Traditional Chinese medicine phellinus igniarius fingerprint as well as detection method and application thereof

Abstract

The invention discloses a traditional Chinese medicine phellinus igniarius fingerprint spectrum, a detection method thereof and application of the traditional Chinese medicine phellinus igniarius fingerprint spectrum in identification and screening of traditional Chinese medicine phellinus igniarius. The detection method comprises the following steps: step 1, preparing multiple batches of traditional Chinese medicine phellinus igniarius test solution; step 2, injecting each batch of traditional Chinese medicine phellinus igniarius test sample solution into an ultra-high performance liquid tandem QE high-resolution mass spectrometer, and recording an ultraviolet absorption chromatogram and a base peak map of each batch of traditional Chinese medicine phellinus igniarius test sample solution to obtain a fingerprint spectrum of each batch of traditional Chinese medicine phellinus igniarius test sample solution; 3, exporting the fingerprint spectrum obtained in the step 2, and determining the nature by using a TCM database; selecting peaks existing in different batches of traditional Chinese medicine phellinus igniarius test sample solutions as common peaks, generating a fingerprint spectrum corresponding to the traditional Chinese medicine phellinus igniarius by using an average value calculation method, and calculating the relative retention time and the relative peak area of each common peak; and comparing the TCM database, and labeling the chemical components of the peaks in the corresponding fingerprint spectrum of the traditional Chinese medicine phellinus igniarius.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine fingerprint spectrum detection, and in particular to a fingerprint spectrum of the traditional Chinese medicine Phellinus linteus (wild mulberry tree Phellinus linteus) and a detection method and application thereof. Background Art

[0002] Traditional Chinese medicine fingerprint technology is a method that uses modern analytical technology to conduct comprehensive chemical composition analysis on traditional Chinese medicines.

[0003] By separating, detecting and quantifying multiple chemical components in Chinese medicinal materials, a characteristic "fingerprint" of the Chinese medicine can be formed to characterize the authenticity, quality and stability of its quality.

[0004] By analyzing the fingerprint of Chinese medicinal materials, researchers can evaluate the quality of Chinese medicinal materials, identify their authenticity, and detect changes in their ingredients. Exploring the application of Chinese medicine fingerprint technology in the quality control of Chinese medicinal materials is of great significance to improving the quality control level of Chinese medicinal materials and promoting the standardization and standardization of the Chinese medicine industry.

[0005] Phellinus igniarius (wild mulberry tree Phellinus igniarius) has extremely high development and utilization value. Studies have shown that it has anti-tumor, hypoglycemic, liver protection, anti-inflammatory, immunomodulatory, and antioxidant pharmacological effects, and has significant effects in liver protection, anti-tumor, and anti-oxidation.

[0006] There is only one kind of true mulberry, which only grows on the trunk of mulberry trees and is rare in the wild. However, there are many varieties that look similar to mulberry, especially Yanghuang (growing on poplar trunks) and Baoma (growing on clove trunks). This type is easier to cultivate than the real mulberry, and also has certain pharmacological effects. Therefore, it is often used as mulberry, but its pharmacological effect and economic value are much lower than the real mulberry.

[0007] At present, it is common to see Yanghuang and Clove Coriolus replacing Coriolus versicolor. This not only makes the essential characteristics and medicinal effects of Coriolus versicolor unclear in scientific research, but also seriously hinders the selection and breeding of Coriolus versicolor varieties and the development of related industries.

[0008] The patent specification with publication number CN111948326A discloses a method for constructing a fingerprint spectrum of Phellinus igniarius, a fingerprint spectrum and an application, which belongs to the technical field of fingerprint spectrum construction and spectrum effect research of edible and medicinal fungi. This patented technology provides a method for constructing a fingerprint spectrum of Phellinus igniarius, comprising the following steps: taking an alcohol extract of Phellinus igniarius to prepare a test sample, performing high-performance liquid chromatography analysis, mobile phase A is 0.01% acetic acid-water solution; mobile phase B is acetonitrile, and the detection wavelength is 254nm; according to the results of high-performance liquid chromatography analysis, a fingerprint spectrum is constructed. The construction method described in this patented technology can realize the efficient screening of different varieties of Phellinus igniarius. More specifically, the fingerprint spectrum obtained by the construction method described in this patented technology can realize the efficient screening of poplar Phellinus igniarius (Sanghuangporousvaninii).

