Epimedium koreanum genuine research method based on traditional Chinese medicine quality markers

Abstract

The invention discloses a traditional Chinese medicine quality marker-based epimedium koreanum geography research method, which comprises the following steps: S1, carrying out spectrum-effect relationship research of epimedium for treating osteoporosis by combining pharmacodynamics research with chemical component analysis to obtain possible components of epimedium for treating osteoporosis; the screened components are subjected to network pharmacology, molecular docking research and the like to screen out the quality marker of the epimedium koreanum for treating osteoporosis; and step S2, finally obtaining the eight flavonoid quality markers of the epimedium koreanum according to the step S1: quercetin, epimedin A, epimedin B, epimedin C, icariin, icariside I, baohuoside I and icaritin. According to the invention, pharmacodynamic research is combined with chemical component analysis to research the spectrum-effect relationship of epimedium for treating osteoporosis, the quality marker of epimedium koreanum for treating osteoporosis is screened out, and a scientific basis is provided for safe and effective clinical medication of traditional Chinese medicine.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine detection, and in particular to a method for studying the authenticity of Korean epimedium based on traditional Chinese medicine quality markers. Background Art

[0002] Traditional Chinese medicine is a complex system. The ecological environment of traditional Chinese medicine and the organism on which traditional Chinese medicine acts are also complex systems. To understand and study these three complex systems, we must adhere to the characteristics of "systemic traditional Chinese medicine" under the guidance of complex system science theories and methods and apply multidimensional evaluation methods. The method of "multidimensional evaluation" of traditional Chinese medicine is to combine system analysis methods with self-organization theory methods according to the characteristics of the traditional Chinese medicine system and under the guidance of the idea of ​​"systemic traditional Chinese medicine" to study the six elements of the traditional Chinese medicine system, namely "variety, quality, pharmaceutical preparation, medicinal properties, efficacy, and application", so as to reveal the elements, structure, function, and changing laws of the traditional Chinese medicine system interacting with the environment. In the process of research on authentic medicinal materials, it is necessary to adhere to the idea of ​​"systematic traditional Chinese medicine", apply the method of "multidimensional evaluation and integrated analysis", adopt multidisciplinary technologies such as omics, chemistry, and pharmacology, conduct comprehensive analysis of complex networks, and evaluate the multidimensional relationship of authentic medicinal materials "drug (environment-genetic-material)-body (whole-target organ-target cell-target molecule)-efficacy (pharmacological effect-health effect-toxic and side effects)", integrate and analyze the multidimensional relationship of authentic medicinal materials "authentic medicinal materials-ecological factors-efficacy", "authentic medicinal materials-material structure-efficacy", and "authenticity-dose time-efficacy", so as to reveal the scientific connotation of the "authenticity-efficacy" relationship of authentic medicinal materials and solve the difficult problem of "authenticity-efficacy" research on authentic medicinal materials.

[0003] At present, the evaluation system for the authenticity of Korean Epimedium is imperfect. It only uses a single evaluation method, and only uses pharmacodynamic research or chemical component determination to find the differences in efficacy indicators or components between different origins of Epimedium. In order to establish a more complete evaluation method for the authenticity of Korean Epimedium, the quality markers are screened by combining chemistry with efficacy. The establishment of this evaluation method will provide certain help for the authenticity evaluation of other similar medicinal materials. Summary of the invention

[0004] The present invention provides a method for studying the authenticity of Korean epimedium based on traditional Chinese medicine quality markers. By establishing and improving a system for evaluating the authenticity of Korean epimedium medicinal materials, it can be judged from aspects such as the types and content differences of chemical components and pharmacological and pharmacodynamic effects, so as to ultimately provide ideas for comprehensive research on the "authenticity" of Korean epimedium, and also provide scientific research guidance for the further development and utilization of Korean epimedium medicinal materials, so as to solve the problem of the relationship between the "authenticity-efficacy" of Korean epimedium medicinal materials.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] The research method of the authenticity of Korean Epimedium based on the quality markers of traditional Chinese medicine includes the following steps:

