method is good in precision.
TABLE 3 characteristic common peak relative retention time
TABLE 4 characteristic common peaks relative peak area
Note: the area of other common peaks accounts for less than 5% of the total peak area, so no calculation is performed.
Test example 3 stability test of ginkgo biloba leaf extract
The sample solution prepared in example 1 was sampled and detected under the same chromatographic conditions as in example 1 at 0, 2, 4, 8, 12, 24h and 48h, 5 μ L was injected for each sample, and the relative retention time, the relative peak area and the similarity of the common peaks were examined. As shown in tables 5 and 6, the RSD of the common peak relative retention time and the relative peak area were less than 5%, indicating that the test solution was stable within 48 hours.
TABLE 5 characteristic common peak relative retention time
TABLE 6 characteristic common peaks relative peak area
Note: the area of other common peaks accounts for less than 5% of the total peak area, so no calculation is performed.
Example 2
The embodiment of the invention provides a method for constructing a terpene lactones fingerprint in a Shuxuening injection, which comprises the following steps: the method comprises the following steps:
step a, precisely measuring 15mL of the product, adding 2 drops of diluted hydrochloric acid, adjusting the pH value to 2, shaking and extracting with ethyl acetate for 4 times (20mL, 10mL and 10mL), combining ethyl acetate solutions, washing with 20mL of 5 wt% sodium acetate solution, separating ethyl acetate solutions, washing sodium acetate solutions with 10mL of ethyl acetate, combining ethyl acetate solutions and washing liquid, washing with water for 2 times, 20mL each time, combining ethyl acetate solutions, evaporating to dryness, dissolving residual liquid with methanol, transferring to a 5mL measuring flask, adding methanol to a scale, shaking uniformly, filtering with a 0.45-micrometer microporous filter membrane, and taking a subsequent filtrate as a sample solution of the Shuxuening injection.
And B, taking bilobalide, bilobalide J, bilobalide C, bilobalide A and bilobalide B as reference substances, and dissolving with methanol to obtain a reference substance solution, wherein the concentrations of the bilobalide, the bilobalide J, the bilobalide C, the bilobalide A and the bilobalide B in the reference substance solution are respectively 0.085mg/mL, 0.025mg/mL, 0.062mg/mL, 0.082mg/mL and 0.063 mg/mL.
And c, respectively carrying out high performance liquid chromatography analysis on the test solution and the reference solution by adopting high performance liquid chromatography to construct a fingerprint, wherein the chromatographic conditions of the high performance liquid chromatography are as follows: the chromatographic column is a YMC-Pack ODS column (250mm multiplied by 4.6mm, 5 mu m), the mobile phase A is methanol, the mobile phase B is formic acid aqueous solution with the volume concentration of 0.2%, the detector is an electrospray detector, and the column temperature is 30 ℃; the flow rate is 0.5mL/min, the sample injection amount is 5 mu L, and the gradient elution conditions are as follows:
0-20min, 20% mobile phase A; 80% mobile phase B;
20-25min, 20% → 30% mobile phase a; 80% → 70%, mobile phase B;
25-39min, 30% → 35% mobile phase a; 70% → 65%, mobile phase B;
39-49min, 35% → 75% mobile phase a; 65% → 25%, mobile phase B;
49-51min, 75% mobile phase A; 25%, mobile phase B;
51-53min, 75% → 40% mobile phase a; 25% → 60%, mobile phase B;
53-60min, 40% mobile phase A; 60% mobile phase B.
The high performance liquid chromatography chromatogram of the control solution is shown in FIG. 1, wherein peak BB is bilobalide; peak GJ is bilobalide J; peak GC is bilobalide C; peak GA is bilobalide A; peak GB is ginkgolide B.
By adopting the construction method of the fingerprint of the Shuxuening injection, the high-efficiency liquid chromatogram of 10 batches of the Shuxuening injection is established, and the fingerprint of 10 batches of samples is synthesized by adopting a Chinese medicine chromatogram fingerprint similarity evaluation system of the State pharmacopoeia Committee, and as shown in figure 3, the HPLC fingerprint of the Shuxuening injection consisting of 13 common peaks is generated, wherein the No. 4 chromatographic peak is ginkgolide C. And calculating the similarity by taking the reference spectra generated by 10 batches of samples as a reference, wherein the similarity is more than 0.90.