[0009] As mentioned above, there is only one kind of real mulberry, that is mulberry mulberry (Sanghuangporussanghuang), also known as mulberry. At present, poplar mulberry is called Yanghuang, which is flooding the market. Its efficacy is far lower than that of mulberry, which has brought interference to scientific research and the market.

[0010] Based on this, the main content of the present invention is to use the fingerprint spectrum of Phellinus igniarius to distinguish the counterfeit products on the market - Yanghuang and Clove Phellinus igniarius. Summary of the invention

[0011] In view of the above technical problems and the shortcomings in the art, the present invention provides a fingerprint spectrum of Chinese medicine Phellinus igniarius (wild mulberry Phellinus igniarius) and its detection method and application. The present invention uses ultra-high performance liquid phase tandem QE high-resolution mass spectrometry, with higher resolution, and can obtain the specific content of effective ingredients. The detection method of the present invention can objectively, comprehensively and accurately evaluate the authenticity and drug quality of Chinese medicine Phellinus igniarius, which is of great significance to controlling the quality of Chinese medicine Phellinus igniarius and ensuring its clinical efficacy.

[0012] The specific technical solutions are as follows:

[0013] [1] A method for detecting the fingerprint spectrum of the traditional Chinese medicine Phellinus igniarius, comprising:

[0014] Step 1, preparing multiple batches of Chinese medicine Phellinus igniarius test sample solutions;

[0015] Step 2, injecting each batch of Chinese medicine Phellinus igniarius test solution into an ultra-high performance liquid phase tandem QE high-resolution mass spectrometer, recording the ultraviolet absorption chromatogram and base peak diagram of each batch of Chinese medicine Phellinus igniarius test solution, and obtaining the fingerprint spectrum of each batch of Chinese medicine Phellinus igniarius test solution;

[0016] Step 3, export the fingerprint spectrum obtained in step 2, and use the TCM database for qualitative analysis; select the peaks present in different batches of Chinese medicine Phellinus igniarius test solution as common peaks, generate the fingerprint spectrum corresponding to the Chinese medicine Phellinus igniarius by the average value calculation method, and calculate the relative retention time and relative peak area of ​​each common peak; compare the TCM database, and mark the chemical composition of the peaks in the fingerprint spectrum corresponding to the Chinese medicine Phellinus igniarius.

[0017] The TCM database is an exclusive Chinese medicine ingredient database established for plant samples. The database contains information on more than 5,000 Chinese medicine ingredient standards, covering 10 major categories of Chinese medicine ingredients, including alkaloids, phenolic acids, flavonoids, coumarins, phenylpropanoids, lignans, and terpenes.

[0018] In the detection method described in the present invention, the Chinese medicinal herb Phellinus linteus is Sanghuangporussanghuang.

[0019] In step 1, methanol can be used to dissolve and extract the Chinese medicine Phellinus linteus sample to obtain the Chinese medicine Phellinus linteus test sample solution.

[0020] In some embodiments, step 1 may specifically include: mixing and grinding the Chinese medicine Phellinus igniarius sample and methanol, ultrasonically extracting and centrifuging to obtain the supernatant, and diluting the obtained supernatant with methanol to obtain the Chinese medicine Phellinus igniarius test solution. Furthermore, the Chinese medicine Phellinus igniarius sample and methanol can be mixed and ground at 100 mg:1 mL. For example, 100 mg of the Chinese medicine Phellinus igniarius sample can be weighed in a 1.5 mL centrifuge tube, 1 mL of methanol can be added, ultrasonically extracted for 60 min, filtered after centrifugation, diluted with methanol, and 200 μL of supernatant can be taken for analysis.