[0007] Step S1: The spectrum-effect relationship of epimedium in treating osteoporosis is studied by combining pharmacodynamics research with chemical component analysis, and possible components of epimedium in treating osteoporosis are obtained. The screened components are used to conduct network pharmacology and molecular docking research to screen out quality markers of epimedium in treating osteoporosis;

[0008] Step S2: According to step S1, eight flavonoid quality markers of Korean epimedium are finally obtained: quercetin, epimedin A, epimedin B, epimedin C, icariin, icariin I, baculoside I and icariin, and then the difference in component content between Korean epimedium and other varieties of epimedium is determined based on the quality markers;

[0009] Step S3: The difference in component content between Epimedium koreana and other original Epimediums is as follows: the content ratio of the eight quality markers is 1:1:1:1:1:1:1:1:1;

[0010] Finally, HPLC and UPLC-QE-MS were used to determine whether the content ratio of the eight flavonoid compounds in the Korean epimedium to be analyzed was 1:1:1:1:1:1:1:1:1, and the authenticity of the Korean epimedium was evaluated.

[0011] As a further improvement of the technical solution: other original epimediums include the dried leaves of the Berberidaceae plants Epimedium, Epimedium sagittatum, Epimedium pubescens, and Epimedium koreana.

[0012] As a further improvement of the present technical solution: the detection method for determining the content of flavonoid components by HPLC is as follows: the chromatographic column is a C18 column, the mobile phase A is acetonitrile, the mobile phase B is water, the detection wavelength is 270nm, the column temperature is 35°C, the injection volume is 10uL, and the flow rate is 1mL / min.

[0013] As a further improvement of the present technical solution: the detection method for determining the content of flavonoid components using UPLC-QE-MS is as follows: the chromatographic column is a Waters UPLC BEH-C18 column (1.7 μm, 2.1 mm × 100 mm, Waters, Ireland), the mobile phase A is a 0.1% formic acid aqueous solution, the mobile phase B is acetonitrile, the detection wavelength is 88 nm, the column temperature is 30 ° C, the injection volume is 2 uL, and the flow rate is 0.3 mL / min.

[0014] As a further improvement of this technical solution: the fingerprint data of epimedium of different origins are imported into the "Chinese Medicine Chromatographic Fingerprint Similarity Software Evaluation System" of the State Pharmacopoeia Committee, and the fingerprints are automatically matched and generated after multi-point correction using the median method, and similarity evaluation is performed.

[0015] As a further improvement of the present technical solution: the 11 common chromatographic peaks of Epimedium brevicornum from different origins were imported into SPSS23.0 statistical software. Based on the HPLC-MS peak area data of 25 batches of samples, the samples were classified and compared using Euclidean distance, inter-group comparison and other methods. The similarity between the samples was calculated, and the analysis results were displayed with the help of a dendrogram.

[0016] As a further improvement of the technical solution: In order to further distinguish the quality differences within the different origin Epimedium groups, the 11 common peak areas of the different origin Epimedium samples were imported into SIMAC14.1 software for orthogonal partial least squares-discriminant method analysis, and the OPLS-DA model was established.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The present invention studies the spectrum-effect relationship of epimedium in treating osteoporosis by combining pharmacodynamics research with chemical component analysis, and obtains possible components of epimedium in treating osteoporosis. The components obtained by screening are used to conduct network pharmacology and molecular docking research to screen out quality markers of epimedium in treating osteoporosis, enrich the connotation of "authenticity" of epimedium, and provide a scientific basis for safe and effective use of traditional Chinese medicine in clinical practice.