As can be seen from fig. 3, the fingerprint spectrum of the shuxuening injection has 13 characteristic common peaks, wherein the chromatographic peak No. 4 is ginkgolide C, the chromatographic peak of ginkgolide C is taken as a reference peak, and the relative retention times of other characteristic common peaks and the reference peak are respectively as follows: peak 1: 0.164 of; peak 2: 0.195; peak 3: 0.888; peak 4: 1.00; peak 5: 1.672 a; peak 6: 1.822; peak 7: 2.238, respectively; peak 8: 2.282, respectively; peak 9: 2.321, respectively; peak 10: 2.399, respectively; peak 11: 2.463, respectively; peak 12: 2.568; peak 13: 2.659.
Test example 4 repeatability test of Shuxuening injection
Taking the same batch of samples, preparing 6 test sample solutions according to the step a in the construction method of the terpene lactones fingerprint in the Shuxuening injection in the example 2, and performing repeatability investigation under the same chromatographic conditions as the example 2, wherein the results are shown in tables 7 and 8, and the RSD of the relative retention time of the common peak and the relative peak area are less than 5%, which indicates that the method has good repeatability.
TABLE 7 characteristic common peak relative retention time
TABLE 8 characteristic common peaks relative peak area
Note: the area of other common peaks accounts for less than 5% of the total peak area, so no calculation is performed.
Test example 5 precision test of Shuxuening injection
The sample solution prepared in example 2 was taken, and sample injection was repeated 6 times under the same chromatographic conditions as in example 2, precision investigation was performed, and the consistency of the relative retention time and the relative peak area of the characteristic common peak was examined, the results are shown in tables 9 and 10, respectively, and RSD of the relative retention time and the relative peak area of the characteristic common peak was less than 5%, indicating that the method was good in precision.
TABLE 9 characteristic common peak relative retention time
TABLE 10 characteristic common peaks relative peak area
Note: the area of other common peaks accounts for less than 5% of the total peak area, so no calculation is performed.
Test example 6 stability test of Shuxuening injection
The sample solution prepared in example 2 was taken and subjected to detection at 0, 2, 4, 8, 12, 24h, and 48h under the same chromatographic conditions as in example 2, 5 μ L was injected each time, and the relative retention time, the relative peak area, and the similarity of the common peaks were examined. As shown in tables 11 and 12, the RSD of the common peak relative retention time and the relative peak area were less than 5%, indicating that the test solution was stable within 48 hours.
TABLE 11 characteristic consensus peak relative retention time
TABLE 12 characteristic common peaks relative peak area
Note: the area of other common peaks accounts for less than 5% of the total peak area, so no calculation is performed.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (9)
1. A method for constructing a terpene lactone fingerprint in a ginkgo leaf extract or a preparation thereof is characterized by comprising the following steps: the fingerprint is constructed by adopting a high performance liquid chromatography, wherein the chromatographic conditions of the high performance liquid chromatography are as follows: the stationary phase is a chromatographic column taking octadecyl bonded silica gel as a filler, a mobile phase A is methanol, a mobile phase B is formic acid aqueous solution with the volume concentration of 0.05-0.3%, a detector is an electrospray detector, and the column temperature is 20-40 ℃; the flow rate is 0.5-1mL/min, and gradient elution is carried out.
2. The method for constructing a fingerprint of terpene lactones contained in the ginkgo biloba leaf extract or the preparation thereof as claimed in claim 1, wherein the method comprises the steps of: the conditions of the gradient elution are as follows:
0-20min, 20% mobile phase A; 80% mobile phase B;
20-25min, 20% → 30% mobile phase a; 80% → 70% mobile phase B;
25-39min, 30% → 35% mobile phase a; 70% → 65% mobile phase B;
39-49min, 35% → 75% mobile phase a; 65% → 25% mobile phase B;
49-51min, 75% mobile phase A; 25% mobile phase B;
51-53min, 75% → 40% mobile phase a; 25% → 60% mobile phase B;
53-60min, 40% mobile phase A; 60% mobile phase B.