[0021] In step 2:

[0022] Chromatographic conditions may include:

[0023] Chromatographic column: ACQUITY UPLC HSS T3, 100 mm × 2.1 mm, 1.8 μm;

[0024] Column temperature: 45°C;

[0025] Mobile phase A: aqueous solution containing 0.1 vol% formic acid;

[0026] Mobile phase B: acetonitrile;

[0027] Flow rate: 0.35 mL / min;

[0028] Injection volume: 5 μL;

[0029] PDA scanning range: 210-400nm;

[0030] Elution gradient information table:

[0031] Time / min Mobile phase A / vol% Mobile phase B / vol% 0 95 5 2 95 5 4 70 30 8 50 50 10 20 80 14 0 100 15 0 100 15.1 95 5 16 95 5

[0032] Mass spectrometry conditions may include:

[0033] Ion source: HESI;

[0034] The sample mass spectrometry signal was collected using positive and negative ion scanning modes, respectively;

[0035] Data acquisition mode: DDA;

[0036] Scan mode: Full MS / dd-MS2;

[0037] Mass spectrometry parameter information table:

[0038]

[0039] The UV absorption wavelength can be selected: UV absorption is measured at 210nm and 254nm wavelength.

[0040] In the fingerprint corresponding to the Chinese medicine Phellinus igniarius in step 3, the top 10 compounds with the highest common peak intensity (the top 10 compounds with the highest average value (relative content) of the ratio of the compound response intensity in the fingerprint to the sum of the response intensity of all compounds in the Chinese medicine Phellinus igniarius) are:

[0041] α-lactose, retention time 0.72min, mass-to-charge ratio 387.1143, detection mode NEG, peak 51;

[0042] Phytosphingosine, retention time 9.6min, mass-to-charge ratio 318.3, detection mode POS, peak 25;

[0043] Milk echinopsine, retention time 5.02min, mass-to-charge ratio 245.0454, detection mode NEG, peak 43;

[0044] Luciferin free acid, retention time 5.49min, mass-to-charge ratio 281.0046, detection mode POS, peak 9;

[0045] melibiose, retention time 0.73min, mass-to-charge ratio 360.1497, detection mode POS, peak 30;

[0046] Sorbitol, retention time 0.72min, mass-to-charge ratio 181.0711, detection mode NEG, peak 17;

[0047] L-glutamic acid, retention time 0.71min, mass-to-charge ratio 148.0603, detection mode POS, peak 29;

[0048] Mogroic acid D, retention time 9.38min, mass-to-charge ratio 479.2763, detection mode POS, peak 164;

[0049] Withanolide C, retention time 8.5min, mass-to-charge ratio 473.2911, detection mode NEG, peak 179;

[0050] Withaferin A, retention time is 7.9min, mass-to-charge ratio is 453.2634, detection mode is POS, and it is peak No. 38.

[0051] [2] The fingerprint of the traditional Chinese medicine Phellinus igniarius obtained according to the detection method described in [1].

[0052] [3] Application of the detection method described in [1] or the fingerprint spectrum of the Chinese medicine Phellinus igniarius described in [2] in the identification and screening of the Chinese medicine Phellinus igniarius.

[0053] The technical solution of the present invention optimizes fingerprint spectrum detection in the following ways:

[0054] 1. Optimize LC-MS conditions

[0055] The present invention adopts a liquid chromatography-mass spectrometry system consisting of an ACQUITY UPLC I-Class HF ultra-high performance liquid phase tandem QE high-resolution mass spectrometer, which has extremely high resolution and excellent mass accuracy and can perform highly selective, high-resolution, and highly sensitive qualitative and quantitative analysis of small and macromolecular compounds.

[0056] 2. Optimization of detection wavelength

[0057] By optimizing the detection wavelength, it is found that saponin components have strong absorption at 210nm, and components containing structures such as benzene rings and double bonds have strong absorption at a wavelength of 254nm. Therefore, the present invention extracts ultraviolet absorption diagrams at wavelengths of 210nm and 254nm.

[0058] 3. Optimization of the standard reference database

[0059] The present invention uses the TCM database to qualitatively characterize the components of traditional Chinese medicine. The TCM database is an exclusive database of traditional Chinese medicine components established for plant samples. The database contains information on more than 5,000 standard products of traditional Chinese medicine components, covering 10 major categories of traditional Chinese medicine components, including alkaloids, phenolic acids, flavonoids, coumarins, phenylpropanoids, lignans, and terpenes.