[0019] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention in conjunction with the accompanying drawings. The specific implementation of the present invention is given in detail by the following embodiments and their accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of this application. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0021] Figure 1 It is a schematic diagram of the structure of the method for studying the authenticity of Epimedium koreanum based on the quality markers of traditional Chinese medicine proposed by the present invention;

[0022] Figure 2 Schematic diagram of the total flavonoids content of each base of Epimedium;

[0023] Figure 3 This is a schematic diagram of the contents of six flavonol components in each base of epimedium;

[0024] Figure 4 This is a schematic diagram of the contents of six flavonols in each of the original Epimedium species;

[0025] Figure 5 HPLC superimposed spectra (S1-S25) and reference spectra (R) of 25 batches of Epimedium;

[0026] Figure 6 is the HPLC spectrum;

[0027] Figure 7 This is a schematic diagram of the contents of eight flavonols in each base of Epimedium;

[0028] Figure 8 It is a liquid-mass spectrometer;

[0029] Fig. 9 This is a schematic diagram of the contents of five flavonols in each base of Epimedium;

[0030] Fig.10 Schematic diagram of OPLS-DA principal component analysis. DETAILED DESCRIPTION

[0031] See also Figure 1 In an embodiment of the present invention, a method for studying the authenticity of Epimedium koreanum based on a traditional Chinese medicine quality marker comprises the following steps:

[0032] Step S1: The spectrum-effect relationship of epimedium in treating osteoporosis is studied by combining pharmacodynamics research with chemical component analysis, and possible components of epimedium in treating osteoporosis are obtained. The screened components are used to conduct network pharmacology and molecular docking research to screen out quality markers of epimedium in treating osteoporosis;

[0033] Step S2: According to step S1, eight flavonoid quality markers of Korean epimedium are finally obtained: quercetin, epimedin A, epimedin B, epimedin C, icariin, icariin I, baculoside I and icariin, and then the difference in component content between Korean epimedium and other varieties of epimedium is determined according to the quality markers. Other original epimediums include dried leaves of Berberidaceae plants Epimedium, sagittatum, pubescent epimedium and Korean epimedium;

[0034] Step S3: The difference in component content between Epimedium koreana and other original Epimediums is as follows: the content ratio of the eight quality markers is 1:1:1:1:1:1:1:1:1;

[0035] Finally, high performance liquid chromatography (HPLC) and ultra high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry (UPLC-QE-MS) combined with multivariate statistical analysis methods were used to determine whether the content ratio of the eight flavonoid compounds in the Korean epimedium to be analyzed was 1:1:1:1:1:1:1:1:1, and the authenticity of the Korean epimedium was evaluated.

[0036] Among them, the experimental method for determining the difference in the content of ingredients between Korean epimedium and other original epimedium is:

[0037] 1.1 Instrument

[0038] High performance liquid chromatograph (Shimadzu LC2030, Japan), freeze dryer (initial code IC-01), rotary evaporator (Shanghai Yarong Biochemical Instrument Factory 5-1-2000), UV spectrophotometer (Shimadzu UV1780, Japan), QE-MS / MS instrument (Thermo Fisher Scientific, USA)

[0039] 1.2 Reagents and Drugs

[0040] Quercetin reference substance (lot number O29HB199514, purity ≥98%), epimedin A reference substance (lot number J12HB184811, purity ≥98%), epimedin B reference substance (lot number J12HB184812, purity ≥98%), epimedin C reference substance (lot number N18GB166250, purity ≥98%), icariin reference substance (lot number T11A11B111118, purity ≥98%), icariin I reference substance (lot number P29O10F101627, purity ≥98%), baohuoside I reference substance ( Batch number A20GB158231, purity ≥ 98%), icariin reference substance (batch number N22GB168780, purity ≥ 98%), arauroside A reference substance (batch number J15HB181573, purity ≥ 98%), rhamnosyl epimedin II reference substance (batch number P28J9S66923, purity ≥ 98%), hyperoside reference substance (batch number J16GB143379, purity ≥ 98%), dehydrated epimedium reference substance (batch number J02HB183847, purity ≥ 98%) were all purchased from Shanghai Yuanye Biotechnology Co., Ltd. Methanol (chromatographic grade), acetonitrile (chromatographic grade), formic acid (chromatographic grade), and ethanol (analytical grade) were all purchased from Merck. The batches and sources of medicinal materials are shown in Table 1. There are 28 batches of samples in total, which were identified by Associate Professor Cai Guangzhi of Changchun University of Chinese Medicine as dried leaves of Epimedium brevicornuMaxim., Epimedium sagittatum (Sieb. et Zucc.) Maxim., Epimedium pubescens Maxim. or Epimedium koreanum Nakai. of the Berberidaceae family.