3. The method for constructing a fingerprint of terpene lactones contained in the ginkgo biloba leaf extract or the preparation thereof as claimed in claim 1, wherein the method comprises the steps of: the chromatographic conditions of the high performance liquid chromatography are as follows: the stationary phase is a chromatographic column taking octadecyl bonded silica gel as a filler, a mobile phase A is methanol, a mobile phase B is a formic acid aqueous solution with the volume concentration of 0.2 percent, a detector is an electrospray detector, and the column temperature is 30 ℃; the flow rate was 0.5 mL/min.
4. The method for constructing fingerprint of terpene lactones contained in ginkgo biloba leaf extract or a preparation thereof according to any one of claims 1 to 3, wherein the method comprises the steps of: the chromatographic column has parameters of 250mm × 4.6mm, 5 μm.
5. The method for constructing fingerprint of terpene lactones contained in ginkgo biloba leaf extract or a preparation thereof according to any one of claims 1 to 3, wherein the method comprises the steps of: the sample injection amount of the high performance liquid chromatography is 2-10 mu L.
6. The method for constructing a fingerprint of terpene lactones contained in the ginkgo biloba leaf extract or the preparation thereof as claimed in claim 1, wherein the method comprises the steps of: the fingerprint spectrum of the ginkgo leaf extract has 13 characteristic common peaks, the chromatographic peak of the ginkgolide C is taken as a reference peak, and the retention time of the chromatographic peak is 1, and the relative retention time of other characteristic common peaks and the reference peak is respectively as follows: peak 1: 0.165 +/-0.005; peak 2: 0.587 plus or minus 0.003; peak 3: 0.742 +/-0.004; peak 4: 0.888 plus or minus 0.002; peak 5: 1.00; peak 6: 1.672 plus or minus 0.005; peak 7: 1.821 +/-0.006; peak 8: 2.281 plus or minus 0.022; peak 9: 2.320 plus or minus 0.018; peak 10: 2.398 +/-0.021; peak 11: 2.462 +/-0.026; peak 12: 2.567 +/-0.029; peak 13: 2.658 + -0.032.
7. The method for constructing a fingerprint of terpene lactones contained in the ginkgo biloba leaf extract or the preparation thereof as claimed in claim 1, wherein the method comprises the steps of: the fingerprint spectrum of the Shuxuening injection has 13 characteristic common peaks, wherein the chromatographic peak of the ginkgolide C is taken as a reference peak, and the retention time of the chromatographic peak is 1, and the relative retention times of other characteristic common peaks and the reference peak are respectively as follows: peak 1: 0.164 +/-0.006; peak 2: 0.195 plus or minus 0.01; peak 3: 0.888 plus or minus 0.002; peak 4: 1.00; peak 5: 1.672 +/-0.006; peak 6: 1.822 +/-0.005; peak 7: 2.238 ± 0.018; peak 8: 2.282 +/-0.020; peak 9: 2.321 +/-0.021; peak 10: 2.399 +/-0.013; peak 11: 2.463 +/-0.025; peak 12: 2.568 plus or minus 0.028; peak 13: 2.659 +/-0.030.
8. The method for constructing a fingerprint of terpene lactones contained in the ginkgo biloba leaf extract or the preparation thereof as claimed in claim 1, wherein the method comprises the steps of: dissolving bilobalide, bilobalide J, bilobalide C, bilobalide A and bilobalide B with methanol to obtain reference solution.
9. The method for constructing a fingerprint of terpene lactones contained in the ginkgo biloba leaf extract or the preparation thereof as claimed in claim 8, wherein the method comprises the steps of: the concentrations of bilobalide, bilobalide J, bilobalide C, bilobalide A and bilobalide B in the reference solution are respectively 0.07-0.10mg/mL, 0.015-0.035mg/mL, 0.05-0.08mg/mL, 0.07-0.10mg/mL and 0.05-0.075 mg/mL.
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