[0060] Compared with the prior art, the present invention has the following beneficial effects:

[0061] According to the properties of the active ingredients contained in the Chinese medicine Phellinus igniarius, the present invention screens out the optimal mobile phase, elution program, flow rate, detection wavelength, column temperature and other analysis conditions through a large number of experiments. Multiple experimental analyses show that the Chinese medicine Phellinus igniarius fingerprint established by the means provided by the present invention can effectively characterize the quality of the Chinese medicine Phellinus igniarius, avoids the one-sidedness of judging its quality due to the determination of individual chemical components of Phellinus igniarius, and can comprehensively, objectively and accurately detect and evaluate the quality of the Chinese medicine Phellinus igniarius, which is of great significance for ensuring the clinical efficacy of Phellinus igniarius.

[0062] The detection method of the fingerprint spectrum of the traditional Chinese medicine Phellinus igniarius provided by the present invention also has the advantages of being stable and reliable, having high precision, and having good repeatability. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] Figure 1 This is the chromatogram of the Chinese medicine Sanghuangporus sanghuang at 210 nm according to the detection method of the present invention.

[0064] Figure 2 This is the chromatogram of the Chinese medicine Sanghuangporus sanghuang at 254 nm according to the detection method of the present invention.

[0065] Figure 3 This is the base peak diagram (BPC) of the Chinese medicine Sanghuangporus sanghuang in the positive ion mode (POS) of the detection method of the present invention.

[0066] Figure 4 This is the base peak chart (BPC) of the Chinese medicine Sanghuangporus sanghuang in the negative ion mode (NEG) of the detection method of the present invention.

[0067] Figure 5 It is the chromatographic and mass spectrometric characteristic peaks of α-lactose in the fingerprint spectrum of the traditional Chinese medicine Phellinus igniarius in the detection method of the present invention.

[0068] Figure 6 It is the chromatographic and mass spectrometric characteristic peaks of phytosphingosine in the fingerprint spectrum of the traditional Chinese medicine Phellinus igniarius in the detection method of the present invention.

[0069] Figure 7 It is the chromatographic and mass spectrometric characteristic peaks of milkvine in the fingerprint spectrum of the traditional Chinese medicine mulberry linterus in the detection method of the present invention.

[0070] Figure 8 It is the characteristic peak of chromatogram and mass spectrum of luciferin free acid in the fingerprint spectrum of Chinese medicine Phellinus igniarius in the detection method of the present invention.

[0071] Fig. 9 It is the characteristic peak of melibiose chromatography and mass spectrometry in the fingerprint spectrum of the traditional Chinese medicine Phellinus igniarius in the detection method of the present invention.

[0072] Fig.10 It is the characteristic peak of sorbitol chromatography and mass spectrometry in the fingerprint spectrum of the traditional Chinese medicine Phellinus igniarius in the detection method of the present invention.

[0073] Fig.11 It is the characteristic peak of L-glutamic acid chromatography and mass spectrometry in the fingerprint spectrum of the traditional Chinese medicine Phellinus igniarius in the detection method of the present invention.

[0074] Fig.12 It is the characteristic peak of the chromatography and mass spectrometry of mogroside acid D in the fingerprint spectrum of the traditional Chinese medicine mulberry linterus in the detection method of the present invention.

[0075] Fig.13 It is the characteristic peak of withanolide C chromatography and mass spectrometry in the fingerprint of the traditional Chinese medicine Phellinus igniarius in the detection method of the present invention.

[0076] Fig.14 It is the characteristic peak of withaferin A chromatography and mass spectrometry in the fingerprint spectrum of the traditional Chinese medicine mulberry linterus in the detection method of the present invention. DETAILED DESCRIPTION

[0077] The present invention will be further described below in conjunction with the accompanying drawings and specific examples. It should be understood that these examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention. The operating methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer.

[0078] The instruments and reagents used in the embodiments are as follows:

[0079] Experimental equipment

[0080] 1.1 Instrument

[0081] High-resolution liquid chromatography-mass spectrometer (Thermo-Obritrap-QE), PDA detector (ACQUITY UPLC), chromatographic column (ACQUITY UPLC HSS T3), high-performance liquid chromatography (ACQUITY UPLC I-Class HF), desktop high-speed refrigerated centrifuge (TGL-16MS), vortex oscillator (TYXH-I), and ultrasonic cleaning machine (F-060SD).