[0041] Table 128 Origin Information of Epimedium Samples

[0042]

[0043]

[0044] 1.4 Preparation of mixed reference substances

[0045] Accurately weigh 5 mg each of 12 flavonoid components including quercetin, epimedin A, epimedin B, epimedin C, icariin, icariin I, baculoside I, icariin, quercetin A, rhamnosyl icariin II, icariin I, and dehydrated icariin in a volumetric flask, dissolve them in methanol, make up to 25 mL, and prepare a 0.2 mg / mL solution. Take 1 mL of each flavonoid component standard solution with a mass concentration of 0.2 mg / mL in a volumetric flask, add methanol to make up to 10 mL, and prepare a mixed standard with a mass concentration of 20 ug / mL.

[0046] 1.5 Preparation of test samples

[0047] Epimedium was crushed and passed through a No. 3 sieve. About 0.2 g was taken out and accurately weighed. The mixture was placed in a stoppered conical flask. 20 mL of dilute ethanol was accurately added and weighed. The mixture was ultrasonically treated (power 400 W, frequency 50 kHz) for 1 h. The mixture was cooled and weighed again. The lost weight was supplemented with dilute ethanol. The mixture was shaken and filtered. The filtrate was obtained for the determination of total flavonoid content and flavonol glycoside content.

[0048] 1.6 Determination of total flavonoids content

[0049] The UV-visible spectrophotometry method was used to detect the content of epimedium medicinal materials in accordance with the "Chinese Pharmacopoeia" (2020 edition).

[0050] 1.7 Determination of flavonoid content

[0051] 1.7.1 Determination of flavonoid content using HPLC

[0052] The HPLC method established by the research group to detect flavonoid components in Epimedium was used for detection. The detection method is: the chromatographic column is C18 column, the mobile phase A is acetonitrile, the mobile phase B is water, the elution program is shown in Table 2, the detection wavelength is 270nm, the column temperature is 35℃, the injection volume is 10uL, and the flow rate is 1mL / min.

[0053] Table 2 HPLC gradient elution program

[0054]

[0055] 1.7.2 Determination of flavonoid content using UPLC-QE-MS

[0056] The chromatographic column was a Waters UPLC BEH-C18 column (1.7 μm, 2.1 mm×100 mm, Waters, Ireland), mobile phase A was 0.1% formic acid in water, mobile phase B was acetonitrile, the elution program was shown in Table 3, the detection wavelength was 88 nm, the column temperature was 30° C., the injection volume was 2 uL, and the flow rate was 0.3 mL / min.

[0057] Table 3 UPLC-QE-MS gradient elution program

[0058]

[0059] The ion source was an electrospray ion source (H-ESI), with an auxiliary gas flow rate of 13arb, an auxiliary heater temperature of 300℃, an ion transfer tube temperature of 320℃, an automatic gain control (AGC) of 1×106, an S-lens radio frequency level of 50, positive ion mode, a sheath gas (N2) flow rate of 35arb, and a spray voltage of 3.5kV; in negative ion mode, a sheath gas (N2) flow rate of 35arb, and a spray voltage of 2.5kV. The scanning mode adopted positive and negative ion Full MS / dd-MS2 mode, with a scanning range of 80-1200 and a collision energy gradient of 10V, 20V, and 40V.

[0060] 1.8 Establishment and similarity evaluation of HPLC fingerprints of different origins of Epimedium

[0061] The fingerprint data of 25 batches of Epimedium with different origins were imported into the "Chinese Medicine Chromatographic Fingerprint Similarity Software Evaluation System" of the National Pharmacopoeia Committee. The median method was used, and after multi-point correction, the S1 spectrum was used as the reference spectrum to automatically match and generate fingerprints, and the similarity evaluation was performed.