[0082] 1.2 Drugs and reagents

[0083] Ten batches (210720; 210721; 230102; 230123; 230510; 230511; 230720; 231223; 231225; 240312; corresponding to S1-S10 respectively) of Chinese medicine Phellinus igniarius samples were from Hangzhou Academy of Agricultural Sciences, methanol (Thermo Fisher, A452-4), acetonitrile (Thermo Fisher, A998-4), (Thermo Fisher, A117-50), and water (pure water).

[0084] Example:

[0085] A method for detecting the fingerprint spectrum of traditional Chinese medicine Phellinus igniarius, comprising:

[0086] Step 1, preparation of Phellinus linteus test solution:

[0087] (1) Weigh approximately 100 mg of sample into a 1.5 mL centrifuge tube;

[0088] (2) Add 1000 μL methanol, vortex for 1 min, and add steel beads;

[0089] (3) After precooling in a -40°C refrigerator for 2 min, grind in a grinder (60 Hz, 2 min);

[0090] (4) Ultrasonic extraction in an ice-water bath for 60 min, and then standing at -40°C for 30 min;

[0091] (5) Centrifugation for 10 min (12000 rpm, 4°C);

[0092] (6) After diluting 20 times with methanol, 200 μL of the supernatant was transferred into an LC-MS injection vial in an inner tube for analysis.

[0093] Step 2, accurately aspirate 10 batches of mulberry linterus test solution of step 1 respectively, inject into ultra-high performance liquid chromatograph, record the ultraviolet absorption chromatogram and base peak diagram of each batch of Chinese medicine mulberry linterus test solution, and obtain the fingerprint spectrum of each batch of Chinese medicine mulberry linterus test solution.

[0094] Chromatographic conditions: chromatographic column: ACQUITY UPLC HSS T3 (100 mm×2.1 mm, 1.8 μm); column temperature: 45°C; mobile phase: A-water (containing 0.1 vol% formic acid), B-acetonitrile; flow rate: 0.35 mL / min; injection volume: 5 μL, PDA scanning range: 210-400 nm.

[0095] Elution gradient information table:

[0096] Time / min Mobile phase A / vol% Mobile phase B / vol% 0 95 5 2 95 5 4 70 30 8 50 50 10 20 80 14 0 100 15 0 100 15.1 95 5 16 95 5

[0097] Mass spectrometry conditions: ion source: HESI; sample mass spectrometry signal acquisition adopts positive and negative ion scanning mode respectively; data acquisition mode: DDA; scanning mode: Full MS / dd-MS2.

[0098] Mass spectrometry parameter information table:

[0099]

[0100] UV absorption wavelength selection: UV absorption is measured at 210nm and 254nm wavelengths. Figure 1 , Figure 2 The chromatograms are the averaged chromatograms of batches S1-S10 of Chinese medicine Phellinus igniarius samples at 210nm and 254nm respectively.

[0101] Figure 3 , Figure 4 They are the base peak graphs in positive ion mode (POS) and negative ion mode (NEG) after averaging of batches S1-S10 of the Chinese medicine Phellinus igniarius samples.

[0102] Step 3, data processing.

[0103] The fingerprint obtained in step 2 was exported, and the data were processed by the metabolomics processing software Progenesis QI v3.0 software (Nonlinear Dynamics, Newcastle, UK) for baseline filtering, peak identification, integration, retention time correction, peak alignment, and normalization. The identification of the compounds was based on accurate mass, secondary fragmentation, and isotopic distribution, and the TCM database was used for qualitative analysis.

[0104] Peaks present in different batches of Chinese medicine Phellinus igniarius test solutions were selected as common peaks, and the corresponding fingerprint spectrum of Chinese medicine Phellinus igniarius was generated by the average value calculation method. The relative retention time and relative peak area of ​​each common peak were calculated; the TCM database was compared to annotate the chemical components of the peaks in the fingerprint spectrum of Chinese medicine Phellinus igniarius.

[0105] Qualitative and quantitative results table (top 20 compounds):

[0106]

[0107] Figures 5 to 14 The chromatographic and mass spectrometric characteristic peaks of the top 10 compounds in the above table are shown respectively.