[0062] 1.9 Cluster Analysis

[0063] The 11 common chromatographic peaks of 25 batches of Epimedium were imported into SPSS23.0 statistical software. Based on the HPLC-MS peak area data of 25 batches of samples, the samples were classified and compared using Euclidean distance, inter-group comparison and other methods. The similarity between samples was calculated, and the analysis results were presented with the help of a dendrogram.

[0064] 1.10 Orthogonal Partial Least Squares-Discriminant Analysis

[0065] In order to further distinguish the quality differences within different origin Epimedium groups, the 11 common peak areas of 25 batches of samples were imported into SIMAC 14.1 software for orthogonal partial least squares-discriminant (OPLS-DA) analysis, and the OPLS-DA model was established.

[0066] 2 Experimental results

[0067] 2.1 Comparison of total flavonoids content in different origins of epimedium

[0068] The total flavonoid content of epimedium was determined according to the method for determining the content of total flavonoids in epimedium specified in the Chinese Pharmacopoeia (2020 edition), as shown in Table 4, and the results were qualified. The data results of the total flavonoid content determination of epimedium UV in each base were analyzed using GraphPad Prism 8 software, as shown in Table 4. Figure 2 According to the data results, it can be seen intuitively that the content of UV total flavonoids in Epimedium sagittatum is the highest, and the range of total flavonoids content in the 10 batches of Epimedium koreanatum is the most stable, with the smallest degree of dispersion.

[0069] Table 4 Total flavonoids content of each base of Epimedium

[0070]

[0071]

[0072]

[0073] 2.2 Comparison of flavonoid glycoside content in different origins of Epimedium

[0074] The total flavonoid content of each base was not significantly different, so the flavonol glycosides were detected by high performance liquid chromatography, and the results showed that they were qualified, see Table 5. The Origin software was used to analyze the data results of the content determination of each base of epimedium (3 samples in each batch: Epb1, Epb2, Epb3, Eps1, Eps2, Eps3, Epk1, Epk2, Epk3, Epp1, Epp2, Epp3). With the components as the horizontal axis, it can be seen intuitively that the content of epimedin C is the highest in different epimediums, the content of icariin also accounts for a relatively large proportion, and the content of baohuoside I is the lowest, see Figure 3 Taking the base as the horizontal axis, it can be seen intuitively that the total flavonoid glycoside content in Epimedium sagittatum is the highest. Figure 4 Icariin and epimedin C are different components of epimedium from different origins.

[0075] Table 5 Content of flavonols in each base of Epimedium 6

[0076]

[0077]

[0078] 2.3 Establishment and similarity evaluation of HPLC fingerprints of different origins of Epimedium

[0079] The similarity of the 25 batches of samples ranged from 0.79 to 0.98, indicating that the different origins of Epimedium had relatively good similarity, but there were differences in their intrinsic quality. Figure 5 , Table 6.

[0080] Table 6 Similarity of each epimedium

[0081]

[0082]

[0083] 2.4 Results of differences in the contents of flavonoid components in different origins of Epimedium based on HPLC

[0084] To further compare the differences in the contents of various flavonoid components in different origin Epimedium samples and to find different compounds, the origin Epimedium samples (number: Epb3, Eps1, Epk1, Epp3) were taken respectively, and the contents of quercetin, epimedin A, epimedin B, epimedin C, icariin, icariin I, baculoside I and icariin were determined by HPLC. The results are shown in Table 1. Figure 7 . The sum of the eight components of Epimedium koreanum was lower than that of Epimedium pubescens, Epimedium pubescens and Epimedium sagittatum, and only the content of quercetin was higher than that of other epimediums. The sum of the eight components of Epimedium pubescens was higher than that of Epimedium koreanum, Epimedium koreanum and Epimedium sagittatum, and the contents of epimedin C, epimedin I and baohuoside were much higher than those of other epimediums. The content of icariin in Epimedium koreanum was 21.96 mg / g, which was much higher than that of other epimediums. Icariin was detected in Epimedium sagittatum, but not in other epimediums. The content of flavonoids in each origin varied. In terms of the proportion of components, the content of Epimedium koreanum was the same, close to 1:1:1:1:1:1:1:1:1.