[0108] According to the above-mentioned detection method of the fingerprint spectrum of the Chinese medicine Phellinus igniarius, the similarity of the above-mentioned 10 batches of Chinese medicine Phellinus igniarius is as follows:

[0109]

[0110] In addition, it should be understood that after reading the above description of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A method for detecting the fingerprint of Chinese medicine Phellinus igniarius, characterized in that: include: Step 1, preparing multiple batches of Chinese medicine Phellinus igniarius test sample solutions; Step 2, injecting each batch of Chinese medicine Phellinus igniarius test solution into an ultra-high performance liquid phase tandem QE high-resolution mass spectrometer, recording the ultraviolet absorption chromatogram and base peak diagram of each batch of Chinese medicine Phellinus igniarius test solution, and obtaining the fingerprint spectrum of each batch of Chinese medicine Phellinus igniarius test solution; Step 3, export the fingerprint spectrum obtained in step 2, and use the TCM database for qualitative analysis; select the peaks present in different batches of Chinese medicine Phellinus igniarius test solution as common peaks, generate the fingerprint spectrum corresponding to the Chinese medicine Phellinus igniarius by the average value calculation method, and calculate the relative retention time and relative peak area of ​​each common peak; compare the TCM database, and mark the chemical composition of the peaks in the fingerprint spectrum corresponding to the Chinese medicine Phellinus igniarius.

2. The detection method according to claim 1, characterized in that: The Chinese medicinal herb Phellinus linteus is Sanghuangporus sanghuang.

3. The detection method according to claim 1, characterized in that: In step 1, the Chinese medicine Phellinus linteus sample is dissolved and extracted with methanol to obtain the Chinese medicine Phellinus linteus test sample solution.

4. The detection method according to claim 3, characterized in that: Step 1 specifically includes: mixing and grinding the Chinese medicine Phellinus igniarius sample and methanol, performing ultrasonic extraction, and centrifuging to obtain the supernatant, and diluting the obtained supernatant with methanol to obtain the Chinese medicine Phellinus igniarius test sample solution.

5. The detection method according to claim 4, characterized in that: The Chinese medicine Phellinus igniarius sample and methanol were mixed and ground at a ratio of 100 mg:1 mL.

6. The detection method according to claim 1, characterized in that: In step 2: Chromatographic conditions included: Chromatographic column: ACQUITY UPLC HSS T3, 100 mm × 2.1 mm, 1.8 μm; Column temperature: 45°C; Mobile phase A: aqueous solution containing 0.1 vol% formic acid; Mobile phase B: acetonitrile; Flow rate: 0.35 mL / min; Injection volume: 5 μL; PDA scanning range: 210-400nm; Elution gradient information table: Mass spectrometry conditions included: Ion source: HESI; The sample mass spectrometry signal was collected using positive and negative ion scanning modes, respectively; Data acquisition mode: DDA; Scan mode: Full MS / dd-MS2; Mass spectrometry parameter information table: UV absorption wavelength selection: UV absorption was measured at 210nm and 254nm wavelengths.

7. The detection method according to claim 1, characterized in that: In the fingerprint of the Chinese medicine Phellinus igniarius in step 3, combined with chromatography and mass spectrometry, the top 10 compounds with the highest common peak intensity are: α-lactose, retention time 0.72 min, mass-to-charge ratio 387.1143, detection mode NEG; Phytosphingosine, retention time 9.6min, mass-to-charge ratio 318.3, detection mode POS; Milk echinopsine, retention time 5.02min, mass-to-charge ratio 245.0454, detection mode NEG; Luciferin free acid, retention time 5.49min, mass-to-charge ratio 281.0046, detection mode POS; melibiose, retention time 0.73min, mass-to-charge ratio 360.1497, detection mode POS; Sorbitol, retention time 0.72min, mass-to-charge ratio 181.0711, detection mode NEG; L-glutamic acid, retention time 0.71min, mass-to-charge ratio 148.0603, detection mode POS; Mogroic acid D, retention time 9.38min, mass-to-charge ratio 479.2763, detection mode POS; Withanolide C, retention time 8.5min, mass-to-charge ratio 473.2911, detection mode NEG; Withaferin A, retention time 7.9min, mass-to-charge ratio 453.2634, detection mode POS.

8. The fingerprint of the traditional Chinese medicine Phellinus igniarius obtained by the detection method according to any one of claims 1 to 7.

9. Use of the detection method according to any one of claims 1 to 7 or the fingerprint of the Chinese medicine Phellinus igniarius according to claim 7 in identifying and screening the Chinese medicine Phellinus igniarius.

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