[0085] 2.5 Results of differences in the contents of flavonoid components in different origins of Epimedium based on UPLC-QE-MS

[0086] On the basis of HPLC determination results, in order to further compare the differences in the content of trace flavonoid components in different origin Epimedium samples, the difference compounds were found. UPLC-QE-MS was used to determine the contents of arbutin A, rhamnosyl icariin II, icariin I, quercetin, and dehydrated icariin in each origin Epimedium sample (No.: Epb1, Epb2, Epb3, Eps1, Eps2, Eps3, Epk1, Epk2, Epk3, Epp1, Epp2, Epp3), and the determination data results were analyzed using Origin software. The results are shown in Fig. 9 . The sum of the five components of Epimedium koreana was lower than that of Epimedium pubescens, Epimedium pubescens and Epimedium sagittatum. The sum of the five components of Epimedium pubescens was higher than that of Epimedium pubescens, Epimedium koreanatum and Epimedium sagittatum, and the content of rhamnosyl epimediside II was much higher than that of Epimedium from other origins. The content of flavonoids in each origin varied. In terms of the proportion of components, the content of Epimedium koreanatum was the same, close to 1:1:1:1:1:1:1:1:1.

[0087] 2.6 Orthogonal Partial Least Squares-Discriminant Analysis

[0088] SIMCA software was used for partial least squares regression analysis to extract the variable importance projection value (VIP) of the comprehensive index model and to find the components that contributed greatly to the differences between the groups (VIP>1.0). The VIP of the peaks of icariin A, icariin, epimedin C, baohuoside, and rhamnosylicariin II were all greater than 1.0. Fig.10 .

[0089] 2.3 Conclusion

[0090] ① The total flavonoids content of each epimedium was measured, and it was found that the total flavonoids content of Korean epimedium was lower than that of sagittaria epimedium, but higher than that of the other two epimediums.

[0091] ②The HPLC analysis of the epimedium samples showed that the content ratios of quercetin, epimedin A, epimedin B, epimedin C, icariin, icariin I, baculoside I and icariin were different for each origin, among which the ratio of Korean epimedium was close to 1:1:1:1:1:1:1:1:1. At the same time, it was found that the content of epimedin C accounted for the highest proportion among the epimediums of different origins, and the content of icariin also accounted for a relatively large proportion.

[0092] ③ Epimedium brevifolium from different origins had relatively good similarity, and the similarity of 25 batches of samples was between 0.79 and 0.98.

[0093] ④The samples of each origin of Epimedium were determined by UPLC-QE-MS, and it was found that the content ratios of icariin A, rhamnosyl icariin II, icariin I, quercetin, and dehydrated icariin were different for each origin, among which the Korean Epimedium was close to 1:1:1:1:1:1:1:1:1.

[0094] In summary, the batches of samples used in this study meet the relevant provisions of the "Chinese Pharmacopoeia" (2020 edition) on the content of total flavonoids and flavonol glycosides, but the total flavonoid content of each base of Epimedium is different. After determination, the fingerprints of each base of Epimedium have good similarity. After HPLC and UPLC-QE-MS determination and analysis, quercetin, epimedin A, epimedin B, epimedin C, icariin, icariin I, baohuoside I, icariin, quercetin, dehydrated epimedin A, rhamnosyl epimedin II, epimedin I, quercetin, dehydrated epimedin and other flavonol compounds were selected as differential compounds of each base of Epimedium for subsequent study.

[0095] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any ordinary technician in the industry can smoothly implement the present invention as shown in the drawings and described above. However, any equivalent changes, modifications and evolutions made by technicians familiar with the profession without departing from the scope of the technical solution of the present invention using the technical content disclosed above are all equivalent embodiments of the present invention. At the same time, any equivalent changes, modifications and evolutions made to the above embodiments based on the essential technology of the present invention are still within the protection scope of the technical solution of the present invention.

Claims

1. A method for studying the authenticity of Epimedium koreanum based on traditional Chinese medicine quality markers, characterized in that: The following steps are involved: Step S1: The spectrum-effect relationship of epimedium in treating osteoporosis is studied by combining pharmacodynamics research with chemical component analysis, and possible components of epimedium in treating osteoporosis are obtained. The screened components are used to conduct network pharmacology and molecular docking research to screen out quality markers of epimedium in treating osteoporosis; Step S2: According to step S1, eight flavonoid quality markers of Korean epimedium are finally obtained: quercetin, epimedin A, epimedin B, epimedin C, icariin, icariin I, baculoside I and icariin, and then the difference in component content between Korean epimedium and other varieties of epimedium is determined based on the quality markers; Step S3: The difference in component content between Epimedium koreana and other original Epimediums is as follows: the content ratio of the eight quality markers is 1:1:1:1:1:1:1:1:1; Finally, HPLC and UPLC-QE-MS were used to determine whether the content ratio of the eight flavonoid compounds in the Korean epimedium to be analyzed was 1:1:1:1:1:1:1:1:1, and the authenticity of the Korean epimedium was evaluated.

2. The method for studying the authenticity of Epimedium koreanum based on traditional Chinese medicine quality markers according to claim 1, characterized in that: Other origin epimediums include the dried leaves of the Berberidaceae plants Epimedium, Epimedium sagittatum, Epimedium pubescens, and Epimedium koreana.

3. The method for studying the authenticity of Epimedium koreanum based on traditional Chinese medicine quality markers according to claim 1, characterized in that: The detection method for determining the content of flavonoid components using HPLC is as follows: the chromatographic column is a C18 column, the mobile phase A is acetonitrile, the mobile phase B is water, the detection wavelength is 270nm, the column temperature is 35°C, the injection volume is 10uL, and the flow rate is 1mL / min.

4. The method for studying the authenticity of Epimedium koreanum based on traditional Chinese medicine quality markers according to claim 1, characterized in that: The detection method for determining the content of flavonoid components using UPLC-QE-MS is as follows: the chromatographic column is a Waters UPLC BEH-C18 column (1.7 μm, 2.1 mm×100 mm, Waters, Ireland), the mobile phase A is a 0.1% formic acid aqueous solution, the mobile phase B is acetonitrile, the detection wavelength is 88 nm, the column temperature is 30°C, the injection volume is 2 uL, and the flow rate is 0.3 mL / min.

5. The method for studying the authenticity of Epimedium koreanum based on traditional Chinese medicine quality markers according to claim 1, characterized in that: The fingerprint data of Epimedium brevicornum with different origins were imported into the "Chinese Medicine Chromatographic Fingerprint Similarity Software Evaluation System" of the National Pharmacopoeia Committee. The fingerprints were automatically matched and generated after multi-point correction using the median method, and the similarity evaluation was performed.

6. The method for studying the authenticity of Epimedium koreanum based on traditional Chinese medicine quality markers according to claim 1, characterized in that: The 11 common chromatographic peaks of Epimedium brevicornum from different origins were imported into SPSS23.0 statistical software. Based on the HPLC-MS peak area data of 25 batches of samples, the samples were classified and compared using Euclidean distance, inter-group comparison and other methods. The similarity between samples was calculated, and the analysis results were displayed with the help of a dendrogram.

7. The method for studying the authenticity of Epimedium koreanum based on traditional Chinese medicine quality markers according to claim 1, characterized in that: In order to further distinguish the quality differences within the different origin Epimedium groups, the 11 common peak areas of the different origin Epimedium samples were imported into SIMAC14.1 software for orthogonal partial least squares-discriminant method analysis, and the OPLS-DA model was established.

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